CN112544781B - Antibiotic-replacement-type deodorizing feeding microecological preparation and preparation method and application thereof - Google Patents
Antibiotic-replacement-type deodorizing feeding microecological preparation and preparation method and application thereof Download PDFInfo
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- CN112544781B CN112544781B CN202011208303.4A CN202011208303A CN112544781B CN 112544781 B CN112544781 B CN 112544781B CN 202011208303 A CN202011208303 A CN 202011208303A CN 112544781 B CN112544781 B CN 112544781B
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Classifications
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses an antibiotic substitution type deodorization feeding microecological preparation and a preparation method and application thereof. The microecological preparation comprises the following components in parts by weight: 200-300 parts of coated lactobacillus plantarum, 200-300 parts of coated enterococcus faecium, 20-50 parts of bacillus subtilis and 500 parts of fructo-oligosaccharide. The microecological preparation of the invention adopts the coated lactobacillus plantarum and the coated enterococcus faecium, and solves the problems of poor stress resistance of the lactobacillus, namely short shelf life under normal temperature storage condition, easy inactivation in the feed granulation process and the like. In addition, the microecological preparation can reduce the content of odor substances in animal wastes, reduces the pollution of livestock and poultry breeding to the environment in the nonreactive breeding process, has the functions of promoting growth and deodorizing, and has important economic and social meanings.
Description
Technical Field
The invention belongs to the technical field of feed additives. More particularly, relates to an antibiotic substitution type deodorization feeding microecological preparation, and a preparation method and application thereof.
Background
For a long time, the addition of antibiotics in the feed has a great promoting effect on the development of animal husbandry, but along with the popularization and application of the feed antibiotics, adverse reactions of the feed antibiotics are gradually prominent, such as: the problems of endogenous infection and cross infection caused by antibiotics, generation of drug resistance, reduction of immunity of livestock and poultry, destruction of normal flora of intestinal tracts, residue of normal flora in livestock products and environment and the like all bring serious threats to the breeding industry, the feed industry and animals, and further endanger the health of human beings through a food chain.
In addition, with the rapid development of livestock breeding industry in China, the side effects of sewage, livestock excrement and malodorous gas of livestock farms on livestock and poultry and the environmental pollution become more and more serious. At present, the prior art mainly focuses on a daily ration formula aiming at reducing the content of indole and skatole in excrement, and has strong specificity aiming at animals and lacks wide usability; certain fermented feeds using complex microorganisms also require specific production conditions and are not widely applicable. Most of the composite microecologics are only applied to the aspects of production performance, immunity performance, intestinal microorganisms and the like, and are not focused on reducing odor substances such as skatole, indole and the like.
Therefore, it is necessary to develop a micro-ecological agent which can be used as an antibiotic substitute, and can effectively deodorize and reduce environmental pollution.
Disclosure of Invention
The invention aims to provide an antibiotic-substituted deodorizing microecological preparation for feed. The microecological preparation can obviously improve the growth performance of animals and can reduce the content of indole and skatole in the excrement of livestock and poultry.
The invention also aims to provide a preparation method of the antibiotic-substituted deodorant microecological preparation for the feed.
The third purpose of the invention is to provide the application of the antibiotic substitute type deodorization feeding microecological preparation in livestock and poultry feeding or the application in preparing livestock and poultry feed.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides an antibiotic-substituted deodorizing microecological preparation for feed, which comprises the following components in parts by weight: 200-300 parts of coated lactobacillus plantarum, 200-300 parts of coated enterococcus faecium, 20-50 parts of bacillus subtilis and 500 parts of fructo-oligosaccharide.
Optionally, the probiotic further comprises yucca extract. According to a specific embodiment of the present invention, in the microecological formulation, the yucca extract is 100-150 parts by mass.
According to the results of earlier researches, the coated lactobacillus plantarum and the coated enterococcus faecium have excellent effects on the aspects of improving the production performance of animals, inhibiting harmful bacteria, improving the immunity and the oxidation resistance of organisms; meanwhile, according to in vitro research results, fructo-oligosaccharide is a prebiotic with the best synergistic effect with lactobacillus plantarum, and can provide energy for intestinal microorganisms and promote the intestinal beneficial microorganisms to play a role; the bacillus subtilis self-synthesizes enzymes such as protease, cellulase and the like, can improve the digestion and utilization rate of nutrient substances and reduce the generation of odor substances such as ammonia gas and the like. The yucca extract has strong adsorption effect, can be directly combined with harmful odor gases such as ammonia gas and hydrogen sulfide, and has synergistic effect in removing odor substances.
Optionally, the viable count contents of the coated lactobacillus plantarum, the coated enterococcus faecium and the bacillus subtilis are (1-5) multiplied by 10 respectively10CFU/g、(1-5)×1010CFU/g and (5-10). times.1010CFU/g。
In a second aspect, the invention provides a preparation method of the antibiotic-substituted deodorant feeding microecological preparation, which comprises the step of uniformly mixing the coated lactobacillus plantarum, the coated enterococcus faecium, the bacillus subtilis and the fructo-oligosaccharide according to the formula proportion to obtain the antibiotic-substituted deodorant feeding microecological preparation.
The invention provides a preparation method of coated lactobacillus plantarum or enterococcus faecium, which comprises the following steps of:
(1) preparing a sodium alginate solution with the concentration of 10g/L-20g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:1-1.5 of the calcium carbonate to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 35-38 ℃, adding lactobacillus plantarum or enterococcus faecium seed liquid at the last logarithmic growth stage until the density of the lactobacillus plantarum or the enterococcus faecium is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as an oil phase, wherein the volume percentage of the span 80 is 0.1-0.3%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.1-0.3mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 400 plus 500rpm according to the volume ratio of the water phase to the oil phase of 1:3-5, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:500 plus 600, immobilizing for 10-20min, stopping stirring, adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:10-20, slowly settling the microcapsule, and then sucking away the oil phase;
(6) adding a culture medium suitable for growth of lactobacillus plantarum or enterococcus faecium into a fermentation tank, adding ammonia water in a flowing manner to control the pH value of the fermentation liquid to be 6.2-6.9 under the conditions of 35-38 ℃ and the stirring speed of 50-100rpm, continuously culturing for 8-22h until more than 80% of the space in the microcapsule is filled with the lactobacillus plantarum or the enterococcus faecium, and centrifuging to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution, and fully mixing to perform film forming reaction to obtain double-layer microcapsule bacterial sludge;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-30-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 5-10 parts of glycerol, 5-10 parts of trehalose, 0.5-1 part of skimmed milk powder, 0.5-1 part of sodium acetate and 300 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.5-1 part of xanthan gum, 0.001-0.005 part of sodium carboxymethylcellulose, 0.2-0.8 part of microcrystalline cellulose and 100 parts of water and 200 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) and (3) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 40-55 ℃, controlling the air outlet temperature to be 30-40 ℃, drying until the water content of the sample is 10-20%, spraying an enteric coating material to form a coating, and sieving the dried and coated sample by using a double-layer sieve to obtain a final product with the particle size of 20-60 meshes.
In the step (7), the chitosan acidic buffer solution is prepared by preparing chitosan with molecular weight of 300-500kDa into a chitosan aqueous solution with mass concentration of 0.2-0.5%, and adjusting the pH value of the aqueous solution to 4.5-5.5.
In the step (9), the enteric coating material is 20-40% of polyacrylic resin latex solution by mass. According to the specific embodiment of the invention, the mass ratio of the enteric coating material to the microcapsule master batch is 1:4-10, and the microcapsule master batch is sprayed at a speed of 25-50 mL/min.
In a second aspect, the invention provides application of the antibiotic substitute type deodorization feeding microecological preparation in livestock and poultry feeding or preparation of livestock and poultry feed.
Further, the application of the antibiotic substitute type deodorization feeding microecological preparation in any one of the following aspects also belongs to the protection scope of the invention:
1) the application in reducing the content of indole and skatole in the livestock and poultry manure;
2) the application in preparing the feed for reducing the content of indole and skatole in the excrement of livestock and poultry;
3) the application in improving the growth performance of livestock and poultry;
4) the application in preparing the feed for improving the growth performance of the livestock and poultry;
5) the application in replacing antibiotics, improving the immunity and the anti-oxidative stress capability of the organism;
6) the application of the compound feed in preparing the feed for replacing antibiotics and improving the immunity and the anti-oxidative stress capability of the organism.
Further, in order to better play the role of the antibiotic substitute type deodorization feeding microecological preparation, in the specific embodiment of the invention, the antibiotic substitute type deodorization feeding microecological preparation is added into the complete feed according to the proportion of 0.01-0.2% of the mass of the feed for feeding.
The invention has the following beneficial effects:
the microecological preparation adopts the coated lactobacillus plantarum and the coated enterococcus faecium, solves the problems of poor stress resistance of the lactobacillus, namely short shelf life under normal-temperature storage conditions, easy inactivation in the feed granulation process and the like, and can better exert the action effect.
The microecological preparation prepared by the formula can improve the growth performance of animals (improve the daily gain of pigs and broilers, improve the laying rate of laying hens), the immunity performance (the IgA content and the IgG content in blood, reduce the content of proinflammatory factors and the content of anti-inflammatory factors), the antioxidant stress capacity (improve the content of glutathione peroxidase, the content of superoxide dismutase, the total antioxidant capacity and the content of malonaldehyde which is an oxidation reaction product) by directly feeding the microecological preparation in the antibiotic-free feeding process, moreover, the content of odor substances in animal wastes can be reduced, so that the indole content in the odor substances is reduced by 20-35%, the skatole content is reduced by 15-50%, the pollution of livestock and poultry cultivation to the environment in the nonresistant cultivation process is reduced, the functions of promoting growth and deodorizing are achieved, antibiotics are effectively replaced, and the method has important economic and social meanings.
Detailed Description
In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.
The materials in the following examples, unless otherwise specified, are all materials commonly used in the art and are commercially available.
Example 1
The antibiotic substitution type deodorization feeding microecological preparation comprises the following components in parts by weight: 200 parts of coated lactobacillus plantarum, 200 parts of coated enterococcus faecium, 20 parts of bacillus subtilis and 300 parts of fructo-oligosaccharide.
The preparation method comprises the following steps:
1. preparation of multilayer coated enterococcus faecium microcapsule product
(1) Preparing a sodium alginate solution with the concentration of 10g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:1 of the calcium carbonate to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 37 ℃, adding enterococcus faecium seed liquid at the last logarithmic growth stage until the density of probiotics is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as oil phase, wherein the volume percentage of the span 80 is 0.1%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.1mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 400rpm according to the volume ratio of the water phase to the oil phase of 1:3, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:500, immobilizing for 10-20min, stopping stirring, then adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:10, slowly settling the microcapsule, and then sucking away the oil phase;
(6) sterilizing at 121 ℃ for 30min, adding a fermentation medium cooled to 37 ℃ into a fermentation tank, adding ammonia water under the conditions of 37 ℃ and a stirring speed of 100rpm, controlling the pH value of the fermentation liquor to be 6.2-6.9, continuously culturing for 8-16h until more than 80% of the space in the microcapsule is filled with probiotics, and centrifuging at 15000r/m to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution for film forming reaction to obtain double-layer microcapsule bacterial sludge, and cleaning with sterile water; wherein the chitosan acid buffer solution is prepared by preparing chitosan with molecular weight of 400kDa into a chitosan aqueous solution with mass concentration of 0.4%, and adjusting the pH value of the aqueous solution to 5.0;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 5 parts of glycerol, 10 parts of trehalose, 0.5 part of skimmed milk powder, 0.5 part of sodium acetate and 150 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.5 part of xanthan gum, 0.001 part of sodium carboxymethylcellulose, 0.2 part of microcrystalline cellulose and 120 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 45 ℃, controlling the air outlet temperature to be 35 ℃, and drying for 30 min; starting the side-spraying coating device, spraying enterococcus faecium microcapsule master batches by using an enteric coating material at the speed of 30mL/min (the enteric coating material is 20% by mass of polyacrylic resin emulsion, and the mass ratio of the enteric coating material to the microcapsule master batches is 1:5), forming a coating, and drying to obtain the enteric coating material.
2. Preparation method of multilayer coated lactobacillus plantarum microcapsule product
(1) Preparing a sodium alginate solution with the concentration of 20g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:5 to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 37 ℃, adding lactobacillus plantarum seed liquid at the last logarithmic growth stage until the probiotic density is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as oil phase, wherein the volume percentage of the span 80 is 0.3%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.3mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 500rpm according to the volume ratio of the water phase to the oil phase of 1:5, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:600, immobilizing for 10-20min, stopping stirring, then adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:20, slowly settling the microcapsule, and then sucking away the oil phase;
(6) sterilizing at 121 ℃ for 30min, adding a fermentation medium cooled to 37 ℃ into a fermentation tank, adding ammonia water under the conditions of 37 ℃ and a stirring speed of 50rpm, controlling the pH value of the fermentation liquor to be 6.2-6.9, continuously culturing for 16-22h until more than 80% of the space in the microcapsule is filled with probiotics, and centrifuging at 15000r/m to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution for film forming reaction to obtain double-layer microcapsule bacterial sludge, and cleaning with sterile water; wherein the chitosan acid buffer solution is prepared by preparing chitosan with molecular weight of 400kDa into a chitosan aqueous solution with mass concentration of 0.5%, and adjusting the pH value of the aqueous solution to 5.0;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 10 parts of glycerol, 10 parts of trehalose, 1 part of skimmed milk powder, 1 part of sodium acetate and 300 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.8 part of xanthan gum, 0.003 part of sodium carboxymethyl cellulose, 0.5 part of microcrystalline cellulose and 200 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 55 ℃, controlling the air outlet temperature to be 40 ℃, and drying for 35 min; starting the side-spraying coating device, spraying enterococcus faecium microcapsule master batches by using an enteric coating material at the speed of 30mL/min (the enteric coating material is polyacrylic resin emulsion with the mass percentage of 40%, and the mass ratio of the enteric coating material to the microcapsule master batches is 1:8), forming a coating, and drying to obtain the enteric coating material.
3. And uniformly mixing the coated lactobacillus plantarum, the coated enterococcus faecium, the bacillus subtilis and the fructo-oligosaccharide according to the formula proportion to obtain the compound.
Example 2
The antibiotic substitution type deodorization feeding microecological preparation comprises the following components in parts by weight: 300 parts of coated lactobacillus plantarum, 300 parts of coated enterococcus faecium, 50 parts of bacillus subtilis and 500 parts of fructo-oligosaccharide.
The preparation method comprises the following steps: the same as in example 1.
Test example 1: the influence of the microecological preparation on growth performance, oxidation resistance, immunity and odor substances in the nonreactive breeding process of the broilers
216 broiler chickens of 1 day old are selected and randomly divided into 3 treatment groups, each treatment group has 6 repetitions, and each repetition has 12 chickens. Group 1 was supplemented with basal diet, group 2 with antibiotics, group 3 with the probiotic of the invention (example 1), with a test period of 42 days.
The influence of the microecological preparation on growth, oxidation resistance and immunity, and the content of odorous substances such as indole and skatole in excrement and urine is evaluated through daily gain, material-to-weight ratio, MDA content in serum, GSH-Px, T-AOC, SOD activity and IL-2, IL-6, IL-10 and TNF-alpha content.
The MDA content, GSH-Px, T-AOC, SOD activity and IL-2, IL-6, IL-10 and TNF-alpha content in serum are detected by an ELISA kit, the indole and skatole content are detected by liquid chromatography-mass spectrometry, and the specific method comprises the following steps:
(1) sample pretreatment: weighing a proper amount of sample, adding a proper amount of distilled water, homogenizing, adding 1ml of acetonitrile after homogenizing, and mixing uniformly. And (5) instantly extracting the supernatant for later use. Taking 50ul of supernatant, adding 200ul of protein precipitator, whirling for 1min, centrifuging for 4min at 13200 r, and taking the supernatant to be tested.
(2) Test parameters are as follows:
liquid phase conditions:
a chromatographic column: MSLab50AA-C18(100 x 4.6mm 5um) column temperature: 50 ℃, flow rate: 1ml/min, mobile phase: a water phase: water (1mmol/l amine acetate) B organic phase: acetonitrile (1mmol/l amine acetate), sample size: 10 ul.
Gradient conditions:
TABLE 1 gradient elution conditions
Mass spectrum conditions:
an ion source: -ESI electrospray ion source;
IS: 4500V (spray voltage);
GS 1: 55psi (atomizing gas);
GS 2: 60psi (assist gas);
the scanning mode is as follows: monitoring MRM multiple reactions;
CAD: medium (collision gas);
TEM: 500 ℃ (atomization temperature);
and (4) CUR: 20psi (air curtain air);
CXP: 2.0 (collision cell ejection voltage);
EP: -10 (injection voltage).
(3) Standard curve:
standard curve for indole: y ═ 0.000269x +0.0152(r ═ 0.9999);
standard curve for skatole: y 0.000204x +0.0102(r 0.9999).
(4) The results of the experiment are shown in the following table:
table 2 influence of adding microecologics in daily ration on growth performance of broiler chickens
1Each treatment was 6 replicates, with 12 chickens per replicate. On the same line, the superscript letters are different and indicate statistical differences, P<0.05。
The results show that: the daily gain of the microecological preparation group is increased by 11.66 percent compared with that of the control group 1-21d, and the daily gain is increased by 15.00 percent and 8.37 percent respectively compared with that of the control group and the antibiotic group 1-42 d.
Table 3 influence of addition of microecologics in daily ration on oxidation resistance and immunity of broiler chickens
On the same line, the superscript letters are different, which indicates that there is a statistical difference, and P is less than 0.05.
The results show that: the content of the microecological preparation group is respectively increased by 26.90 percent, 23.85 percent and 27.80 percent compared with the GSH-PX, SOD and T-AOC content of a control group, 16.19 percent, 13.46 percent and 15.28 percent compared with the content of the antibiotic group, the content of MDA is reduced by 15.08 percent and 12.00 percent compared with the control group, the content of the microecological preparation group is reduced by 25.68 percent, 20.72 percent and 19.04 percent compared with the TNF-alpha, IL-6 and IL-2 content of the control group, the content of IL-10 is increased by 24.81 percent, the content of TNF-alpha, IL-6 and IL-2 is reduced by 16.11 percent, 13.82 percent and 9.34 percent compared with the TNF-alpha, IL-6 and IL-2 content of the antibiotic group is increased by 19.62 percent. The content of indole and skatole in the microecological preparation group is respectively reduced by 26.9 percent and 26.5 percent compared with the content of indole and skatole in the control group, and the content of indole and skatole in the microecological preparation group is respectively reduced by 26.5 percent and 30.8 percent compared with the content of antibiotic in the antibiotic group.
Therefore, the microecological preparation provided by the invention can be used for remarkably improving the growth performance, the immunity and the oxidation resistance of the broiler chicken and remarkably reducing the content of skatole and indole in the excrement of the broiler chicken.
Test example 2: application effect evaluation of microecologics in laying hens
300 kalanchoe brown laying hens aged 25 weeks are selected and randomly divided into 5 treatment groups, each treatment group is 5 times of repetition, each treatment group is 10 times of repetition, and the test period is 12 weeks.
The experiments were grouped into the following basal diet group, probiotic group (example 1), antibiotic group. The deodorization performance of the microecological preparation is evaluated by odor substances indole and skatole in excrement. The methods for detecting skatole and indole are shown in test example 1.
The test results are given in the following table:
TABLE 4 influence of the Microecological preparation on the malodorous substances in the excrements of egg-laying hens
The result shows that compared with the control group, the addition of the microecological preparation in the daily ration reduces indole by 28.81 percent and reduces skatole by 24.99 percent; compared with the control group, the antibiotic group has increased the content of skatole in feces.
Test example 3 evaluation of application Effect of Microecological preparation in pig farming
The experimental piglets are randomly divided into 3 treatments according to 108 piglets of the Du multiplied by long multiplied by big ternary hybrid 28-day-old weaned piglets which have similar weight, similar nest age and good health condition, wherein each treatment is 8 repetitions (4 repetitions of each male and female), and each repetition is 5 pigs. The three treatments were: (1) corn-soybean meal type basal ration; (2) basal diet + 0.03% chlortetracycline; (3) basal diet + microecologics group. All pigs were fed and drunk freely. The test period ranged from day 1 post-weaning to day 50 post-weaning.
The test result shows that:
compared with a control group, the microecological preparation group (example 2) reduces the diarrhea rate of weaned piglets and obviously improves the daily gain and feed intake of the weaned piglets; meanwhile, compared with a control group, the material weight ratio is reduced by 15%. Compared with the antibiotic group, the growth performance of the microecological group has no obvious difference, but the daily gain is improved by 20 percent compared with the antibiotic group, the material-weight ratio is reduced by 7.3 percent, and the diarrhea rate is reduced by 12 percent. In addition, compared with the control group, the microecological group has 32.9% and 30.6% reduced indole and skatole respectively.
TABLE 5 influence of the Microecological preparations on the growth behavior of piglets and the content of malodorous substances in faeces
On the same line, the superscript letters are different, which indicates that there is a statistical difference, and P is less than 0.05.
Test example 4 evaluation of application Effect of Microecological preparation in Large-Scale layer Chicken Breeding
The popularization test of the deodorizing microecological preparation (examples 1 and 2) is completed in a livestock farm in the Beijing valley region, laying hens aged 183 days are selected as test animals under the non-resistant condition, and the feeding period is 183-one 239-day-old (57 d).
The test was divided into 3 groups of about 10000 layers per group, three groups were a control group (basal diet group), a test group 1 (example 1) and a test group 2 (example 2), respectively, and the test groups were fed with the deodorizing probiotic for a feeding period of 3 months.
Production performance data are collected, the contents of skatole and indole in excrement are detected by using the excrement of the laying hens, and the detection method of skatole and indole is shown in example 1. The specific results are shown in the following table:
TABLE 6 Effect of the Microecological Agents on Large-Scale egg hen Breeding
On the same line, the superscript letters are different, which indicates that there is a statistical difference, and P is less than 0.05.
The results show that: the production performance of the laying hens is obviously improved by adding the microecologics 1 and 2, including the improvement of the laying rate and the daily egg weight, and the reduction of the feed-egg ratio and the death rate; compared with a control group, the content of indole and skatole in the excrement of the microecological preparation 1 is respectively reduced by 30 percent and 47.07 percent; compared with the control group, the content of indole and skatole in the feces of the microecological preparation 2 is respectively reduced by 31.35 percent and 49.15 percent.
It should be understood that the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention, and it will be obvious to those skilled in the art that other variations or modifications may be made on the basis of the above description, and all embodiments may not be exhaustive, and all obvious variations or modifications may be included within the scope of the present invention.
Claims (8)
1. The antibiotic-substituted deodorizing feed microecological preparation is characterized by comprising the following components in parts by weight: coating lactobacillus plantarum 200-300 parts, coating enterococcus faecium 200-300 parts, bacillus subtilis 20-50 parts and fructo-oligosaccharide 300-500 parts;
the preparation method of the coated lactobacillus plantarum or the coated enterococcus faecium specifically comprises the following steps:
(1) preparing a sodium alginate solution with the concentration of 10g/L-20g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:1-1.5 of the calcium carbonate to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 35-38 ℃, adding lactobacillus plantarum or enterococcus faecium seed liquid at the last logarithmic growth stage until the density of the lactobacillus plantarum or the enterococcus faecium is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as an oil phase, wherein the volume percentage of the span 80 is 0.1-0.3%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.1-0.3mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 400 plus 500rpm according to the volume ratio of the water phase to the oil phase of 1:3-5, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:500 plus 600, immobilizing for 10-20min, stopping stirring, adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:10-20, slowly settling the microcapsule, and then sucking away the oil phase;
(6) adding a culture medium suitable for growth of lactobacillus plantarum or enterococcus faecium into a fermentation tank, adding ammonia water in a flowing manner to control the pH value of the fermentation liquid to be 6.2-6.9 under the conditions of 35-38 ℃ and the stirring speed of 50-100rpm, continuously culturing for 8-22h until more than 80% of the space in the microcapsule is filled with the lactobacillus plantarum or the enterococcus faecium, and centrifuging to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution, and fully mixing to perform film forming reaction to obtain double-layer microcapsule bacterial sludge;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-30-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 5-10 parts of glycerol, 5-10 parts of trehalose, 0.5-1 part of skimmed milk powder, 0.5-1 part of sodium acetate and 300 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.5-1 part of xanthan gum, 0.001-0.005 part of sodium carboxymethylcellulose, 0.2-0.8 part of microcrystalline cellulose and 100 parts of water and 200 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) and (3) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 40-55 ℃, controlling the air outlet temperature to be 30-40 ℃, drying until the water content of the sample is 10-20%, spraying an enteric coating material to form a coating, and sieving the dried and coated sample by using a double-layer sieve to obtain a final product with the particle size of 20-60 meshes.
2. The antibiotic-replacement deodorant feed probiotic of claim 1, further comprising yucca extract.
3. The antibiotic-replacement-type deodorant feeding micro-ecological preparation as claimed in claim 2, wherein the micro-ecological preparation comprises 150 portions of yucca extract.
4. The antibiotic-replacement-type deodorant feed microecological preparation according to claim 1, wherein the viable cell count contents of the coated lactobacillus plantarum, the coated enterococcus faecium and the bacillus subtilis are (1-5) x 1010CFU/g、(1-5)×1010CFU/g and (5-10). times.1010CFU/g。
5. The method of preparing the antibiotic-replacement-type deodorant feed probiotic of claim 1, comprising: and uniformly mixing the coated lactobacillus plantarum, the coated enterococcus faecium, the bacillus subtilis and the fructo-oligosaccharide according to a formula ratio to obtain the compound.
6. Use of the antibiotic-replacement-type deodorant feeding microecological preparation according to any one of claims 1 to 4 in the feeding of livestock and poultry or in the preparation of livestock and poultry feed.
7. Use of the antibiotic-replacement deodorant feed probiotic of any one of claims 1 to 4 in any one of:
1) the application in reducing the content of indole and skatole in the livestock and poultry manure;
2) the application in preparing the feed for reducing the content of indole and skatole in the excrement of livestock and poultry;
3) the application in improving the growth performance of livestock and poultry;
4) the application in preparing the feed for improving the growth performance of the livestock and poultry;
5) the application in replacing antibiotics, improving the immunity and the anti-oxidative stress capability of the organism;
6) the application of the compound feed in preparing the feed for replacing antibiotics and improving the immunity and the anti-oxidative stress capability of the organism.
8. The use as claimed in claim 6 or 7, wherein the antibiotic-substituting type deodorant feeding microecological preparation is added in an amount of 0.01-0.2% of the mass of the livestock and poultry feed.
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| CN105567669A (en) * | 2016-01-08 | 2016-05-11 | 魏永刚 | Probiotics microcapsule preparation and preparation method thereof |
| CN108669298A (en) * | 2018-04-26 | 2018-10-19 | 九江牧威利元科技中心（普通合伙） | A kind of feeding probiotic microcapsule and its preparation method and application |
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| CN103275966B (en) * | 2013-06-04 | 2014-10-22 | 国家粮食局科学研究院 | Method for preparing coating microcapsule before microbial fermentation by utilizing common fermentation tank |
| CN104381619A (en) * | 2014-10-21 | 2015-03-04 | 邵素英 | Lactobacillus plantarum capsule for poultry and application thereof |
| CN104814276B (en) * | 2015-05-13 | 2020-02-28 | 济南益邦生物科技有限公司 | Biological deodorant for feeding animals |
| CN105746931A (en) * | 2016-03-01 | 2016-07-13 | 广东省农业科学院动物科学研究所 | Feed additive premix capable of reducing excrement stink of broilers and application of feed additive premix |
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