CN112790287A - Preparation technology and application of environment-friendly comprehensive deodorization biological feed for fattening pigs - Google Patents

Preparation technology and application of environment-friendly comprehensive deodorization biological feed for fattening pigs Download PDF

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CN112790287A
CN112790287A CN202011446043.4A CN202011446043A CN112790287A CN 112790287 A CN112790287 A CN 112790287A CN 202011446043 A CN202011446043 A CN 202011446043A CN 112790287 A CN112790287 A CN 112790287A
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feed
accc
fermentation
culture medium
seeds
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李习龙
孟昆
蒋显仁
杨培龙
蔡红英
梁友
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Jilin Yinong Agricultural Biotechnology Co ltd
Feed Research Institute of Chinese Academy of Agricultural Sciences
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Feed Research Institute of Chinese Academy of Agricultural Sciences
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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
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • 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
    • 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
    • A23V2400/175—Rhamnosus

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Abstract

The invention provides a composite fermented biological feed and a preparation method thereof. The composite fermented biological feed comprises the following raw materials in parts by weight: corn 65-72%, soybean meal 5-12%, rice bran meal 9-12%, bran 7-10%, stone powder 1.0-1.5%, calcium hydrogen phosphate 0-0.2%, lysine 0.5-0.9%, methionine 0-0.1%, threonine 0.1-0.2%, tryptophan 0-0.1%, salt 0.2-0.5%, phytase 0-0.05%, multi-vitamin 0.01-0.03%, and multi-mineral 0.2-0.3%. The 6 fermentation strains adopted were: lactobacillus rhamnosus, lactobacillus plantarum, bacillus licheniformis, bacillus subtilis, saccharomyces cerevisiae and candida utilis. The invention realizes the production of high-quality biological fermentation feed by using conventional complete feed, has balanced nutrition and good palatability, can obviously reduce the concentration of ammonia gas and hydrogen sulfide in the excrement odor of the fattening pigs by more than 60 percent, and improves the air environment of a pigsty; meanwhile, the feed conversion ratio can be reduced by 14.5%, the production performance is effectively improved, and the feed cost is obviously reduced.

Description

Preparation technology and application of environment-friendly comprehensive deodorization biological feed for fattening pigs
Technical Field
The invention relates to a compound fermentation biological feed and a preparation method thereof, belonging to the technical field of biology.
Background
China currently becomes a world big country for live pig breeding. According to the data of the national statistical bureau, the marketing amount of domestic pigs in 2014-2018 is about 7 hundred million, and the annual pigs affected by African swine fever in 2019 are reduced by 21.6 percent compared with the pigs in the last year, but the marketing amount also reaches 5.44 hundred million. The pig breeding brings great economic and social benefits, and increasingly highlights the problems of shortage of protein feed resources and serious pig manure pollution in China. Among the environmental pollution caused by pig manure, odor pollution is an important aspect of livestock manure pollution. The malodorous substance has complex components, and various components in the malodorous substance have strong toxicity, such as ammonia gas and hydrogen sulfide which can poison people and livestock when the concentration is high. As one of the environmental pollution, the stink directly harms the health of people and restricts the sustainable development of the pig industry. Therefore, how to control the offensive odor generated by the pig farm is an environmental problem which is very important for the current breeding enterprises, and is also one of the important subjects for the research of domestic and foreign stockman workers.
The pig manure deodorization method mainly includes a physical method (adsorption method, masking method, etc.), a chemical method (oxidation method, catalytic oxidation method, etc.), a biological method, and the like. The physical and chemical methods have higher equipment requirements and cost, and can bring secondary pollution. The biological deodorization method mainly utilizes microorganisms to degrade, convert and utilize substances causing stink in animal wastes or inhibits the growth of other microorganisms generating stink substances, so the biological deodorization method has wide prospect. At present, the pig manure biological deodorization method is most widely applied to compost, and odor substances in pig manure are eliminated through the microbial fermentation. But the biological compost also has the defects of lagged processing of the odor in the pigsty environment, large occupied compost field, soil and water source pollution, long processing time, different processing conditions, unstable effect and the like. These defects are all the difficult problem of avoiding in the actual production, if can reduce the stink concentration of pig manure itself from the source promptly, then pig house environment stink material can be effectively controlled to effectively improve the stink and to the negative effect of pig breeding, also reduce the cost of pig manure discharge back deodorization processing simultaneously by a wide margin.
Aiming at the problems, the invention adopts the microorganism to ferment the low-protein daily ration to obtain the biological fermentation feed containing the probiotic live bacteria. The low-protein daily ration can reduce environmental pollution, and in the growth fattening stage, the total nitrogen excretion can be reduced by about 10 percent when the protein level of the feed is reduced by 1 percent, the ammonia gas released in the air is reduced by 13 percent, the water intake is reduced by 3 percent, and the volume of the excrement is reduced by 5 percent. The fermented low-protein daily ration is used for feeding pigs, so that the intestinal health of the pigs can be effectively adjusted, the ammonia-producing microbial flora in the pig manure is inhibited, the ammonia nitrogen substances in the pig manure are effectively reduced, the substance basis for forming the odor of the pig manure is reduced from the source, and the odor coefficient of the discharged manure is greatly reduced. Compared with various methods for deodorizing pig manure in the existing pig breeding, the method effectively breaks through the limitation of secondary pollution of a physical and chemical deodorizing method, solves the problems of treatment lag, occupied treatment field, long treatment period, secondary air pollution and the like in the pig manure biological compost deodorizing method, and achieves the effect that the odor substances in the manure are not discharged and removed. Therefore, the application advantages of the strategy of the invention for deodorizing the feces directly without discharging feces are incomparable with the above methods.
On the other hand, various complex microbial odor eliminating microbial inoculums are on the market at present. These microbial inoculum products can be divided into two main categories: the deodorant is directly used for deodorizing the livestock and poultry manure compost, and can be used for composting and reducing the manure odor concentration by directly feeding livestock and poultry. Because the two products have the key function of achieving the deodorization effect through compost, the microbial inoculum compositions of the products contain a large amount of nitrobacteria, denitrifying bacteria, nitrogen fixing fungi, sulfur reducing bacteria or other photosynthetic bacteria. The above strains are not in the probiotic strain category specified in the feed additive variety catalog (No. 2045 bulletin of Ministry of agriculture), so that feeding the non-feeding strains to livestock and poultry has potential high risk to animals, violates the feed management standards and regulations, and is also the main reason why no effective feeding type fecal deodorizing bactericide product is actually used in domestic large livestock and poultry farms. This current situation reflects that the content of pig manure odor substances formed in the intestinal tract cannot be efficiently reduced only by the metabolic digestion of probiotics reaching the intestinal tract, as fed by micro-ecological probiotic agents. Therefore, there is a need for a feed which can significantly reduce the concentration of ammonia and hydrogen sulfide in pig manure and is beneficial to fattening pigs.
Disclosure of Invention
According to the invention, a fermented feed for fattening pigs is produced by combining a feed probiotic strain capable of effectively digesting excrement smelly substances through a large amount of screening verification with a complete feed low-protein daily ration formula for fattening pigs. The breeding test of the fattening pig proves that the biological fermentation feed provided by the invention can obviously reduce the concentration of ammonia gas and hydrogen sulfide in pig manure, provides a healthy and friendly breeding environment for the live pig, and has the additional effect of obviously improving various production performances of the fattening pig. The invention breaks through various limitations of the prior art or products on pig manure deodorization, and provides an effective and feasible way for biological deodorization and air pollution reduction.
Specifically, the present invention provides in a first aspect a complex bacterial species combination comprising: saccharomyces cerevisiae (ACCC 21162), Candida utilis (ACCC 20060), Bacillus subtilis (ACCC 19373), Bacillus licheniformis (ACCC 19372), Lactobacillus rhamnosus (ACCC 05450), and Lactobacillus plantarum (CGMCC 17955).
The present invention provides in a second aspect a method for preparing a complex fermented biological feed, the method comprising:
1) calcium hydrogen phosphate, lysine, methionine, threonine, tryptophan, salt, phytase, multi-vitamin and multi-mineral are premixed, then the premix, corn, bean pulp, rice bran meal, bran and stone powder are put into a mixer to be fully mixed, and a fermentation raw material is prepared; wherein the fermented raw material comprises 65-72 wt% of corn, 5-12 wt% of soybean meal, 9-12 wt% of rice bran meal, 7-10 wt% of bran, 1.0-1.5 wt% of stone powder, 0-0.2 wt% of calcium hydrophosphate, 0.5-0.9 wt% of lysine, 0-0.1 wt% of methionine, 0.1-0.2 wt% of threonine, 0-0.1 wt% of tryptophan, 0.2-0.5 wt% of salt, 0-0.05 wt% of phytase, 0.01-0.03 wt% of multivitamins and 0.2-0.3 wt% of polyminerals, based on 100 wt% of the fermented raw material;
2) expanding and culturing seeds of Saccharomyces cerevisiae (ACCC 21162), Candida utilis (ACCC 20060), Bacillus subtilis (ACCC 19373), Bacillus licheniformis (ACCC 19372), Lactobacillus rhamnosus (ACCC 05450) and Lactobacillus plantarum (CGMCC17955), and mixing;
3) inoculating 0.5-1 wt% of liquid secondary seeds based on the fermentation raw material in the step 1);
3) uniformly mixing the seeds of the fermentation strain in the step 2) with sterile water, and then inoculating the mixture into the fermentation raw material obtained in the step 1, wherein the seeds of the fermentation strain account for 0.5-1% of the total weight of the material, and the water content of the material reaches 30-50%;
4) performing anaerobic fermentation culture on the materials in the step 3) at the temperature of 20-40 ℃ for 3-5 days to obtain a fermentation product, namely the finished feed after the fermentation culture is finished.
In one embodiment, the seed propagation comprises performing propagation of primary and secondary seeds.
In a preferred embodiment, wherein the expanding culture of the primary and secondary seeds is: inoculating single colony of picked strain in 30ml of corresponding liquid culture medium in 100ml triangular flask, and culturing at 20-40 deg.C and 100-Obtaining a primary seed solution after 24 hours; inoculating the first-class seeds according to 1-3% of the volume of the culture medium into 200ml of corresponding liquid culture medium in a 1000ml triangular flask, and culturing at 20-40 ℃ for 18-48h at 100-200r/min to obtain the strain with the concentration of more than or equal to 1 × 109CFU/mL bacterial liquid is the secondary seed liquid.
In one embodiment, the medium to which the species corresponds is a Lactobacillus plantarum-MRS medium; lactobacillus rhamnosus-MRS medium; bacillus subtilis-LB culture medium; bacillus licheniformis-LB culture medium; saccharomyces cerevisiae-YPD medium; candida utilis-YPD medium.
In one embodiment, the primary or secondary seed solution may be one or a mixture of two or more of the strains of the present invention.
In one embodiment, the anaerobic fermentation process is bagged, vacuum sealed, and left at room temperature for 3-5 days.
In a third aspect of the invention, there is provided a complex fermented biological feed prepared according to the preparation method of the invention.
The invention realizes the production of high-quality biological fermentation feed by using conventional complete feed, has balanced nutrition and good palatability, can obviously reduce the ammonia concentration in the excrement odor of the fattening pigs by over 60 percent, and improves the air environment of a pigsty. Meanwhile, the feed conversion ratio can be reduced by 14.5%, the production performance is effectively improved, and the feed cost is obviously reduced.
Detailed Description
The preparation process of the compound feed is as follows.
1. Preparation of fermentation feedstock
The fermentation raw materials comprise: corn 65-72%, soybean meal 5-12%, rice bran meal 9-12%, bran 7-10%, stone powder 1.0-1.5%, calcium hydrogen phosphate 0-0.2%, lysine 0.5-0.9%, methionine 0-0.1%, threonine 0.1-0.2%, tryptophan 0-0.1%, salt 0.2-0.5%, phytase 0-0.05%, multi-vitamin 0.01-0.03%, and multi-mineral 0.2-0.3%. The preparation process of the fermentation raw material comprises the following steps: calcium hydrogen phosphate, lysine, methionine, threonine, tryptophan, salt, phytase, multi-vitamin and multi-mineral are premixed, and then the premixed material, corn, bean pulp, rice bran meal, bran and stone powder are put into a mixer to be fully mixed to prepare the finished feed.
2. Liquid fermentation process of strain
2.1 selection of fermentation strains and media
Bacterium A-plant lactobacillus, MRS culture medium
Bacterium B-Lactobacillus rhamnosus, MRS medium.
And C, bacillus subtilis and LB culture medium.
D, bacillus licheniformis and LB culture medium.
E bacteria-Saccharomyces cerevisiae, YPD medium.
F strain, candida utilis and YPD culture medium.
2.2 activation of Glycerol tube preserving strains
Dissolving the strain glycerol storage tube on ice, dipping bacterial liquid from the storage tube by using inoculating loops respectively, and streaking on a plate containing corresponding solid culture medium until a single bacterial colony is 1-2mm for later use.
2.3 preparation of first and second seed
Selecting single bacterial colony of the strain, inoculating the single bacterial colony into 30ml of corresponding liquid culture medium in a 100ml triangular flask, and culturing at 20-40 ℃ and 100-200r/min for 18-24h to obtain first-stage seed liquid; inoculating the first-class seeds in 2L of corresponding liquid culture medium in a 5L triangular flask according to 1-3% of the volume of the culture medium, and culturing at 20-40 ℃ for 18-48h at 100-200r/min to obtain the strain with the concentration of more than or equal to 1 × 109The CFU/mL bacterial liquid, namely the secondary seed liquid, accounts for 46L-92L, and the seed liquid can be one or more than two mixed bacterial liquids in the above strains.
3. Solid state fermentation process of raw materials
And (3) uniformly mixing the fermentation strain seed liquid prepared in the step (2.3) with sterile water, inoculating the mixture into the raw material mixture obtained in the step (1), enabling the fermentation strain seeds to be 0.5-1% of the total weight of the materials, enabling the water content of the materials to reach 30-50%, carrying out anaerobic fermentation culture at the temperature of 20-40 ℃ for 3-5 days, and obtaining a fermentation product after the fermentation culture is finished.
4. Detection of application effect of animal experiment
4.1 Experimental methods
The feed formula comprises:
control group: daily ration for fattening pig with conventional protein
Test groups: 3, obtaining the low-protein daily ration fermented feed
200 fattening pigs with the average body weight of 93.7 kilograms are selected for the test, the fattening pigs are divided into 2 treatment groups according to the body weights of a control group and a test group, the control group is fed with conventional fattening pig feed, and the test group is fed with biological fermented feed. The control group pigs are fed in the same pigsty, the test group pigs are fed in an adjacent pigsty, 10 repetitions (pigsty) are arranged in each pigsty, 10 fattening pigs are arranged in each repetition (pigsty), and the test period is 28 days. The fattening pigs are raised in the cement crack ground fence and are repeatedly and uniformly distributed in a room with controlled temperature. The area of each column is 10 square meters, a fan connected with temperature control is arranged for keeping ventilation, and the temperature in the house is kept at 23-25 ℃. All fattening pigs eat and drink water freely.
4.2 analysis of results
And (3) measuring the production performance: on days 0 and 28, the weight was in columns. The charge was recorded for each replicate day and the balance weighed on test day 28. And calculating the average daily gain, the average feed intake and the material weight ratio.
Ammonia (NH) in air in pigsty3) And hydrogen sulfide (H)2S) measurement: days 2, 5, 14 and 28 of the test period, 06: 00 and 18: 00, collecting NH of pigsty3And H2And S. The total length of the pigsty is 20 meters, the intervals between sampling points are 5 meters, 4 sampling points are arranged totally, and each point is sampled and arranged in 2 parallels. NH (NH)3And H2The S detector used a portable MS400 ammonia/hydrogen sulfide detector.
Compared with the prior art, the invention has the following advantages:
the method of the invention does not need to establish and purchase equipment required by a chemical deodorization method of a physical deodorization method at a large investment cost; the method of the invention avoids the defect that the active ingredients and beneficial microorganisms of the raw materials are easy to damage by the traditional processing technologies such as high-temperature granulation, puffing, curing and the like in the traditional feed production. Specifically, the method comprises the following steps:
1) according to the invention, the low-protein daily ration raw material is fermented by the feed probiotics, so that the high-efficiency environment-friendly fermented feed for the fattening pigs can be obtained. The ammonia gas removal rate of the fermented feed obtained by the invention can reach 60-80% after feeding fattening pig house.
2) The feed conversion ratio of the fermented feed obtained by the invention after being fed to fattening pigs is reduced by 14.5 percent compared with that of a control group, and other production performance indexes
3) The method has simple and easily controlled process, can better meet the practical conditions of industrial production, and is easy to realize industrialization.
4) The invention can reduce two percentage points of the crude protein of the feed, save the use amount of the soybean meal and relieve the problem of shortage of protein feed resources.
Examples
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.
In the following examples, YPD medium, MRS medium and LB medium were used in accordance with the conventional recipes in the art. The saccharomyces cerevisiae (ACCC 21162), the candida utilis (ACCC 20060), the bacillus subtilis (ACCC 19373), the bacillus licheniformis (ACCC 19372) and the lactobacillus rhamnosus (ACCC 05450) used by the invention are all purchased from China agricultural microbial culture collection management center (ACCC); the lactobacillus plantarum (CGMCC17955) has a preservation number of CGMCC17955, is preserved in China general microbiological culture Collection center (CGMCC) in 2019, 6 months and 18 days by the inventor, and has the preservation center address: the institute of microbiology, national academy of sciences, west road No. 1, north Chen, Chaozhou, Chaoyang.
Example 1 preparation of composite fermented feed
A preparation method of a composite fermented biological feed comprises the following steps:
1) preparation of low-protein daily ration raw material for fattening pigs
Weighing the following raw materials in proportion: corn 65-72%, soybean meal 5-12%, rice bran meal 9-12%, bran 7-10%, stone powder 1.0-1.5%, calcium hydrogen phosphate 0-0.2%, lysine 0.5-0.9%, methionine 0-0.1%, threonine 0.1-0.2%, tryptophan 0-0.1%, salt 0.2-0.5%, phytase 0-0.05%, multi-vitamin 0.01-0.03%, and multi-mineral 0.2-0.3%. The fermented feed raw material is obtained by premixing small materials (calcium hydrophosphate, lysine, methionine, threonine, tryptophan, salt, phytase, multi-vitamin and multi-mineral) and then fully mixing the premix with large materials (corn, bean pulp, rice bran meal, bran and stone powder).
2) Preparation of fermented seed liquid
Preparing a bacillus licheniformis and bacillus subtilis fermentation seed solution:
single colony separation: streaking the strains of the bacillus licheniformis and the bacillus subtilis glycerin tube preserved at low temperature on a culture dish containing an LB solid culture medium, and culturing in a constant temperature incubator at 30 ℃ for 18 hours to obtain single colony seeds; culturing single colony in 30ml LB liquid culture medium in 100ml triangular flask at 30 deg.C and 200rpm for 12 hr to obtain bacteria concentration of 1 × 10 or more8CFU/mL of first-order seed liquid; inoculating 1% of the first-class seed solution into 200ml glucose beef extract peptone medium (or 2L glucose beef extract peptone medium in 5L triangular flask) in 1L triangular flask, and culturing at 30 deg.C and 200rpm for 18 hr to obtain strain concentration of 1 × 109CFU/mL of secondary seed liquid;
preparing lactobacillus rhamnosus and lactobacillus plantarum fermented seed liquid:
single colony separation: marking strains of lactobacillus rhamnosus and lactobacillus plantarum glycerinum preservation pipe which are preserved at low temperature on a culture dish containing MRS solid culture medium, and performing static culture in a constant temperature incubator at 30 ℃ for 30 hours to obtain single colony seeds; inoculating single colony in 30ml MRS liquid culture medium in 100ml triangular flask, and standing at 30 deg.C in constant temperature incubator for 30 hr to obtain bacteria concentration of 1 × 10 or more8CFU/mL of first-order seed liquid; inoculating the first-stage seed solution into 1L triangular flask 200ml MRS culture medium (or 5L triangular flask 2L MRS culture medium), and standing at 30 deg.C for 48 hr in a constant temperature incubator to obtain a bacterial concentration of 1 × 10 or more9CFU/mL of secondary seed liquid;
preparing a saccharomyces cerevisiae and candida utilis fermentation seed liquid:
single colony separation: the low-temperature preserved saccharomyces cerevisiae is subjected to fermentation,Scribing a candida utilis glycerinum tube storage tube strain on a culture dish containing a YPD solid culture medium, and performing static culture in a constant-temperature incubator at 30 ℃ for 36 hours to obtain a single colony seed; placing single colony in 30ml YPD liquid culture medium in 100ml triangular flask, and standing at 30 deg.C in constant temperature incubator for 36 hr to obtain bacteria concentration of 1 × 10 or more8CFU/mL of first-order seed liquid; inoculating 1% of the first-stage seed solution into 200ml YPD medium (5L of 2L YPD medium) in 1L triangular flask, and standing at 30 deg.C for 48 hr in constant temperature incubator to obtain the final product with a bacterial concentration of 1 × 10 or more9CFU/mL of secondary seed solution.
Mixing secondary seed liquids of bacillus licheniformis, bacillus subtilis, lactobacillus rhamnosus, lactobacillus plantarum, saccharomyces cerevisiae and candida utilis to obtain a fermented seed liquid.
3) Low-protein daily ration solid fermentation for fattening pigs
The beneficial microbial flora comprises the following components in parts by weight: 2 parts of bacillus subtilis seed liquid, 2 parts of bacillus licheniformis seed liquid, 2 parts of lactobacillus rhamnosus, 2 parts of lactobacillus plantarum, 1 part of saccharomyces cerevisiae and 1 part of candida utilis; the weight ratio of the fermentation raw material to the seed liquid is 1000 parts of the fermentation raw material, and the seed liquid is 10-40 parts according to different combinations of strains. Fully mixing 280 plus 800 parts by weight of sterile water with the seed liquid, adding the fermentation raw materials, uniformly stirring, and filling 50kg of sterile water into a special anaerobic fermentation bag with a breather valve, wherein the air in the bag is normally reserved and is not extruded. Placing the charged fermented feed in an anaerobic fermentation culture at 20-40 ℃ for 3-5 days, and obtaining the low-protein daily ration fermented feed for fattening pigs after the fermentation culture is finished.
Example 2 fattening pig raising with fermented biological feed
200 fattening pigs with the average weight of 93.7 kilograms and healthy physique are selected for the test, the fattening pigs are randomly divided into a control group and 100 test groups according to the principle of similar body weight, the fattening pigs in the control group are fed with conventional protein feed, the test groups are fed with low-protein fermented feed, each treatment is carried out in a pigsty with the same conditions and divided into 10 rows, each row is marked as 1 repetition, and each 10 rows are repeated to carry out the feeding test for 28 days. The fattening pigs are raised in the cement crack ground fence and are repeatedly and uniformly distributed in a room with controlled temperature. The area of each column is 10 square meters, a fan connected with temperature control is arranged for keeping ventilation, and the temperature in the house is kept at 22-25 ℃. All fattening pigs eat and drink water freely. And observing the feeding, health and activity conditions of the test pigs every day, and making a record of the date and the weight. The pigs are marked after the sick pigs are found, the fattened pigs with serious diseases need to be eliminated, and the eliminated pigs or dead pigs and the residual materials need to be weighed and recorded.
Example 3 Effect of fermented biological feed on Ammonia and Hydrogen sulfide concentration in fattening pig House
Determination of ammonia gas and hydrogen sulfide in piggery air: days 2, 5, 14 and 28 of the test period, 6: 00 and 18: 00, collecting NH of pigsty3And H2And S. The window is closed in the evening before the measurement day, and the window is opened for ventilation after the air sample is collected in the morning. The total length of the pigsty is 20 meters, the intervals between sampling points are 5 meters, 4 sampling points are arranged totally, and each point is sampled and arranged in 2 parallels. NH (NH)3And H2The S detector used a portable MS400 ammonia/hydrogen sulfide detector.
TABLE 1 fermented feed vs. Ammonia and Hydrogen sulphide concentrations (mg/m) in the pig house3) Influence of (2)
Figure BDA0002824427450000111
a,bThe different shoulder marks in the same row represent significant differences, P<0.05
*,
Figure BDA0002824427450000112
The difference is marked by different shoulder marks in the same column, P<0.05
According to the test results, after 5 days of the test, the concentrations of ammonia and hydrogen sulfide in the test group are obviously lower than those in the control group; the concentrations of ammonia gas and hydrogen sulfide in the test group are gradually reduced along with the development of the test, the concentrations of ammonia gas and hydrogen sulfide after the fermentation material is fed for 14 days are basically stable, and the concentrations are respectively reduced by 70 percent and 63 percent compared with the concentrations in the 2 nd day.
Example 4 Effect of fermented biological feed on growth of fattening pigs
On days 0 and 28, the weight was in columns. The charge was recorded for each replicate day and the balance weighed on test day 28. And calculating the average daily gain, the average feed intake and the material weight ratio.
Table 2 shows the effect of fermented feed on growth performance of the finishing pigs, the fermented group fed with low protein diet increased the body weight by 3.8% (P ═ 0.088), increased the average daily gain by 16.8% (P ═ 0.039), and decreased the feed conversion by 14.5% (P ═ 0.054) compared to the control group.
TABLE 2 influence of Low-protein daily ration fermented feed on growth performance of fattening pig
Figure BDA0002824427450000121
*Feed intake was calculated on an air-dry basis
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (8)

1. A composite bacterial combination, comprising: saccharomyces cerevisiae (ACCC 21162), Candida utilis (ACCC 20060), Bacillus subtilis (ACCC 19373), Bacillus licheniformis (ACCC 19372), Lactobacillus rhamnosus (ACCC 05450), and Lactobacillus plantarum (CGMCC 17955).
2. A method of preparing a composite fermented biological feed, the method comprising:
1) calcium hydrogen phosphate, lysine, methionine, threonine, tryptophan, salt, phytase, multi-vitamin and multi-mineral are premixed, then the premix, corn, bean pulp, rice bran meal, bran and stone powder are put into a mixer to be fully mixed, and a fermentation raw material is prepared; wherein the fermented raw material comprises 65-72 wt% of corn, 5-12 wt% of soybean meal, 9-12 wt% of rice bran meal, 7-10 wt% of bran, 1.0-1.5 wt% of stone powder, 0-0.2 wt% of calcium hydrophosphate, 0.5-0.9 wt% of lysine, 0-0.1 wt% of methionine, 0.1-0.2 wt% of threonine, 0-0.1 wt% of tryptophan, 0.2-0.5 wt% of salt, 0-0.05 wt% of phytase, 0.01-0.03 wt% of multivitamins and 0.2-0.3 wt% of polyminerals, based on 100 wt% of the fermented raw material;
2) expanding and culturing seeds of Saccharomyces cerevisiae (ACCC 21162), Candida utilis (ACCC 20060), Bacillus subtilis (ACCC 19373), Bacillus licheniformis (ACCC 19372), Lactobacillus rhamnosus (ACCC 05450) and Lactobacillus plantarum (CGMCC17955), and mixing;
3) uniformly mixing the seeds of the fermentation strain in the step 2) with sterile water, and then inoculating the mixture into the fermentation raw material obtained in the step 1), so that the seeds of the fermentation strain account for 0.5-1% of the total weight of the material, and the water content of the material reaches 30-50%;
4) performing anaerobic fermentation culture on the materials in the step 3) at the temperature of 20-40 ℃ for 3-5 days to obtain a fermentation product, namely the finished feed after the fermentation culture is finished.
3. The method of claim 2, wherein said expanding of seeds comprises expanding of primary and secondary seeds.
4. The method of claim 3, wherein the expanding culture of the primary and secondary seeds is: selecting single bacterial colony of the strain, inoculating the single bacterial colony into 30ml of corresponding liquid culture medium in a 100ml triangular flask, and culturing at 20-40 ℃ and 100-200r/min for 18-24h to obtain first-stage seed liquid; inoculating the first-class seeds according to 1-3% of the volume of the culture medium into 200ml of corresponding liquid culture medium in a 1L triangular flask or 2L of corresponding liquid culture medium in a 5L triangular flask, and culturing at 20-40 ℃ and 100-200r/min for 18-48h to obtain the strain with the concentration of more than or equal to 1 × 109CFU/mL bacterial liquid is the secondary seed liquid.
5. The production method according to any one of claims 2 to 4, wherein the culture medium corresponding to the species is Lactobacillus plantarum, MRS medium; lactobacillus rhamnosus, MRS medium; bacillus subtilis and LB culture medium; bacillus licheniformis and LB culture medium; saccharomyces cerevisiae, YPD medium; candida utilis, YPD medium.
6. The method according to any one of claims 2 to 4, wherein the seed liquid is one or a mixture of two or more of the above bacterial species.
7. The method of claim 2, wherein the anaerobic fermentation process is performed in a bag, sealed under vacuum, and allowed to stand at room temperature for 3-5 days.
8. A complex fermented biological feed produced by the production method according to any one of claims 2 to 7.
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