CN106260540B - Biological feed for creep feed and creep feed - Google Patents

Biological feed for creep feed and creep feed Download PDF

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CN106260540B
CN106260540B CN201510503967.6A CN201510503967A CN106260540B CN 106260540 B CN106260540 B CN 106260540B CN 201510503967 A CN201510503967 A CN 201510503967A CN 106260540 B CN106260540 B CN 106260540B
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feed
creep
biological
bacillus
fish meal
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CN106260540A (en
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张宗伟
刘明忠
胡忠宏
张静
沈美英
孙建晨
孟庆军
姜倩倩
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Harbin Dabei Farming And Animal Husbandry Technology Co ltd
JILIN DABEINONG AGRICULTURE ANIMAL HUSBANDRY TECHNOLOGY CO LTD
Shenyang Yingda Technology Development Co ltd
Beijing Dabeinong Biotechnology Co Ltd
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Harbin Dabei Farming And Animal Husbandry Technology Co ltd
JILIN DABEINONG AGRICULTURE ANIMAL HUSBANDRY TECHNOLOGY CO LTD
Shenyang Yingda Technology Development Co ltd
Beijing Dabeinong Technology Group Co Ltd
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Abstract

The invention discloses a biological feed for creep feed, which is prepared by hermetically fermenting feed raw materials with the water content of 20-50% at the temperature of 15-35 ℃ for 3-10 days, wherein the number of viable bacteria contained in a fermentation product is more than or equal to 1 multiplied by 109The feed additive is characterized in that the feed additive contains organic acid more than or equal to 2 percent per gram, and the feed raw materials contain the following components in parts by weight: 0.05-0.2g/kg of microzyme, 0.05-0.2g/kg of lactobacillus, 0.05-0.2g/kg of bacillus, 800g/kg of puffed peeled soybean meal, 50-100g/kg of corn flour and 300g/kg of wheat bran. The 5-10% of the invention is used for the complete feed production of the creep feed, can replace fish meal, has balanced nutrition and high digestibility of the prepared creep feed, reduces the application of antibiotics, can effectively reduce the diarrhea rate and the death rate of piglets, and has positive significance for the daily gain of the piglets and the improvement of the environment.

Description

Biological feed for creep feed and creep feed
Technical Field
The invention belongs to the field of animal feed, and particularly relates to biological feed for creep feed and creep feed added with the biological feed.
Background
The biological feed, namely the fermented feed, is a hotspot for research and development of livestock workers at home and abroad at present, and shows good prospects in various aspects such as reasonable application of feed raw material resources in China, enhancement of feed and livestock product safety production, promotion of livestock farm emission reduction, reduction of environmental pollution and the like, so that the development of the biological feed has great strategic significance for the livestock industry at home and abroad. The study of biological feeds abroad was said to have started from the 80 s of the 20 th century, but was actually developed after the 90 s of the 20 th century. The research of China on biological feed starts late, beginning at the time of recent years. The research and development of the bioactive feed additive for foreign organisms are rapidly advanced, the overall level in the industry is higher than that in China at present, and the level of foreign researchers is relatively higher particularly in the aspect of physiological and biochemical research of the bioactive feed additive. The research of biological feed in China generally has the problems of incomplete production technology level, few varieties, unmatched application technology and the like except a few traditional fermented products. However, with the development and progress of the industry, the development of biological feeds in China is in a situation of high-speed development no matter theoretical research or practical application. With the extensive application of modern biotechnology, the breeding of zymocyte of enzyme preparation, the improvement of fermentation process and the improvement of production level have caught up to the corresponding level of developed countries even in some fields, for example, the production technology of high temperature resistant phytase naked enzyme has gone ahead of the world. At present, research and development on probiotics, prebiotics, natural plant extracts and the like serving as antibiotic substitutes are also vigorous, and certain achievement is achieved at present. With the further improvement of the safety concern of livestock products in the whole society, the research and application of fermentation feed raw materials, even complete feed fermentation, become a great trend of industrial development. However, there are many problems in the application techniques such as those of probiotics, those of antibacterial peptides, and the like. The fermented feed is a feed raw material which is characterized in that under the condition of manual control, the microorganism decomposes or converts anti-nutritional factors in plant, animal and mineral substances through the metabolic activity of the microorganism, so that nutrients which can be better eaten, digested and absorbed by livestock and poultry are generated, and the fermented feed has the functions of improving the palatability of the feed, stimulating the eating of the livestock and poultry and improving the digestibility and utilization rate of the nutrient substances in the feed, is beneficial to the intestinal health of the livestock and poultry, enhancing the immunity, degrading toxic substances in the feed, generating growth promoting factors, and effectively inhibiting the propagation and growth of escherichia coli, salmonella, some pathogenic bacteria and pathogens in the digestive tract.
The creep feed is a feed product in the piglet stage, and is a special feed product containing high nutritional ingredients in the stage from the birth to 4 weeks of age of the piglet. The high-quality creep feed at least has the advantages of being suitable for digestion and nutrient absorption; the palatability is good; the nutrition is comprehensive and balanced; the incidence of diseases of the pigs is low; has no weaning stress phenomenon. The conventional creep feed usually contains a plurality of protein source feeds and has high cost, so that the creep feed with low cost, good palatability and excellent creep effect is urgently needed.
Disclosure of Invention
In order to solve the problems, the invention aims to provide a biological feed for a creep feed and the creep feed.
The biological feed for creep feed provided by the invention is prepared by fermenting feed raw materials with the water content of 20-50% at the temperature of 15-35 ℃ for 3-10 days, wherein the number of viable bacteria in the active feed is more than or equal to 1 multiplied by 109The feed additive is characterized in that the feed additive contains organic acid more than or equal to 2 percent per gram, and the feed raw materials contain the following components in parts by weight: 0.05-0.2g/kg of microzyme, 0.05-0.2g/kg of lactobacillus, 0.05-0.2g/kg of bacillus, 800g/kg of puffed peeled soybean meal, 50-100g/kg of corn flour and 300g/kg of wheat bran.
In one embodiment of the invention, the biological feed for the creep feed is prepared by hermetically fermenting 72 feed raw materials with the water content of 30% at the temperature of 30 ℃, wherein the feed raw materials comprise the following components in parts by weight: 0.1g/kg of microzyme, 0.1g/kg of lactobacillus, 0.1g/kg of bacillus, 525g/kg of puffed peeled soybean meal, 75g/kg of corn flour, 225g/kg of wheat bran and 175g/kg of water.
Wherein the viable count of the yeast, the lactic acid bacteria and the bacillus is 20-30 hundred million/g.
Wherein the yeast is Saccharomyces cerevisiae CGMCC No.6560 (disclosed in CN201210430095.1), the bacillus is Bacillus subtilis CGMCC No.8148, and the lactic acid bacteria is Lactobacillus plantarum CGMCC No. 8150.
Wherein the puffed peeled soybean meal is crushed before fermentation, the crushing sieve sheet is 0.8 multiplied by 0.8mm, the corn crushing sieve sheet is 0.8 multiplied by 0.8mm, and the wheat bran crushing sieve sheet is 0.8 multiplied by 0.8 mm.
The invention also provides a pig creep feed, wherein the complete feed of the creep feed uses biological feed accounting for 5-10% of the complete feed by mass percent to replace fish meal as a protein raw material.
In a preferred embodiment of the invention, the pig creep feed comprises the following components in parts by weight: 30% of corn, 20% of puffed corn, 17% of soybean meal, 3% of cane sugar, 3% of glucose, 6% of biological feed, 8% of fermented soybean meal, 6% of plasma protein powder, 2% of calcium dihydrogen phosphate, 2% of stone powder, 1% of feed salt, 0.3% of 98.5% of lysine, 0.2% of methionine, 0.01% of tryptophan, 0.5% of acidifying agent and 1.5% of porket premix.
In the invention, the Bacillus subtilis and Lactobacillus plantarum have good gastric acid resistance, intestinal juice resistance, cholate resistance (high temperature resistance of Bacillus) and other stress resistance and probiotic functions of acid production, enzyme production, pathogenic bacterium inhibition and the like, are obtained by separating and breeding from healthy animal intestinal tracts for an applicant, and are preserved in China general microbiological culture Collection center (CGMCC) in 2013, 9, 11, and the addresses are as follows: the microbial research institute of the national academy of sciences No. 3, Xilu No. 1, Beijing, Chaoyang, the preservation numbers are respectively: CGMCC No.8148 and CGMCC No. 8150.
The selected Bacillus subtilis CGMCC No.8148 is treated in artificial simulated gastric juice (pH2.0-4.0) for 3 hours, and the survival rate is more than 65.8 percent; the intestinal juice is treated in the artificial simulated intestinal juice for 3 hours, and the survival rate is 74.7 percent; treating the pig gall with a pig gall salt solution (the concentration is 0.3-3 g/kg) for 24 hours, wherein the survival rate is over 51.2 percent; treating at 85-100 ℃ for 3min, wherein the survival rate is more than 11.2%; the antibiotic has drug resistance to 3 antibiotics of streptomycin, polymyxin B and lincomycin; can produce a certain amount of protease, lipase, amylase, cellulase and xylanase, and has better effect than the bacillus subtilis CGMCC No.4628 used in the application No. 201110116641X.
The lactobacillus plantarum CGMCC No.8150 bred by the invention is treated in artificial simulated gastric juice (pH2.0-4.0) for 3 hours, and the survival rate is above 31.3 percent; the intestinal juice is treated in the artificial simulated intestinal juice for 3 hours, and the survival rate is 33.0 percent; treating the pig gall with a pig gall salt solution (the concentration is 0.3-3 g/kg) for 24 hours, wherein the survival rate is more than 3.7%; the antibiotic has drug resistance to 7 antibiotics of cefepime, gentamicin, spectinomycin, streptomycin, polymyxin B, ciprofloxacin and vancomycin; has good inhibition effect on Escherichia coli K88 and K99, and the diameters of the inhibition zones are 18.60mm and 15.43mm respectively.
The technical scheme of the invention has the following advantages:
1. the lactobacillus in the invention has strong activity and can tolerate high temperature and acid environment, thus inhibiting the growth of bacteria in the fermentation process, producing a large amount of organic acid and small peptide, improving the palatability of piglets, allowing a large amount of live bacteria to enter the intestinal tract of the piglets, playing an antibacterial function in the gastrointestinal environment of the piglets, improving the immunity of the piglets, reducing the diarrhea rate of the piglets and improving the survival rate of the piglets.
2. The yeast in the invention can generate a large amount of B vitamins and small peptides, and improve the immunocompetence of piglets.
3. The bacillus in the invention can produce a large amount of protease, amylase and the like, and improves the digestibility of the piglet on nutrients after entering the gastrointestinal tract of the piglet.
4. The anti-nutritional factors in the invention are greatly reduced, and the waste of feed is reduced.
5. The protein solubility of the invention is improved, and the digestion utilization rate of the true protein is improved.
Detailed Description
The present invention will be more specifically illustrated by the following examples, but it should be understood that the examples are only for illustrating the present invention and do not limit the scope of the present invention in any way.
Example 1 isolation, screening, selection and identification of superior strains
1) And (3) strain propagation: collecting a fresh intestinal content sample of a healthy pig, inoculating the sample into an LB liquid culture medium, and culturing and propagating the inoculated culture medium in a 37 ℃ incubator for 48 hours;
2) separation and purification of bacillus subtilis: heating the expanded propagation culture obtained in the step 1) in a water bath kettle at 80 ℃ for 20min, then taking out a treatment solution, diluting and coating the treatment solution in an LB (LB) culture medium, culturing the culture medium in a constant-temperature incubator at 37 ℃ for 24h, selecting each bacterial colony with a characteristic shape from the culture medium, performing streak culture and separation until a pure bacterial colony is obtained, performing gram staining microscopic examination on the result, inoculating the separated suspected pure bacterial colony into a test tube inclined plane, expanding propagation, and storing the test tube inclined plane in a refrigerator at 4 ℃ for later use;
colony morphology: the bacterial colony is round, the surface is smooth, the bacterial colony is milky white, and the size of the bacterial colony is 1-3 mm; the cells are round or oval, the diameter is 0.5-1.0 um, most of the cells are short-chain or paired, and gram staining is positive.
The separated strain is identified to be bacillus subtilis and is preserved in China general microbiological culture Collection center (CGMCC) in 2013, 9 and 11 months, and the address is as follows: the collection number of the microorganism research institute of China academy of sciences No. 3, Xilu No. 1 of Beijing, Chaoyang, is CGMCC No. 8148.
3) And (3) separating and purifying lactobacillus plantarum: diluting the enrichment culture medium in the step 1) in a multiple ratio, coating the enrichment culture medium in an MRS culture medium, culturing for 36-48 h at 37 ℃, selecting each bacterial colony with a characteristic shape from the enrichment culture medium, performing streak culture and separation until a pure bacterial colony is obtained, performing gram staining microscopy, inoculating the separated suspected pure bacterial colony into a test tube inclined plane for proliferation, and then storing in a refrigerator at 4 ℃ for later use;
colony morphology: the bacterial colony is soft and moist, cheese color, luster, flat or slightly convex, and regular edge; the cells are spherical or ovoid, 5-10 μm in diameter, and are budded in a propagation manner.
The separated strain is lactobacillus plantarum and is preserved in China general microbiological culture Collection center (CGMCC) in 2013, 9 and 11 months, and the address is as follows: the collection number of the microorganism research institute of China academy of sciences No. 3, Xilu No. 1 of Beijing, Chaoyang, is CGMCC No. 8150.
EXAMPLE 22 determination of stress resistance and biological Properties of Excellent strains
2 strains: the biological performance and stress resistance of the bacillus subtilis CGMCC No.8148 and the lactobacillus plantarum CGMCC No.8150 are determined:
1) preparation of Bacillus subtilis CGMCC No.8148 and Bacillus licheniformis CGMCC No.2383 cultures: inoculating slant strains preserved in a refrigerator into an LB seed culture medium for activation, and culturing at 35 ℃ and 100rpm for 16h to obtain the strain;
2) preparation of Lactobacillus plantarum CGMCC No.8150 culture: inoculating slant strains preserved in a refrigerator into an MRS seed culture medium for activation, and culturing at 30 ℃ and 150rpm for 16h to obtain the strain;
3) and (3) acid resistance measurement: inoculating the culture with 10% inoculum size into artificial simulated gastric juice with pH of 2.0, 3.0, 4.0, adjusting pH to 2.0, 3.0, 4.0, counting for 0h as control, sampling for 3h, diluting with 0.85% normal saline in 10 times series, counting viable bacteria on plate, and calculating survival rate.
The invention breeds 2 strains: the results of the survival rates of the bacillus subtilis CGMCC No.8148 and the lactobacillus plantarum CGMCC No.8150 in gastric acid are shown in table 1.
Preparing artificial simulated gastric juice: measuring 16.4 mL of 9.5-10.5% concentrated hydrochloric acid, adding distilled water to 1000 mL to obtain basic artificial gastric juice, adjusting pH to 2.0, 3.0 and 4.0 with hydrochloric acid or sodium hydroxide, performing steam sterilization at 121 ℃ for 15 minutes, adding pepsin according to 1% of the addition amount, passing through a 0.22 mu m sterile filter membrane to obtain the artificial gastric juice, and subpackaging 5mL (4.5mL) in test tubes.
Survival rate of table 12 strains in simulated gastric fluid
Figure BDA0000781438290000061
4) And (3) intestinal juice resistance determination: inoculating the above prepared culture in artificial simulated intestinal fluid at an inoculation amount of 5%, counting for 0h as control, sampling for 3h, diluting with 0.85% physiological saline in 10 times, performing flat plate viable bacteria counting, and calculating survival rate.
The invention breeds 2 strains: the results of the survival rates of the bacillus subtilis CGMCC No.8148, the lactobacillus plantarum CGMCC No.8150 and the laboratory-preserved bacillus licheniformis CGMCC No.2383 in intestinal juice are shown in table 2.
Preparing artificial simulated intestinal juice: weighing 6.8g, adding distilled water to 1000 mL to obtain basic artificial intestinal juice, steam sterilizing at 121 deg.C for 15 min, adding trypsin according to 1% addition amount, passing through 0.22 μm sterile filter membrane to obtain artificial gastric juice, and packaging 5mL (4.5mL) in test tubes.
Survival rate of strain in artificial simulated intestinal fluid of Table 22
Survival rate/%)
Time of treatment 3h
Bacillus subtilis CGMCC No.8148 74.7
Lactobacillus plantarum CGMCC No.8150 33.0
5) And (3) bile salt resistance determination: the 3 strain cultures prepared above are respectively inoculated in LB solution of pig bile salt with different concentrations of 0.03%, 0.1%, 0.2% and 0.3% according to the inoculum size of 10%, counting is carried out for 0h as a contrast, sampling is carried out for 24h, the counting is carried out by 10 times serial dilution with 0.85% normal saline, the counting of flat plate viable bacteria is carried out, and the survival rate is calculated.
The 2 strains bred by the invention are as follows: the results of the survival rates of the bacillus subtilis CGMCC No.8148, the lactobacillus plantarum CGMCC No.8150 and the laboratory-preserved bacillus licheniformis CGMCC No.2383 in the bile salts are shown in Table 3.
Preparation of bile salt: adding pig bile salt with different concentrations of 0.03%, 0.1%, 0.2% and 0.3% into prepared LB liquid culture medium, steam sterilizing at 121 deg.C for 30 min, and sub-packaging 5mL pig bile salt solution with different concentrations of 0.03%, 0.1%, 0.2% and 0.3% into test tubes under aseptic condition.
Survival rate of Table 32 strains treated in pig bile salt of different concentrations for 24h
Figure BDA0000781438290000071
6) High-temperature resistance determination of bacillus: the 2 Bacillus cultures were treated in water bath at 85 deg.C, 90 deg.C, 95 deg.C, and 100 deg.C for 3min, respectively, and the survival rate was calculated.
TABLE 4 survival/% of Bacillus subtilis treated at different high temperatures for 3min
Figure BDA0000781438290000072
Figure BDA0000781438290000081
7) And (3) drug resistance determination: adopts a drug sensitive paper sheet method.
Table 52 strains of drug resistance determination results
Figure BDA0000781438290000082
Figure BDA0000781438290000091
8) Enzyme production assay of bacillus:
a flat transparent ring method is adopted. Respectively dotting separated bacillus on a cellulase culture medium, a protease culture medium, an amylase culture medium, a xylanase culture medium and a lipase culture medium, setting 3 times for each treatment, culturing for 36H at 37 ℃, observing the existence and the size of a transparent ring of a flat plate, if a transparent colony or a fuzzy ring is formed around the colony, proving that the bacillus can produce corresponding enzyme, respectively measuring the diameter (H) of the transparent ring and the diameter (C) of the colony by using vernier calipers, and calculating the ratio (H/C) of the two. The ratio of H/C of the above 5 enzymes was determined simultaneously by using Bacillus subtilis CGMCC No.4628 used in application No. 201110116641X as a control.
TABLE 6 enzyme production assay results for Bacillus subtilis strains
Figure BDA0000781438290000092
Figure BDA0000781438290000101
9) Acid production measurement:
adopting a calcium carbonate plate to measure the yield of the organic acid of the lactobacillus plantarum CGMCC No. 8150;
TABLE 7 acid production assay results for Lactobacillus plantarum strains
Figure BDA0000781438290000102
10) And (3) bacteriostatic test: and (3) carrying out bacteriostatic determination on common pathogenic bacteria by adopting an oxford cup method.
a. Two pathogens were activated in a test tube containing 10mL of nutrient broth: culturing at 37 deg.C for 20 hr under K88 and K99;
b. preparing a double-layer flat plate: taking a flat plate with the diameter of 90mm, injecting 15-20 mL of sterilized nutrient agar, horizontally placing the flat plate to be solidified to be used as a bottom layer, uniformly mixing the nutrient agar (cooled to about 50 ℃) with a proper amount of an indicator bacterium solution (about 50mL of culture medium and 8 mL) cultured at 37 ℃ for 24 hours, sucking 10mL of the indicator bacterium solution, pouring the indicator bacterium solution on the bottom layer culture medium, horizontally placing the indicator bacterium solution to be solidified to be used as a bacterium layer;
c. adding a sample: the sterilized oxford cup was gripped with sterile forceps, the lid opened, and placed on the culture medium. The same amount of fermentation broth supernatant (about 200uL) was topped up in an oxford cup, 2 replicates per sample. Carefully placing the double dishes with the samples in a constant temperature box at 37 ℃, culturing for 16-18 h, and taking out to measure the size of the bacteriostatic zone.
TABLE 8 bacteriostatic results of Lactobacillus plantarum on common pathogenic bacteria
Figure BDA0000781438290000103
EXAMPLE 3 screening of fermentation temperature and fermentation time
Under laboratory conditions, the fermentation temperature of the biological feed is screened. The fermentation raw materials comprise: 0.1g/kg of microzyme, 0.1g/kg of lactobacillus, 0.1g/kg of bacillus, 525g/kg of puffed peeled soybean meal, 75g/kg of corn flour, 225g/kg of wheat bran and 175g/kg of water. Wherein the puffed peeled soybean meal is pulverized before fermentation, the pulverizing sieve sheet is 0.8 × 0.8mm, the corn pulverizing sieve sheet is 0.8 × 0.8mm, and the wheat bran pulverizing sieve sheet is 0.8 × 0.8 mm. Taking 200g of 10 uniformly mixed samples, uniformly spreading the 10 uniformly mixed samples in a large culture dish, sealing the large culture dish by using an adhesive tape, respectively adjusting the temperature of a mould culture box to be below 15 ℃,20 ℃, 25 ℃, 30 ℃ and 35 ℃, putting the two culture dishes into the large culture dish for fermentation culture under each temperature condition, and observing the fermentation progress every day, wherein if the surface of the culture dish begins to generate water and gas, the fermentation is proved to have started. From the first day, 5g of the sample was taken every 24 hours for observation and measurement.
In the results, as the laboratory temperature increased from 15 ℃,20 ℃, 25 ℃, 30 ℃ and 35 ℃, the time required for completion of fermentation was gradually shortened, with 15 ℃ requiring fermentation for 10 days, 20 ℃ requiring fermentation for 8 days, 25 ℃ requiring fermentation for 5 days, 30 ℃ requiring fermentation for 3 days, and 35 ℃ requiring fermentation for 3 days. In the industrial production, increasing the temperature means increasing the energy consumption, and prolonging the fermentation time means reducing the production efficiency, so that after various factors are comprehensively considered, the optimal condition of fermentation for 3d, namely 72h at 30 ℃ is selected.
EXAMPLE 4 screening of fermentation moisture content
Under plant conditions, a screen of water addition was performed. The fermentation raw materials comprise: 0.1g/kg of microzyme, 0.1g/kg of lactobacillus, 0.1g/kg of bacillus, 525g/kg of puffed peeled soybean meal, 75g/kg of corn flour and 225g/kg of wheat bran. Wherein the puffed peeled soybean meal is pulverized before fermentation, the pulverizing sieve sheet is 0.8 × 0.8mm, the corn pulverizing sieve sheet is 0.8 × 0.8mm, and the wheat bran pulverizing sieve sheet is 0.8 × 0.8 mm. Taking 14 plastic barrels, calculating the water content of the uniformly mixed dry materials according to 12%, adding tap water to ensure that the water content of the dry materials respectively reaches 20%, 25%, 30%, 35%, 40%, 45% and 50%, and filling each water content into two barrels to be used as a parallel group. Compacting and sealing after filling, and fermenting for 72h at 30 ℃.
In the results, the fermentation effect is better and better with the continuous increase of the water content, but the fluidity of the biological feed is poorer and poorer with the increase of the water content, and the poor fluidity means that the biological feed cannot be added into the complete feed. Therefore, the water content of 30% is the best group, namely the percentage of water added is 25%, after the fluidity and the fermentation effect are considered comprehensively.
EXAMPLE 5 assay of biological feed produced under optimal conditions according to the invention
Under factory conditions, fermenting for 72h by using a biological feed fermentation bag under the conditions that the optimal screening condition temperature is 30 ℃ and the water content is 30%, wherein the fermentation raw materials comprise: 0.1g/kg of microzyme, 0.1g/kg of lactobacillus, 0.1g/kg of bacillus, 525g/kg of puffed peeled soybean meal, 75g/kg of corn flour and 225g/kg of wheat bran. Wherein the puffed peeled soybean meal is pulverized before fermentation, the pulverizing sieve sheet is 0.8 × 0.8mm, the corn pulverizing sieve sheet is 0.8 × 0.8mm, and the wheat bran pulverizing sieve sheet is 0.8 × 0.8 mm. After fermentation, three parallel samples were taken and sent to the Beijing Dabei agricultural biological feed detection center for detection, and the detection results are shown in Table 9.
TABLE 9 results of testing biological feeds produced under optimal conditions
Figure BDA0000781438290000121
The detection result shows that the number of the living bacteria contained in the fermentation product is more than or equal to 1 multiplied by 109Each g contains more than or equal to 2 percent of organic acid
Example 6 feeding effect of the invention for creep feed production
Materials and methods
Test samples: a fish meal-free creep was formulated with 6% of the biological feed prepared in example 5 of the present invention.
Control group: 30% of corn, 20% of puffed corn, 17% of soybean meal, 3% of cane sugar, 3% of glucose, 6% of super-import fish meal, 8% of fermented soybean meal, 6% of plasma protein powder, 2% of calcium dihydrogen phosphate, 2% of stone powder, 1% of feed salt, 0.3% of 98.5% of lysine, 0.2% of methionine, 0.01% of tryptophan, 0.5% of an acidifier and 1.5% of Dabei farm piglet premix.
Test groups: 30% of corn, 20% of puffed corn, 17% of soybean meal, 3% of cane sugar, 3% of glucose, 6% of the biological feed prepared in example 5, 8% of fermented soybean meal, 6% of plasma protein powder, 2% of calcium dihydrogen phosphate, 2% of stone powder, 1% of feed salt, 0.3% of 98.5% of lysine, 0.2% of methionine, 0.01% of tryptophan, 0.5% of an acidifier and 1.5% of a premix for Dabei farm pigs.
The daily ration is produced by adopting a cold granulation mode.
Test time: 11/month 7/11/month 28/2014
And (3) experimental design:
24 litters of 1-week-old Du-growing piglets are selected, a single-factor completely random experimental design is adopted, and the piglets are divided into two treatment groups, each group has 12 repetitions, and each repetition has 8-12 heads. Initial body weights were statistically analyzed and the differences between groups were not significant (P > 0.05).
Feeding management
From day 7 of birth, piglets start to feed creep feed, feed artificially, feed a few meals, drink water freely, feed food freely, the temperature is 26 ℃, the diarrhea condition of the piglets is observed every morning, the feed intake is calculated, the piglets stay on a delivery bed for 7 days after weaning at the age of 21 days, the test is finished, and the piglets are weighed at the end of the test.
Test determination indexes are as follows: and recording the litter weight, the number of dead people, the number of diarrhea people, the feed consumption and the like by taking the repetition as a unit, and calculating the average weight, the daily feed consumption, the death rate, the feed-meat ratio and the like.
Statistical analysis
Statistical analysis was performed on the test data using statistical software SPSS 13.0 software.
2 the results are shown in table 10.
As can be seen from table 10: compared with the control group, the piglet daily gain and feed intake were not significantly different from the beginning of the test to the age of 21 days (P > 0.05). The piglet feed intake and daily gain of the test group were higher than those of the control group in the growth period from 7 days to 28 days of age, but the difference was not significant (P > 0.05). The diarrhea rate of piglets is remarkably different (P <0.05) and the mortality rate of piglets is remarkably different (P < 0.05).
3 conclusion
Test results show that the creep feed produced by using the biological feed disclosed by the invention is balanced in nutrition and high in digestibility, can effectively replace expensive animal and plant protein raw materials such as fish meal, and the average litter weight is obviously greater than that of a control group, so that the diarrhea rate and the death rate of piglets are obviously reduced, and the healthy and rapid growth of the piglets is ensured.
TABLE 10 Effect of the invention on piglet production Performance
Item Control group Test group P value
Average weight of 7 day old nest (kg) 23.6 22.8 P>0.05
Average weight of 28 day old nest (kg) 57.6 65.9 P<0.05
Average material consumption (kg) 19.64 24.13 P<0.05
Diarrhea Rate (%) 6.01 2.39 P<0.05
Mortality (%) 1.9 0.72 P<0.05
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (5)

1. The biological feed for replacing fish meal in creep feed is characterized in that the feed raw materials comprise the following components in parts by weight: 0.1g/kg of yeast, 0.1g/kg of lactic acid bacteria, 0.1g/kg of bacillus, 525g/kg of puffed peeled soybean meal, 75g/kg of corn flour and 225g/kg of wheat bran, wherein the yeast is saccharomyces cerevisiae CGMCC number 6560, the bacillus is bacillus subtilis CGMCC No.8148, and the lactic acid bacteria are lactobacillus plantarum CGMCC No. 8150;
the biological feed is prepared by fermenting feed raw materials with the water content of 30% for 72 hours in a sealing way at the temperature of 30 ℃;
the viable count of the yeast, the lactic acid bacteria and the bacillus is 20-30 hundred million/g.
2. The biological feed as claimed in claim 1, wherein the puffed peeled soybean meal is pulverized before fermentation, and the pulverized sieve sheet is 0.8 x 0.8mm, the corn pulverized sieve sheet is 0.8 x 0.8mm, and the wheat bran pulverized sieve sheet is 0.8 x 0.8 mm.
3. The use of the biological feed of claim 1 or 2 as a protein raw material in pig creep feed complete feed instead of fish meal, wherein the biological feed accounts for 5% -10% of the complete feed in the pig creep feed complete feed, and the biological feed is used for completely replacing the fish meal in the creep feed so as to prepare the fish meal-free creep feed.
4. A pig creep feed without fish meal, which is characterized in that the biological feed of claim 1 or 2 accounting for 5 to 10 percent of the complete feed is used as a protein raw material to replace the fish meal in the complete feed of the creep feed.
5. The fish meal-free pig creep feed according to claim 4, which comprises the following components in parts by weight: 30% of corn, 20% of puffed corn, 17% of soybean meal, 3% of cane sugar, 3% of glucose, 6% of biological feed, 8% of fermented soybean meal, 6% of plasma protein powder, 2% of calcium dihydrogen phosphate, 2% of stone powder, 1% of feed salt, 0.3% of 98.5% of lysine, 0.2% of methionine, 0.01% of tryptophan, 0.5% of acidifying agent and 1.5% of porket premix.
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