CN115462460B - Preparation method and application of liquid bacterial enzyme synergistic feed additive for pigs - Google Patents
Preparation method and application of liquid bacterial enzyme synergistic feed additive for pigs Download PDFInfo
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- CN115462460B CN115462460B CN202211134581.9A CN202211134581A CN115462460B CN 115462460 B CN115462460 B CN 115462460B CN 202211134581 A CN202211134581 A CN 202211134581A CN 115462460 B CN115462460 B CN 115462460B
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- pigs
- enzyme
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Classifications
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- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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Abstract
The invention relates to a preparation method and application of a liquid bacterial enzyme synergistic feed additive for pigs, which can effectively solve the problem of preparing liquid bacterial enzyme synergistic feedThe additive for pig feed has the application problems, and the preparation method of the additive comprises the steps of taking sterile water as a carrier, adding 20-50% of bacterial protectant by mass of the additive, adding 0.1-0.3% of enzyme protectant by mass of the additive, and adding 0.05-0.25% of stabilizer by mass of the additive to prepare additive base solution; then adding beneficial bacteria to make beneficial bacteria concentration not less than 4×10 8 And (3) CFU/mL, finally adding active enzyme to enable the activity of the active enzyme in the feed additive to be 80-1800U/L, and adjusting the pH to 5-6 by using phosphate buffer solution. The invention has the advantages of rich raw materials, simple preparation method, low cost and good use effect, develops a new way for preparing the pig feed additive, is effectively used as the additive of pig feed, obviously improves the feed intake and daily gain of live pigs, reduces the meat-to-feed ratio, improves the feed utilization rate and promotes the growth of fattening pigs.
Description
Technical Field
The invention relates to the technical field of feed additives, in particular to a preparation method and application of a liquid bacterial enzyme synergistic feed additive for pigs.
Background
At present, the pig raising industry in China is in a key phase of transformation and upgrading, and a plurality of problems are faced, namely a series of problems of pig growth performance reduction, mortality rate increase and the like caused by the 'post-age' forbidden; secondly, the problem that the conflict of 'people and livestock contending for grain' is aggravated and the cost of feed is higher and higher is faced with the global grain crisis prominently. In particular, in order to seek green and safe 'anti-replacement' products, the pig feed industry is provided with serious challenges in the face of multiple pressures such as no resistance, environmental protection, shortage of feed raw materials, food safety and the like, and new technologies, products and modes are needed to drive the sustainable development of the industry. The biological feed represented by the fermented feed has the advantages of improving the utilization rate and palatability of animal feed, and reducing or eliminating anti-nutritional factors and mycotoxins in feed raw materials; improving the intestinal barrier function of pigs, maintaining the microecological balance of intestinal flora, enhancing the immunity of organisms, improving the disease resistance of animals, reducing the content of harmful substances and odor substances in excrement, eliminating dust generated in the feeding process of conventional feeds, avoiding the immunity decline caused by the continuous stimulation of animal respiratory systems by the dust, regulating and controlling the lipid metabolism of pigs, improving the meat color redness, intramuscular fat content and muscle tenderness, improving the pork fatty acid (oleic acid, linoleic acid, linolenic acid and the like), the flavor amino acid (glutamic acid, aspartic acid and the like), the volatile flavor substances (alcohol, aldehyde, ketone) and improving the pork quality and the like; the potential effects of the feed replacement, the reduction of the use of antibiotics, the improvement of the immunity of a host, the resistance to disease occurrence and the like in the pig breeding process are highlighted, and the feed replacement additive has been developed in numerous challenges and choices. The bacterium enzyme synergistic fermentation feed has better effect on the feed and the feed raw material than the single bacterium or enzyme treatment effect due to the consideration of bacterium, enzyme and fermented elements, and the former shows obvious advantages in improving the feed nutritive value and feeding effect.
Although the synergistic fermentation of the bacterial enzymes is the most widely used production mode of the fermented feed at present, the method still stays in the theoretical research stage at present, and lacks stable effect and perfect application scheme in the aspect of processing complex raw materials, and also does not see the public report of the application of the liquid bacterial enzyme synergistic fermentation feed additive in the pig feed.
Disclosure of Invention
Aiming at the situation, the invention aims to overcome the defects of the prior art, and the invention aims to provide a preparation method and application of the liquid bacterial enzyme synergistic feed additive for pigs, which can effectively solve the problem of preparing the liquid bacterial enzyme synergistic feed additive and realizing the application in pig feeds.
The technical scheme is that the liquid bacterial enzyme synergistic feed additive for pigs is prepared by uniformly mixing beneficial bacteria, active enzyme, enzyme protecting agent, bacterial protecting agent, stabilizing agent and sterile water, and comprises the following steps:
(1) Preparing seed liquid:
inoculating beneficial bacteria into a seed liquid culture medium, culturing at 34-37 ℃ for 18-24 h for rejuvenation, and preparing the effective viable count of the bacterial strain is more than or equal to 1 multiplied by 10 8 Seed solution of CFU/mL;
the beneficial bacteria are one or a mixture of any ratio of lactobacillus plantarum, pediococcus acidilactici, enterococcus faecalis, bacillus licheniformis, bacillus subtilis, bacillus coagulans, clostridium butyricum and saccharomyces cerevisiae;
the seed liquid culture medium is MRS culture medium, and is prepared by adding 10g of protein, 5g of beef powder, 4g of yeast powder, 2g of glucose, 1mL of Tween 80, 2g of dipotassium hydrogen phosphate, 5g of sodium acetate, 2g of tri-ammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and 15g of agar powder into 1000mL of distilled water, heating and dissolving, adjusting pH to 6, and sterilizing at 121 ℃ under high pressure for 15-20 minutes;
(2) Preparing a fermentation liquid:
inoculating the seed solution into an optimized fermentation medium, wherein the inoculum size is 2-5% of the volume of the fermentation medium, culturing for 16-24 h at 34-37 ℃, the residual sugar in the fermentation liquid is 0-3 g/L, and the viable count of beneficial bacteria is more than or equal to 2.4X10) 9 /mL；
The optimized fermentation medium consists of the following components in percentage by weight: 2 to 5 percent of brown sugar or molasses, 1 to 1.8 percent of yeast extract powder, 0.18 to 0.25 percent of dipotassium hydrogen phosphate, 0.001 to 0.003 percent of magnesium sulfate and 0.08 to 0.1 percent of tween-80, and adding sterile water to 100 percent;
(3) Preparing active enzyme:
the active enzyme is one or a mixture of more than two of cellulase, xylanase, beta-glucanase, phytase, pectase, beta-mannanase and alpha-galactosidase, and the mixture is respectively added with sterile water to prepare 800-1000U/L cellulase, 1500-1800U/L xylanase, 500-800U/L beta-glucanase, 500-800U/L phytase, 800-1500U/L pectase, 400-1000U/L beta-mannanase and 80-200U/L alpha-galactosidase;
(4) Selection of enzyme protectant:
the enzyme protecting agent is EDTA (ethylene diamine tetraacetic acid);
(5) And (3) selecting a bacterial protectant:
the research and experiment show that the glycerol is colorless, odorless and sweet, has a clear and viscous liquid appearance, is an organic matter, can absorb moisture from air, can also absorb hydrogen sulfide, hydrogen cyanide and sulfur dioxide, protects the biological molecular structure from being damaged, and is effectively applied to food sweeteners; trehalose is a typical stress metabolite, and can form a unique protective film on the surface of cells under severe environmental conditions such as high temperature, high cold, high osmotic pressure, drying water loss and the like, so that the biological molecular structure is effectively protected from being damaged, and the life process and biological characteristics of a living body are maintained; the chitosan has a plurality of unique properties such as biodegradability, cell affinity, biological effect and the like, is natural polysaccharide, has good affinity to cells, and does not generate rejection reaction; the fructo-oligosaccharide has double lattices with definite health care function and excellent food ingredients, promotes bacterial proliferation, promotes the colonization of beneficial bacteria such as lactobacillus, bifidobacterium, streptococcus and the like in the digestive tract, inhibits the growth of harmful bacteria, improves intestinal flora, and indirectly achieves the nutrition and growth promoting effect on animal bodies; the isomaltooligosaccharide can promote the remarkable proliferation of beneficial bacteria in the body, has the function of water-soluble dietary fiber, improves the taste of food, does not prevent the fermentation of normal lactobacillus, has low sweetness and low heat, and basically does not increase blood sugar and blood fat. Therefore, glycerol, trehalose, chitosan, fructo-oligosaccharide or isomaltooligosaccharide are selected as bacterial protectants in the invention;
(6) Selection of a stabilizer:
the research and experiment show that sodium alginate or xanthan gum has good physical and chemical properties, is effectively used as a stabilizer in food production, and is natural microbial polysaccharide; guar gum is an extract of guar beans of leguminous plants, has superior dispersion capability in cold water and hot water, does not generate caking phenomenon, and has no biotoxicity; the carrageenan is a hydrophilic colloid extracted from red algae seaweed such as eucheuma, agar, carrageen and the like, and stability is improved, so that sodium alginate, xanthan gum, guar gum or carrageenan is selected as a stabilizer;
(7) Mixing to prepare the liquid bacterial enzyme synergistic feed additive for pigs:
the beneficial bacteria, the active enzyme, the enzyme protective agent, the bacterial protective agent and the stabilizing agent are added into sterile water and are uniformly mixed to prepare the liquid bacterial enzyme synergistic feed additive for pigs, and the method comprises the following steps:
firstly, taking sterile water as a carrier, adding a bacterial protective agent accounting for 20-50% of the mass of a feed additive, then adding an enzyme protective agent accounting for 0.1-0.3% of the mass of the feed additive, and adding a stabilizing agent accounting for 0.05-0.25% of the mass of the feed additive to prepare a feed additive base solution; then adding beneficial bacteria to make beneficial bacteria concentration not less than 4×10 8 Adding active enzyme into CFU/mL to ensure that the activity of the active enzyme in the feed additive is 80-1800U/L, and regulating the pH to 5-6 by using phosphate buffer solution to obtain the liquid bacterial enzyme synergistic feed additive for pigs;
the phosphate buffer solution is a well-known commercial PBS buffer solution, and the pH value is 7.0-8.0.
The liquid bacterial enzyme synergistic feed additive for pigs, which is prepared by the method, is added into the feed for pigs, wherein the addition amount is that 500mL of feed additive is added into each ton of basic ration, 3 times of water is added into 1 ton of basic ration to prepare the liquid feed, 500mL of feed additive is added, and the feed additive is predigested and fermented for 12 hours at room temperature to prepare the fermented feed, and the quality guarantee period of the feed additive is 45 days.
The feed additive has the advantages of rich raw materials, simple preparation method, low cost and good use effect, develops a new way for preparing the pig feed additive, is effectively used as the additive of pig feed, obviously improves the feed intake and daily gain of live pigs, reduces the meat-to-feed ratio, improves the feed utilization rate, promotes the growth of fattening pigs, and has obvious economic and social benefits.
Detailed Description
The following describes in detail the embodiments of the present invention with reference to specific cases and examples.
The invention may be embodied by the following examples.
Example 1
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: mixing 20% glycerol, 0.2% sodium alginate, 0.05% xanthan gum and 0.2% EDTA, and adding effective bacteria number not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, then 900U/L of cellulase, 650U/L of phytase, 700U/L of beta-glucanase and 1600U/L of xylanase are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is regulated to 5.5 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is obtained.
Example 2
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: mixing 20% glycerol, 30% trehalose, 0.1% sodium alginate, 0.05% xanthan gum and 0.1% EDTA, adding effective viable count not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, 700U/L of beta-mannase, 1100U/L of pectase and 140U/L of alpha-galactosidase are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is regulated to 6.0 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is prepared.
Example 3
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: mixing 50% glycerol, 0.1% sodium alginate and 0.0.3% EDTA, adding effective bacteria with number not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, xylanase 1800U/L, alpha-galactosidase 100U/L, beta-mannase 500U/L and phytase 750U/L are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is adjusted to 5.0 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is prepared.
Example 4
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: mixing 20% trehalose, 0.2% sodium alginate, 0.1% xanthan gum and 0.15% EDTA, and adding effective bacterial count not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, 1000U/L of cellulase, 600U/L of beta-glucanase and 1400U/L of pectinase are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is adjusted to 5.5 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is prepared.
Example 5
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: mixing 30% glycerol, 20% trehalose, 0.1% sodium alginate, 0.05% xanthan gum and 0.15% EDTA, adding effective viable count not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, then 900U/L of cellulase, 650U/L of phytase, 700U/L of beta-glucanase and 1600U/L of xylanase are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is adjusted to 5.0 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is prepared.
Example 6
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: mixing 50% trehalose, 0.2% sodium alginate, 0.1% xanthan gum and 0.15% EDTA, and adding effective bacterial count not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, adding cellulose900U/L of enzyme, 650U/L of phytase, 700U/L of beta-glucanase and 1600U/L of xylanase, adding sterile water to 100%, uniformly mixing, and regulating the pH value to 6.0 by using phosphate buffer solution to prepare the liquid bacterial enzyme synergistic feed additive for pigs.
Example 7
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: 22% chitosan, 35% isomaltooligosaccharide, 0.15% guar gum, 0.06% carrageenan and 0.15% EDTA, and adding effective bacteria number not less than 1×10 8 CFU/mL seed solution of Bacillus licheniformis, bacillus subtilis and Bacillus coagulans to make viable count reach 4×10 8 CFU/mL, 800U/L of cellulase, 550U/L of phytase, 800U/L of beta-glucanase and 1550U/L of xylanase are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is adjusted to 5.5 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is prepared.
Example 8
The invention relates to a preparation method of a liquid bacterial enzyme synergistic feed additive for pigs, which takes sterile water as a carrier and adds the total weight of the additive: 45% fructo-oligosaccharide, 0.15% sodium alginate, 0.06% carrageenan and 0.15% EDTA are mixed together, and the effective bacterial count is more than or equal to 1 multiplied by 10 8 CFU/mL Clostridium butyricum and Saccharomyces cerevisiae seed solution to make viable count reach 4×10 8 CFU/mL, xylanase 1800U/L, alpha-galactosidase 100U/L, beta-mannase 500U/L and phytase 750U/L are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is adjusted to 5.5 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is prepared.
The examples given above are only for further illustrating the specific embodiments of the present invention, but the examples are not limited thereto, and in the technical solutions given in the examples, if no special description is given, the present invention is tested in the field, the effect is very good, and the relevant experimental data are as follows:
1. effect of bacterial protectant and stabilizer on lactic acid bacteria survival and sedimentation
Glycerol and trehalose are selected as lactic acidThe composition and the dosage of the bacterial protectant, sodium alginate and xanthan gum, which are used as stabilizers, are respectively changed in the formula, and the effective viable count and the sedimentation degree are used as indexes, and the better combination 6 groups are obtained through test and repeated measurement, and the results are shown in Table 1. In the test, the final concentration of the lactobacillus plantarum, the pediococcus acidilactici and the enterococcus faecalis is not less than 1 multiplied by 10 8 CFU/mL, effective viable count of lactic acid bacteria measured by plate count method was 4.3X10 8 CFU/mL. The components in the system are expressed in weight percent.
The results show that the effective viable count of the experimental groups 1 to 6 is 1 multiplied by 10 at 15 days 8 Order of magnitude, still can reach 1X 10 when left for 45 days 6 The order of magnitude is 1000 times of that of a control group, and the experimental groups can effectively delay the settlement of bacterial liquid.
TABLE 1 survival and anti-sedimentation Properties of lactic acid bacteria of different formulations
Note that: the degree of sedimentation + indicates that there is a small amount of sedimentation, ++ and representation of and (3) complete precipitation.
2. Enzyme protectant concentration and pH Effect on enzyme Activity
The active enzymes are liquid feeding enzymes, and are purchased from Wuhan Xinhua Yangshan biological Co., ltd. Cellulase, phytase, beta-glucanase and xylanase are selected to prepare mother liquor with high enzyme activity, EDTA with different concentrations is set in the system, phosphate buffer solutions with different pH values are used for diluting the compound enzyme solution, the enzyme activity is measured at intervals of 45d, and finally, a better formula is obtained, and the results are shown in Table 2. (initial enzyme activities of cellulase, phytase, beta-glucanase and xylanase are 2500U/L, 2000U/L and 5000U/L respectively)
TABLE 2 influence of different formulations on enzyme Activity under EDTA
Note that: relevant enzyme activity assays were referenced to the following criteria: cellulase reference NY/T912, phytase reference GB/T18634, beta-glucanase reference NY/T911, xylanase reference GB/T18634.
Pre-digestion treatment of corn and soybean meal type feed by liquid bacterial enzyme synergistic fermentation feed additive
3.1 preparation of liquid bacterial enzyme synergistic feed additive
According to the test results 1 and 2, preparing a liquid bacterial enzyme synergistic feed additive, wherein the formula comprises 20% of glycerin, 0.2% of sodium alginate, 0.05% of xanthan gum, 0.2% of EDTA and 4 multiplied by 10 of lactobacillus concentration 8 CFU/mL contains lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis at concentration not less than 1×10 8 CFU/mL, the feed enzymes are cellulase, phytase, beta-glucanase and xylanase, and the pH value of the feed additive is regulated to 5.5 by phosphate buffer solution.
3.2 pretreatment of corn and soybean meal type feed
Commercial feed (basic ration) of corn bean pulp is purchased from Henan Tian Kang Hongzhan commercial limited company to breed pig compound feed-Huimin No. 2, the raw materials comprise corn, wheat, puffed bean pulp, fish meal, calcium hydrophosphate, stone powder, sodium chloride, compound premixed feed, L-lysine hydrochloride, DL-methionine and the like, and the product component analysis assurance values are shown in Table 3.
TABLE 3 product composition analysis assurance value (%)
| Crude protein is not less than | Coarse fiber is less than or equal to | Coarse ash content is less than or equal to | The calcium is more than or equal to | The total phosphorus is more than or equal to | Sodium chloride | Moisture is less than or equal to | Lysine is not less than |
| 15.5 | 6.0 | 8.0 | 0.5 | 0.4 | 0.2～1.5 | 14.0 | 1.0 |
Adding water according to the ratio of 1:3 to obtain liquid feed, adding feed additive into the prepared liquid feed, adding 500 mL/ton (dry material) of the feed additive into the liquid feed, pre-digesting for 12h, and measuring pH, soluble total sugar and acid soluble protein, wherein the measurement result is shown in Table 4, and CK is the group without feed additive.
TABLE 4 corn and soybean meal feed Pre-digestion treatment results
| Detection index | CK | A | B | C | D |
| pH | 6.39±0.01 | 4.42±0.19 | 4.63±0.11 | 4.85±0.13 | 5.02±0.16 |
| Soluble total sugar (μg/g) | 0.27±0.03 | 1.07±0.03 | 1.03±0.06 | 1.13±0.17 | 1.03±0.09 |
| Acid soluble protein (%) | 1.02±0.09 | 1.46±0.13 | 1.38±0.06 | 1.36±0.09 | 1.33±0.10 |
Note that: a to D are respectively the 1 st day, 15 th day, 30 th day and 45 th day of the completion of the preparation of the bacteria enzyme synergistic feed additive; the method for measuring the soluble total sugar adopts a sulfuric acid extraction anthrone colorimetric method; the method for measuring the acid soluble protein refers to the measurement of the acid soluble protein in the feed raw material NY/T3801-2020.
The result shows that the liquid bacterial enzyme synergistic feed additive can effectively ferment basic ration within 45 days, and is specifically characterized by reducing pH and increasing the content of soluble total sugar and acid soluble protein of the feed. Compared with the control group, the total soluble sugar is averagely improved by 2.9 times, and the percentage of acid soluble protein is averagely improved by 35.5 percent.
4 influence of feed additive predigestion on growing and fattening pig growth performance
4.1 feeding test protocol
72 binary growing pigs with good health conditions, about 38kg, are selected for test, and randomly divided into 3 groups (see table 5):
table 5 test group
Each group had 3 replicates, 8 pigs per replicate. Group A is a control group, and dry feeding is adopted; B. the group C is an experimental group, the group B adopts basic ration and liquid feeding, and the water-to-material ratio of the liquid feeding is 3:1; group C is fed by adopting basic ration and bacteria enzyme to cooperate with liquid fermentation feed, and the ratio of liquid feed water to feed is 3:1. The test period was 28 days. The feed intake and the diarrhea head number are recorded every day, the weight is weighed on an empty stomach in the morning respectively at the beginning and the end of the test, and the daily feed intake, the daily gain and the feed conversion ratio are calculated respectively.
4.2 feeding management
The feeding management and epidemic prevention program is carried out according to daily standards in the field, and the feed is fed three times a day during the test period, 8:00 a.m., 13:00 a.m., and 17:00 a.m. respectively, pigs eat and drink water freely, and no residue is left after half an hour of feeding as a feeding standard. The test results are shown in Table 6.
Table 6 results of test on growth performance of growing pigs
In terms of daily gain: the daily gain of the liquid feeding group B is improved by 13.4 percent compared with that of the control group A, and the daily gain of the liquid fermentation group C is improved by 26.8 percent compared with that of the control group A; in terms of feed intake: the average daily feed intake of the liquid feeding group B is improved by 11.8 percent compared with that of the control group A, and the daily feed intake of the liquid fermentation group C is improved by 24.1 percent compared with that of the control group A; in terms of feed conversion: the feed conversion ratio of the liquid feeding group B is reduced by 2.53% compared with that of the control group A, and the feed conversion ratio of the liquid fermentation group C is reduced by 3.66%. Therefore, after the feed is fermented by the liquid bacterial enzyme synergistic feed additive, the feed intake and daily gain of the live pigs can be obviously improved, the feed-meat ratio is obviously reduced, the feed utilization rate is improved, and the feed has a promoting effect on the growth of fattening pigs.
In conclusion, the liquid bacterial enzyme synergistic feed additive for pigs has the advantages of rich raw materials, simple preparation method, low cost and good use effect, develops a new way for preparing the feed additive for pigs, effectively improves daily feed intake of live pigs by more than 24 percent, improves daily weight gain by more than 26 percent, reduces meat ratio by more than 3 percent, obviously improves feed utilization rate, promotes growth of fattening pigs, and has obvious economic and social benefits.
Claims (2)
1. The preparation method of the liquid bacterial enzyme synergistic feed additive for pigs is characterized in that the additive takes sterile water as a carrier, and the total weight of the additive is added: mixing 20% glycerol, 0.2% sodium alginate, 0.05% xanthan gum and 0.2% EDTA, and adding effective bacteria number not less than 1×10 8 CFU/mL seed solution of Lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis for achieving viable count of 4×10 8 CFU/mL, then 900U/L of cellulase, 650U/L of phytase, 700U/L of beta-glucanase and 1600U/L of xylanase are added, sterile water is added to 100%, the mixture is uniformly mixed, and pH is adjusted to 5.5 by phosphate buffer solution, so that the liquid bacterial enzyme synergistic feed additive for pigs is obtained, and the method specifically comprises the following steps:
(1) Preparing seed liquid:
inoculating beneficial bacteria into a seed liquid culture medium, culturing at 34-37 ℃ for 18-24 h for rejuvenation, and preparing the effective viable count of the bacterial strain is more than or equal to 1 multiplied by 10 8 Seed solution of CFU/mL;
the beneficial bacteria are lactobacillus plantarum, pediococcus acidilactici and enterococcus faecalis;
the seed liquid culture medium is MRS culture medium, and is prepared by adding 10g of protein, 5g of beef powder, 4g of yeast powder, 2g of glucose, 1mL of Tween 80, 2g of dipotassium hydrogen phosphate, 5g of sodium acetate, 2g of tri-ammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and 15g of agar powder into 1000mL of distilled water, heating and dissolving, adjusting pH to 6, and sterilizing at 121 ℃ under high pressure for 15-20 minutes;
(2) Preparing a fermentation liquid:
inoculating the seed solution into an optimized fermentation medium, wherein the inoculum size is 2-5% of the volume of the fermentation medium, culturing for 16-24 h at 34-37 ℃, the residual sugar in the fermentation liquid is 0-3 g/L, and the viable count of beneficial bacteria is more than or equal to 2.4X10) 9 /mL；
The optimized fermentation medium consists of the following components in percentage by weight: 2 to 5 percent of brown sugar or molasses, 1 to 1.8 percent of yeast extract powder, 0.18 to 0.25 percent of dipotassium hydrogen phosphate, 0.001 to 0.003 percent of magnesium sulfate and 0.08 to 0.1 percent of tween-80, and adding sterile water to 100 percent;
(3) Preparing active enzyme:
the active enzyme is cellulase, xylanase, beta-glucanase and phytase;
(4) Mixing to prepare the liquid bacterial enzyme synergistic feed additive for pigs:
adding beneficial bacteria, active enzyme, enzyme protective agent, bacterial protective agent and stabilizing agent into sterile water, uniformly mixing, and preparing the liquid bacterial enzyme synergistic feed additive for pigs, wherein the method comprises the following steps:
firstly, taking sterile water as a carrier, adding a bacterial protective agent accounting for 20% of the mass of a feed additive, then adding an enzyme protective agent accounting for 0.2% of the mass of the feed additive, and adding a stabilizing agent accounting for 0.25% of the mass of the feed additive to prepare a feed additive base solution; adding beneficial bacteria to make beneficial bacteria concentration not less than 4×10 8 CFU/mL, then adding active enzyme to make the activity of the active enzyme in the feed additive be 80-1800U/L, and regulating pH to 5.5 with phosphate buffer solution to obtain the liquid bacterial enzyme synergistic feed additive for pigs;
the enzyme protecting agent is EDTA;
the bacterial protectant is glycerol;
the stabilizer is sodium alginate or xanthan gum.
2. The application of the liquid bacterial enzyme synergistic feed additive for pigs prepared by the method of claim 1 in pig feed, wherein 1 ton of basic ration is added with 3 times of water to prepare liquid feed, 500mL of feed additive is added, and the liquid feed additive is subjected to pre-digestion and fermentation for 12 hours at room temperature to prepare fermented feed, and is used for raising fattening pigs, and the feed additive has the best use effect in 45 days.
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