CN112205513A

CN112205513A - Combined feed additive for palm meal type feed and preparation method and application thereof

Info

Publication number: CN112205513A
Application number: CN202011007582.8A
Authority: CN
Inventors: 刘金银; 孙霞
Original assignee: Hunan Joy Biotechnology Co ltd
Current assignee: Hunan Joy Biotechnology Co ltd
Priority date: 2020-09-23
Filing date: 2020-09-23
Publication date: 2021-01-12

Abstract

The invention discloses a combined feed additive for palm meal type feed, which consists of 34-48 parts by mass of a complex enzyme preparation, 6.8-11.2 parts by mass of a compound microorganism and 40.8-59.2 parts by mass of a multi-strain solid fermentation product. The feed additive prepared by combining the complex enzyme preparation, the complex microorganism and the multi-strain solid fermentation product is added into the animal compound feed, so that the utilization rate of the feed is improved, the use cost of the feed is reduced, and the breeding benefit is increased. The invention also provides a preparation method of the combined feed additive for the palm meal type feed, and the preparation method is simple in preparation process, strong in operability and convenient to popularize and apply to small feed enterprises and farms. The invention also relates to the application of the combined feed additive for the palm meal type feed in the palm meal-containing feed, and the usage amount of the palm meal can be increased by 3-5 percent by adopting the combined feed additive, so that the usage amount of the palm meal in the compound feed is effectively increased, and the feed cost is obviously reduced.

Description

Combined feed additive for palm meal type feed and preparation method and application thereof

Technical Field

The invention relates to the technical field of feed additives, in particular to a combined feed additive for palm meal type feed. The invention also relates to a preparation method of the combined feed additive for the palm meal type feed. The invention also relates to application of the combined feed additive for the palm meal type feed in the palm meal-containing feed.

Technical Field

Palm meal is produced primarily in malaysia and indonesia and is a pressing by-product of palm oil plants. The content of crude protein is more than or equal to 14 percent, the content of crude fat is more than or equal to 8 percent, the deficiency is that the content of anti-nutritional factor mannan is as high as 30 to 35 percent, the nutrient digestion and absorption rate of palm meal is reduced, and the using amount and the application effect of the palm meal in the livestock and poultry feed are reduced.

The shortage of feed resources forces us to find all possible substitutes, and despite the characteristic of high anti-nutritional factors, the palm meal has certain cost performance due to low price, and is widely applied to the breeding industry in China. The key to the rational use of palm meal is to eliminate its anti-nutritional effects, particularly to reduce the anti-nutritional effects of mannan and cellulose, while at the same time, there is a need to improve the palatability of palm meal rations and to coordinate the nutritional balance of palm meal with other feed components.

The feed enzyme preparation and the microbial preparation are added into the feed, which are two technical means for improving the feeding value of the palm meal, and the digestive enzyme can decompose mannan in the palm meal, eliminate the anti-nutritional effect and improve the nutrient digestibility. The microbial preparation has the functions of regulating intestinal flora balance, secreting antibacterial peptide and digestive enzyme. Chinese patent CN101983579A discloses a feed additive consisting of xylanase, glucanase, pectinase, mannase and cellulase. CN106417897A discloses a palm meal fermented feed, which is fermented for 2-3 days by using bacillus subtilis and yeast at a controlled temperature, and simultaneously added with protease, arabinase, mannase and xylanase. Although the additive can improve the performance of the palm meal feed and improve the production performance of chicks and piglets, the palm kernel meal is rich in amino acid, and the palm kernel meal is rich in fiber, so that the compound enzyme added into the feed can decompose part of crude fiber of the palm kernel meal, but the digestibility is still lower, and the production cost of the feed is still higher.

Patent application with publication number CN101248835A discloses a method for degrading palm pulp by enzyme, which comprises mixing palm pulp and mannase solution, and reacting at 40-100 deg.C for 4-80 hr. The method only aims at the palm meal, but not the whole palm meal type daily ration, the proportion of the palm meal in the compound feed for pigs and poultry is not more than 10%, the palm meal is only one of the targets, and the anti-nutritional factor characteristics and the nutrient digestibility of the compound feed are comprehensively analyzed by considering the whole compound feed, so that the application methods of the enzyme and the microorganism are determined.

The patent of the publication No. CN103636921B discloses a microbial feed additive and a preparation method thereof, and particularly discloses a microbial feed additive which is prepared by taking a liquid fermentation culture medium left in the production of biological cellulose as a raw material, adding animal blood powder, sterilizing, inoculating compound beneficial microorganisms, fermenting, freezing, drying, crushing into powder, and adding one or more of vitamins, minerals, amino acids and compound enzyme preparations with the total amount of 2-15 wt%; the compound enzyme preparation comprises trypsin and cellulase in a ratio of (1-3) to (2-4), wherein the compound beneficial microorganisms comprise bacillus subtilis, lactobacillus rhamnosus and aspergillus niger, and the ratio of the inoculation amount is (3-8): (1-2): (0.5-1), wherein the addition amount of the animal blood powder is 30-50 g/L. The liquid fermentation culture medium adopted in the patent is greatly different in addition proportion and components, so that the patent cannot be applied to palm meal type feed. The invention application of publication number CN104397351A discloses a biological antibiotic-free feed additive and a preparation method thereof, and specifically discloses a biological antibiotic-free feed additive which comprises the following raw material components in parts by weight: 28-35 parts of yeast cell wall polysaccharide, 23-30 parts of bacillus licheniformis, 28-45 parts of bacillus subtilis, 17-19 parts of lactobacillus acidophilus, 17-19 parts of clostridium butyricum, 5-10 parts of yeast selenium, 60-70 parts of a coated slow-release acidifier, 160 parts of bran 150-containing materials, 150 parts of stone powder 120-containing materials and a complex enzyme preparation, wherein the complex enzyme preparation comprises 7-10 parts of phytase, 7-10 parts of glucanase, 10-15 parts of xylanase and 7-10 parts of cellulase. The application adopts various fermentation processes, the processes are complex, in addition, the addition proportion and the components have great differences, and the application is not suitable for palm meal type feeds.

Disclosure of Invention

In order to solve the technical problems that the utilization rate of the feed is low, the cost is high and the popularization and the application of small feed enterprises and farms are not facilitated due to the application of the additive in the palm meal type compound feed in the prior art, the invention provides the combined feed additive for the palm meal type feed and the preparation method and the application thereof.

The invention provides a combined feed additive for palm meal type feed, which consists of 34 to 48 mass parts of complex enzyme preparation, 6.8 to 11.2 mass parts of compound microorganism and 40.8 to 59.2 mass parts of multi-strain solid fermentation product;

the compound enzyme preparation consists of 8-12 parts by mass of 50000U/g enzyme-activity beta-mannase, 3-5 parts by mass of 100000U/g enzyme-activity xylanase, 15-19 parts by mass of 7000U/g enzyme-activity alpha-amylase, 3-5 parts by mass of 50000U/g enzyme-activity acid protease and 5-7 parts by mass of 50000U/g enzyme-activity neutral protease, wherein the xylanase is formed by mixing yeast type xylanase and trichoderma type xylanase according to the proportion of 7: 3;

the multi-strain solid fermentation product is prepared by solid fermentation of a substrate, 1000 hundred million viable bacteria/g of bacillus subtilis and 100 hundred million viable bacteria/g of bacillus coagulans.

Further, the mass percent of the bacillus subtilis is 0.5-1.0% of the substrate, and the mass percent of the bacillus coagulans is 1.0-1.4% of the substrate.

Further, the substrate consists of 29-33.6 parts by mass of corn flour, 20-30 parts by mass of bran, 23-32 parts by mass of palm meal, 12-16 parts by mass of soybean meal, 0.7-1.5 parts by mass of urea and 0.7-1.5 parts by mass of glucose.

Furthermore, the compound microorganism consists of 0.8 to 1.2 parts by mass of bacillus subtilis with 1000 hundred million/g live bacteria, 3 to 5 parts by mass of enterococcus faecalis with 50 hundred million/g live bacteria and 3 to 5 parts by mass of bacillus coagulans with 100 hundred million/g live bacteria.

Has the advantages that:

1. the feed additive is prepared by combining the complex enzyme preparation, the compound microorganism and the multi-strain solid fermentation product, the complex enzyme preparation and the compound microorganism which are main components come from two production modes of liquid fermentation and solid fermentation, and have complementary action, the characteristics of high activity and high content of the liquid fermentation are fully utilized, the characteristic of strong pertinence of the solid fermentation is utilized, the use cost of the feed is reduced, and the breeding benefit is increased; in addition, the compound fertilizer is directly added into palm meal type compound feed, is convenient to use and wide in adaptability, and a user only needs to simply mix and prepare the compound feed without special fermentation complete equipment and complex procedures.

2. The complex enzyme preparation can fully eliminate the anti-nutritional factors of the palm meal by controlling the added beta-mannase and the adding proportion thereof, and can improve the overall utilization rate of the feed by controlling the adding proportion of xylanase, alpha-amylase, acid protease and neutral protease; the invention adopts the yeast type xylanase and the trichoderma type xylanase, can increase the pH adaptability and the temperature adaptability of the xylanase in the animal digestive tract, improves the activity of the xylanase in the animal digestive tract, and promotes the decomposition of xylan.

3. The solid state fermentation of the invention takes plant raw materials as fermentation substrates, the plant raw materials and feed raw materials have homology, and the fermentation product has strong pertinence and high titer and contains a large amount of other active ingredients. In addition, the bacillus subtilis and the bacillus coagulans are used for co-culture in the solid fermentation stage, so that the complementarity of the bacillus subtilis and the bacillus coagulans in the aspects of enzyme production, flora balance regulation and stress resistance can be utilized, the bacillus subtilis and the bacillus coagulans can be adapted to a solid fermentation culture medium similar to a feed, the corresponding strains are induced to propagate, and the final product can be more suitable for the animal intestinal environment and the specific palm meal type feed.

The invention also provides a preparation method of the combined feed additive for the palm meal type feed, which comprises the following steps:

s5.1, preparing the complex enzyme preparation

Mixing 50000U/g enzyme activity beta-mannase 8-12 parts by mass, 100000U/g enzyme activity xylanase 3-5 parts by mass, 7000U/g enzyme activity alpha-amylase 15-19 parts by mass, 50000U/g enzyme activity acid protease 3-5 parts by mass and 50000U/g enzyme activity neutral protease 5-7 parts by mass for later use;

s5.2, preparation of composite microorganism

0.8-1.2 parts by mass of bacillus subtilis with 1000 hundred million viable bacteria per gram, 3-5 parts by mass of enterococcus faecalis with 50 hundred million viable bacteria per gram and 3-5 parts by mass of bacillus coagulans with 100 hundred million viable bacteria per gram are mixed for standby;

s5.3, preparing multi-strain solid fermentation product

S5.3.1, pretreatment of the substrate: crushing and sterilizing a substrate for later use;

s5.3.2, activating strains: adding a certain amount of mixed strains of 1000 hundred million viable bacteria/g of bacillus subtilis and 100 hundred million viable bacteria/g of bacillus coagulans into sterile water with the mass 5-10 times that of the mixed strains for mixing, then adding glucose with the mass 0.5-1 time that of the mixed strains, fully stirring and preserving heat to prepare seed bacteria liquid for later use, wherein the sterile water is water which is boiled and then placed for cooling to 30-35 ℃;

s5.3.3, fermentation: inoculating S5.3.2 prepared seed bacterial liquid into S5.3.1 pretreated substrate, adding sterile water in batches for fermentation culture, wherein the water content of the substrate is controlled to be 41-56% in the fermentation process;

s5.3.4, drying and crushing: s5.3.3 drying the fermented product at temperature not higher than 60 deg.C under ventilation condition, and pulverizing;

and S5.4, mixing products obtained from S5.1, S5.2 and S5.3 in proportion.

Further, in step S5.3, the mass percentage of the bacillus subtilis is 0.5% -1.0% of the substrate, and the mass percentage of the bacillus coagulans is 1.0% -1.4% of the substrate.

Further, in step S5.3, the substrate is composed of 29-33.6 parts by mass of corn flour, 20-30 parts by mass of bran, 23-32 parts by mass of palm meal, 12-16 parts by mass of soybean meal, 0.7-1.5 parts by mass of urea and 0.7-1.5 parts by mass of glucose.

Further, in step S5.4, the following components and contents are contained after mixing: the enzyme activity of beta-mannase is more than or equal to 4000U/g, the enzyme activity of xylanase is more than or equal to 3000U/g, the enzyme activity of alpha-amylase is more than or equal to 1050U/g, the enzyme activity of acid protease is more than or equal to 1500U/g, the enzyme activity of neutral protease is more than or equal to 2500U/g, the viable count of bacillus subtilis is more than or equal to 8 hundred million/g, the viable count of enterococcus faecalis more than or equal to 1.5 hundred million/g, and the viable count of bacillus coagulans is more than or equal to 3 hundred million.

Further, in step S5.4, the mixing time is 2-4 minutes.

The preparation method disclosed by the invention is simple in preparation process and strong in operability, and is convenient to popularize and apply to small-sized feed enterprises and farms.

The invention also provides application of the combined feed additive for the palm meal type feed in the palm meal-containing feed, when the dosage of the palm meal in the compound feed is 3-6%, the addition amount of the feed additive is 300 g/ton of the compound feed; when the dosage of the palm meal in the compound feed is 6-10%, the addition amount of the feed additive is 300-500 g/ton of the compound feed, and the mixture is uniformly mixed for use. After the additive prepared by the invention is added into the palm meal-containing feed, the usage amount of the palm meal can be increased by 3-5%, the usage amount of the palm meal in the compound feed is effectively increased, and the feed cost is obviously reduced.

Drawings

FIG. 1 shows the effect of different fermentation strains on laying rate, feed intake and feed-egg ratio of laying hens;

FIG. 2 shows the effect of different fermentation strains on the laying rate of laying hens in different growth periods;

FIG. 3 is the effect of different treatment groups on the growth performance of cherry valley meat ducks;

FIG. 4 shows the effect of different treatment groups of the present invention on milk production of cows.

Detailed Description

The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the scope of protection of the present invention.

Example 1

In the embodiment, the additive comprises a complex enzyme preparation, a compound microorganism and a multi-strain solid fermentation product with corresponding quality, the complex enzyme preparation and the compound microorganism as main components come from two production modes of liquid fermentation and solid fermentation, and have complementary action, so that the characteristics of high activity and high content of the liquid fermentation are fully utilized, the characteristic of strong pertinence of the solid fermentation is utilized, the utilization rate of the feed is improved, the use cost of the feed is reduced, and the breeding benefit is increased. Wherein the beta-mannase is a product produced by submerged fermentation of Bacillus lentus, is neutral beta-mannase, and has relative enzyme activity of more than 90% at pH6-7 and 56% in 30 deg.C water bath; the alpha-amylase is obtained by liquid submerged fermentation of bacillus subtilis, the relative enzyme activity is more than 50% when the optimal pH value is 6.0 and the pH value is 4.3-7.7, and the activity can be kept more than 50% in a water bath at the temperature of 28-78 ℃; the neutral protease is obtained by liquid submerged fermentation of bacillus subtilis; the acid protease is produced by aspergillus niger through a solid fermentation process; the xylanase can obviously reduce the molecular size of the araban, and decomposes the xylan into xylooligosaccharide with smaller polymerization degree, thereby improving the feed performance and eliminating the anti-nutritional effect of the xylan in the feed. The yeast xylanase and the trichoderma xylanase have a narrow pH adaptation range, but have poor enzyme activity in the animal digestive tract temperature range, and the pH adaptation range can be widened in the animal digestive tract temperature range by combining the two types of xylanase and trichoderma xylanase.

In the embodiment, the mass percent of the bacillus subtilis is 0.5-1.0% of the substrate, and the mass percent of the bacillus coagulans is 1.0-1.4% of the substrate; under the proportion, the bacillus subtilis and the bacillus coagulans are fermented in a combined manner, so that the bacillus subtilis and the bacillus coagulans can be fully propagated, the viable count is rapidly increased, and digestive enzymes such as amylase, protease and the like with higher activity are generated in the fermentation process; the beneficial bacteria and digestive enzyme are different from the complex enzyme preparation and the complex microorganism added in the subsequent mixing process in source, but have the same effect, have additive effect and more thoroughly decompose the feed.

As a preferred embodiment, the substrate consists of 29-33.6 parts by mass of corn flour, 20-30 parts by mass of bran, 23-32 parts by mass of palm meal, 12-16 parts by mass of soybean meal, 0.7-1.5 parts by mass of urea and 0.7-1.5 parts by mass of glucose.

In a preferred embodiment, the composite microorganism comprises 0.8-1.2 parts by mass of Bacillus subtilis with 1000 hundred million viable bacteria/g, 3-5 parts by mass of enterococcus faecalis with 50 hundred million viable bacteria/g and 3-5 parts by mass of Bacillus coagulans with 100 hundred million viable bacteria/g. Bacillus subtilis is aerobic bacteria, Bacillus coagulans is facultative anaerobe, and the Bacillus coagulans can be cultured together under aerobic conditions; moreover, the two are widely applied in the feed and breeding industries, and the effect is prominent. The bacillus subtilis mainly has the function of producing enzyme and can improve the digestibility of the palm meal type feed; secondly, the feed has a balancing effect on intestinal flora, and can relieve flora disturbance caused by mannan viscosity in palm meal. The bacillus coagulans and the enterococcus faecalis mainly used for producing acid, can jointly maintain the low pH environment of the animal digestive tract, and reduce the risk of the disturbance of animal intestinal flora caused by palm meal.

s5.1, preparing the complex enzyme preparation

s5.2, preparation of composite microorganism

s5.3, preparing multi-strain solid fermentation product

s5.3.2, activating strains: adding a certain amount of mixed strains of 1000 hundred million viable bacteria/g of bacillus subtilis and 100 hundred million viable bacteria/g of bacillus coagulans into sterile water which is 5-10 times of the mixed strains by mass, then adding glucose which is 0.5-1 times of the mixed strains by mass, fully stirring and preserving heat to prepare seed bacteria liquid for later use, wherein the sterile water is water which is boiled and then placed for cooling to 30-35 ℃;

and S5.4, mixing products obtained from S5.1, S5.2 and S5.3 in proportion.

In this embodiment, in step S5.3, the mass percentage of bacillus subtilis is 0.5% to 1.0% of the substrate, and the mass percentage of bacillus coagulans is 1.0% to 1.4% of the substrate;

as a preferred embodiment, in step S5.3, the substrate consists of 29 to 33.6 parts by mass of corn flour, 20 to 30 parts by mass of bran, 23 to 32 parts by mass of palm meal, 12 to 16 parts by mass of soybean meal, 0.7 to 1.5 parts by mass of urea and 0.7 to 1.5 parts by mass of glucose.

In a preferred embodiment, step S5.4 comprises, after mixing, the following ingredients in amounts: the enzyme activity of beta-mannase is more than or equal to 4000U/g, the enzyme activity of xylanase is more than or equal to 3000U/g, the enzyme activity of alpha-amylase is more than or equal to 1050U/g, the enzyme activity of acid protease is more than or equal to 1500U/g, the enzyme activity of neutral protease is more than or equal to 2500U/g, the viable count of bacillus subtilis is more than or equal to 8 hundred million/g, the viable count of enterococcus faecalis more than or equal to 1.5 hundred million/g, and the viable count of bacillus coagulans is more than or equal to 3 hundred million.

In a preferred embodiment, in step S5.4, the mixing time is 2 to 4 minutes.

In this embodiment, the method for producing the multi-strain solid fermentation product is preferably:

s1, pretreatment of a substrate: weighing 29-33.6 parts of corn flour, 20-30 parts of bran, 23-32 parts of palm meal, 12-16 parts of soybean meal, 0.7-1.5 parts of urea and 0.7-1.5 parts of glucose in parts by mass, uniformly mixing, crushing and sieving with a 30-mesh sieve; according to the material-liquid ratio of 1: 0.35-0.85 of water is added and mixed evenly again; high-pressure steam sterilization, the sterilization parameters are as follows: 0.15-0.3 Mpa for 20-30 min;

s2, strain activation: according to the mass percentage of the air-dried substances of the substrate, 0.5 to 1.0 percent of bacillus subtilis with 1000 hundred million living bacteria/g and 1.0 to 1.4 percent of bacillus coagulans with 100 hundred million living bacteria/g are weighed; mixing the mixed strains of the bacillus subtilis and the bacillus coagulans with 5-10 times of sterile water with the mass of 30-35 ℃; simultaneously, adding glucose with the mass of 0.5-1 time of that of the mixed strain, and fully stirring; keeping the temperature for 2-4 hours, wherein the sterile water is boiled and then cooled to the required temperature to obtain seed bacterial liquid;

s3, fermentation: uniformly spraying the activated seed bacterial liquid in the S2 into the substrate treated by the S1, adding sterile water in batches, uniformly mixing, calculating and adjusting the specific amount of the sterile water to control the water content of the fermentation substrate to be 41-56% in the fermentation process, loading into a tray, fermenting for 60-72 hours in a constant temperature and humidity cabinet at the temperature of 35-45 ℃, turning over once every 2 hours, and carrying out fermentation culture;

s4, drying and crushing: and (3) drying the fermented product obtained after fermentation culture in the S3 in a ventilated drying oven at the temperature of less than or equal to 60 ℃ until the water content is less than or equal to 15%, crushing and sieving by a 30-mesh sieve.

In the embodiment, the compound enzyme preparation, the compound microorganism and the multi-strain solid fermentation product are prepared firstly, then the compound enzyme preparation, the compound microorganism and the multi-strain solid fermentation product are respectively weighed according to the proportion and mixed, the mixing time is 2-4 minutes, and the combined feed additive for the palm meal type feed is obtained, so that the utilization rate of the feed is improved, the use cost of the feed is reduced, and the breeding benefit is increased; the preparation method is simple in preparation process and strong in operability, and is convenient to popularize and apply to small-sized feed enterprises and farms.

Example 2

The present embodiment provides a method for preparing a combined feed additive for palm meal type feed, which aims to manufacture 500kg of the combined feed additive for palm meal type feed, and comprises the following steps:

s1, preparing a complex enzyme preparation: respectively weighing 50000U/g beta-mannase with enzyme activity 53kg, 100000U/g xylanase with enzyme activity 18kg, 7000U/g alpha-amylase with enzyme activity 80kg, 50000U/g acid protease with enzyme activity 20kg and 50000U/g neutral protease with enzyme activity 28kg, sequentially putting into a mixer, and mixing for 3 minutes;

s2, preparing a compound microorganism: respectively weighing 5.5kg of bacillus subtilis with 1000 hundred million/g of live bacteria, 18kg of enterococcus faecalis with 50 hundred million/g of live bacteria and 21kg of bacillus coagulans with 100 hundred million/g of live bacteria, sequentially putting into a drum mixer, and mixing for 3 minutes;

s3, preparation of multi-strain solid fermentation:

s3.1, fermentation substrate pretreatment: respectively weighing 154kg of corn flour, 125kg of wheat bran, 70kg of soybean meal, 140kg of palm meal, 5.5kg of urea and 5.5kg of glucose, placing the materials in a double-screw mixer, mixing for 3 minutes, and then crushing by a 30-mesh crusher; steam sterilizing at 0.25Mpa for 20 min to obtain fermentation substrate;

s3.2, strain activation: weighing 230kg of sterilized water, adding the sterilized water into a fermentation substrate in batches, and uniformly mixing; respectively weighing 0.8kg of bacillus subtilis with 1000 hundred million CFU/g, 1.2kg of bacillus coagulans with 100 hundred million CFU/g and 2kg of glucose, adding into a heat-insulating bucket containing 20kg of sterilized water, uniformly stirring, and cooling the sterilized water which is boiled boiling water to 30-35 ℃; keeping the temperature to activate the strains for 4 hours;

s3.3, fermentation: sprinkling the activated strain liquid into the fermentation substrate b in batches, and stirring uniformly, wherein the actual measurement moisture content is 49.6%; the wet fermentation substrate is batched, placed in an automatic temperature and humidity control fermentation box, fermented at the temperature of 37 ℃ for 72 hours, turned over once every 2 hours, and properly sprayed with supplementary water according to the change condition of the water content of the material;

s3.4, drying and crushing: drying the fermented product with hot air of a fluidized bed until the moisture content is actually measured to be 13.7%, crushing, and sieving with a 30-mesh sieve;

s4, weighing 256.5kg of the multi-strain solid fermentation product prepared in the step S3, putting the multi-strain solid fermentation product into a double-screw mixer, sequentially putting the complex enzyme preparation and the complex microorganism prepared in the step S1 and the step S2, starting the machine, mixing for 3 minutes, and bagging to obtain the combined feed additive for the palm meal type feed.

The content of each component in the prepared feed additive is detected in a laboratory, and the result is as follows: the enzyme activity of beta-mannase is 6739U/g, the enzyme activity of xylanase is 4077U/g, the enzyme activity of alpha-amylase is 1443U/g, the enzyme activity of acid protease is 3204U/g, the enzyme activity of neutral protease is 4394U/g, the viable count of bacillus subtilis is 13.7 hundred million CFU/g, the viable count of enterococcus faecalis 1.7 hundred million CFU/g, and the viable count of bacillus coagulans is 5.2 hundred million/g. The content of each component in the result accords with the value specified in the prepared additive, wherein the measured value of most indexes is obviously larger than the theoretical calculated value because the bacillus subtilis and the bacillus coagulans are propagated in the fermentation process of multi-strain solid fermentation products, and simultaneously, a plurality of enzymes are produced in the fermentation process, and the digestibility of the palm pulp type feed is improved through the synergistic effect of the enzymes and the bacteria from a plurality of sources.

Experimental example 3

The experimental example is used for measuring the enzymological characteristics of the combined xylanase mixed by the yeast type xylanase and the trichoderma type xylanase according to the proportion of 7:3, and the enzymological characteristics are compared with the enzymological characteristics of the independent yeast type xylanase and the trichoderma type xylanase respectively to illustrate the advantages of the combined xylanase, and specifically comprises the following steps:

respectively weighing 70g of 100000U/g yeast type xylanase and 30g of 100000U/g trichoderma type xylanase, and uniformly mixing to obtain the combined xylanase for later use.

The enzyme activities of the yeast type xylanase, the trichoderma type xylanase and the combined xylanase in water baths with different pH values and different temperatures are respectively determined by a method specified in GB/T23874-2009 feed additive xylanase activity determination spectrophotometry, and the relative enzyme activities under each determination condition are respectively calculated by taking the highest enzyme activity as 100%.

And (3) enzyme activity determination at different pH: according to the GB/T23874-2009 method, a reaction solution with the pH value of 3.0-8.5 (the pH interval is 0.5) is prepared by using a corresponding buffer solution of the enzyme, the reaction solution is subjected to water bath heat preservation for 4 hours at a corresponding optimal temperature, the enzyme activity is respectively determined, and the highest enzyme activity is taken as the optimal pH value of the enzyme; the pH range with relative enzyme activity greater than 50% is the proper pH range.

And (3) measuring enzyme activity under different temperature treatment conditions: referring to the GB/T23874-containing method, the temperature of a reaction liquid is respectively adjusted to 25-70 ℃ in a constant-temperature water bath kettle (the temperature interval is 5 ℃), the temperature is kept for 30 minutes, the enzyme activity of each sample is immediately measured, and the temperature with the highest enzyme activity is the optimal reaction temperature of the enzyme; the temperature range with relative enzyme activity of more than 50 percent is taken as the suitable temperature range.

The effects of pH and temperature on xylanase activity are shown in tables 1 and 2, respectively.

TABLE 1 Effect of pH on xylanase Activity

TABLE 2 Effect of temperature on xylanase Activity

pH adaptability and temperature adaptability are two important indexes for evaluating enzyme quality. The ideal feed enzyme preparation should have the following characteristics: the optimum action temperature is close to the animal digestive tract temperature (37-40 ℃), and the enzyme preparation has wide pH adaptation range and wide temperature adaptation range so as to play a role in more parts of the animal digestive tract.

As can be seen from tables 1 and 2, the Trichoderma xylanase has the optimum pH of 5, the applicable pH range is very wide, and the relative enzyme activity is more than 50 percent in the range of pH3.5-8.5, which is obviously superior to the yeast xylanase (pH 4.5-7); but the adaptability to temperature is poor (45-60 ℃), particularly, the enzyme activity is lower than 50% in the range of 37-40 ℃ of the animal digestive tract, and is obviously lower than that of yeast type xylanase (40-65 ℃), the pH adaptation of the yeast type xylanase is very narrow, the pH adaptation range is only 4.5-7, and the relative enzyme activity is more than 50%.

However, the combined xylanase has a wider pH adaptation range (4-8) and a wider temperature adaptation range (40-65 ℃), relatively overcomes the defect of singly using the yeast type xylanase or the trichoderma type xylanase, and enables the enzyme preparation to play a better role in the digestive tract of animals.

Experimental example 4

The experimental example is to verify the influence of the selection of different fermentation strains in the multi-strain solid fermentation stage on the final product feeding effect through a layer feeding test.

The test is carried out by setting 3 groups, namely a bacillus subtilis group, a bacillus coagulans group and a compound bacteria group, wherein each group of 1200 Hailan brown laying hens aged 36 weeks is correspondingly fed with 3 types of daily rations with completely same basic compositions, the test period is 70 days, and the formula and the nutrient composition table of the daily rations are respectively shown in tables 3 and 4.

The difference between the compound bacterium group and the bacillus subtilis group and the bacillus coagulans group is that: 300 g/ton of the feed additive prepared in the embodiment 2 is added into the compound bacterium group;

300 g/ton of feed additive is added into the bacillus subtilis group, and the difference of the preparation method of the feed additive of the bacillus subtilis and the preparation method of the feed additive in the embodiment 2 is as follows: in the multi-strain solid fermentation stage, only bacillus subtilis is used for fermentation, and other methods are the same as those in the example 2;

300 g/ton of feed additive is added into the bacillus coagulans group, and the preparation method of the feed additive of the bacillus coagulans is different from the preparation method of the feed additive in the embodiment 2 in that: in the multi-strain solid fermentation stage, only bacillus coagulans is used for fermentation, and other methods are the same as those in example 2;

TABLE 3 prescription of daily ration

TABLE 4 nutrient composition table of daily ration

The influence of different strains of fermentation products on the laying rate, feed intake and feed-egg ratio of the laying hens is shown in figure 1, and the influence of different strains of fermentation products on the laying rate of the laying hens in different growth periods is shown in figure 2.

As can be seen from the graph 1, the egg laying rates of the laying hens of the bacillus subtilis group and the bacillus coagulans group are similar, but the egg laying rate of the compound bacterium group is respectively improved by 0.77 percent and 0.82 percent compared with that of the former two groups, and the egg laying rate improvement is difficult and expensive in the egg laying peak period of the laying hens, and as can be seen from the graph 2, the egg laying rate stability curve analysis in the 70-day test period shows that the egg laying rate of the compound bacterium group is relatively more stable, so that the effect of the combined fermentation of the bacillus subtilis and the bacillus coagulans in the multi-strain solid fermentation stage is better than that of the single bacterium fermentation, the combined fermentation has better egg laying rate, and the change of the egg laying rate to the growth period is more stable.

Experimental example 5

The experimental example is the application of the additive in the palm meal type compound feed for meat ducks, and the application effect of the additive is verified through a feeding test for reducing the feed cost.

360 cherry valley meat ducks are divided into 3 groups, and each group comprises 120 ducks which are respectively a positive control group, a negative control group and a test group. The same daily ration is fed for 0-14 days old in the early stage, and 3 different daily rations are correspondingly fed for 15-38 days old in the later stage. At the later 15-38 days of age, the ration fed to the positive control group is unchanged; on the basis of the daily ration of the positive control group, the formula of the negative control group is adjusted, 8% of palm meal is added, the metabolic energy design level is reduced by 50kcal/kg, and the formula cost is correspondingly reduced; test groups: 400 g/ton of the additive prepared in the example 2 was added on the basis of the daily ration of the negative control group. The formula table and the nutrient composition table of the test daily ration in the later period are respectively shown in tables 5 and 6:

table 5 formula table of experimental diets in later stage

TABLE 6 nutritional composition of test diets at later stages

The test group adds 400g/T of the additive on the basis of negative control daily ration, the cost is increased by 28 yuan/T, and the formula cost is 2358.5 yuan/ton.

And (3) test period: the average body weight and the material weight ratio were counted at the end of the test period for 38 days, and the results are shown in fig. 3.

As can be seen from FIG. 3, compared with the positive control group, when 8% of palm meal is added into the later-stage compound feed and the metabolic energy is reduced by 50kcal/kg, the growth speed of the meat ducks is obviously reduced, compared with the negative control group, the production performance of the meat ducks can be recovered by adding the additive, the weight-to-material weight ratio of the meat ducks is similar to that of the positive control group, the feed cost is reduced, and compared with the positive control group, the feed cost per ton of the test group is reduced by 16.7 yuan. Therefore, the additive can be added into the feed to increase the content of the palm meal in the feed and reduce the use cost of the feed.

Experimental example 6

The experimental example is a feeding effect verification that the additive is added into a cow feed and palm meal is used for replacing part of corn and soybean meal.

267 holstein lactating cows, mean number of lactating days 175d, in three groups: positive control group, negative control group, test group.

The formula of the concentrate of the positive control group is as follows: 55% of corn, 10% of bran, 15% of corn, 10% of soybean meal, 3.5% of cottonseed meal, 4% of oil cake, 0.5% of salt, 0.7% of monocalcium phosphate, 0.8% of baking soda and 0.5% of ruminant premix.

Concentrate of negative control group: on the basis of the refined material of the positive control group, 10 percent of corn and 2 percent of soybean meal are replaced by 12 percent of palm meal.

Test group concentrate: on the basis of the positive control group concentrate, 10% of corn and 2% of soybean meal are replaced by 12% of palm meal; 500 g/ton of the additive prepared in example 2 were added simultaneously.

The daily milk yield of each group of cows is counted once every two days in the test period of 24 days. In 1 week before the test, the dosage of the palm meal is gradually increased by taking 3% as a gradient in the negative control group and the test group, so that the test cattle are adapted to new feed.

TABLE 6 milk production scale for dairy cows

As can be seen from table 6, 12% palm meal instead of 10% corn and 2% soybean meal slightly decreased milk production by cows, and the average milk production per day was decreased by 0.16kg, but the difference was not significant. Compared with a negative control group, after the additive is added in an amount of 500 g/ton while 12% of palm meal is used, the average milk yield of the dairy cow per day is improved by 0.87kg/d, and even compared with a positive control group without the palm meal, the average balance milk yield is improved by 0.71 kg/d. As can be seen from the milk yield curve in FIG. 4, the milk yield of the cows is obviously more balanced and stable after the additive of the invention is added, which is probably the regulating effect of the microorganisms in the milk on the rumen of the cows. In general, this example demonstrates that the additive of the present invention has a significant effect in the application of the palm meal type feed for dairy cows, and can improve the daily milk yield of dairy cows and make the milk yield more stable.

Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. A combined feed additive for palm meal type feed is characterized in that: is composed of 34 to 48 mass portions of complex enzyme preparation, 6.8 to 11.2 mass portions of complex microorganism and 40.8 to 59.2 mass portions of multi-strain solid fermentation product;

2. The combined feed additive for palm meal type feed according to claim 1, wherein: the mass percent of the bacillus subtilis is 0.5-1.0% of the substrate, and the mass percent of the bacillus coagulans is 1.0-1.4% of the substrate.

3. The combined feed additive for palm meal type feed as claimed in claim 2, wherein: the substrate consists of 29-33.6 parts by mass of corn flour, 20-30 parts by mass of bran, 23-32 parts by mass of palm meal, 12-16 parts by mass of soybean meal, 0.7-1.5 parts by mass of urea and 0.7-1.5 parts by mass of glucose.

4. The combined feed additive for palm meal type feed according to any one of claims 1 to 3, wherein: the compound microorganism consists of 0.8 to 1.2 parts by mass of bacillus subtilis with 1000 hundred million/g live bacteria, 3 to 5 parts by mass of enterococcus faecalis with 50 hundred million/g live bacteria and 3 to 5 parts by mass of bacillus coagulans with 100 hundred million/g live bacteria.

5. A preparation method of a combined feed additive for palm meal type feed is characterized by comprising the following steps: the method comprises the following steps:

s5.1, preparing the complex enzyme preparation

s5.2, preparation of composite microorganism

s5.3, preparing multi-strain solid fermentation product

and S5.4, mixing products obtained from S5.1, S5.2 and S5.3 in proportion.

6. The method for preparing a combined feed additive for palm meal type feed according to claim 5, wherein the method comprises the following steps: in the step S5.3, the mass percent of the bacillus subtilis is 0.5-1.0% of the substrate, and the mass percent of the bacillus coagulans is 1.0-1.4% of the substrate.

7. The method for preparing a combined feed additive for palm meal type feed according to claim 5, wherein the method comprises the following steps: in step S5.3, the substrate consists of 29-33.6 parts by mass of corn flour, 20-30 parts by mass of bran, 23-32 parts by mass of palm meal, 12-16 parts by mass of soybean meal, 0.7-1.5 parts by mass of urea and 0.7-1.5 parts by mass of glucose.

8. The method for preparing a combined feed additive for palm meal type feed according to claim 5, wherein the method comprises the following steps: in step S5.4, the components and contents after mixing are as follows: the enzyme activity of beta-mannase is more than or equal to 4000U/g, the enzyme activity of xylanase is more than or equal to 3000U/g, the enzyme activity of alpha-amylase is more than or equal to 1050U/g, the enzyme activity of acid protease is more than or equal to 1500U/g, the enzyme activity of neutral protease is more than or equal to 2500U/g, the viable count of bacillus subtilis is more than or equal to 8 hundred million/g, the viable count of enterococcus faecalis more than or equal to 1.5 hundred million/g, and the viable count of bacillus coagulans is more than or equal to 3 hundred million.

9. The method for preparing a combined feed additive for palm meal type feed according to claim 5, wherein the method comprises the following steps: in step S5.4, the mixing time is 2-4 minutes.

10. Use of the additive according to any one of claims 1 to 4 in a palm meal-containing feed, wherein: when the dosage of the palm meal in the compound feed is 3% -6%, the addition amount of the feed additive is 300 g/ton of the compound feed; when the dosage of the palm meal in the compound feed is 6-10%, the addition amount of the feed additive is 300-500 g/ton of the compound feed, and the mixture is uniformly mixed for use.