CN109007402B

CN109007402B - Laying hen feed additive and preparation method thereof

Info

Publication number: CN109007402B
Application number: CN201810629988.6A
Authority: CN
Inventors: 杨建平; 赵志军; 姚国佳; 常选妞; 李梦云; 聂芙蓉
Original assignee: Henan University of Animal Husbandry and Economy
Priority date: 2018-06-19
Filing date: 2018-06-19
Publication date: 2020-09-25
Anticipated expiration: 2038-06-19
Also published as: CN109007402A

Abstract

The invention discloses a laying hen feed additive and a preparation method thereof. The laying hen feed additive is composed of the following raw materials in parts by weight: 15-25 parts of microbial fermentation products of boxthorn leaves, boxthorn stalks, corn flour and sesame seed meal, 70-80 parts of potato flour, 2-6 parts of medlar, 0.5-1 part of anise and 0.1-0.3 part of thyme. And designs a preparation method of the laying hen feed additive. The laying hen feed additive disclosed by the invention can improve the oxidation resistance of laying hens, enhances the immunity of the laying hens, is beneficial to improving the production performance and the egg quality, and is simple and convenient to operate and controllable in cost.

Description

Laying hen feed additive and preparation method thereof

Technical Field

The invention relates to the technical field of livestock and poultry feed, in particular to a laying hen feed additive and a preparation method thereof.

Background

With the rapid development of livestock and poultry breeding industry, the livestock and poultry feed and feed additive industry is also rapidly expanded, and various feeds and feed additives for reducing the livestock and poultry raising cost, improving the immunity and the production performance of the livestock and poultry and the like appear in the market. However, due to the growth characteristics, optimized nutrient supply and precise breeding mode of the oriented breeding of the livestock and poultry, the digestion and absorption frequency of the livestock and poultry is in a high-intensity state, so that the integrity of intestinal mucosa is seriously damaged by the livestock and poultry, the intestinal health of the livestock and poultry is damaged, and even intestinal diseases are caused. Meanwhile, the flora imbalance and the immunity in the intestinal tract are reduced, and finally, the production performance of the livestock and poultry is reduced.

Because the laying hen breeding industry in China has the problems of high-density breeding environment, frequent immunity, physiological characteristics in the later period of egg laying and the like, the laying hen production performance and the egg laying quality in China cannot be well guaranteed. And the egg loss caused by the quality problem of the eggshell in China is up to more than 10 percent every year, which seriously limits the development of the laying hen breeding industry. Therefore, the technical problems of enhancing the immunity of the laying hens and improving the egg laying production performance and the egg quality of the laying hens are urgently to be solved in the breeding industry at present.

Disclosure of Invention

The invention aims to provide a laying hen feed additive and a preparation method thereof, so that healthy growth of laying hens can be promoted, and the laying performance and egg quality of the laying hens can be improved.

In order to solve the technical problems, the invention adopts the following technical scheme:

the laying hen feed additive is designed and comprises the following raw materials in parts by weight: 15-25 parts of microbial fermentation products of boxthorn leaves, boxthorn stalks, corn flour and sesame seed meal, 70-80 parts of potato flour, 2-6 parts of medlar, 0.5-1 part of anise and 0.1-0.3 part of thyme.

Wherein the preparation method of the microbial fermentation product comprises the following steps:

pulverizing folium Lycii, fructus Lycii stalk, corn flour and semen Sesami cake, sieving, and adding microorganism; and drying after culturing to obtain a fermentation product.

Preferably: the mass ratio of the boxthorn leaves to the boxthorn stalks to the corn flour to the sesame seed meal is 0.5-1.5: 0.5-1.5: 0.1-0.3: 0.05 to 0.2.

Preferably: the microorganism is lactobacillus plantarum BNCC187897 and/or bacillus subtilis AS 1.9086.

Preferably: the mass ratio of the microorganisms to the fermentation liquor is 5: 1.

preferably: the boxthorn leaves, the boxthorn stalks, the corn flour, the sesame meal and the like are crushed and sieved by a sieve of 50-70 meshes.

The preparation method of the laying hen feed additive comprises the following steps:

(1) taking the boxthorn leaves, the boxthorn stalks, the corn flour and the sesame seed meal, crushing and sieving, adding microorganisms, culturing and drying to obtain a fermentation product;

(2) mixing the potato powder and the obtained fermentation product, stirring until the mixture is uniformly mixed, then sequentially adding the star anise powder, the thyme powder and the wolfberry fruit powder, and stirring until the mixture is uniformly mixed to obtain the potato milk.

Compared with the prior art, the invention has the beneficial technical effects that:

1. the laying hen feed additive is designed according to the growth and development characteristics of laying hens, all components are matched with each other for synergism, the cecal flora structure of the laying hens can be greatly improved, the digestion and absorption capacity of birds can be enhanced, the egg laying performance of the laying hens can be effectively improved, the egg quality can be improved, and the immunity of the laying hens can be improved.

2. Furthermore, the fermentation product prepared and added by the method contains a large amount of crude protein, crude fiber, crude ash, total amino acid, water, acetic acid, propionic acid, lactic acid, crude polysaccharide, polyphenol and crude flavone, wherein the polyphenol is an important bioactive substance, has strong antioxidant property and can enhance the antioxidant capacity of livestock and poultry; the crude flavone has strong effects of resisting oxidation, liver toxicity and inflammation, can enhance the immunity of livestock and poultry, promotes the healthy growth of livestock and poultry, and is beneficial to improving the production performance of livestock and poultry and the egg quality; researches find that the aniseed and the thyme contain rich plant essential oil, have the functions of resisting bacteria, fungi, viruses, parasites and the like, and can play a role in replacing antibiotics; the added wolfberry powder can improve the immunity of livestock and poultry, can also enhance the reproductive capacity of the livestock and poultry and improve the laying rate; the laying hen feed additive disclosed by the invention is reasonably matched with crude protein, crude fiber and total amino acid, and can enhance the digestion and absorption capacity of livestock and poultry.

3. The invention has the advantages of wide raw material source, synergistic combination of all components, lower production cost and obvious effect, and is beneficial to wide application in livestock production.

Drawings

FIG. 1 is a bar graph of the relative abundance of species at the gate level, where C is the control group and S is the test group;

FIG. 2 is a histogram of the relative abundance of species at genus level, where C is the control group and S is the test group;

FIG. 3 is a diagram showing the analysis of the principal components of the cecal bacteria group in the later period of egg laying by the phylum level layer, wherein the abscissa of the diagram represents the first principal component, and the percentage represents the contribution value of the first principal component to the difference of the sample; the ordinate represents the second principal component, and the percentage represents the contribution of the second principal component to the difference of the sample; each point in the graph represents a sample, and samples of the same group are represented using the same color, as follows;

FIG. 4 is a diagram of analysis of the main components of the cecal bacteria group of the laying hens belonging to the horizontal later period of laying.

Detailed Description

The following examples are intended to illustrate the present invention in detail and should not be construed as limiting the scope of the present invention in any way.

The instruments and devices referred to in the following examples are conventional instruments and devices unless otherwise specified; the related products are all conventional products sold in the market if not specifically indicated; the test methods or analytical methods referred to are, unless otherwise indicated, conventional.

The first embodiment is as follows: preparation of laying hen feed additive

1. Screening of fermentation Strain

Through a large number of experimental researches, the lactobacillus plantarum BNCC187897 freeze-dried powder (purchased from China microbial culture collection center, BNCC) and the bacillus subtilis AS1.9086 (purchased from China academy of sciences and microbiological research institute strain collection center) suitable for the mixture of the boxthorn leaves, the boxthorn stalks, the corn flour and the sesame seed meal are preferably selected AS the fermentation microorganisms, the fermentation product has comprehensive and rich nutrient substances, the materials such AS the boxthorn leaves, the boxthorn stalks, the sesame seed meal and the like can be fully degraded, and more effective components suitable for the poultry are generated.

2. Culture medium

Nutrient Broth (NB): 3g of beef powder, 10g of peptone, 5g of sodium chloride and 1000mL of distilled water. The pH value is 6.8-7.0, and sterilization is carried out for 15min at 121 ℃.

Nutrient agar medium: on the basis of a nutrient broth culture medium, 15-20 g of agar powder is added into each liter of culture solution.

MRS broth culture medium: 10g of peptone, 4g of yeast powder, 5g of beef powder, 20g of glucose, 5g of sodium acetate, 2g of triammonium citrate, 2g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and 801 mL of tween.

MRS agar medium: on the basis of an MRS broth culture medium, 15-20 g of agar powder is added into each liter of culture solution.

3. Seed liquid preparation

B, bacillus subtilis: selecting a small amount of frozen bacillus subtilis strain, streaking and inoculating the bacillus subtilis strain to a sterilized nutrient agar culture medium, culturing at a constant temperature of 37 ℃ for 24h for strain activation, then inoculating the activated strain to the sterilized nutrient broth culture medium, placing in a constant temperature shaking table, and culturing at a temperature of 37 ℃ and a rotating speed of 180r/min for 24h for later use.

Lactobacillus plantarum: selecting a small amount of frozen lactobacillus plantarum strain, streaking and inoculating the strain on a sterilized MRS agar culture medium, culturing at the constant temperature of 37 ℃ for 24h for strain activation, then inoculating the activated strain in the sterilized MRS broth culture medium, and culturing in a constant temperature incubator at the temperature of 37 ℃ for 24h for later use.

4. Composition and preparation of fermentation substrate

Pulverizing folium Lycii, fructus Lycii stalk, corn flour and semen Sesami cake, and sieving with 60 mesh sieve. The raw materials of 4 kinds are as follows: 1: 0.2: mixing at 0.1 mass ratio for use.

5. Solid fermentation

Fermentation conditions are as follows:

taking a fermentation substrate, and mixing feed liquid according to the ratio of 10: 1, wherein the ratio of the lactobacillus plantarum to the bacillus subtilis is 5: 1.

the mass ratio of the initial water content is 35 percent, the culture is carried out for 72 hours at 37 ℃, and the fermentation sample is sent to Henan Hairui positive detection technology company Limited for detection.

The conventional nutrient components in the fermentation product are shown in table 1:

TABLE 1 conventional nutrient content in fermentation product (dry matter basis)

	Crude protein%	Crude fiber%	Coarse ash content%	Total amino acid%
					Content (wt.)	18.11	8.81	23.86	13.08
According to	GB/T 6432-1994	GB/T 6434-2006	GB/T 6438-2007	GB/T18246-2000

The bioactive components in the fermentation product are shown in table 2:

TABLE 2 bioactive components in the fermentation product

Acetic acid mg/kg

Propionic acid mg/kg

Lactic acid mg/kg

Crude polysaccharide%

Polyphenol%

Crude flavone%

Content (wt.)

2296.18

102.21

974.52

2.40

0.98

0.91

According to

Gas chromatography

GB/T 23877-2009

SN/T4260-2015

Spectrophotometry

6. Drying the fermentation product

Drying the fermentation product at 50 ℃ for later use.

7. Preparation of laying hen feed additive

The method specifically comprises the following steps:

mixing 75 parts of potato powder and 20 parts of fermentation product, stirring for 3-5 min until the mixture is uniformly mixed, then sequentially adding 0.8 part of star anise powder, 0.2 part of thyme powder and 4 parts of medlar powder, and stirring for 5min until the mixture is uniformly mixed to obtain the potato milk.

Example two: preparation of laying hen feed additive

The difference from the first embodiment is that:

1. crushing the boxthorn leaves, the boxthorn stalks, the corn flour and the sesame seed meal in the fermentation substrate according to the weight ratio of 0.5: 1.5: 0.1: mixing at 0.2 mass ratio for use.

2. Preparation of laying hen feed additive

Mixing 70 parts of potato powder and 15 parts of fermentation product, stirring for 3-5 min until the mixture is uniformly mixed, then sequentially adding 0.5 part of star anise powder, 0.1 part of thyme powder and 2 parts of medlar powder, and stirring for 5min until the mixture is uniformly mixed to obtain the potato milk.

Example three: preparation of laying hen feed additive

The difference from the first embodiment is that:

1. crushing the boxthorn leaves, the boxthorn stalks, the corn flour and the sesame seed meal in the fermentation substrate according to a proportion of 1.5: 0.5: 0.3: mixing at 0.05 mass ratio for use.

2. Preparation of laying hen feed additive

Mixing 80 parts of potato powder and 25 parts of fermentation product, stirring for 3-5 min until the mixture is uniformly mixed, then sequentially adding 1 part of star anise powder, 0.3 part of thyme powder and 6 parts of medlar powder, and stirring for 5min until the mixture is uniformly mixed to obtain the potato milk.

Example four: research on influence of laying hen feed additive on egg quality

1. Test animal

Animal experiments were carried out in Beijing Xin Sheng breed, Inc., and the layer variety was Hailan brown layer.

2. Design and management of experiments

The test adopts single-factor completely random design, and 5000 healthy, active, laying rate and weight similar 406-day-old kalanchoe brown laying hens are randomly selected and divided into 2 treatment groups which are respectively a control group and a test group. The pre-feeding period is set for 7 days, the hailan brown laying hens are subjected to epidemic prevention, feeding and drinking in the pre-feeding period, and then the formal test period is started. The control group was fed a conventional basal diet and the test group was fed a diet supplemented with the feed additive of example one at 0.5% of its weight in a conventional basal diet for a test period of 49 days.

3. Daily management

The test is carried out by adopting a cage-breeding mode in a house, natural illumination, free drinking and ingestion of all layers, disinfection and immunization according to a conventional method. Feed intake and egg production weight were weighed and recorded daily at 16:00 pm. The manure is cleaned once every two days. The mental state and behavior of the laying hens are observed daily during the test period, the daily feed consumption, egg production and egg breaking amount of each group are recorded, and the test period is 49 days.

4. Data processing

Data were analyzed for variance and multiple comparisons (LSD method) using SPSS statistical software 18.0.

5. Measurement index

Egg quality analysis was tested in the egg quality analysis laboratory of the institute of economic sciences of husbandry in Henan province.

(1) Production Performance

And (4) counting the feed consumption, the egg yield and the egg weight in the whole period according to the egg yield and the weighing, and calculating the average egg yield, the average egg weight, the average daily feed intake, the feed-egg ratio and the average broken egg rate in the whole period.

The results are shown in Table 3:

TABLE 3 comparison of the results of the production Performance tests

Item	Control group	Test group
			Laying rate (%)	82.04±0.12^b	84.81±0.16^a
Egg weight	62.58±1.78^b	66.84±4.71^a
			Feed intake	125.22±2.10	123.08±0.47
Material to egg ratio	2.34±0.05^a	2.21±0.02^b
			Percentage of broken eggs (%)	1.70±0.06^a	0.67±0.04^b

Note: different lower case letters after the same column of values indicate significant differences at the 5% level, as follows.

(2) Egg quality

The results are shown in Table 4:

table 4 comparison of egg quality test results

Item	Control group	Test group
			Egg shell weight (g)	7.93±0.24^b	8.81±0.91^a
Strength of egg shell	4.21±0.35	4.33±0.37
			Yolk color	6.46±0.57^b	7.50±0.29^a
Protein height (mm)	7.29±0.56	7.93±0.81
			Protein weight (g)	37.30±1.16^b	39.95±2.37^a
Yolk weight (g)	16.84±1.47	17.54±1.01
			Ha's unit	84.50±6.22^b	85.81±6.95^a

As can be seen from tables 3 and 4:

in the layer feeding experiment, the layer added with the feed additive has the following changes: the laying rate and the egg weight are both obviously increased, the feed intake, the feed-egg ratio and the egg breaking rate are all obviously reduced, the numerical values of the eggshell weight, the eggshell strength, the yolk color, the egg white height, the egg white weight and the yolk weight are all increased, namely the laying quality of the laying hens is obviously improved, and the numerical value of the Haugh unit is also obviously changed, namely the freshness of the laid eggs is obviously improved.

Example five: detecting influence of laying hen feed additive on laying hen intestinal flora

1. Experimental animals and experimental design same as example four

2. Index and method of measurement

Measurement indexes are as follows: the relative abundance of the caecal flora of the laying hens and the structure of the caecal flora.

The determination method comprises the following steps: and (3) carrying out high-throughput sequencing on the 16S rDNA gene V3-V4 region of the bacteria by using an Illumina HiSeq sequencing platform, and carrying out bioinformatics analysis on a sequencing result.

Detection unit: the detection of the cecal bacteria of the laying hens is carried out by Beijing Nuo He biogenic bioinformatics science and technology company, and relevant data are provided by the company.

3. Relative abundance of caecum flora in later egg laying period of laying hens

To investigate the species composition diversity of the samples, Tags were clustered for all samples, sequences were clustered into OTUs with 97% identity, and then species annotations were made for representative sequences of OTUs. And according to the species annotation result, selecting the species with the highest abundance ranking at the gate and genus classification level of each group, and generating a species relative abundance column accumulation chart so as to visually check the species with higher relative abundance and the proportion thereof of each sample at different classification levels.

At the phylogenetic level, the structural composition of ceca content flora with abundance >1% is shown in fig. 1.

As can be seen from FIG. 1, the abundance of the fungus of Geotrichum at greater than 1: (Firmicutes) Bacteroidetes gate (Bacteroidetes) Shuttle bacteria door (Fusobacteria) (iii) Proteobacteria (Proteobacteria) Actinomycetes door (Actinobacteria) And (5) waiting for bacteria. The relative abundance of bacteroidetes in the test group is significantly increased compared with that in the control group (P<0.05) The relative abundance of firmicutes and fusobacteriales is significantly reduced (P<0.05) (iv) Proteobacteria and Actinomycetes are not significantly different: (P>0.05）。

At the genus classification level, the genus composition of the caecal content in abundance >1% is shown in figure 2.

As can be seen from FIG. 2, there were 5 statistically significant differences in the genus between the control group and the test group (S) ((S))P<0.05) Bacteroides of the test group compared with the control group: (Bacteroides) Andunidentified_Ruminococcaceaethe relative abundance is remarkably increased, and the genus Clostridium (Fusobacterium）、RuminococcaceaeUCG-005, Anaerococcus (A. sp.), (B. sp.)Anaerotruncus) The abundance is remarkably reduced.

4. Cecal flora structure of laying hens in later egg laying period

Principal Component Analysis (PCA), a method for extracting the most dominant elements and structures in data by applying variance decomposition to perform dimensionality reduction on multidimensional data. By applying PCA analysis, two coordinate axes which reflect the difference between samples to the maximum extent can be extracted, so that the difference of multi-dimensional data is reflected on a two-dimensional coordinate graph, and further a simple rule under a complex data background is disclosed. The closer they are in the PCA plot if the colony compositions of the samples are more similar. PCA analysis used ade4 package and ggplot2 software package for R software.

At the phylogenetic level, the analysis of differences in microbial flora between groups is shown in FIG. 3.

As can be seen from fig. 3, principal component 1 (PC1) and principal component 2 (PC2) are the 2 largest difference features responsible for the samples, with contributions of 32.06% and 20.85%, respectively. The main components of the control group and the test group are obviously different, the contribution rate of the main component 2 of the control group is higher, and the contribution rate of the main component 1 of the test group is higher. The 6 samples in the control group were close and relatively concentrated, indicating that the species similarity of the 6 samples in the control group was high. The 6 samples in the test group are far apart, indicating that the species similarity of the 6 samples in the test group is greatly different.

At the genus classification level, the analysis of differences in microbial flora between groups is shown in FIG. 4.

As can be seen from fig. 4, the contributions of principal component 1 (PC1) and principal component 2 (PC2) are 18.40% and 14.13%, respectively. The main components of the control group and the test group are obviously different, the contribution rate of the main component 2 of the control group is higher, and the contribution rate of the main component 1 of the test group is higher.

Although the present invention has been described in detail with reference to the embodiments, it will be understood by those skilled in the art that various changes in the specific parameters of the embodiments may be made without departing from the spirit of the present invention, and a plurality of specific embodiments are formed, which are common variations of the present invention, and will not be described in detail herein.

Claims

1. A laying hen feed additive is characterized in that: the composition is characterized by comprising the following raw materials in parts by weight: 15-25 parts of microbial fermentation products of boxthorn leaves, boxthorn stalks, corn flour and sesame seed meal, 70-80 parts of potato flour, 2-6 parts of medlar, 0.5-1 part of anise and 0.1-0.3 part of thyme;

the preparation method of the microbial fermentation product comprises the following steps:

taking the mixture with the mass ratio of 0.5-1.5: 0.5-1.5: 0.1-0.3: 0.05-0.2 of boxthorn leaves, boxthorn stalks, corn flour and sesame seed meal, crushing and sieving, and adding microorganisms; drying after culturing to obtain a fermentation product;

the microorganism is lactobacillus plantarum BNCC187897 and bacillus subtilis AS 1.9086;

the mass ratio of the microorganisms to the fermentation product is 5: 1.

2. the laying hen feed additive of claim 1, wherein: the number of the sieved meshes is 50-70 meshes.

3. A method for preparing the laying hen feed additive of claim 1, which comprises the following steps: