Ganoderma lucidum fruiting body fermented feed additive for improving production performance of laying hens and use thereof

Abstract

A method for preparing a Ganoderma lucidum fruiting body fermented feed additive for improving the production performance of laying hens and a use of the Ganoderma lucidum fruiting body fermented feed additive. Main components are Ganoderma lucidum fruiting body, microbial inoculant 1, fermented brown sugar, soybean meal, and distilled water. The weight proportions of the Ganoderma lucidum fruiting body, microbial inoculant 1 (EM bacterium:white rot fungus Z10210:Phanerochaete chrysosporium 154637=(1:1:1)), fermented brown sugar, soybean meal, and distilled water are as follows: Ganoderma lucidum fruiting body: 100; microbial inoculant: 13; fermented brown sugar: 5.2; soybean meal: 8.2; and water: 295. The preparation method comprises: (1) screening for a Ganoderma lucidum fruiting body wall-breaking complex microbial inoculant; (2) optimizing a Ganoderma lucidum fruiting body wall-breaking and fermentation process; and (3) optimizing a response surface. When the Ganoderma lucidum fruiting body fermented feed additive is applied to the feeding process of laying hens, 0.3% of fermented Ganoderma lucidum fruiting body is added into feeds for laying hens, so that the oxidation resistance and immunity of the laying hens can be improved.

Description

Ganoderma lucidum fruiting body fermented feed additive for improving laying hen production performance and application thereof Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine fermented feed additives, and more particularly to a ganoderma lucidum fruiting body fermented feed additive for improving the production performance of laying hens and a preparation method thereof. Background Art

[0002] After decades of rapid development, my country's laying hen industry has led the world in egg production for 21 consecutive years. Traditionally, antibiotics were often added to laying hen feed to maintain health, but long-term antibiotic use can lead to drug resistance in animals. This not only impacts the ecological balance of organisms but also pollutes the environment. Traditional Chinese medicine (TCM) contains a variety of nutrients essential to life, such as proteins and carbohydrates. These nutrients not only enhance livestock and poultry growth but also increase feed palatability and feed intake. However, most TCMs are composed of plants and fungi, which have dense cell walls, making it difficult for animals to effectively utilize the nutrients within. Bio-cell wall disruption can utilize enzymes produced by microorganisms to degrade the cell walls, hydrolyzing macromolecules within the cell walls and releasing active substances, thereby increasing their utilization. Ganoderma lucidum, a medicinal fungus of the genus Ganoderma in the phylum Basidiomycetes, contains a variety of nutrients with anti-tumor, immune-modulating, and free radical-scavenging properties. Through bio-cell wall disruption, it can be made into a fermented TCM feed additive, replacing antibiotics in livestock and poultry farming.

[0003] The present invention features and advantages that Ganoderma lucidum fruiting bodies contain multiple active ingredients, including polysaccharides and triterpenes, that function similarly to feed additives. The fermented Ganoderma lucidum fruiting body product, when used as an additive, is less likely to develop drug resistance, is easily metabolized, and offers environmental benefits, significantly promoting the development of the livestock and poultry industries. Large-scale production of Ganoderma lucidum fruiting bodies as feed also yields significant economic, social, and environmental benefits.

[0004] In summary, how to provide a Ganoderma lucidum fruiting body fermentation feed additive is a problem that those skilled in the art urgently need to solve.

[0005] Summary of the Invention

[0006] In view of this, the present invention provides a Ganoderma lucidum fruiting body fermented feed additive and a preparation method and application thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] The invention provides a ganoderma lucidum fruiting body fermented feed additive. The ganoderma lucidum fruiting body fermented feed additive is obtained by taking 100 ganoderma lucidum fruiting bodies, 13 g of bacterial agent, 5.2 g of fermented brown sugar, 8.2 g of soybean meal and 295 g of water as raw materials and fermenting the feed at 28°C for 6 days.

[0009] A method for preparing a Ganoderma lucidum fruiting body fermented feed additive comprises the following steps:

[0010] (1) Screening of composite microbial agents for breaking the fruiting body wall of Ganoderma lucidum: Microbial agent 1 (EM bacteria: white rot fungus Z10210: Phanerochaete chrysosporium 154637 = (1:1:1)) and microbial agent 2 (EM bacteria) were selected. The dried Ganoderma lucidum fruiting body was crushed, and 5% microbial agent, 5% brown sugar, 5% soybean meal and distilled water with a solid-liquid ratio of 5:3 were added according to the weight ratio, and the mixture was allowed to ferment at 28°C. Samples were taken every 3 days, and the microbial agents suitable for Ganoderma lucidum fruiting body fermentation were screened according to the content of Ganoderma lucidum polysaccharides in the samples;

[0011] (2) Screening of factors affecting the fermentation process of Ganoderma lucidum fruiting body wall breaking: Using the bacterial agent screened in step (1), the fermentation temperature, initial pH value of fermentation, inoculation amount, soybean meal addition amount, brown sugar addition amount and solid-liquid ratio were used as screening targets, and Ganoderma lucidum polysaccharide was quantitatively analyzed to screen out three factors that significantly affected the wall breaking of Ganoderma lucidum fruiting body;

[0012] (3) Response surface optimization: Based on the factors affecting the wall breaking of Ganoderma lucidum fruiting bodies screened in step (2), a response surface experiment was designed. Taking polysaccharide content as an indicator, the effects of soybean meal addition, brown sugar addition, and solid-liquid ratio on polysaccharide release were investigated. The interaction and optimal levels of these three factors were investigated. The response surface model was fitted to determine the optimal conditions and adjusted and verified based on actual conditions. The optimal fermentation process of Ganoderma lucidum fruiting bodies obtained from the above experiments was used for subsequent fermentation.

[0013] Furthermore, in the above step (1), the screened bacterial agent is: bacterial agent 1; in steps (2) and (3), the optimal fermentation process is: bacterial agent inoculation amount 3%, soybean meal addition amount 8.2%, brown sugar addition amount 5.2%, solid-liquid ratio 5:5.9, fermentation temperature 28°C, and fermentation time 6 days.

[0014] The present invention also seeks to protect the use of the Ganoderma lucidum fruiting body fermented feed additive as described above or the Ganoderma lucidum fruiting body fermented feed additive prepared by the above preparation method in laying hen breeding.

[0015] It can be seen from the above technical solution that compared with the existing technology, the beneficial effects achieved by the present invention are: after feeding the fermented Ganoderma fruiting body feed additive to laying hens, the daily feed intake, egg production rate, feed-to-egg ratio, horizontal and vertical diameters of the eggs, and the antibody concentration of the laying hens are all improved. DETAILED DESCRIPTION

[0016] The reagents required for the present invention are conventional experimental reagents, purchased from commercial channels; the experimental methods not mentioned are conventional experimental methods and will not be described in detail here.

[0017] Example 1

[0018] Screening of compound bacterial agents for breaking the fruiting body wall of Ganoderma lucidum

[0019] The dried Ganoderma lucidum fruiting bodies were crushed and then added with 5% bacterial inoculum, 5% brown sugar, 5% soybean meal, and distilled water at a solid-to-liquid ratio of 5:3 by weight. The mixture was then fermented at 28°C. Samples were collected every three days to determine the polysaccharide content in the fruiting bodies after fermentation with different bacterial inoculums.

[0020] Table 1 Determination of Ganoderma lucidum polysaccharide content in Ganoderma lucidum fruiting body fermentation

[0021] Both agents 1 and 2 were able to disrupt the fruiting bodies of Ganoderma lucidum for fermentation. With increasing fermentation time, the polysaccharide content initially increased and then decreased. Agent 1 had the highest polysaccharide content on the sixth day of fermentation, showing a significant difference from agent 2, indicating that it was more effective in disrupting the fruiting bodies.

[0022] Example 2

[0023] Screening of factors affecting the fermentation process of Ganoderma lucidum fruiting body wall breaking

[0024] The bacterial agent 1 was selected to carry out the wall-breaking fermentation of Ganoderma lucidum fruiting bodies. The fermentation temperature, initial pH value of fermentation, inoculation amount, soybean meal addition amount, brown sugar addition amount and solid-liquid ratio were used as screening targets. The Ganoderma lucidum polysaccharides in Ganoderma lucidum fruiting bodies were quantitatively analyzed to screen out the factors that significantly affected the fermentation of Ganoderma lucidum fruiting bodies.

[0025] As fermentation temperature increased, polysaccharide content showed a trend of first decreasing, then increasing, and then decreasing again. Polysaccharide content reached its highest level at 28°C, which was selected as the optimal fermentation temperature for Ganoderma lucidum fruiting bodies. Initial pH had little effect on polysaccharide content, and the initial pH was not varied in subsequent experiments. Polysaccharide content showed an initial increase followed by a decrease with increasing inoculum level, soybean meal addition, brown sugar addition, and solid-to-liquid ratio. Polysaccharide content reached its highest level at an inoculum level of 3%, significantly different from the other groups. Considering practical production costs, an inoculum level of 3% was selected. Polysaccharide content reached its highest level at a soybean meal addition level of 9%, with a highly significant difference compared to the untreated group. Therefore, soybean meal addition levels of 5%-11% were selected for subsequent fermentation experiments. Polysaccharide content reached its highest level at a brown sugar addition level of 5%, with a highly significant difference compared to the untreated group. Therefore, brown sugar addition levels of 3%-9% were selected for subsequent fermentation experiments with Ganoderma lucidum fruiting bodies. The polysaccharide content was highest when the solid-liquid ratio was 5:6, and the difference was extremely significant compared with the untreated group. The solid-liquid ratio of 5:4-5:7 was selected for subsequent fermentation tests.

[0026] Example 3

[0027] Response surface optimization experimental design

[0028] Based on the screening of parameters influencing the fermentation process, significant differences in the polysaccharide content of Ganoderma lucidum fruiting body fermentations obtained under different fermentation conditions were analyzed. Among all conditions, soybean meal addition, brown sugar addition, and solid-to-liquid ratio had the greatest impact on the wall-breaking fermentation process of Ganoderma lucidum fruiting body. Therefore, appropriate levels of soybean meal addition, brown sugar addition, and solid-to-liquid ratio were selected to optimize the fermentation conditions. Regression analysis of the experimental results was performed using Design-Expert software, resulting in a quadratic polynomial regression equation for the polysaccharide content on the coded variables A, B, and C, which was then validated against the fermentation results.

[0029] Table 2 Factor levels in response surface experiment

[0030] Table 3 Response surface experimental design and results

[0031] Table 4 Response surface variance analysis

[0032] The above results show that soybean meal addition and solid-liquid ratio have significant effects on polysaccharide content, but their interaction has no significant effect on polysaccharide content. The interaction between soybean meal addition and brown sugar addition has a significant impact on the results, with soybean meal addition having a more significant effect on polysaccharide content than brown sugar addition. The solid-liquid ratio has a more significant effect on polysaccharide content than brown sugar addition, and the interaction between solid-liquid ratio and brown sugar addition also has a significant effect on polysaccharide content.

[0033] The optimal fermentation process was predicted to be 8.16% soybean meal addition, a solid-liquid ratio of 5:5.9, and 5.23% brown sugar addition. For practical purposes, the fermentation conditions were set to 8.2% soybean meal addition, a solid-liquid ratio of 5:5.9, and 5.2% brown sugar addition. Fermentation verification under these conditions yielded a polysaccharide content of 4.95 mg / g, which was not significantly different from the predicted value of 5.05 mg / g. This demonstrates that the optimal process parameters are accurate and reliable, and that the model can effectively predict the extracted polysaccharide content.

[0034] Example 4

[0035] Observation on the microstructure of Ganoderma lucidum fruiting body

[0036] Distilled water (CK) and inoculant 1 were added to Ganoderma lucidum fruiting bodies and fermented using the optimal fermentation conditions described above. Samples were taken every three days for a total of five times. Finally, the samples were dried and metal-coated, and the cell wall microstructure of each sample was observed using a scanning electron microscope.

[0037] Figure 1 Microstructure of the cell wall of Ganoderma lucidum fruiting body fermentation product

[0038] Before fermentation, the mycelium of the Ganoderma lucidum fruiting bodies' cell walls remained intact. After fermentation with the composite inoculant, the mycelium gradually broke with increasing fermentation time, with most of the mycelium already broken by day 9. Naturally fermented Ganoderma lucidum fruiting bodies showed no noticeable mycelium breakage over time. Inoculant 1 had a more pronounced and effective cell wall-breaking effect on the Ganoderma lucidum fruiting bodies.

[0039] Analysis of the Activities of Enzymes Related to Cell Wall Degradation in Fermented Products of Ganoderma Lucidum Fruiting Bodies

[0040] The fermentation products of Ganoderma lucidum fruiting bodies at different stages (0, 3, 6, 9, 12 and 15 days) were taken as samples, and their cellulase activities were determined using an endo-β-1,4-glucanase / cellulase kit.

[0041] Table 5 Results of determination of enzyme activities related to cell wall degradation in Ganoderma lucidum fruiting bodies

[0042] With the increase of fermentation time, the enzyme activity first increased and then decreased. On the 9th day, the enzyme activity reached the highest level of 1802.43 μg / h / g.

[0043] Example 5

[0044] Feeding of Ganoderma lucidum fruiting body fermented feed additive

[0045] A total of 560 healthy laying hens aged 50 weeks were randomly divided into seven groups, with eight replicates per group and ten hens per replicate. Fermented and unfermented Ganoderma fruiting bodies were added to the basal diet at varying doses for a 28-day trial.

[0046] Table 6 Specific feeding trial groups for laying hens

[0047] Effects of Ganoderma lucidum fruiting body fermented feed additive on production performance of laying hens

[0048] The number of eggs laid by each replicate was recorded daily, the egg production rate was calculated, and the eggs were weighed using an electronic balance to calculate the average egg weight to the nearest 0.01 g. The feed intake of each replicate during the experiment was calculated to calculate the feed-to-egg ratio (feed-to-egg ratio = feed material / egg mass).

[0049] Table 7 Effects of Ganoderma lucidum fruiting bodies on laying hen production performance

[0050] The test results showed that after adding fermented Ganoderma lucidum fruiting bodies, the daily feed intake of laying hens increased significantly, and the egg production rate, average egg weight and feed-to-egg ratio all increased, but the differences were not significant.

[0051] Effects of Ganoderma lucidum fruiting body fermented feed additive on egg quality

[0052] After the feeding period, three eggs were randomly selected from each group. The vertical and horizontal diameters of the eggs were measured with a vernier caliper, and the egg shape index (egg shape index = horizontal diameter / vertical diameter) was calculated. The ends and middle of the eggs were measured with a spiral micrometer, and the average value was used to calculate the eggshell thickness to the nearest 0.01 mm. Eggshell strength was measured using an eggshell strength meter. Yolk color, albumen height, and Haugh units were measured using a multifunctional egg quality tester.

[0053] Table 8 Effects of Ganoderma lucidum fruiting bodies on egg quality

[0054] As shown in the table, the addition of Ganoderma lucidum fruiting bodies increased the albumen height, Haugh value, and eggshell weight, while the yolk weight decreased, but the difference was not significant compared with the control. Adding 0.3% fermented Ganoderma lucidum fruiting bodies significantly increased the yolk color and egg shape index.

[0055] Effects of fermented products from Ganoderma lucidum fruiting bodies on antioxidant activity in laying hens

[0056] After feeding, one chicken from each group was taken, blood was drawn from the wing vein, and the blood was centrifuged at 4000 rpm. The serum was aliquoted. T-AOC and glutathione peroxidase (GPx) activity assays were used to measure T-AOC and GPx in the laying hen serum.

[0057] Table 9 Effects of Ganoderma lucidum fruiting body fermentation on antioxidant performance of laying hens

[0058] The results showed that the addition of Ganoderma lucidum fruiting bodies to the feed significantly improved the total antioxidant capacity of laying hens, and the total antioxidant activity was highest in treatment 4. Compared with the control, the addition of Ganoderma lucidum fruiting bodies increased the activity of glutathione peroxidase slightly, but not significantly, indicating that Ganoderma lucidum fruiting bodies have no effect on glutathione peroxidase.

[0059] Effects of fermented products from Ganoderma lucidum fruiting bodies on immune function of laying hens

[0060] The concentrations of infectious rhinitis antibodies, IgA, IgM and IgY in laying hen serum were determined using chicken infectious rhinitis antibody, chicken immunoglobulin IgA, immunoglobulin IgM and immunoglobulin IgY detection kits.

[0061] Table 10 Effects of Ganoderma lucidum fruiting body fermentation on immunoglobulin levels in laying hens serum

[0062] Adding fruiting bodies can increase the immune capacity of laying hens. Adding 0.3% fermented Ganoderma lucidum fruiting bodies can significantly increase the antibody concentration of laying hens, among which the concentrations of infectious rhinitis, IgA, IgM and IgY antibodies increased by 1.31 times, 2.05 times, 0.94 times and 1.08 times respectively.

[0063] In summary, the fermented Ganoderma lucidum fruiting body in this application has the efficacy of a microbial feed additive. Adding 0.3% of the fermented Ganoderma lucidum fruiting body to the diet of laying hens can improve the laying performance, egg quality, antioxidant activity and immunity of laying hens, and can replace traditional antibiotics to feed laying hens.

Claims

1. A Ganoderma lucidum fruiting body fermented feed additive, characterized in that: The invention comprises the following raw materials in parts by weight: 100 parts of ganoderma lucidum fruiting body, 13 parts of bacterial agent, 5.2 parts of fermented brown sugar, 8.2 parts of soybean meal and 295 parts of water.

2. The Ganoderma lucidum fruiting body fermented feed additive according to claim 1, characterized in that: The specific composition of the microbial agent 1 is: EM bacteria, white rot fungus Z10210 and Phanerochaete chrysosporium 154637, and the ratio of each component is 1:1:

1.

3. A method for preparing a Ganoderma lucidum fruiting body fermented feed additive, characterized in that: The specific steps include: (1) Screening of composite bacterial agents for breaking the fruiting body wall of Ganoderma lucidum; (2) Screening of factors affecting the fermentation process of Ganoderma lucidum fruiting body wall breaking; (3) Response surface optimization.

4. The method for preparing a Ganoderma lucidum fruiting body fermented feed additive according to claim 3, characterized in that: In step (1), the screened bacterial agent is bacterial agent 1; in steps (2) and (3), the optimal fermentation process is: bacterial agent inoculation amount 3%, soybean meal addition amount 8.2%, brown sugar addition amount 5.2%, solid-liquid ratio 5:5.9, fermentation temperature 28°C, and fermentation time 6 days.

5. Use of the Ganoderma lucidum fruiting body fermented feed additive according to claim 1 or the Ganoderma lucidum fruiting body fermented feed additive prepared by the preparation method according to claims 3-4 in laying hen breeding.

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