Apple pomace fermented feed and method for preparing the same

Abstract

The present application belongs to the technical field of fermented feed, and particularly relates to an apple residue fermented feed and a preparation method thereof. The apple residue fermented feed provided by the present application is obtained by first fermenting apple residue by Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1 and Lactobacillus bulgaricus, and then by Aspergillus chevalieri CTCF-AC-1. The present application has the advantages that, on the one hand, the apple residue waste is reused and a high-quality protein feed is obtained by fermenting the apple residue by multiple probiotics; on the other hand, the content of crude fiber and tannin anti-nutritional factors and other components in the feed is greatly reduced by applying the compound lactobacillus and Aspergillus chevalieri CTCF-AC-1 to the preparation process of the apple residue feed in a staged fermentation manner, the crude fiber in the apple residue is converted into crude protein and other nutrients, and the crude protein in the fermented feed reaches 33 g / kg, and the tannin content is only 0.16%.

Description

Technical Field

[0001] This invention belongs to the field of fermented feed technology, specifically relating to an apple pomace fermented feed and its preparation method. Background Technology

[0002] Apples are plants belonging to the genus Malus of the family Rosaceae in the order Rosales. They are one of the most famous fruits in the world. China is the world's largest apple producer. While processing apples, a large amount of apple pomace waste is also generated. This waste apple pomace is rich in nutrients such as pectin, starch, and cellulose, but has a low protein content. Furthermore, due to its high water content (about 80% or more), it is extremely prone to microbial growth, leading to rotting and foul odors, making it difficult to utilize.

[0003] Protein feed generally refers to a type of feed with a moisture content of less than 45%, a crude fiber content of less than 18% in dry matter, and a crude protein content of not less than 20%. According to the different sources of feed, protein feed can be divided into four categories: plant protein feed, animal protein feed, single-cell protein feed, and non-protein nitrogen feed.

[0004] Given the high water content, high fiber content, and low protein content of apple pomace, resulting in low utilization rates, using waste apple pomace to produce protein feed is a good way to utilize this waste at present. Patents such as CN103250874A, CN105661008A, CN110144317A, and CN103976147A have disclosed research on using single or multiple strains selected by them to ferment apple pomace to prepare protein feed. However, due to differences in the types and properties of the selected strains, as well as the different fermentation process conditions controlled during fermentation, the content of crude protein, crude fiber, and other nutrients in the protein feed obtained by various technologies varies greatly.

[0005] In addition, apple pomace contains anti-nutritional factors such as tannins, which can easily cause astringency. Although some studies have shown that the fermentation process of microorganisms can reduce the content of anti-nutritional factors such as tannins, in reality, different types of microorganisms have a large difference in the degree of utilization of components such as tannins, resulting in significantly different degrees of reduction in tannin content in fermented feed products, and thus a large difference in taste.

[0006] Therefore, screening one or more microbial strains that can efficiently ferment apple pomace and effectively reduce the tannin content in fermented apple pomace feed, and providing a more efficient fermentation process for apple pomace feed, is not only conducive to the full utilization of apple pomace waste, but also to the industrialization of fermented apple pomace feed. Summary of the Invention

[0007] To address the aforementioned technical problems, this invention provides an apple pomace fermented feed and its preparation method.

[0008] During their research on the fermentation of apple pomace waste to prepare animal feed, the inventors discovered that if only Lactobacillus was used for fermentation, although the crude protein content of the obtained animal protein feed was improved, the feed had a strong sour taste and a strong fermented taste, and the astringent taste was also more severe, meaning that the protein feed had a poor taste.

[0009] To address this, the inventors took a different approach, screening out a completely different species of fermentation strain, Aspergillus serrata, and applying it to the fermentation process of apple pomace. This greatly improved the taste and quality of the protein feed. Furthermore, there are currently no reports of using Aspergillus serrata for fermentation to prepare protein feed.

[0010] Aspergillus chevalieri, belonging to the class Hyphomycetes, order Hyphomyales, family Phyllostachyaceae, genus Aspergillus, is a multicellular mold with septate hyphae. Aspergillus chevalieri possesses functions such as phosphorus solubilization, potassium release, and nitrogen fixation. Furthermore, it produces various nutrients during its growth and reproduction, including organic acids, amino acids, polysaccharides, and natural growth factors. Therefore, Aspergillus chevalieri is often used to control bacterial or fungal soil-borne diseases in plants, or to promote increased plant yields and income.

[0011] The apple pomace fermented feed provided by this invention is specifically obtained by fermenting fresh apple pomace in stages using *Lactobacillus plantarum* CTCF-LP-1, *Lactobacillus casei* CTCF-LC-1, *Lactobacillus bulgaricus*, and *Aspergillus chevalieri* CTCF-AC-1. The *Lactobacillus plantarum* CTCF-LP-1 was deposited on September 1, 2023, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 28329 and classification name *Lactobacillus plantarum*. The *Lactobacillus casei* CTCF-LC-1 was also deposited on September 1, 2023, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC. No. 28328, classified and named: Lactobacillus casei; Aspergillus chevalieri CTCF-AC-1, was deposited on September 1, 2023, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 40816, classified and named: Aspergillus chevalieri.

[0012] Unless otherwise specified, the strains described above were used in the following embodiments of the present invention.

[0013] The Aspergillus schwanniferus CTCF-AC-1 selected in this invention can efficiently decompose fibrous materials in apple pomace. After fermentation with Aspergillus schwanniferus CTCF-AC-1, the content of crude fiber and tannin in the protein feed is significantly reduced, while the crude protein content of the feed is high, which greatly improves the taste and quality of animal protein feed obtained from apple pomace fermentation.

[0014] The above-mentioned method for preparing fermented apple pomace feed includes the following steps:

[0015] S1 Raw Material Pretreatment: Select clean, mold-free, fresh apple pomace, with a total bacterial count controlled to ≤2.5×10⁻⁶. 3 cfu / g, dried apple pomace;

[0016] S2 Mixing: Take corn residue, soybean meal, and apple residue treated in S1 and mix them evenly. Then adjust the moisture content of the mixed raw materials to 35-45%.

[0017] S3 strain activation: Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus were activated in liquid MRS medium, while Aspergillus chevalieri CTCF-AC-1 was activated in PDB medium.

[0018] S4 Stage 1 Fermentation: The activated Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus from S3 are added to the mixed raw materials obtained in S2 and fermented to obtain Stage 1 feed;

[0019] S5 Second-stage fermentation: Add the activated Aspergillus schwanniferus CTCF-AC-1 from S3 to the first-stage feed obtained in S4 for fermentation. After fermentation, the feed is turned over to mix the mycelia of Aspergillus schwanniferus CTCF-AC-1 evenly, dried, and crushed into granules to obtain apple pomace feed.

[0020] In the above steps, preferably, during the pretreatment of raw material S1, a heat pump dryer is used to dry the apple pomace at 50-65°C for 8-12 hours.

[0021] Since fresh apple pomace has a high moisture content, generally above 80%, if it is not dried, it is very easy for bacteria to grow due to the high moisture content, causing the pomace to rot and deteriorate. Therefore, this invention dries the apple pomace before processing it to obtain dry apple pomace raw material.

[0022] Preferably, in the S2 mixing process, the weight parts of each raw material are: 10-20 parts corn grits, 10-30 parts soybean meal, and 60-70 parts apple pomace treated in S1. After mixing the raw materials, the total weight parts of each component are 100 parts.

[0023] Although apple pomace is rich in sugars such as pectin, starch, and cellulose, which can provide carbon sources for microbial fermentation and reproduction, it is low in protein. Therefore, in order to provide more abundant nutrients for microbial fermentation, this invention adds soybean meal, corn residue, and other ingredients to the pomace. Abundant nutrients also help to enrich the products of microbial fermentation.

[0024] Preferably, in S3, *Lactobacillus plantarum* CTCF-LP-1, *Lactobacillus casei* CTCF-LC-1, and *Lactobacillus bulgaricus* are activated in liquid MRS medium for 48 hours, and *Aspergillus chevaleri* CTCF-AC-1 is activated in PDB medium for 7–10 days, so that the total colony count of each lactobacillus after activation is not less than 1.8 × 10⁻⁶. 7 CFU / mL medium, with a total colony count of *Aspergillus chevaleri* CTCF-AC-1 not less than 3 × 10⁻⁶. 8 CFU / mL culture medium.

[0025] The total number of bacterial colonies has a direct impact on the fermentation effect of apple pomace. If the number of fermentation colonies is low, not only will the fermentation rate be low, but the substrate may not be fermented thoroughly, resulting in waste of raw materials. At the same time, the protein and other nutrient content in the fermented feed product will not be high. However, if the number of colonies is too high during fermentation, the nutrients in the substrate may be consumed too quickly due to the rapid reproduction rate of microorganisms, and there may even be an accumulation of toxic components, which will also affect the quality of the fermented feed.

[0026] Preferably, during the S4 stage of fermentation, the total inoculation mass of *Lactobacillus plantarum* CTCF-LP-1, *Lactobacillus casei* CTCF-LC-1, and *Lactobacillus bulgaricus* accounts for 5-15% of the total mass of the mixed raw materials, wherein the mass ratio of *Lactobacillus plantarum* CTCF-LP-1: *Lactobacillus casei* CTCF-LC-1: *Lactobacillus bulgaricus* is 1:1:1.

[0027] Preferably, during the first stage of fermentation in S4, the fermentation temperature is 30–40°C, and the fermentation time is 12–96 hours.

[0028] Preferably, during the S5 two-stage fermentation, the inoculation mass of Aspergillus sieboldii CTCF-AC-1 accounts for 8-20% of the mass of the added first-stage feed, the temperature of the second-stage fermentation is 25-35℃, and the fermentation time is 6-15 days.

[0029] Preferably, in S5, a heat pump dryer is used to dry the apple pomace fermented feed at 50-65℃ for 8-12 hours.

[0030] Preferably, in step S5, the obtained apple pomace fermented feed is crushed into particles of 2-6 mm, screened to remove impurities, and then sealed in bags to produce the finished apple pomace fermented feed.

[0031] Furthermore, the preparation steps of the apple pomace fermented feed provided by the present invention are as follows:

[0032] S1 Raw Material Pretreatment: Select clean, mold-free, fresh apple pomace, with a total bacterial count controlled to ≤2.5×10⁻⁶. 3 cfu / g, dried apple pomace;

[0033] S2 Mixing: Take the following parts by weight of raw materials: 10-20 parts corn grits, 10-30 parts soybean meal, and 60-70 parts apple pomace treated in S1, mix them evenly, and then adjust the moisture content of the mixed raw materials to 35-45%.

[0034] S3 strain activation: *Lactobacillus plantarum* CTCF-LP-1, *Lactobacillus casei* CTCF-LC-1, and *Lactobacillus bulgaricus* were activated in liquid MRS medium, while *Aspergillus chevaleri* CTCF-AC-1 was activated in PDB medium, ensuring that the total colony count of each lactobacillus strain after activation was not less than 1.8 × 10⁻⁶. 7 CFU / mL medium, with a total colony count of *Aspergillus chevaleri* CTCF-AC-1 not less than 3 × 10⁻⁶. 8 CFU / mL culture medium;

[0035] S4 First-stage fermentation: The activated Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus from S3 are added to the mixed raw materials obtained in S2. The total volume of the added lactobacilli accounts for 5-15% of the total volume of the mixed raw materials. Then, fermentation is carried out at 30-40℃ for 12-96 hours to obtain the first-stage feed.

[0036] S5 Second-stage fermentation: The activated Aspergillus schwanniferus CTCF-AC-1 from S3 is inoculated into the first-stage feed obtained from S4 and fermented at 25-35℃ for 6-15 days. The inoculation mass of Aspergillus schwanniferus CTCF-AC-1 accounts for 8-20% of the first-stage feed mass. After fermentation, the feed is turned over to mix the Aspergillus schwanniferus CTCF-AC-1 mycelium evenly, dried, and crushed into granules to obtain apple pomace fermented feed.

[0037] Furthermore, the application of the strains provided in this invention—Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, Lactobacillus bulgaricus, and Aspergillus chevalieri CTCF-AC-1—in the preparation of fermented feed also falls within the scope of protection of this invention. Specifically, the application is carried out in a staged fermentation manner, that is, the fruit pomace is first fermented with a compound of Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus, and then a second stage of fermentation is carried out with Aspergillus chevalieri CTCF-AC-1 to prepare fermented feed.

[0038] The beneficial effects of this invention are as follows:

[0039] (1) Microorganisms that can efficiently ferment apple pomace waste were screened out, including: Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Aspergillus sieboldii CTCF-AC-1, etc. By using the above-mentioned probiotics to ferment apple pomace waste, the utilization rate of apple pomace waste resources was improved, and high-quality animal protein feed was also obtained.

[0040] (2) A compound of lactobacilli such as Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus were used to ferment apple pomace in one stage, followed by a second stage of fermentation with Aspergillus sieboldii CTCF-AC-1. Through the two-stage fermentation of specific microorganisms, the dry matter recovery rate of the feed reached 103.8%, and the crude fiber content of the feed was low, only 50g / kg, the tannin content was only 0.11-0.16%, the crude protein content of the feed reached up to 33g / kg, the acidity of the feed was suitable, and the nutrients were rich.

[0041] (3) An application of Aspergillus schwannii CTCF-AC-1 in the fermentation preparation of apple pomace feed is provided. The use of Aspergillus schwannii CTCF-AC-1 in this invention results in a crude protein content in the feed that is about 80.3% higher than that of the feed that was not fermented with Aspergillus schwannii CTCF-AC-1 in the second stage (18.3 g / kg), and a tannin content (0.16%) that is nearly 3 times lower than that of the feed that was not fermented with Aspergillus schwannii CTCF-AC-1 in the second stage (0.62%). Detailed Implementation

[0042] To enable those skilled in the art to better understand the present invention, the present invention will now be further described in conjunction with specific embodiments.

[0043] Example 1

[0044] A fermented apple pomace feed is prepared as follows:

[0045] S1 Raw Material Pretreatment: Select clean, mold-free, fresh apple pomace, with a total bacterial count controlled to ≤2.5×10⁻⁶. 3 cfu / g, dried at 60℃ for 10h using a heat pump dryer;

[0046] S2 Mixing: Take 15 parts corn grits, 25 parts soybean meal, and 60 parts apple pomace treated in S1, mix them evenly, and then adjust the moisture content of the mixed raw materials to 45%.

[0047] S3 strain activation: *Lactobacillus plantarum* CTCF-LP-1, *Lactobacillus casei* CTCF-LC-1, and *Lactobacillus bulgaricus* (commercially available common strain) were activated in liquid MRS medium for 48 h, while *Aspergillus chevaleri* was activated in PDB medium for 7 days, resulting in a total colony count of 1.8 × 10⁻⁶ for each type of lactobacillus after activation. 7 The total colony count of *Aspergillus schwanniferus* CTCF-AC-1 in CFU / mL medium was 3 × 10⁻⁶. 8 CFU / mL culture medium;

[0048] S4 Stage 1 Fermentation: The activated Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus from S3 are added to the mixed raw materials obtained in S2. The total mass of the added lactobacilli accounts for 8% of the total mass of the mixed raw materials. Then, fermentation is carried out at 32℃ for 60 hours to obtain Stage 1 feed.

[0049] S5 Second-stage fermentation: Add 15% of the activated Aspergillus schwanniferus CTCF-AC-1 from S3 to the first-stage feed obtained from S4 and ferment at 30℃ for 7 days. After fermentation, turn the feed over to mix the mycelia of Aspergillus schwanniferus CTCF-AC-1 evenly, dry it, and crush it into 5mm particles to obtain apple pomace fermented feed.

[0050] Example 2

[0051] Unlike Example 1, in S4-S5, the fermentation time of the first stage was adjusted to 12 hours; in the second stage fermentation, the fermentation time of Aspergillus sieboldii CTCF-AC-1 was adjusted to 6 days, and the rest were the same as in Example 1.

[0052] Example 3

[0053] Unlike Example 1, in S4-S5, the fermentation time of the first stage was adjusted to 96 hours, and in the second stage fermentation, the fermentation time of Aspergillus sieboldii CTCF-AC-1 was adjusted to 15 days. The rest were the same as in Example 1.

[0054] Example 4

[0055] Unlike Example 1, in S4-S5, during the first stage of fermentation, the total mass of added Lactobacillus was 5% of the total mass of the mixed raw materials, and during the second stage of fermentation, the mass of added Aspergillus sieboldii CTCF-AC-1 accounted for 8% of the mass of the first stage feed added. The rest were the same as in Example 1.

[0056] Example 5

[0057] Unlike Example 1, in S4-S5, during the first stage of fermentation, the total mass of added Lactobacillus is 15% of the total mass of the mixed raw materials, and during the second stage of fermentation, the mass of added Aspergillus sieboldii CTCF-AC-1 accounts for 20% of the mass of the first stage feed added. The rest are the same as in Example 1.

[0058] Comparative Example 1

[0059] Unlike Example 1, only Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus were used for the first stage of fermentation of apple pomace, and Aspergillus chevalieri CTCF-AC-1 was no longer used for the second stage of fermentation.

[0060] Comparative Example 2

[0061] Unlike Example 1, commercially available Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus bulgaricus were used for the first stage of fermentation of apple pomace, instead of using Aspergillus sieboldii CTCF-AC-1 for the second stage of fermentation.

[0062] Comparative Example 3

[0063] Unlike Example 1, in S4, only Lactobacillus plantarum CTCF-LP-1 and Lactobacillus casei CTCF-LC-1 were used for the first stage of fermentation of the mixed raw materials, while in S5, Aspergillus schwannii CTCF-AC-1 was used for the second stage of fermentation.

[0064] Comparative Example 4

[0065] Unlike Example 1, in S2, only apple pomace and corn pomace are used as raw materials for fermentation, and the ratio of apple pomace to corn pomace is 65:35.

[0066] The quality of the protein feeds obtained in the above embodiments and comparative examples is shown in Table 1 below, and the sensory evaluation of the feeds is shown in Table 2.

[0067] Table 1. Quality of protein feed prepared under different conditions

[0068]

[0069] Table 2 Sensory evaluation results of fermented feed from different embodiments

[0070]

[0071] The results in Table 1-2 show:

[0072] In Example 1, Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus were used to carry out a first-stage fermentation of apple pomace raw materials, followed by a second-stage fermentation using Aspergillus chevaleri CTCF-AC-1. The resulting protein feed had a crude protein content of 33 g / kg, a crude fiber content of 50 g / kg, and a significantly reduced tannin content of only 0.16%. The feed was golden yellow, had suitable acidity, a feed aroma, and was rich in nutrients.

[0073] Compared to Example 1, Example 2 shortened the fermentation time for the first and second stages. Obviously, the total acid and crude protein content in the prepared protein feed were reduced, while the tannin content was higher. It can be seen that when fermentation is insufficient, the tannin components in the fruit pomace cannot be well utilized, thus affecting the taste of the protein feed. The fruit pomace flavor is stronger, and the feed is relatively dry and loose overall, resulting in a slight reduction in feed quality.

[0074] Example 3, based on Example 1, increased the fermentation time for the first and second stages. Although the tannin content in the obtained feed was further reduced, the total acid content was high and the pH of the feed was only 3.2. Over-fermentation made the feed have a pungent sour smell and poor taste.

[0075] In addition, in Examples 4-5, the inoculation amount of the microorganisms in the first and second stages of fermentation were adjusted respectively. As can be seen from the results in Table 1, if the inoculation amount is low, the lactic acid fermentation effect is poor, the total acid content is low, and the dry matter recovery rate is reduced, thereby affecting the crude protein content in the feed. In addition, tannins cannot be fully decomposed. If the inoculation amount of the microorganisms is high, the total acid content of the feed after fermentation is high, which has a greater impact on the sensory characteristics of the feed, making the feed have a certain sour smell. Furthermore, continuous fermentation will also affect the crude protein and crude fiber content in the feed.

[0076] Furthermore, in Comparative Example 1, only Lactobacillus probiotics were used for the first stage of fermentation of apple pomace, without the use of Aspergillus schwanniferus CTCF-AC-1 for the second stage of fermentation. As can be seen from the feed quality parameter table, although the acidity of the feed was not significantly different from that in Example 1, the content of crude fiber, tannins and other components was much higher than that of the feed product in Example 1. It is evident that it is difficult for Lactobacillus alone to decompose and transform the crude fiber and tannins in the pomace. The second stage of fermentation with Aspergillus schwanniferus CTCF-AC-1 played a crucial role in the decomposition and transformation of crude fiber and tannins. Almost the same conclusion can be drawn from Comparative Example 3.

[0077] Furthermore, in Comparative Example 3, only Lactobacillus plantarum CTCF-LP-1 and Lactobacillus casei CTCF-LC-1 were used for a single fermentation stage, without adding Lactobacillus bulgaricus. It can be seen that the total acid content after fermentation was higher than that of the feed in Example 1. The main reason is that Lactobacillus bulgaricus produces less acid and generates exogenous polysaccharides during fermentation. Therefore, adding Lactobacillus bulgaricus during the actual fermentation process would relatively inhibit the growth of the two types of lactobacilli. Consequently, the crude fiber conversion rate of the fermented feed in Comparative Example 3 was poor, and the crude protein content was reduced.

[0078] In addition, to verify the high efficiency of the strains screened in this invention, commercially available Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus bulgaricus strains were used to ferment apple pomace in Comparative Example 2. Compared with the feed obtained in Example 1, the total acid content of the feed was reduced. It can be seen that even under the same substrate and fermentation conditions, the fermentation efficiency of commercially available conventional strains is relatively limited. In addition, the crude fiber content in the feed was as high as 66.1 g / kg, while the crude protein content was low, at only 20.1 g / kg.

Claims

1. A method for preparing fermented apple pomace feed, characterized in that, The steps include the following: S1 Raw Material Pretreatment: Select clean, mold-free, fresh apple pomace, with a total bacterial count controlled to ≤2.5×10⁻⁶. 3 The apple pomace was dried using a heat pump dryer at 60°C for 10 hours (cfu / g). S2 Mixing: Take 15 parts corn grits, 25 parts soybean meal, and 60 parts apple pomace treated in S1, mix them evenly, and then adjust the moisture content of the mixed raw materials to 45%; S3 strain activation: *Lactobacillus plantarum* CTCF-LP-1 (CGMCC No. 28329), *Lactobacillus casei* CTCF-LC-1 (CGMCC No. 28328), and *Lactobacillus bulgaricus* were activated in liquid MRS medium for 48 h. *Aspergillus chevaleri* CTCF-AC-1 (CGMCC No. 40816) was activated in PDB medium for 7 days. The total colony count of each lactobacillus after activation was 1.8 × 10⁻⁶. 7 The total colony count of *Aspergillus chevaleri* CTCF-AC-1 on CFU / mL medium was 3 × 10⁻⁶. 8 CFU / mL culture medium; S4 First-stage fermentation: The activated Lactobacillus plantarum CTCF-LP-1, Lactobacillus casei CTCF-LC-1, and Lactobacillus bulgaricus from S3 are added to the mixed raw materials obtained in S2. The total mass of the added lactobacilli accounts for 8% of the total mass of the mixed raw materials. Then, fermentation is carried out at 32℃ for 60 h to obtain the first-stage feed. S5 Second-stage fermentation: Add 15% of the activated Aspergillus schwanniferus CTCF-AC-1 from S3 to the first-stage feed obtained from S4 and ferment at 30℃ for 7 days. After fermentation, the feed is turned over to mix the mycelia of Aspergillus schwanniferus CTCF-AC-1 evenly, dried, and crushed into 5mm particles to obtain apple pomace fermented feed.

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