Improved palatability reed leaf fungus enzyme synergistic segmented fermentation feed and preparation method thereof

Abstract

The present application belongs to the field of unconventional forage resource utilization and microbial fermentation feed technology, and particularly relates to a reed leaf bacteria-enzyme synergistic segmented fermentation feed for improving palatability and a preparation method thereof. The feed takes dry and crushed reed leaves as the core raw material, is supplemented with corn flour, wheat bran and molasses, and is added with a compound fermentation enzyme preparation composed of Lactobacillus plantarum, Candida utilis, Aspergillus niger, cellulase and xylanase, and the total moisture of the material is regulated to 48% to 52%. The present application adopts an "aerobic fiber lysis + anaerobic acidification energy storage" segmented coupling fermentation process, directionally lyses lignocellulose in the early stage to realize flexible modification of forage, rapidly produces acid to inhibit bacteria in the later stage, and enriches organic acid and aromatic flavor substances. The obtained feed is soft in texture, aromatic in smell, has a significant degradation of crude fiber, and has a significant increase in crude protein and dry matter digestibility, can eliminate the oral mucosa damage of beef cattle, and improve the feed intake, daily weight gain and feed conversion rate, and is suitable for large-scale feeding of beef cattle.

Description

Technical Field

[0001] This invention belongs to the field of unconventional forage resource utilization and microbial fermentation feed technology, specifically relating to a segmented fermentation feed with synergistic enzymes in reed leaf extract to improve palatability and its preparation method. Background Technology

[0002] With the continuous improvement of the scale and intensification of herbivorous animal husbandry, the market supply gap for high-quality roughage resources is constantly widening. Exploring and developing low-cost, high-reserve unconventional forage resources is not only an important direction for alleviating the supply and demand contradiction of forage and reducing the cost of livestock production, but also an effective path to achieve high-value utilization of agricultural and forestry waste and promote the green development of the industry. Reed is a perennial grass widely distributed in river channels, tidal flats, wetlands, and other habitats in my country. It has large natural resource reserves, a wide distribution range, and low raw material acquisition costs, possessing the potential for development as an unconventional roughage resource. However, limited by the structure and nutritional characteristics of reed leaves themselves, as well as the insufficient adaptability of existing processing technologies, the large-scale conversion of reed resources into feed has always faced significant technical bottlenecks.

[0003] First, the physical structure of natural reed leaves directly leads to poor palatability and feeding safety risks. Natural reed leaves have a high degree of cutinization, with numerous sharp, hard fibers distributed along the leaf edges and main veins. The lignin fibers have a dense cross-linked structure, resulting in an overall hard and sharp physical texture. Current industry-standard pretreatment methods only involve mechanical crushing. This process can only shorten the material length but cannot destroy the three-dimensional cross-linked network of lignin fibers at the structural level. Even after processing, the material still retains a large number of sharp, thorny fiber structures. During consumption by ruminants such as beef cattle, these hard fibers can easily scratch the oral cavity, tongue, and esophageal mucosa, causing mucosal redness, superficial damage, and other injuries. This leads to problems such as picky eating, refusal to eat, and decreased feeding enthusiasm in animals, making it impossible to support large-scale continuous feeding. This is the primary physical obstacle limiting the promotion of reed leaf as feed.

[0004] Secondly, the nutritional composition of reed leaves makes fermentation processing difficult and results in poor quality stability. Reed leaves are a typical low-sugar, high-buffered roughage with low soluble sugar content, leading to insufficient substrate for microbial fermentation and difficulty in overcoming the material's high buffering capacity. Under natural conditions, the silage fermentation start-up cycle is long, making it difficult for beneficial microorganisms to quickly colonize and form a dominant flora, resulting in a slow acidification process. Existing conventional reed silage processes mostly use single lactic acid bacteria fermentation schemes, which are not well-suited to the low-sugar, high-buffered reed raw materials. They generally suffer from incomplete fermentation and insufficient acidification, making the fermented products prone to rancidity, mold, and unpleasant odors. The product has a short storage period, and the fermentation quality fluctuates greatly between batches, resulting in poor stability and making it difficult to meet the quality control requirements of large-scale production.

[0005] Meanwhile, existing processing technologies cannot improve the nutritional quality of reed leaves, resulting in low feed utilization efficiency. Natural reed leaves have a high proportion of structural crude fiber and lignin, which are difficult for rumen microorganisms in ruminants to fully degrade, leading to low rumen degradation rate, poor animal digestibility, and low nutritional added value. Current reed forage processing technologies can only achieve basic material preservation; they cannot target the improvement of the raw material's nutritional structure or convert indigestible structural fibers into small-molecule nutrients that can be utilized by animals. Overall, the feed value is low, failing to bring significant economic benefits to aquaculture production.

[0006] In summary, existing reed forage processing technologies generally suffer from the shortcomings of "only preserving without improving quality, and only fermenting without modifying properties," failing to simultaneously address the three core industry pain points: physical feeding damage, unstable fermentation quality, and low nutrient utilization. Currently, no specialized microbial-enzyme synergistic segmented fermentation process suitable for highly lignified reed leaves has been developed in this field. There is a lack of integrated preparation technology that can simultaneously achieve fiber softening, improved feeding safety, stable and controllable fermentation quality, and synergistic improvement of nutritional indicators, severely restricting the resource utilization and large-scale application of reed waste for feed. Summary of the Invention

[0007] The purpose of this invention is to provide a segmented fermentation feed with synergistic enzymes in reed leaves to improve palatability and its preparation method. This feed can realize the resource recycling of wetland reed waste, reduce the cost of purchasing commercial feed for livestock farming, and take into account both ecological and environmental protection value and the need for cost reduction and efficiency improvement in beef cattle farming. It has extremely high value for large-scale promotion and application.

[0008] The objective of this invention is achieved through the following technical solution: This invention provides a palatable, segmented fermented feed made from reed leaves with synergistic microbial and enzyme fermentation, prepared by the following raw materials in parts by weight: 100 parts dried and crushed reed leaves, 10-15 parts corn flour, 5-10 parts wheat bran, 3-5 parts molasses, and 0.1-0.3 parts compound fermentation microbial enzyme preparation; the total moisture content of the fermented materials is adjusted to 48%-52%; the compound fermentation microbial enzyme preparation comprises compound live bacteria strains and compound degradation enzyme preparations; the compound live bacteria strains consist of *Lactobacillus plantarum*, *Candida utilis*, and *Aspergillus niger*, with a live bacteria ratio of (2-3):(1-1.5):(0.5-1), and the total effective live bacteria count of the compound live bacteria strains is ≥1×10⁻⁶. 9 CFU / g; The compound degrading enzyme preparation is composed of cellulase and xylanase, with an enzyme activity ratio of 1:0.5, cellulase activity ≥15000 U / g, and xylanase activity ≥8000 U / g.

[0009] Furthermore, the dried and pulverized reed leaves are clean reed leaves that have been naturally sun-dried to a moisture content of ≤12%, and are pulverized and cut into sections of 1–3 cm in length, removing any moldy, blackened, or rotten leaves; the molasses is selected from sugarcane molasses or beet molasses.

[0010] Furthermore, the weight ratio of the raw materials is as follows: 100 parts dried and crushed reed leaves, 12 parts corn flour, 8 parts wheat bran, 4 parts molasses, and 0.2 parts compound fermentation bacteria enzyme preparation; the ratio of live Lactobacillus plantarum, Candida utilis, and Aspergillus niger in the compound live bacteria strain is 2.5:1.2:0.8; and the moisture content of the fermentation material is 50%.

[0011] The present invention also provides a method for preparing the palatability-improving reed leaf fungus enzyme-co-fermented feed, comprising the following steps: ① Raw material pretreatment: Collect clean, mold-free reed leaves, air dry them naturally until the moisture content is ≤12%, crush and cut them into 1-3 cm sections, and remove impurities for later use; ② Premixing: Add the pretreated reed leaves, corn flour, wheat bran, and molasses into a mixing device and mechanically mix for 3-5 minutes to obtain the basic mixture. ③ Enzyme activation and moisture control: The compound fermentation enzyme preparation is activated with warm water at 30-40℃ for 10-15 minutes to obtain the enzyme activation solution, which is then sprayed onto the basic mixture while being stirred simultaneously to control the total moisture content of the material to 48%-52%; ④ Layered filling, compaction and degassing: After mixing, the material is filled into the fermentation tank in layers, with a single layer thickness of 20-30 cm. Each layer is compacted, and the entire filling process takes ≤6 hours. ⑤ Segmented sealed fermentation: The fermentation tank is sealed with a double-layer impermeable membrane and fermented in a sealed environment of 25-35℃ for 15-30 days; the fermentation is divided into two stages: fermentation 1-5 days is the enzymatic softening stage, and fermentation 6-30 days is the anaerobic acidification and energy storage stage. ⑥ Finished product handling and storage: After fermentation is complete, take out the material in layers, with a single take-out thickness of ≤30 cm. After taking out the material, immediately reseal and compact it, and store it in a cool, dark place.

[0012] Furthermore, the fermentation environment temperature is 30℃, the fermentation cycle is 25 days, and the total moisture content of the fermentation material is 50%.

[0013] Furthermore, the material is yellowish-brown and loose, with a soft texture and no hard thorns. It has a sweet and sour fermented aroma, no moldy or rotten odor, and can be stored stably for more than 6 months under cool and dark conditions.

[0014] The present invention also provides an application of the aforementioned palatability-improving reed leaf fungus enzyme-synergistic segmented fermentation feed in beef cattle farming.

[0015] Furthermore, the nutritional indicators of the reed leaf fungus enzyme-co-fermented feed are as follows: crude protein content ≥9.0%, crude fiber ≤30%, and lactic acid content ≥2.5%.

[0016] Furthermore, the beef cattle farming mentioned is large-scale Simmental fattening cattle farming.

[0017] The beneficial effects of this invention are as follows: This invention pioneers a segmented modification mechanism combining enzymatic softening with anaerobic acidification and energy storage. It utilizes a complex degradation system formed by Aspergillus niger, Candida utilis, and exogenous cellulase and xylanase to directionally cleave the cross-linked structure of the lignocellulose within the reed during the early fermentation stage, completely disintegrating the spiky, hard fibers at the leaf edges and main veins. The fiber softening effect, which cannot be achieved by mechanical crushing alone, is fully realized through the synergistic action of bacteria and enzymes. This fundamentally improves feeding safety by reducing the risk of mucosal abrasions, redness, and superficial damage in beef cattle during feeding. Beef cattle exhibit no avoidance, refusal to eat, or picky eating behavior. Feeding trial data shows that the average oral mucosal damage score of Simmental fattening cattle fed with the fermented feed of this invention was significantly lower than that of the control group, reaching as low as 0 points, completely solving the feeding obstacles caused by the physical irritation of natural reed leaves.

[0018] This invention addresses the fermentation challenges of reed leaves, which are characterized by low soluble sugar content, high buffering capacity, and difficulty in colonizing beneficial bacteria. It utilizes a gradient carbon-nitrogen synergistic buffering fermentation system, combining corn flour as a fast-acting carbon source, wheat bran as a slow-release nitrogen source, and molasses as a rapid probiotic sugar source. This system rapidly drives the proliferation and colonization of a complex of beneficial bacteria, significantly accelerating acidification. The pH value of the material decreases from 6.85 in the raw material state to 3.98 after fermentation, entering a significantly acidic antibacterial range. This effectively inhibits the growth of various harmful microorganisms such as molds, butyric acid bacteria, and putrefactive bacteria. It avoids the shortcomings of traditional reed silage fermentation, such as incomplete fermentation, rancidity, mold, strong odor, and short storage period. The fermented product has stable and controllable quality and can be stably stored for more than six months in a cool, dark environment, significantly extending the shelf life of the forage.

[0019] This invention simultaneously improves the nutritional quality of forage by completing the physical modification of fiber and fermentation stabilization. During fermentation, the structural crude fiber in reeds that is difficult for beef cattle to digest and utilize in the rumen is degraded into small-molecule usable sugars. At the same time, microbial protein, lactic acid, acetic acid, and ester aromatic flavor substances are enriched. According to the test comparison, the crude protein content after fermentation is 21.12% higher than that of unfermented reed leaves, while the crude fiber, neutral detergent fiber, and acid detergent fiber decrease by 24.23%, 22.06%, and 23.55%, respectively. The dry matter digestibility is significantly improved, lactic acid is enriched to 2.85%, acetic acid is enriched to 0.68%, and the previously undetectable ester aromatic flavor substances reach 86%. With a concentration of 35mg / kg, the finished product possesses a naturally sweet and sour fermented aroma, free from any putrid odor. It not only optimizes the rumen microecological environment of beef cattle but also further enhances the palatability of the forage through flavor compounds. Large-scale beef cattle feeding trials have shown that, in actual feeding comparisons, replacing untreated reed leaves entirely with the fermented feed of this invention increased the average daily dry matter intake of Simmental fattening cattle by 8.67%, increased the average daily weight gain by 6.86% over a 60-day trial period, and optimized the feed conversion ratio by 5.21%. Feed conversion efficiency and beef cattle growth performance improved simultaneously. Through the synergistic effect of fermentation modification and scientific nutritional formulation, it truly achieves the high-value transformation of low-value reed waste into high-quality beef cattle-specific forage.

[0020] The overall production process of this invention is compatible with existing conventional silage fermentation equipment and TMR mixing equipment. There is no need to purchase high-end precision processing equipment. The production operation process is simple and easy to implement. The entire filling time can be controlled within six hours to complete the material layering, compaction and sealing operation. The threshold for industrial production is low and the industrialization adaptability is strong. The raw material is waste reed leaves widely distributed in rivers, tidal flats and wetlands. The resource reserves are large and the material cost is low. Only a small increase in auxiliary materials, bacterial enzyme preparations and labor processing costs can complete the forage quality improvement and transformation. The overall economic benefits of breeding are significantly improved. It can realize the resource recycling of wetland reed waste, reduce the cost of purchasing commercial forage at the breeding end, take into account the ecological and environmental protection value and the need for cost reduction and efficiency improvement in beef cattle breeding, and has extremely high value for large-scale promotion and application. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 This is the activated bacterial enzyme mixture used in this invention; Figure 2 The raw material for fermented reed leaves in this invention; Figure 3This is the experimental sample of fermented reed leaves used in this invention; Figure 4 This is the fermentation process of reed leaves in this invention; Figure 5 This describes the process of filling fermented reed leaves in this invention. Detailed Implementation

[0023] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0024] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0025] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0026] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0027] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0028] The palatability-improving reed leaf microbial enzyme-co-synergistic segmented fermented feed of this invention uses dried and pulverized reed leaves as the core raw material, combined with carbon and nitrogen adjuvants and molasses, and is prepared through a segmented coupled fermentation process of "enzymatic softening + anaerobic acidification and energy storage" mediated by a compound fermentation microbial enzyme preparation. The compound fermentation microbial enzyme preparation is composed of a compound of live bacterial strains and a compound of degrading enzyme preparations: the live bacterial strains include *Lactobacillus plantarum*, *Candida utilis*, and *Aspergillus niger*; the degrading enzyme preparations include cellulase and xylanase.

[0029] In the following examples, the dried and pulverized reed leaves used were all clean reed leaves that had been naturally sun-dried to a moisture content of ≤12%, then pulverized and cut into 1-3 cm sections, removing moldy, blackened, and rotten impurities; sugarcane molasses or beet molasses could be used, which is beneficial for the enrichment of flavor substances; the total effective viable count of the compound live bacteria strain was ≥1×10⁻⁶. 9 CFU / g; the compound degradative enzyme preparation contains cellulase activity ≥15000 U / g and xylanase activity ≥8000 U / g.

[0030] Example 1 The palatability-improving reed leaf microbial enzyme-co-fermented feed described in this embodiment comprises the following raw materials by weight: 100 parts dried and crushed reed leaves, 12 parts corn flour, 8 parts wheat bran, 4 parts molasses, and 0.2 parts compound fermentation microbial enzyme preparation. The effective viable count ratio of *Lactobacillus plantarum*: *Candida utilis*: *Aspergillus niger* in the compound live bacteria strain is 2.5:1.2:0.8; the activity ratio of cellulase: xylanase in the compound degradation enzyme preparation is 1:0.5. The total moisture content of the fermentation material is controlled to 50%, the fermentation temperature is 30℃, and the fermentation cycle is 25 days.

[0031] The preparation method in this embodiment specifically includes the following steps: ① Raw material pretreatment: Collect clean, mold-free reed leaves, air dry them naturally until the moisture content is ≤12%, and then use a straw crusher to cut and crush them into sections, controlling the length of the material to 1-3 cm, removing impurities, and reducing the initial sharpness of the raw materials to obtain pretreated reed leaf raw materials for later use.

[0032] ② Material premixing: Put the pretreated reed leaves into the silage fermentation tank or TMR mixing equipment, add corn flour and wheat bran in sequence according to the ratio, and mechanically stir for 3-5 minutes. Dilute the molasses with an appropriate amount of warm water (which can be included in the total moisture content), and add it together with the bacterial enzyme activation solution by atomization spray to obtain a uniformly mixed basic mixture.

[0033] ③ Enzyme activation and moisture control: The compound enzyme preparation is activated in warm water at 30-40℃ for 10-15 min to prepare a homogeneous enzyme activation solution; for example... Figure 1 As shown, the activated bacterial enzyme mixture was evenly dispersed without obvious clumping or sedimentation. The activation solution was evenly sprayed onto the surface of the mixture using an atomizing spray method, with simultaneous and continuous stirring. The moisture content of the material was monitored in real time, and the total moisture content was ultimately controlled to 50%. Figure 4 As shown, during the fermentation of reed straw, mechanical stirring combined with atomized spraying ensures that the enzyme activation solution and the materials are fully in contact and mixed evenly.

[0034] ④ Layered filling, compaction, and deoxygenation: The prepared materials are filled into the fermentation tank in batches and layers, with each layer controlled to a thickness of 20-30 cm. Each layer is compacted layer by layer to fully remove air from the gaps between materials. The entire filling process should be controlled within 6 hours to minimize aerobic loss of the materials. Figure 5 As shown, the fermentation process of reed straw is carried out using a layered filling and compaction method to ensure dense material and low residual oxygen content. Figure 2 As shown, the fermented reed straw raw material before filling is uniform in shape and meets the feeding requirements.

[0035] ⑤ Segmented Sealed Fermentation: After filling, the entire structure is sealed with a double-layer impermeable film, and the edges are pressed tightly to isolate it from air and rainwater. Anaerobic fermentation is carried out at 30℃ for 25 days. The fermentation process is divided into two stages: (1) Enzymatic softening stage (fermentation days 1-5): After the material is sealed, the trace oxygen remaining in the gaps is rapidly consumed within a few hours. This stage mainly relies on exogenously added cellulase and xylanase to efficiently enzymatically hydrolyze the reed fiber, directionally cleaving the cross-linked structure of the lignocellulose and disintegrating the sharp, hard fibers; at the same time, Candida utilis undergoes limited proliferation under microaerobic conditions, and Aspergillus niger survives in spore form. The three work together to achieve physical and flexible modification of the forage; (2) Anaerobic acidification and energy storage stage (from day 6 of fermentation to the end of fermentation): After the oxygen in the system is exhausted, it enters an anaerobic environment. Lactobacillus plantarum proliferates and produces acid, which rapidly reduces the pH value of the material, inhibits the growth of harmful microorganisms such as mold, butyric acid bacteria, and putrefactive bacteria, and enriches organic acids and aromatic flavor substances, thus completing the nutritional quality improvement and stable preservation.

[0036] ⑥ Finished Product Processing and Storage: After fermentation, the finished fermented feed is obtained, such as... Figure 3 As shown, the fermented reed straw test sample was yellowish-brown and loose, with a soft texture and no hard thorns, emitting a sweet and sour fermented aroma. The finished product was obtained by taking material from the end in layers, with a single material thickness ≤30 cm. After taking the material, it was immediately resealed and compacted, and stored in a cool, dark environment, where it could be stably preserved for more than 6 months.

[0037] The characteristics of the fermented product in this embodiment are as follows: the material is yellowish-brown and loose, with a soft texture and no hard thorns, emitting a sweet and sour fermented aroma, without mold or putrid odor, and the pH value of the finished product is 3.98, indicating excellent fermentation quality.

[0038] Comparative Example 1 The raw material weight ratio, microbial enzyme system parameters, fermentation temperature and cycle of this comparative example are the same as those of Example 1, except that the total moisture content of the fermented material is adjusted to 48%. The test results after 25 days of fermentation showed that the material was relatively dry, the metabolic rate of the microbial strain was slow, the fiber degradation was incomplete, the lactic acid accumulation was lower than that of Example 1, and the softness and palatability of the forage were weaker than those of Example 1.

[0039] Comparative Example 2 The raw material weight ratio, bacterial enzyme system parameters, fermentation temperature and cycle of this comparative example are the same as those of Example 1, except that the total moisture content of the fermentation material is adjusted to 52%. After 25 days of fermentation, the test results showed that the material was prone to leakage and adhesion, and localized breeding of miscellaneous bacteria resulted in slight "micro-rot," leading to a decline in fermentation quality and weaker storage stability than in Example 1.

[0040] Comparative Example 3 The weight ratio of raw materials, moisture content, fermentation temperature, and cycle in this comparative example are consistent with those in Example 1. Only a single *Lactobacillus plantarum* was used for fermentation, and a compound degrading enzyme preparation with the same activity as in Example 1 was added, but *Aspergillus niger* and *Candida utilis* were not added. After fermentation, the results showed that the reed leaves still retained a large amount of hard, sharp fibers, the pH value of the material was 4.6, the acidification effect was poor, and mold growth was likely to occur later. Fiber softening and palatability improvement could not be achieved, and the feeding safety and nutritional quality were significantly lower than in Example 1.

[0041] Experimental Example 1 To verify the technical effectiveness of the fermented feed of this invention, unfermented dried reed leaves were used as the blank control group, and the fermented product prepared in Example 1 was used as the experimental group. A series of index tests and feeding trials were conducted. All experimental data were statistically analyzed using SPSS software, and the results are expressed as "mean ± standard deviation". P <0.05 indicates a significant difference. P >0.05 indicates no significant difference.

[0042] 1. Comparison of conventional nutritional components before and after fermentation Routine nutritional indicators were tested in the blank group and the experimental group respectively, and the results are shown in Table 1.

[0043] Table 1 Comparison of conventional nutrients before and after fermentation

[0044] As shown in Table 1, the crude protein content of fermented reed leaves increased by 21.12% compared to unfermented leaves. P <0.05); the levels of crude fiber, neutral detergent fiber, and acid detergent fiber were significantly reduced after fermentation compared to unfermented fiber. P <0.05), which decreased by 24.23%, 22.06%, and 23.55%, respectively; the dry matter digestibility was significantly improved after fermentation ( P <0.05), indicating that the process of the present invention can effectively degrade the structural fibers of reeds, thereby improving the nutritional quality and digestibility of feed.

[0045] 2. Comparison of functional active ingredients The functional active ingredients of the blank group and the experimental group were detected separately, and the results are shown in Table 2.

[0046] Table 2 Comparison of Functional Active Ingredients

[0047] As shown in Table 2, the lactic acid content in the reed leaves increased significantly to 2.85% after fermentation. P <0.05%, acetic acid content significantly enriched to 0.68% ( P <0.05); ester aromatic substances increased from undetectable to 86.35 mg / kg, which are flavor substances specifically generated during fermentation; the effective viable count increased from naturally occurring bacteria to ≥1.0×10⁻⁵. 9 CFU / g, enriched with highly active beneficial bacteria; the pH value of the material decreased significantly from 6.85 to 3.98 ( P <0.05), a decrease of 41.90%, successfully entering the acidic antibacterial zone, ensuring the stability of feed storage.

[0048] 3. Cost Accounting Analysis Based on the calculation of one ton of finished feed, and according to the dry basis ratio of Example 1 (100 parts reed leaves, 12 parts corn flour, 8 parts wheat bran, 4 parts molasses, and 0.2 parts bacterial enzyme preparation), a total of 124.2 parts of feed were used to prepare 1 ton of finished product. Combining the market price of raw materials (corn flour 2340 yuan / t, wheat bran 1800 yuan / t, molasses 1280 yuan / t), the costs of harvesting, drying, crushing, storage pretreatment, raw materials, and labor were comprehensively calculated, and the results are shown in Table 3.

[0049] Table 3 Cost Accounting Table

[0050] The calculation results show that the fermented feed of this invention has achieved a significant improvement in the quality of reed forage while slightly increasing the processing cost. It still has a significant cost advantage compared with high-quality commercial forage, and its industrialization and promotion value is outstanding.

[0051] 4. Feeding Effect Trial on Beef Cattle Twenty healthy Simmental fattening cattle weighing (343±12.45) kg were randomly divided into a control group and an experimental group, with 10 cattle in each group. The experimental period was 60 days. The feeding environment, drinking water, and concentrate ratio were completely identical in all groups. The control group was fed unfermented pure reed leaves (simulating the actual situation in current production where pure reed leaves are used as roughage), while the experimental group was fed fermented reed feed prepared in Example 1 (containing corn flour, bran, molasses, and other supplementary ingredients). This comparison aims to verify the comprehensive application effect of using the product of this invention to completely replace pure reed leaves in actual feeding. The safety of feeding was evaluated using a five-level scoring standard for oral mucosal damage in beef cattle, as shown in Table 4.

[0052] Table 4 Evaluation Criteria for Oral Mucosal Injury

[0053] After the experiment, the growth performance and oral damage of each group of beef cattle were statistically analyzed, and the results are shown in Table 5.

[0054] Table 5. Effects of fermented feed on the growth performance of beef cattle

[0055] As shown in Table 5, compared with the unfermented reed leaf control group, the average daily dry matter intake of beef cattle in the fermented reed leaf experimental group of this invention was significantly increased by 8.67% ( P <0.05), the average daily weight gain over 60 days increased significantly by 6.86% ( P <0.05), the overall material weight ratio was significantly optimized by 5.21% ( P <0.05); the oral mucosal injury score decreased from 1.60 to 0.00, and the physical injury from feeding was completely eliminated.

[0056] In summary, the fermentation process of this invention significantly improves the palatability and digestibility of reed leaves through the synergistic modification of fiber structure softening and organic acids and aromatic esters. While completely eliminating the physical damage to the oral mucosa of beef cattle caused by the sharp fibers of reeds, it effectively promotes the simultaneous increase in feed intake and daily weight gain, realizing the functional transformation of low-quality reed forage into safe, efficient and high-quality forage. It can be widely used in the TMR diet of large-scale Simmental fattening cattle.

[0057] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A segmented fermented feed with synergistic enzymes and mycotoxins from reed leaves to improve palatability, characterized in that, This product is prepared by staged fermentation of the following raw materials in parts by weight using a synergistic fermentation process involving bacteria and enzymes: 100 parts dried and pulverized reed leaves, 10-15 parts corn flour, 5-10 parts wheat bran, 3-5 parts molasses, and 0.1-0.3 parts compound fermentation enzyme preparation; the total moisture content of the fermentation materials is adjusted to 48%-52%; the compound fermentation enzyme preparation comprises compound live bacteria strains and compound degradation enzyme preparations; the compound live bacteria strains consist of *Lactobacillus plantarum*, *Candida utilis*, and *Aspergillus niger*, with a live bacteria ratio of (2-3):(1-1.5):(0.5-1), and the total effective live bacteria count of the compound live bacteria strains is ≥1×10⁻⁶. 9 CFU / g; The compound degrading enzyme preparation is composed of cellulase and xylanase, with an enzyme activity ratio of 1:0.5, cellulase activity ≥15000 U / g, and xylanase activity ≥8000 U / g.

2. The reed leaf mycelium enzyme-co-fermented feed with improved palatability according to claim 1, characterized in that, The dried and pulverized reed leaves are clean reed leaves that have been naturally sun-dried to a moisture content of ≤12%, and are pulverized and cut into sections of 1-3 cm in length, removing any moldy, blackened, or rotten leaves; the molasses is selected from sugarcane molasses or beet molasses.

3. The reed leaf mycelium enzyme-co-fermented feed with improved palatability according to claim 1 or 2, characterized in that, The raw materials are formulated in the following weight proportions: 100 parts dried and pulverized reed leaves, 12 parts corn flour, 8 parts wheat bran, 4 parts molasses, and 0.2 parts compound fermentation enzyme preparation; the ratio of live Lactobacillus plantarum, Candida utilis, and Aspergillus niger in the compound live bacteria strain is 2.5:1.2:0.8; and the moisture content of the fermentation material is 50%.

4. A method for preparing a segmented fermented feed with synergistic enzymes and improved palatability of reed leaf fungi as described in any one of claims 1 to 3, characterized in that, Includes the following steps: ① Raw material pretreatment: Collect clean, mold-free reed leaves, air dry them naturally until the moisture content is ≤12%, crush and cut them into 1-3cm sections, and remove impurities for later use; ② Premixing: Add the pretreated reed leaves, corn flour, wheat bran, and molasses into a mixing device and mechanically mix for 3-5 minutes to obtain the basic mixture. ③ Enzyme activation and moisture control: The compound fermentation enzyme preparation is activated with warm water at 30-40℃ for 10-15 minutes to obtain the enzyme activation solution, which is then sprayed onto the basic mixture while being stirred simultaneously to control the total moisture content of the material to 48%-52%; ④ Layered filling, compaction and degassing: After mixing, the material is filled into the fermentation tank in layers, with a single layer thickness of 20-30 cm. Each layer is compacted, and the entire filling process takes ≤6 hours. ⑤ Segmented sealed fermentation: The fermentation tank is sealed with a double-layer impermeable membrane and fermented in a sealed environment of 25-35℃ for 15-30 days; the fermentation is divided into two stages: fermentation 1-5 days is the enzymatic softening stage, and fermentation 6-30 days is the anaerobic acidification and energy storage stage. ⑥ Finished product handling and storage: After fermentation is complete, take out the material in layers, with a single take-out thickness of ≤30 cm. After taking out the material, immediately reseal and compact it, and store it in a cool, dark place.

5. The preparation method according to claim 4, characterized in that, The fermentation environment temperature was 30℃, the fermentation cycle was 25 days, and the total moisture content of the fermentation material was 50%.

6. The preparation method according to claim 4, characterized in that, The material is yellowish-brown and loose, with a soft texture and no hard thorns. It has a sweet and sour fermented aroma, and is free from mold, rot, or other off-odors. It can be stored stably for more than 6 months under cool, dark conditions.

7. The application of the palatability-improving reed leaf fungus enzyme-synergistic segmented fermented feed as described in claim 1 in beef cattle farming.

8. The application according to claim 7, characterized in that, The nutritional indicators of the reed leaf fungus enzyme-synergistic segmented fermented feed are as follows: crude protein content ≥9.0%, crude fiber ≤30%, and lactic acid content ≥2.5%.

9. The application according to claim 8, characterized in that, The beef cattle farming mentioned refers to the large-scale breeding of Simmental fattening cattle.

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