Fattening feed additive composition for old-piece cattle and fattening method

Abstract

The invention discloses a fattening feed additive composition for old cattle and a fattening method, and belongs to the technical field of feed. The additive composition comprises the following components in parts by weight: 13-20 parts of a rumen function regulator, 4-10 parts of an intestinal repair agent, 10-20 parts of a rumen bypass compound enzyme preparation, 5-10 parts of a muscle-increasing growth-promoting composition, 5.5-7.3 parts of a functional trace element composition and 32.7-62.5 parts of a carrier. Wherein the rumen function regulator is compounded by a coated saccharomyces cerevisiae culture and bacillus subtilis; the rumen bypass compound enzyme preparation comprises acid protease, neutral protease and cellulase; the muscle-building and growth-promoting combination is coated guanidinoacetic acid. Through regulation and control of a rumen-intestinal tract-tissue three-level linkage network, the rumen fermentation function of the old cattle is remarkably improved, the feed conversion rate is increased, and meanwhile muscle hyperplasia and meat tenderization are achieved. By means of the fattening method, the average daily gain of the old cattle is increased by 22.8% or above, the muscle shear force is reduced by 29.6% or above, the feed conversion ratio is reduced by 17.1% or above, and green and efficient fattening is achieved.

Description

Technical Field

[0001] This invention relates to the field of feed technology, and in particular to a fattening feed additive composition for old cattle and a fattening method. Background Technology

[0002] With the increasingly refined development of the beef cattle industry, the fattening and reuse of older, thinner adult cows (referring to adult cows culled due to declining reproductive performance or the end of their working years) has become a crucial link in improving the overall efficiency of beef cattle farming. However, compared with young cattle (18-24 months old), older cattle exhibit significant differences in physiological metabolism, not only in degree but also in nature. Based on the preliminary experiments and literature review presented in this invention, older cattle mainly exhibit the following three quantitative characteristics:

[0003] (1) Rumen function decline: The rumen microbiota ages, and the abundance of core fiber-decomposing bacteria (such as Bacteroides succinate and Rumenococcus albus) decreases by more than 50% compared with young cattle, resulting in a 30%-50% reduction in crude fiber utilization.

[0004] (2) Hardening of muscle connective tissue: The degree of cross-linking of collagen in the muscle is significantly increased, and the proportion of heat-soluble collagen decreases by about 40%, resulting in a muscle shear force value that is more than 40% higher than that of young cattle;

[0005] (3) Imbalance between fat deposition and muscle growth: The secretion level of endogenous hormones (such as IGF-1 and growth hormone) decreases by 30%-50%, which leads to a slowdown in protein synthesis rate and energy metabolism tends to deposit fat rather than synthesize muscle.

[0006] The specificity of the problem: This means that conventional fattening techniques effective for young cattle (such as adding guanidinoacetic acid alone to promote growth, or adding enzyme preparations alone to aid digestion) will be largely ineffective or significantly less effective in older cattle due to "substrate limitation" (excessive cross-linking of collagen locks in the growth space of muscle fibers) and "insufficient energy supply" (low rumen fermentation efficiency). Therefore, the fattening problem of older cattle is a highly specific and complex systems engineering problem, rather than a simple application of general fattening techniques.

[0007] Currently, fattening technologies for ruminants are mostly focused on calves and growing cattle, while research and development of additives specifically for older cattle is still lacking.

[0008] 1. Analysis of existing technical solutions and their shortcomings

[0009] To more clearly demonstrate the differences between this invention and existing technologies, relevant domestic and international patents and non-patent literature were searched and analyzed. The following three most representative examples were selected: Existing technology 1: Antibiotic growth promoters, with monensin and other ion carrier antibiotics as the main components, which increase propionic acid production by inhibiting Gram-positive bacteria in the rumen and changing the rumen fermentation pattern; Existing technology 2: Monoguanidinoacetic acid, with monoguanidinoacetic acid as the main component, which acts as a creatine precursor to promote muscle growth and increase daily weight gain; Existing technology 3: Ordinary compound enzyme preparations, composed of single enzymes such as cellulase and xylanase, which are directly added to the diet to break down fiber in roughage.

[0010] 2. Summary of existing problems with the technology:

[0011] Existing technologies, when addressing the specific technical problem of "fatting old cattle," mainly suffer from the following technical gaps and drawbacks:

[0012] (1) Lack of targeted technology – the “triple superposition” problem of fattening old cattle: no integrated solution was provided for the triple superposition physiological dilemma of “rumen microbiota aging + muscle connective tissue hardening + fat metabolism slowdown” in old cattle. There are significant differences in physiological metabolism between old cattle (culled cows or retired draft cattle) and young cattle, mainly manifested as: rumen function decline: rumen microbiota aging, fiber-decomposing bacteria abundance decreased by 30%-50%, crude fiber utilization rate significantly reduced; muscle connective tissue hardening: collagen cross-linking intensified, muscle shear force value more than 40% higher than that of young cattle; difficulty in fat deposition: insufficient secretion of endogenous hormones, decreased fat synthesis capacity, difficulty in intramuscular fat deposition.

[0013] These three deficiencies are interconnected and mutually causal, forming a complex network of physiological imbalances. However, existing technologies 1, 2, and 3 all employ a linear "supplement what is lacking" approach: Existing technology 1 uses antibiotics to inhibit Gram-positive bacteria in the rumen, targeting only a single point of regulation in the energy metabolism pathway; Existing technology 2 uses guanidinoacetic acid alone to promote muscle growth, targeting only weight gain; and Existing technology 3 adds cellulase and xylanase to improve fiber digestibility, targeting only digestion.

[0014] Those skilled in the art have long been bound by this linear thinking, viewing older cattle as a "simplified version of younger cattle" rather than a complex system requiring systematic intervention. This cognitive limitation has resulted in a long-standing technological gap in the field—despite the decades-long existence of the problem of fattening older cattle, it has never been effectively resolved.

[0015] (2) Low utilization rate of active ingredients - a long-standing technical challenge in the field of "rumen protection".

[0016] Functional proteins (such as enzymes) or live bacteria are often largely degraded or inactivated by rumen microorganisms when passing through the rumen. This has long been a technical challenge in the field of "rumen protection," specifically manifested as follows:

[0017] ① Low rumen clearance rate of enzyme preparations: Ordinary compound enzyme preparations are not coated and are easily degraded by microorganisms in the rumen. Studies have shown that the residual rate of unprotected enzyme preparations in the rumen is less than 10%, and the effective dose that actually reaches the small intestine to exert its effect is seriously insufficient, resulting in a significant reduction in the technical efficacy.

[0018] ② Low survival rate of probiotics: Probiotics (such as Bacillus and yeast) face multiple pressures when passing through the rumen, including a low pH environment, hydrolysis by digestive enzymes, and competition from other microorganisms, resulting in a significantly reduced survival rate. Conventionally added probiotics are unlikely to effectively colonize the gut and perform their functions.

[0019] ③ Rumen loss of guanidinoacetic acid: As a precursor to creatine, guanidinoacetic acid will also be degraded by microorganisms in the rumen if it is not protected, resulting in low bioavailability and affecting its growth-promoting effect.

[0020] ④ Limitations of existing protection technologies: Conventional enteric coating technology can only solve the problem of gastric acid degradation, but cannot effectively protect active ingredients from rumen microbial degradation. The residual rate of single-layer coated enzyme preparations in the rumen is only about 30%, which is still difficult to meet the needs of efficient delivery.

[0021] Therefore, how to achieve efficient rumen protection for multiple functional components (enzymes, probiotics, guanidinoacetic acid) so that they remain active in the rumen and are released at specific points in the small intestine is a long-standing technical problem that has never been effectively solved in this field.

[0022] (3) Disconnect between meat quality improvement and weight gain – lack of systematic solutions

[0023] Existing technologies lack a systematic technical solution that can simultaneously achieve "weight gain, tenderization, and fat deposition," specifically manifested in the following ways:

[0024] ① Weight gain and meat quality are difficult to achieve simultaneously: It is well known in the field that rapid weight gain often leads to insufficient fat deposition and a decline in meat quality; excessive pursuit of tenderness may also affect weight gain. The "Technical Specifications for Beef Cattle Fattening" (NY / T 2665-2016) states: "In the later stages of fattening, a balance needs to be sought between weight gain and fat deposition, and the two are difficult to optimize simultaneously." The Beef Cattle Association's "Fattening Handbook" (2020) also clearly states: "Measures to increase daily weight gain usually reduce marbling scores, and the two are negatively correlated."

[0025] ② Limitations of single-component functions: Antibiotics only act on energy metabolism pathways, which can improve weight gain, but do not take into account muscle tenderness and cannot improve meat quality; compound enzyme preparations only improve fiber digestibility and have no direct effect on muscle growth and fat deposition; single guanidinoacetic acid only promotes muscle growth and does not improve meat tenderness, and unprotected guanidinoacetic acid has low bioavailability.

[0026] ③ Lack of temporal synergistic design: Existing technologies do not consider the roles of different functional components at different stages of fattening. In the fattening process of older cattle, there is a temporal relationship between the degradation of muscle connective tissue and the synthesis of muscle protein—it is necessary to first degrade aged collagen to "make room" before promoting the synthesis of new muscle protein to "fill the space." Existing technologies lack this "destruction before construction" temporal synergistic design.

[0027] Therefore, how to design a systematic technical solution that can simultaneously optimize multiple indicators such as "weight gain, tenderness, and fat deposition" is a technical problem that urgently needs to be solved in this field.

[0028] 3. Incompatible with the trend of green development – ​​the urgent need for antibiotic alternatives:

[0029] There is an urgent need in the industry to develop a green and safe fattening program that does not rely on antibiotics at all and can simultaneously optimize multiple indicators such as weight gain, tenderization, and fat deposition.

[0030] 4. Technical bottlenecks in the combined application of probiotics and enzyme preparations:

[0031] In addition to the issues mentioned above, existing technologies also face technical bottlenecks in the combined application of probiotics and enzyme preparations. How to achieve synergistic effects between probiotics and enzyme preparations while ensuring their activity, and to obtain comprehensive results superior to those of a single component, is also a technical problem that needs to be solved in this field.

[0032] ④ Reverse Verification from Market Practice: Besides warnings in academic literature, experimental "probiotic + enzyme" combination products have appeared on the market. However, due to the failure to effectively address the hydrolytic inactivation of probiotic cell wall proteins by enzymes during storage, and the conflicting release sequences in the gut after rumen passage, the product efficacy was extremely unstable, even leading to adverse reactions such as increased diarrhea when used at high doses. These products largely withdrew from the Chinese market around 2018. The market's "voting with its feet" further reinforced the industry consensus that "probiotics and enzymes are incompatible," leading those skilled in the art to generally prefer "choosing one or the other" or alternating between the two when designing feed formulations, rather than risking combining them in the same product.

[0033] In conclusion, our research found that there is currently no dedicated fattening additive product in China specifically designed for cattle over 35 months of age. Existing products all target young or growing cattle, and their design concepts and technical parameters are completely unsuitable for the unique physiological characteristics of older cattle. Summary of the Invention

[0034] The technical solution of this invention to solve the above-mentioned technical problems is to provide a composition of fattening feed additive for old cattle, comprising the following raw materials in parts by weight:

[0035] Rumen function modifier: 13-20 parts;

[0036] Intestinal repair agent: 4-10 parts;

[0037] Rumen-protected complex enzyme preparation: 10-20 parts;

[0038] Muscle-building and growth-promoting combination: 5-10 servings;

[0039] Functional trace element combination: 5.5-7.3 parts;

[0040] Carrier: 32.7-62.5 copies;

[0041] The rumen function regulator is composed of coated Saccharomyces cerevisiae culture and Bacillus subtilis.

[0042] The intestinal repair agent is Bacillus licheniformis;

[0043] The rumen-protected complex enzyme preparation contains acidic protease (Aspergillus niger), neutral protease (Bacillus subtilis), and cellulase.

[0044] The muscle-building and growth-promoting combination is coated with guanidinoacetic acid;

[0045] The functional trace element combination contains only three elements: zinc, iron, and selenium.

[0046] The carrier is one or more of defatted rice bran, zeolite powder, or maifanite powder.

[0047] Furthermore, in the rumen function regulator, the coated Saccharomyces cerevisiae culture comprises 10-15 parts, and Bacillus subtilis comprises 3-5 parts; the coated Saccharomyces cerevisiae culture is coated using a fluidized bed spraying process, wherein a composite coating material of hydrogenated vegetable oil and ethyl cellulose is coated on the surface of the yeast culture particles, and the coating thickness is 20-50 μm.

[0048] Furthermore, the rumen-protected complex enzyme preparation is composed of the following enzymes in weight percentage: 40-50% acidic protease, 30-40% neutral protease, and 20-30% cellulase; the rumen-protected complex enzyme preparation is treated with a double-layer coating technology, wherein the double-layer coating is: an inner layer of pH-sensitive polyacrylic acid resin coating and an outer layer of hydrogenated vegetable oil coating.

[0049] Furthermore, the enzyme preparation treated with the double-layer coating technology has a residual rate of ≥85% after 24 hours of simulated rumen culture in vitro, and a release rate of ≥90% after 2 hours of simulated digestion in the small intestine.

[0050] Furthermore, the functional trace element combination consists of the following components: 3-3.6 parts of methionine hydroxy analog chelated zinc, 1-2 parts of glycine chelated iron, and 1.5-1.7 parts of yeast selenium.

[0051] Furthermore, the carrier is a mixture of defatted rice bran and zeolite powder in a weight ratio of 3:1.

[0052] A method for fattening older cattle using the additive composition described above is also proposed, wherein the additive composition is added to the total mixed ration at a mass ratio of 0.5%-1.0%.

[0053] (1) Early fattening stage: 1-15 days, the additive composition in the total mixed ration is added at a rate of 0.5%;

[0054] (2) Mid-finishing stage: 16-60 days, the additive composition in the total mixed ration is added at a rate of 1.0%;

[0055] (3) Late fattening period: 61-130 days, the amount of additive composition added to the total mixed diet is 1.0%.

[0056] The total mixed ration comprises roughage and concentrate, wherein the concentrate comprises 1% of the additive composition.

[0057] Compared with the prior art, the technical solution of the present invention has the following significant advantages:

[0058] (1) Significantly improves feed conversion efficiency and reshapes rumen microecology:

[0059] By leveraging the synergistic effect of coated Saccharomyces cerevisiae culture and Bacillus subtilis, this invention directionally optimizes the rumen microbiota of aging cattle. The relative abundance of fiber-decomposing bacteria is significantly increased, crude fiber digestibility is improved by 15%-20%, and the feed conversion ratio is reduced by more than 12%.

[0060] (2) Simultaneously improve meat tenderness and carcass quality to achieve meat quality improvement of "muscle-enhancing and tenderizing":

[0061] This invention achieves simultaneous improvement in meat quality through the synergistic effect of a double-coated complex enzyme preparation and a coated guanidinoacetic acid:

[0062] Improved tenderness: After being released in the small intestine, the rumen-protected complex protease (acidic + neutral) effectively degrades the excessively cross-linked collagen in feed protein and muscle connective tissue, reducing muscle shear force by 29.6% and significantly improving meat tenderness.

[0063] Muscle building and shaping: By adding the recommended amount to the total mixed ration (TMR) (such as 1.0% in the mid-to-late fattening stage, about 1000 g / ton of TMR) to precisely regulate energy metabolism and guide energy to be deposited in muscles, the eye muscle area increased by 17.6% and the lean meat percentage of the carcass increased significantly.

[0064] (3) Achieve antibiotic-free fattening throughout the entire process and enhance the body's antioxidant capacity:

[0065] Replacing traditional antibiotics with a probiotic combination (Bacillus subtilis + Bacillus licheniformis) meets the requirements of green farming and has no withdrawal period. At the same time, the antioxidant effect of yeast selenium increases the total antioxidant capacity of serum by more than 65%, effectively ensuring the health of older cattle during the fattening period.

[0066] (4) Shorten the fattening cycle and improve the overall breeding benefits:

[0067] Targeting the slower metabolism of older cattle, a three-stage precise nutritional regulation method was used to shorten the traditional 6-8 month fattening cycle to 4-5 months, a reduction of over 30%. Experimental data showed that the experimental group achieved an average daily weight gain of 1.45 ± 0.15 kg, 22.8% higher than the control group, resulting in a significant improvement in overall breeding efficiency. Detailed Implementation

[0068] This invention proposes a fattening feed additive composition and fattening method for older cattle, aiming to design a special fattening additive product specifically for "older cattle over 35 months of age".

[0069] The following will describe the composition of the fattening feed additive for old-fashioned cattle proposed in this invention through specific embodiments:

[0070] Example 1:

[0071] A composition of fattening feed additive for old-fashioned cattle, comprising the following raw materials in parts by weight:

[0072] Rumen function modifier: 13-20 parts;

[0073] Intestinal repair agent: 4-10 parts;

[0074] Rumen-protected complex enzyme preparation: 10-20 parts;

[0075] Muscle-building and growth-promoting combination: 5-10 servings;

[0076] Functional trace element combination: 5.5-7.3 parts;

[0077] Carrier: 32.7-62.5 copies;

[0078] The rumen function regulator is composed of coated Saccharomyces cerevisiae culture and Bacillus subtilis.

[0079] The intestinal repair agent is Bacillus licheniformis;

[0080] The rumen-protected complex enzyme preparation contains acidic protease (Aspergillus niger), neutral protease (Bacillus subtilis), and cellulase.

[0081] The muscle-building and growth-promoting combination is coated with guanidinoacetic acid;

[0082] The functional trace element combination contains only three elements: zinc, iron, and selenium.

[0083] The carrier is one or more of defatted rice bran, zeolite powder, or maifanite powder.

[0084] Furthermore, in the rumen function regulator, the coated Saccharomyces cerevisiae culture comprises 10-15 parts, and Bacillus subtilis comprises 3-5 parts; the coated Saccharomyces cerevisiae culture is coated using a fluidized bed spraying process, wherein a composite coating material of hydrogenated vegetable oil and ethyl cellulose is coated on the surface of the yeast culture particles, and the coating thickness is 20-50 μm.

[0085] Understandably, the coated Saccharomyces cerevisiae culture "promotes fermentation and provides energy": It employs a fluidized bed spraying process to coat the yeast culture with a composite coating of hydrogenated vegetable oil and ethyl cellulose, with the coating thickness controlled at 20-50 μm. The yeast culture is slowly released in the rumen, consuming residual oxygen and creating a suitable environment for strictly anaerobic fiber-decomposing bacteria (Bacteroides succinate-producing and Rumenococcus albus), while also providing growth factors such as small peptides and oligosaccharides to promote the proliferation of fiber-decomposing bacteria and improve crude fiber digestibility; Bacillus subtilis has a viable count ≥1×10⁻⁶. 9CFU / g. "Oxygen-consuming enzyme production" consumes oxygen in the rumen and anterior intestinal tract, secreting digestive enzymes such as proteases and amylases to promote feed digestion and reduce harmful fermentation in the hindgut; simultaneously, it inhibits the colonization of harmful bacteria through competitive inhibition. *Bacillus licheniformis*, "synergistically protects the gut": working synergistically with *Bacillus subtilis*, it further consumes oxygen in the intestine, inhibiting aerobic harmful bacteria (such as *Escherichia coli* and *Salmonella*), stabilizing the intestinal microecological balance; its metabolites can enhance the intestinal epithelial barrier function and reduce diarrhea rates. In the aforementioned double-layer coating technology, the outer layer uses hydrogenated vegetable oil as the coating material. This material is a processing aid that forms a physical barrier in the rumen to protect the inner active ingredients. It can be digested and absorbed in the small intestine without residue or safety risks. Using hydrogenated vegetable oil as a coating material complies with the usage specifications for feed processing aids.

[0086] Bacillus licheniformis, viable count ≥1×10 9 The dosage is 4-10 CFU / g. Its function is to work synergistically with Bacillus subtilis to further consume oxygen in the intestine, inhibit aerobic harmful bacteria, stabilize the intestinal microecological balance, and enhance the intestinal barrier function through metabolites.

[0087] Furthermore, the rumen-protected complex enzyme preparation is composed of the following enzymes in weight percentage: 40-50% acidic protease, 30-40% neutral protease, and 20-30% cellulase; the rumen-protected complex enzyme preparation is treated with a double-layer coating technology, wherein the double-layer coating is: an inner layer of pH-sensitive polyacrylic acid resin coating and an outer layer of hydrogenated vegetable oil coating.

[0088] Understandably, the product is treated with a double-layer coating technology: the inner layer is a pH-sensitive polyacrylic acid resin to ensure that the enzyme preparation does not dissolve in the acidic environment of the rumen (pH 5.5-6.8) and protects the enzyme activity from degradation by rumen microorganisms; the outer layer is hydrogenated vegetable oil to prevent heat damage during the feed pelleting process. Acidic protease (from Aspergillus niger) comprises 40-50% by mass, with an enzyme activity ≥50,000 U / g, and is used to initiate protein hydrolysis in the acidic environment of the abomasum. Neutral protease (from Bacillus subtilis) comprises 30-40% by mass, with an enzyme activity ≥50,000 U / g, and is used to continue protein hydrolysis in the neutral environment of the small intestine. Cellulase comprises 20-30% by mass, with an enzyme activity ≥20,000 U / g, and is used to degrade cellulose in the diet, releasing encapsulated starch and protein. These three components synergistically improve protein and fiber digestibility, while the protease also degrades collagen in muscle connective tissue, improving meat tenderness.

[0089] Furthermore, guanidinoacetic acid undergoes a double-coating process to ensure it is not degraded in the rumen and is released and absorbed in the small intestine. Its function is to act as a precursor to creatine, promoting creatine synthesis, regulating energy metabolism, directing more energy towards muscle growth, and improving daily weight gain, lean meat percentage, and feed conversion ratio. The coated guanidinoacetic acid exhibits a 24-hour residual rate of ≥90% in the rumen and a 2-hour release rate of ≥95% in the small intestine. The enzyme preparation treated with this double-coating technology exhibits a ≥85% residual rate after 24 hours of simulated rumen culture in vitro and a ≥90% release rate after 2 hours of simulated digestion in the small intestine.

[0090] Furthermore, the functional trace element combination consists of the following components: 3-3.6 parts of methionine hydroxy analog chelated zinc, 1-2 parts of glycine chelated iron, and 1.5-1.7 parts of yeast selenium. It contains only the three functional trace elements of zinc, iron, and selenium, and does not contain copper, iodine, or cobalt (provided in the mixed feed).

[0091] Understandably, methionine hydroxy analogue chelated zinc is used for "strengthening muscles and bones": with moderate chelation strength, it participates in protein synthesis and keratinization, promotes hoof health, and prevents hoof diseases during the fattening period; zinc, as a cofactor of many enzymes, participates in nucleic acid and protein metabolism; glycine chelated iron is used for "color enhancement and oxygenation": with strong chelation strength, it increases hemoglobin content, improves meat color redness, and prevents the appearance of "white muscle" (PSE meat); it also participates in the composition of cytochromes and respiratory chain enzyme systems, enhancing tissue oxygenation; yeast selenium is used for "antioxidant protection of meat": bioavailable selenium, as a component of glutathione peroxidase (GSH-Px), scavenge free radicals, reduce oxidative stress, and extend the shelf life of meat products; it also enhances the body's immunity.

[0092] Furthermore, the carrier is a mixture of defatted rice bran and zeolite powder in a weight ratio of 3:1. For "carrier adsorption": defatted rice bran provides mixing uniformity and flowability, ensuring uniform distribution of trace components in the premix; zeolite powder has a porous structure and ion exchange capacity, which can adsorb ammonia nitrogen in the rumen, improve the rumen environment, and slowly release the adsorbed trace elements, improving utilization.

[0093] Preparation method of additive composition:

[0094] The method for preparing the additive composition includes the following steps:

[0095] (1) Raw material preparation: Prepare or purchase raw materials for each component according to the proportions to ensure that all indicators meet the requirements.

[0096] (2) Stepwise mixing: In order to avoid mutual interference of active ingredients, the probiotic components (saccharitomyces culture, Bacillus subtilis, Bacillus licheniformis) and enzyme components (rumen-protected complex enzymes) are premixed with a portion of the carrier, and then added to the V-type mixer with the remaining components and mixed thoroughly for 15 minutes until the coefficient of variation of the mixing uniformity is ≤5%.

[0097] (3) Packaging and storage: The well-mixed additive premix is ​​packaged in 25 kg bags and stored in a cool, dry and dark place.

[0098] This invention achieves efficient fattening of older cattle (over 35 months old) through a three-level regulatory network of "rumen-gut-tissue":

[0099] (1) Rumen dimension (upstream): Coated Saccharomyces cerevisiae culture promotes the proliferation of fiber-decomposing bacteria and increases energy supply; Bacillus subtilis consumes oxygen and secretes digestive enzymes, laying the foundation for subsequent digestion.

[0100] (2) Intestinal dimension (midstream): Bacillus licheniformis and Bacillus subtilis work together to form a triple effect of "oxygen consumption-enzyme production-harm inhibition" to maintain intestinal health; double-layer coating technology ensures that enzyme preparations and guanidinoacetic acid are released at specific points after passing through the rumen.

[0101] (3) Tissue dimension (downstream): Rumen-exposed complex enzyme preparations degrade feed protein and muscle collagen, improving tenderness; coated guanidinoacetic acid guides energy deposition to muscle, increasing eye muscle area; functional trace elements provide basic nutritional support and antioxidant support.

[0102] Example 2:

[0103] A method for fattening older cattle using the additive composition described above is also proposed, wherein the additive composition is added to the total mixed ration at a mass ratio of 0.5%-1.0%.

[0104] (1) Early fattening stage: 1-15 days, the additive composition in the total mixed ration is added at a rate of 0.5%;

[0105] (2) Mid-finishing stage: 16-60 days, the additive composition in the total mixed ration is added at a rate of 1.0%;

[0106] (3) Late fattening period: 61-130 days, the amount of additive composition added to the total mixed diet is 1.0%.

[0107] The total mixed ration comprises roughage and concentrate, wherein the concentrate comprises 1% of the additive composition.

[0108] Early fattening stage (concentrate to roughage ratio 40:60, TMR additive composition added at 0.5%).

[0109] The additive composition, by weight, comprises 12 parts coated Saccharomyces cerevisiae culture, 4 parts Bacillus subtilis as a rumen function regulator, 4 parts Bacillus licheniformis as an intestinal repair agent, 4.5 parts Aspergillus niger-derived acidic protease, 3.5 parts Bacillus subtilis-derived neutral protease, and 2.0 parts cellulase as a rumen-protected complex enzyme preparation, 10 parts coated guanidinoacetic acid as a muscle-building and growth-promoting combination, 3.3 parts methionine hydroxy analog chelated zinc, 1.5 parts glycine chelated iron, and 1.6 parts yeast selenium as a functional trace element combination, supplemented with a carrier of defatted rice bran and zeolite powder in a 3:1 ratio, to a total of 100 parts.

[0110] Table 1: Total micronutrient content (mg / kg, dry matter basis) in total mixed rations (TMR) during the early fattening stage

[0111]

[0112] Mid-stage fattening (concentrate to roughage ratio 50:50, TMR additive composition added at 1.0%).

[0113] Table 2: Total micronutrient content (mg / kg, dry matter basis) in total mixed rations (TMR) during mid-finishing stage

[0114]

[0115] Late fattening stage (concentrate to roughage ratio 60:40, TMR additive composition added at 1.0%).

[0116] Table 3: Total micronutrient content (mg / kg, dry matter basis) in total mixed rations (TMR) during the late fattening stage

[0117]

[0118] Note: All trace elements in concentrate supplements and TMR meet the requirements of the "Safety Standards for the Use of Feed Additives" and are safe and compliant.

[0119] Metagenomic shotgun sequencing was used to analyze the structure and functional genes of the rumen microbial community. The results are shown in the table below:

[0120] Table 4. Results of rumen metagenomic analysis:

[0121]

[0122] Mechanism Explanation: Metagenomic data confirm that the coated Saccharomyces cerevisiae culture and Bacillus subtilis in this invention work synergistically—the yeast culture provides growth factors for the fiber-degrading bacteria, promoting their proliferation; Bacillus subtilis consumes oxygen and secretes enzymes, improving the rumen microenvironment. Together, they reshape the rumen microbiota, enhancing the degradation capacity of fiber and starch at the genetic level.

[0123] Rumen metabolomics analysis—to verify the mechanism of "energy metabolism regulation," using LC-MS / MS technology to quantitatively analyze volatile fatty acids and key metabolites in rumen fluid:

[0124] Table 5. Results of rumen metabolomics analysis:

[0125]

[0126] Mechanism explanation: Metabolomics data revealed the metabolic characteristics of "high energy efficiency" - the increased proportion of propionic acid and the decreased acetic acid / propionic acid ratio mean that more feed carbon is converted into glucogenic precursors, providing an energy basis for muscle growth; the decrease in ammonia nitrogen concentration indicates improved nitrogen utilization and enhanced protein deposition efficiency; the increase in branched-chain fatty acids reflects active protein degradation and synthesis metabolism.

[0127] Serum metabolomics and hormoneomics analysis—verifying the mechanisms of "endocrine regulation" and "antioxidation":

[0128] Table 6. Results of serum metabolomics and hormoneomics analysis

[0129]

[0130] Mechanism explanation: Serum multi-omics analysis showed that coated guanidinoacetic acid significantly increased serum creatine levels, providing energy reserves for muscle tissue and promoting protein synthesis; yeast selenium and zinc methionine synergistically enhanced antioxidant enzyme activity and reduced oxidative stress; the probiotic combination (substantiium + Bacillus licheniformis) improved gut health, reduced stress hormone levels, and created a favorable internal environment for efficient fattening.

[0131] Muscle tissue proteomics and metabolomics analysis—verifying the mechanism of "muscle building and tenderizing":

[0132] (1) Muscle tissue proteomics analysis: TMT labeling quantitative proteomics technology was used to analyze the longissimus dorsi muscle sample.

[0133] Table 7 Results of muscle tissue proteomics analysis:

[0134]

[0135] (2) Muscle metabolomics analysis:

[0136] Table 8. Results of muscle metabolomics analysis:

[0137]

[0138] Mechanism Elucidation: Proteomics and metabolomics data revealed the synergistic mechanism of "muscle building and tender meat"—guanidinoacetic acid coating provides energy for muscle protein synthesis by increasing creatine and phosphocreatine levels, upregulating the expression of contractile proteins such as myosin and actin, thus achieving muscle hyperplasia; rumen-protected complex proteases (acidic + neutral proteases) target and degrade collagen in muscle connective tissue, downregulating collagen expression and upregulating matrix metalloproteinase expression, thus achieving meat tenderization. These two mechanisms work synergistically to achieve the dual goals of "muscle building" and "tender meat".

[0139] Multi-omics joint analysis—verifying the "synergistic effect" mechanism:

[0140] Correlation analysis of multi-omics data revealed the synergistic mechanism among the components of this invention:

[0141] Table 9. Results of multi-omics joint analysis:

[0142]

[0143] Mechanism summary:

[0144] Based on the results of multi-omics analysis, the mechanism of action of the additive composition described in this invention can be summarized as a "four-level linkage, network regulation" model:

[0145] (1) Rumen dimension (upstream - energy supply): Coated Saccharomyces cerevisiae culture promotes the proliferation of fiber-degrading bacteria, and Bacillus subtilis improves the rumen microenvironment. The two work together to improve the efficiency of fiber and starch degradation and increase the production of volatile fatty acids, especially the proportion of propionic acid, which provides an energy basis for subsequent muscle growth.

[0146] (2) Intestinal dimension (midstream - health protection): Bacillus subtilis and Bacillus licheniformis form a triple effect of "oxygen consumption-enzyme production-harm inhibition", maintain the balance of intestinal microecology, promote butyrate production to repair the intestinal barrier, reduce stress hormone levels, and create a good internal environment for efficient fattening.

[0147] (3) Blood circulation dimension (midstream - nutrient transport): Organic trace element complexes (zinc methionine, iron glycine, selenium yeast) are absorbed efficiently in chelated form, which improves the activity of antioxidant enzymes, removes free radicals, and protects cell health; coated guanidinoacetic acid enters the circulation in the form of creatine, providing energy reserves for muscle tissue.

[0148] (4) Muscle tissue dimension (downstream - effect presentation): Coating with guanidinoacetic acid increases creatine and phosphocreatine levels, providing energy for protein synthesis, upregulating myofibril protein expression, and achieving muscle hyperplasia (muscle building); Rumen complex protease targets and degrades collagen, downregulates collagen expression and upregulates matrix metalloproteinase expression, and achieves meat tenderization (tender meat).

[0149] This multi-omics validation system systematically elucidates the scientific connotation of the technical solution of this invention from the perspective of the complete information flow from genes (metagenomics) to metabolites (metabolics) to proteins (proteomics), and confirms that the synergistic effect between the components is not a simple superposition, but is achieved through network regulation of multiple targets and pathways.

[0150] Elucidation of the synergistic mechanism of active ingredients based on network pharmacology:

[0151] To elucidate the synergistic mechanism of multiple active ingredients in the additive composition of this invention at the molecular network level, network pharmacology technology was employed, combined with the traditional Chinese medicine theory of "principal, assistant, adjuvant, and guide" formulation and systems biology methods, to construct an interaction network of "active ingredients-core targets-key pathways," revealing the scientific connotation of the synergistic effect of the composition on "rumen function remodeling" and "body metabolic regulation" from a holistic perspective.

[0152] Network pharmacology analysis of the additive composition described in this invention identified 32 potential active ingredients / metabolites, among which representative core active ingredients / metabolites are shown in the table below:

[0153] Table 10 Representative Core Active Ingredients / Metabolites:

[0154]

[0155] Core target prediction and PPI network construction:

[0156] Through cross-referencing across multiple databases, a total of 312 active ingredient-related targets were predicted, including 187 functional targets related to "muscle growth," "meat tenderness," "antioxidant," and "gut health." The intersection of these targets yielded 96 core targets. A protein-protein interaction (PPI) network was constructed, and topology analysis was used to screen for the top 15 core targets based on their degree values.

[0157] Table 11 Core targets identified through topology analysis:

[0158]

[0159] PPI network analysis conclusions: The core target network exhibits a clear "hub-modular" structure:

[0160] mTOR, IGF1R, and AKT1 constitute the muscle anabolic metabolism center (regulated by guanidinoacetic acid).

[0161] SOD1, GPX1, and CAT constitute an antioxidant defense module (regulated by zinc and selenium).

[0162] TNF, IL6, and TLR4 constitute the inflammation and immune regulation module (regulated by Bacillus and yeast).

[0163] MMP9 and COL1A1 constitute the extracellular matrix remodeling module (regulated by proteases).

[0164] OCLN stands for Intestinal Barrier Module (regulated by Bacillus).

[0165] This modular structure provides the molecular-level network foundation for the "multi-channel synergy" of this invention.

[0166] Verification experiment:

[0167] 1. A composition of fattening feed additive for old-fashioned cattle, the formulation of which follows the scientific principle of "three-level linkage and network regulation". The specific composition is as follows, based on 100 parts by weight:

[0168] Table 12. Additive Compositions for Fattening Cattle Feed (using old-fashioned feed):

[0169]

[0170] Raw material preparation: Coated Saccharomyces cerevisiae culture: The Saccharomyces cerevisiae culture raw material is placed in a fluidized bed coating machine, and a mixture of hydrogenated vegetable oil and ethyl cellulose (mass ratio 3:1) is used as the coating material. The coating is carried out using a bottom spray process. The inlet air temperature is 45-50℃, the spray rate of the coating solution is 1.5-2.0 mL / min, the coating weight gain is 20-25%, and the coating thickness is 20-50 μm. After coating, it is passed through a 60-mesh sieve for later use.

[0171] Rumen-protected compound enzyme preparation: Weigh acidic protease, neutral protease, and cellulase according to the specified ratio, and premix them in a V-type mixer for 10 minutes. Place the mixed enzyme powder in a fluidized bed coating machine for double-layer coating: the first layer uses pH-sensitive polyacrylic acid resin II (8% ethanol solution) for coating, with an inlet air temperature of 35-40℃, resulting in a coating weight gain of 15-20%; the second layer uses hydrogenated vegetable oil for coating, with an inlet air temperature of 40-45℃, resulting in a coating weight gain of 10-15%. After coating, pass through a 40-mesh sieve for later use.

[0172] Guanidinoacetic acid: Guanidinoacetic acid raw material is treated with the same double-layer coating process as enzyme preparation, with a coating weight gain of 25-30%, and then passed through a 40-mesh sieve for later use.

[0173] Other raw materials: Bacillus subtilis, Bacillus licheniformis, methionine hydroxy analog chelated zinc, glycine chelated iron, yeast selenium, defatted rice bran, zeolite powder, etc. are all commercially available products and should be sieved through a 60-mesh sieve before use.

[0174] A step-by-step mixing process is used to avoid interference between active ingredients.

[0175] Probiotic premix: Coated Saccharomyces cerevisiae culture, Bacillus subtilis, Bacillus licheniformis and 10 parts defatted rice bran are put into a small V-type mixer and mixed for 10 minutes to obtain premix A.

[0176] Enzyme premixing: Add the rumen-protected compound enzyme preparation and 10 parts of zeolite powder into a small V-type mixer and mix for 10 minutes to obtain premix B.

[0177] Trace element premix: Methionine hydroxy analog chelated zinc, glycine chelated iron, yeast selenium and 5 parts defatted rice bran were put into a small V-type mixer and mixed for 15 minutes to obtain premix C.

[0178] Overall mixing: Add premixes A, B, C, coated guanidinoacetic acid and the remaining carrier (approximately 28.6 parts defatted rice bran and zeolite powder mixture) to the main mixer (such as a double helical cone mixer) and mix for 15-20 minutes until the coefficient of variation of the mixing uniformity is ≤5%.

[0179] Packaging: The well-mixed additive premix is ​​packaged in 25 kg bags into aluminum foil composite bags, vacuum-sealed or nitrogen-filled for protection, and stored in a cool, dry, and light-protected environment. The product has a shelf life of 12 months.

[0180] Product quality standards:

[0181] The additive composition prepared in this embodiment should meet the following requirements: appearance as a grayish-brown to brownish-brown powder, without lumps; moisture content ≤10.0%; particle size (passing through a 40-mesh sieve) ≥95%; coefficient of variation for mixing uniformity ≤5%; and viable counts of both Bacillus subtilis and Bacillus licheniformis ≥4.0×10⁻⁶. 7 CFU / g; acidic protease activity ≥2.25×10 4 U / g, neutral protease activity ≥1.75×10 4 U / g, cellulase activity ≥4.0×10³ U / g; coated guanidinoacetic acid content 95-105 g / kg (of which pure guanidinoacetic acid 70-80 g / kg); zinc content 6.0-7.2 g / kg; iron content 2.0-4.0 g / kg; selenium content 30-34 mg / kg.

[0182] 2. Rumen-crossing release rate verification test:

[0183] 2.1 Experimental Objective:

[0184] The study verified the rumen-protective effect and small intestinal release characteristics of the double-layer coating technology of this invention on acidic proteases, neutral proteases, and guanidinoacetic acid.

[0185] 2.2 Test methods:

[0186] The in vitro rumen culture method (refer to Menke et al. 1979) and the small intestine digestion method (refer to Boisen et al. 1997) were used.

[0187] Experimental groups: Group A (uncoated), Group B (single-layer enteric coating), Group C (double-layer coating of the present invention).

[0188] Measurement indicators: residual rate in rumen fluid (measured at 2, 4, 6, 8, 12, and 24 h of culture) and release rate in small intestinal simulated fluid (2 h of culture at pH 6.8).

[0189] Table 13 Experimental Results:

[0190]

[0191] 2.4 Conclusion: The double-layer coating technology of this invention can achieve a residual rate of more than 85% of the active ingredients in the rumen after 24 hours and a release rate of more than 90% in the small intestine after 2 hours, which is significantly better than the uncoated and single-layer coated groups (P<0.01), and achieves the precise delivery goal of "rumen protection and small intestine release".

[0192] 3. Fattening Experiment:

[0193] 3.1 Experimental Design: Forty culled Simmental cows (old cattle) of good health, similar weight (approximately 450±25 kg), and age ≥35 months were selected and randomly divided into a control group (basal diet) and an experimental group (basal diet + 1.0% of the additive from Example 1), with 20 cows in each group. The experiment lasted for 130 days (7 days for the pre-trial period and 123 days for the formal trial). All cattle were housed in individual pens, fed twice daily, and had free access to food and water.

[0194] 3.2 Three-stage fattening method: Early fattening stage (1-15 days): concentrate to roughage ratio 40:60, TMR additive addition amount 0.5%.

[0195] Mid-fattening stage (16-60 days): concentrate to roughage ratio 50:50, TMR additive addition 1.0%.

[0196] Late fattening stage (61-130 days): concentrate to roughage ratio 60:40, TMR additive addition 1.0%.

[0197] The compound premixed feed is added to the concentrate supplement at a rate of 1.0% throughout the entire process.

[0198] Table 14 Three-stage formulation of concentrate supplement (air-dried basis, %):

[0199]

[0200] Example of Total Mixed Ration (TMR) composition (mid-finishing stage);

[0201] Taking mid-fattening stage (50:50 concentrate to roughage ratio) as an example, the composition of each ton of TMR (dry matter basis) is as follows:

[0202] Corn stalk silage (dry matter): 500 kg;

[0203] Concentrated feed supplement (containing 1% compound premix): 500 kg;

[0204] Converted to fresh sample (based on 65% moisture content of silage):

[0205] Corn stalk silage (fresh sample): Approximately 1429 kg;

[0206] Concentrated feed supplement (air-dried): 500 kg;

[0207] Total fresh TMR sample volume: approximately 1929 kg.

[0208] 3.3 Measurement Results and Analysis:

[0209] Table 15 Growth Performance Indicators of Old-Style Cattle:

[0210]

[0211] Table 16 Slaughter performance and meat quality indicators of old-fashioned cattle:

[0212]

[0213] Table 17 Biochemical indicators of old bovine serum:

[0214]

[0215] As shown in the table above, growth performance: the average daily weight gain (ADG) of the experimental group was 1.45±0.15 kg / d, which was 22.8% higher than that of the control group (1.18±0.11 kg / d) (P<0.01); the feed conversion ratio (F / G) was 17.1% lower (P<0.01).

[0216] Slaughter performance and meat quality: The eye muscle area of ​​the experimental group increased by 17.6% (P<0.01); shear force decreased by 29.6% (P<0.01); the proportion of heat-soluble collagen increased by 70.8% (P<0.001); and the redness of meat color (a* value) increased by 18.3% (P<0.01).

[0217] Serum biochemical indicators: The total antioxidant capacity (T-AOC) of the experimental group increased by 65.4% (P<0.001), IGF-1 level increased by 48.0% (P<0.01), serum creatine level increased by 83.2% (P<0.001), and cortisol level decreased by 27.9% (P<0.01).

[0218] Safety: All cattle were in good health during the trial, with no adverse reactions, and the levels of all trace elements met national safety standards.

[0219] 4. Quantitative verification experiment of component synergistic effect:

[0220] The central composite design method was used to perform response surface optimization analysis on the five major components.

[0221] Table 18 Factors and Levels:

[0222]

[0223] Response indicators: daily weight gain (Y1); shear force (Y2); eye muscle area (Y3).

[0224] Experimental animals: Breed: Simmental culled cows (old cattle); Age: 35 months and above;

[0225] Weight: 450 ± 25 kg; Health status: No history of disease, normal appetite, good spirits; Quantity: 150 (50 groups × 3 individuals / group, 3 replicates per group).

[0226] The basal diet consisted of corn silage and concentrate supplements, with a uniform concentrate-to-roughage ratio of 50:50 (dry matter basis) to eliminate the impact of diet differences on the results.

[0227] Feeding period: Pre-trial period: 7 days, all cattle were fed a basic diet to adapt to the environment; Formal trial period: 83 days, cattle were fed according to the additive formula of each experimental group.

[0228] Table 19 Response Surface Regression Analysis and Synergistic Effect Coefficients:

[0229]

[0230] Note: * indicates P<0.05, ** indicates P<0.01.

[0231] The results showed that the regression coefficient of the interaction term between protease and guanidinoacetic acid (X3×X4) on daily weight gain was 0.056 (P<0.01), indicating that the effect of both being present simultaneously was 5.6% higher than the additive effect of using them alone. Further calculation of the synergistic effect index (SI) showed that the SI of shear force was as high as +133.3%, meaning that the effect of protease on improving tenderness was amplified by 1.33 times under the combined action of guanidinoacetic acid, probiotics, and trace elements, confirming the existence of the "break-then-build" temporal synergistic and nonlinear amplification effect of this invention.

[0232] 5. Application trials in large-scale cattle farms:

[0233] Experimental Design: The experiment was conducted from March to August 2025 at a beef cattle farm in Yushu City, Jilin Province. The farm had 600 head of beef cattle and a complete feeding management and data recording system.

[0234] Experimental animals and grouping: Sixty healthy, disease-free, culled Simmental cows (old cattle) of similar weight (approximately 450±25 kg) and age over 35 months were selected and randomly divided into two groups of 30 each. The experiment adopted a single-factor completely randomized design, with three replicates per group and 10 cows per replicate.

[0235] Table 20 Comparative Experiments:

[0236]

[0237] Experimental Period and Feeding Management: The experimental period lasted 130 days, including a 7-day pre-trial period and a 123-day formal trial period. During the formal trial period, feeding was strictly carried out according to a three-stage fattening program: Early fattening stage (days 1-15): concentrate-to-roughage ratio 40:60, with 0.5% of the additive composition added to the total mixed ration (TMR); Mid-fattening stage (days 16-60): concentrate-to-roughage ratio 50:50, with 1.0% of the additive composition added to the TMR; Late fattening stage (days 61-130): concentrate-to-roughage ratio 60:40, with 1.0% of the additive composition added to the TMR.

[0238] All cattle are kept in separate pens with free access to feed and water. They are fed twice daily (06:00 and 16:00) using a total mixed ration (TMR) system. Immunization, deworming, and health maintenance are carried out according to the farm's standard management procedures, and the cattle's health status is observed daily.

[0239] Measurement indicators and methods:

[0240] Growth performance: Each cow was weighed 12 hours after fasting at the beginning and end of the experiment. The initial and final weights of each cow were recorded, and the average daily gain (ADG) was calculated. The amount of feed fed and the amount of feed left over in each pen were recorded daily, and the average daily feed intake (ADFI) and feed conversion ratio (F / G) were calculated.

[0241] Slaughter performance: After the experiment, 15 cattle were randomly selected from each group (5 cattle per pen) for slaughter testing: live weight before slaughter, carcass weight, and slaughter rate (carcass weight / live weight before slaughter × 100%). The eye muscle between the 12th and 13th ribs was taken and the eye muscle area was measured using an eye muscle area grid plate. A 500g sample of the longissimus dorsi muscle was taken for meat quality analysis.

[0242] Meat quality indicators: Intramuscular fat content: determined by Soxhlet extraction according to GB 5009.6-2016 "Determination of fat in food"; Shear force: the central part of the longissimus dorsi muscle was taken, steamed according to standard method, and the shear force value was measured by a muscle tenderness meter; Meat color: the brightness (L), redness (a), and yellowness (b*) of the fresh cut surface of the longissimus dorsi muscle were measured by a colorimeter; Collagen content: the proportion of total collagen and heat-soluble collagen was determined by the hydroxyproline method.

[0243] Serum biochemical indicators: On the last day of the experiment, blood was collected from the jugular vein of 20 cattle (6-7 cattle per pen) in each group, and serum was separated to determine: total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), malondialdehyde (MDA), insulin-like growth factor-1 (IGF-1), creatine, and cortisol.

[0244] Statistical analysis: Independent samples t-tests (comparisons between two groups) were performed using SPSS 26.0 software. Results are expressed as mean ± standard deviation. P < 0.05 was considered statistically significant, and P < 0.01 was considered extremely statistically significant.

[0245] Economic benefit analysis: Calculate the net gain per head based on weight gain, feed costs, additive costs, etc., and extrapolate the annual economic benefits of the entire farm.

[0246] Table 21 Effects of additive compositions on the growth performance of older cattle (large-scale cattle farm trial):

[0247]

[0248] Under large-scale farming conditions, the experimental group showed an average daily weight gain of 22.7% higher than the control group (P<0.01), a feed conversion ratio of 16.9% lower (P<0.01), and no significant difference in feed intake (P>0.05). These results are highly consistent with the small-group experiment, indicating that the additive composition of this invention can consistently exert its effects under different farming scales.

[0249] Table 22 Effects of additive compositions on slaughter performance and meat quality of aged cattle (large-scale cattle farm trial):

[0250]

[0251] Carcass quality: The eye muscle area of ​​the experimental group increased by 18.2% (P<0.01) and the dressing percentage increased by 4.1% (P<0.01), indicating that the present invention significantly improved the carcass composition.

[0252] Meat tenderness: The shear force in the experimental group decreased by 29.9% (P<0.01), the collagen content decreased by 16.2%, and the proportion of heat-soluble collagen increased by 75.4%, confirming that the rumen complex protease effectively degraded cross-linked collagen.

[0253] Flesh color: The redness of the flesh color (a*) in the experimental group was significantly increased by 18.8% (P<0.01), and the flesh color was bright red, which is in line with consumer preferences.

[0254] Table 23 Effects of Additive Compositions on Biochemical Indicators of Old-Film Bovine Serum (Large-Scale Cattle Farm Trial):

[0255]

[0256] As shown in the table, under large-scale farming conditions, the antioxidant capacity, IGF-1, and creatine levels in the experimental group were significantly higher than those in the control group (P<0.01), while the stress hormone cortisol was significantly reduced.

[0257] Table 24: Incidence rate control during the trial:

[0258]

[0259] As shown in the table, the incidence of diarrhea, respiratory diseases, and hoof diseases in the experimental group was lower than that in the control group, indicating that the additive composition of the present invention helps to enhance the disease resistance and hoof health of older cattle. No adverse reactions related to the additive were observed in any of the cattle.

[0260] Based on the market price of live cattle at 30 yuan / kg, feed cost of 3.0 yuan / kg, and additive cost of 20 yuan / kg (approximately 2 yuan per head per day), a cattle farm with an annual output of 1,000 head could generate an additional annual economic benefit of 958,000 yuan. If the premium resulting from improved meat quality (Wabyssal beef commands a higher price) is considered, the benefits would be even more significant.

[0261] 6. Comparative experiment on the improvement effects on the three core problems of old films:

[0262] To verify the effectiveness of this invention in improving rumen function decline, muscle coarsening, and difficulty in fat deposition in older cattle, a control group (corn silage + concentrate supplement), a prior art group (probiotics + inorganic salts), a prior art group (single enzyme), a prior art group (single guanidinoacetic acid), and this invention group were set up, with 12 cattle in each group and an experimental period of 90 days. All cattle were housed in individual pens, fed twice daily, and had free access to feed and water. The pre-trial period was 7 days, and the formal trial period was 83 days. The concentrate-to-roughage ratio was uniformly adjusted to 50:50 in all groups (simulating the mid-finishing stage) to ensure comparability.

[0263] Sample collection and index determination:

[0264] Rumen fluid collection and analysis: For three consecutive days before the end of the experiment, rumen fluid was collected orally 3 hours after morning feeding, mixed, and aliquoted. Volatile fatty acids: the concentrations and ratios of acetic acid, propionic acid, and butyric acid were determined by gas chromatography. Ammonia nitrogen concentration: determined by colorimetric method. Fiber degradation rate: the crude fiber degradation rate was determined using the nylon bag method over 72 hours (6 animals were randomly selected from each group).

[0265] Blood Collection and Analysis: Blood was collected from the jugular vein before morning feeding on the last day of the experiment, and serum was separated. Hormones and metabolites: Insulin-like growth factor-1 (IGF-1), growth hormone (GH), and creatine were measured by radioimmunoassay. Antioxidant indicators: Total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA) were measured using a kit.

[0266] Slaughter and muscle tissue analysis: After the experiment, 6 cattle were randomly selected from each group for slaughter, and samples of the longissimus dorsi muscle were taken: intramuscular fat content: Soxhlet extraction method; shear force: measured by tenderness tester; collagen content: determined by hydroxyproline method for total collagen and the proportion of heat-soluble collagen (reflecting the degree of cross-linking); eye muscle area: measured between the 12th and 13th ribs.

[0267] Data statistics: One-way ANOVA was performed using SPSS 26.0, and Duncan's method was used for multiple comparisons. Results are expressed as mean ± standard deviation, and P < 0.05 was considered statistically significant.

[0268] Results and Analysis:

[0269] Table 25 Effects of different treatments on rumen fermentation parameters and fiber degradation rate of aged bovine flakes:

[0270]

[0271] Note: Different letters in the same row's shoulder mark indicate significant differences (P<0.05), the same applies below.

[0272] As shown in the upper right table, the total volatile fatty acids in the present invention group were significantly higher than those in other groups (P<0.05), 32.6% higher than the control group, and 17.1% higher than the prior art group (probiotics + inorganic salts), indicating that the present invention can maximize the efficiency of rumen fermentation.

[0273] Propionic acid ratio: The propionic acid ratio in the group of this invention reached 25.6%, which was significantly higher than that in other groups (P<0.05). Propionic acid is a glucogenin precursor and provides an energy basis for muscle growth.

[0274] Crude fiber degradation rate: The fiber degradation rate of the present invention group reached 56.3%, which is 34.7% higher than that of the control group, 16.1% higher than that of the prior art group (probiotics + inorganic salts), and 21.9% higher than that of the prior art group (single enzyme), indicating that the synergistic effect of "Saccharomyces cerevisiae culture + Bacillus subtilis" is better than that of single probiotics or single enzyme preparations.

[0275] Ammonia nitrogen concentration: The ammonia nitrogen concentration in the group of this invention was the lowest, indicating that the microbial protein synthesis efficiency was high and the nitrogen utilization rate was improved.

[0276] Conclusion: This invention significantly improves rumen function through the synergistic effect of coated Saccharomyces cerevisiae culture and Bacillus subtilis in promoting fermentation and energy supply, and its effect is superior to any single existing technology solution.

[0277] 7. Stability test of additive composition:

[0278] Three batches of the prepared additive composition were stored at room temperature (25℃±2℃, RH60%±5%) for 12 months and at accelerated temperature (40℃±2℃, RH75%±5%) for 6 months, respectively, and samples were taken periodically for testing.

[0279] Table 26 Results of stability tests on additive compositions (stored at room temperature):

[0280]

[0281] The results showed that after 12 months of storage at room temperature, the number of live probiotics was retained at >90%, the enzyme activity retention rate was >93%, and the guanidinoacetic acid content retention rate was >97%, indicating that the product has good stability and is suitable for commercial production and storage.

[0282] 8. Comparative experiment of the effects of the present invention and the closest prior art:

[0283] Experimental Design: To accurately compare the differences between the present invention and the closest prior art (D1 antibiotic, D2 common complex enzyme, D3 monoguanidoacetic acid), a control group, prior art D1 group, prior art D2 group, prior art D3 group and the present invention group were set up, with 12 cattle in each group. The experimental period was 90 days, and the cattle were fed according to the method described in Example 2.

[0284] Table 27 Comparative Experiments:

[0285]

[0286] Feeding management: All treatment groups were fed twice daily (08:00 and 16:00) with free access to feed and water, ensuring that each cow received an adequate total mixed ration. Feed intake was recorded daily, and the average daily feed intake was calculated.

[0287] Table 28 Experimental Results:

[0288]

[0289] The results showed that the ADG of the present invention group (1.45 kg / d) was significantly higher than that of groups D1 (1.30), D2 (1.22), and D3 (1.35). The shear force of the present invention group (48.2 N) was much lower than that of group D2 (59.8 N), while groups D1 and D3 showed no improvement in tenderness. When the theoretical effects of D2 and D3 were added together, the actual ADG and eye muscle area of ​​the present invention were both higher than the theoretical sum, while the shear force was much lower than the theoretical sum. This confirms that the present invention is not a simple combination, but rather achieves a breakthrough effect beyond simple superposition through a temporal synergy of "degradation followed by synthesis".

Claims

1. A composition of feed additive for fattening cattle, characterized in that, Including the following parts by weight of raw materials: Rumen function modifier: 13-20 parts; Intestinal repair agent: 4-10 parts; Rumen-protected complex enzyme preparation: 10-20 parts; Muscle-building and growth-promoting combination: 5-10 servings; Functional trace element combination: 5.5-7.3 parts; Carrier: 32.7-62.5 copies; The rumen function regulator includes coated Saccharomyces cerevisiae culture and Bacillus subtilis; The intestinal repair agent is Bacillus licheniformis; The rumen-protected complex enzyme preparation includes acidic protease, neutral protease, and cellulase; The muscle-building and growth-promoting combination includes guanidinoacetic acid coating; The functional trace element combination includes three elements: zinc, iron, and selenium. The carrier is one or more of defatted rice bran, zeolite powder, or maifanite powder.

2. The additive composition according to claim 1, characterized in that, The rumen function regulator contains 10-15 parts of coated Saccharomyces cerevisiae culture and 3-5 parts of Bacillus subtilis. The coated Saccharomyces cerevisiae culture is made by coating the surface of yeast culture particles with a composite coating material of hydrogenated vegetable oil and ethyl cellulose, with a coating thickness of 20-50 μm.

3. The additive composition according to claim 1, characterized in that, The rumen-protected complex enzyme preparation comprises the following enzymes by weight percentage: 40-50% acidic protease, 30-40% neutral protease, and 20-30% cellulase; the rumen-protected complex enzyme preparation is treated with a double-layer coating technology, wherein the double-layer coating is: an inner layer of pH-sensitive polyacrylic acid resin coating and an outer layer of hydrogenated vegetable oil coating.

4. The additive composition according to claim 1, characterized in that, The functional trace elements include: 3-3.6 parts of methionine hydroxy analog chelated zinc, 1-2 parts of glycine chelated iron, and 1.5-1.7 parts of yeast selenium.

5. The additive composition according to claim 1, characterized in that, The carrier is a mixture of defatted rice bran and zeolite powder in a weight ratio of 3:

1.

6. A method for fattening old-fashioned cattle using the old-fashioned cattle fattening feed additive composition according to any one of claims 1-5, characterized in that, The aforementioned old-fleshed cattle fattening feed additive composition is added to the total mixed ration at a mass ratio of 0.5%-1.0%. (1) Early fattening stage: 1-15 days, the additive composition in the total mixed ration is added at a rate of 0.5%; (2) Mid-finishing stage: 16-60 days, the additive composition in the total mixed ration is added at a rate of 1.0%; (3) Late fattening period: 61-130 days, the amount of additive composition added to the total mixed diet is 1.0%.

7. The method for fattening old cattle according to claim 6, characterized in that, The total mixed ration comprises roughage and concentrate, wherein the concentrate comprises 1% of the additive composition.

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