Composition for rapidly relieving ruminant stress and promoting growth and application thereof

Abstract

The invention relates to the technical field of ruminant stress resistance, in particular to a composition for rapidly relieving ruminant stress and promoting growth and application of the composition. The composition comprises gamma-aminobutyric acid and a preparation containing bacillus subtilis and lactobacillus acidophilus. The composition can be used for preparing products for resisting ruminant stress, improving growth performance and promoting growth. The bacillus subtilis and the lactobacillus acidophilus are used as a symbiotic bacterium system and are combined with the gamma-aminobutyric acid, so that rapid colonization of rumen bacteria is realized, oxidative stress and inflammatory response of ruminants are relieved, and the effects of rapidly relieving stress of the ruminants and promoting growth are achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of anti-stress for ruminants, and in particular to a composition for rapidly relieving stress and promoting growth of ruminants and an application thereof. Background Art

[0002] Huyang (Huyang) sheep have become a preferred breed for intensive sheep farming due to their rapid early growth, high reproductive capacity, and delicious meat. However, intensive farming also significantly increases the potential risk of disease for Huyang. However, with the widespread adoption of intensive farming practices and the intensification of global warming, heat stress has become a key environmental stressor that restricts the production and health of Huyang sheep. The negative impact of heat stress on Huyang sheep severely restricts their large-scale development.

[0003] When Huyang are exposed to high temperatures, the body's nonspecific physiological response to the heat produces a stress response. Due to underdeveloped or absent sweat glands, animals struggle to dissipate heat effectively in hot environments, leading to elevated body temperature, rapid breathing, and increased heart rate. Currently, heat stress has become a bottleneck that urgently needs to be addressed in animal production, especially in southern regions where environmental stress can last up to six months. Huyang have become a preferred breed for intensive sheep farming due to their rapid early growth, high fertility, and delicious meat. However, intensive farming also significantly increases the potential risk of disease in Huyang. High-density housing, in particular, leads to poor ventilation in pens and difficulty regulating temperature and humidity. Especially in summer, heat stress can induce metabolic disorders, oxidative damage, and immunosuppression, leading to decreased growth performance and reduced meat quality.

[0004] Conventional response methods involve physical cooling measures, such as water spraying, increased ventilation, and nutritional adjustments to alleviate stress, such as supplementing nutrients like electrolytes and vitamins. However, these are limited in high-humidity environments and may cause hoof disease or mastitis due to slippery pen floors. In addition, frequent physical cooling measures increase energy consumption and raise breeding costs, making them difficult to apply on a large scale in open pastures. While nutritional adjustments can partially alleviate symptoms, long-term use can easily lead to antibiotic dependence or residue problems. Currently, there are no effective methods for ruminants such as cattle and sheep to combat heat stress during the fattening stage. Summary of the Invention

[0005] The purpose of the present invention is to provide a composition for rapidly relieving stress of ruminants and promoting growth. The composition can be used to prepare products for resisting stress of ruminants, improving growth performance and promoting growth.

[0006] The technical solution adopted by the present invention to solve its technical problem is:

[0007] The invention discloses an application of gamma-aminobutyric acid (GABA) in the preparation of a product for resisting stress and promoting growth of ruminants.

[0008] The invention discloses an application of gamma-aminobutyric acid in the preparation of a product for regulating the neuroendocrine system of ruminants, inhibiting oxidative stress and / or optimizing lipid metabolism.

[0009] A composition for rapidly relieving stress and promoting growth in ruminants, comprising gamma-aminobutyric acid and a preparation containing Bacillus subtilis and Lactobacillus acidophilus. The present invention uses a commercially available bacterial agent for testing the preparation containing Bacillus subtilis and Lactobacillus acidophilus, which is hereinafter referred to as FAM. The combined use of gamma-aminobutyric acid, Bacillus subtilis, and Lactobacillus acidophilus has outstanding performance in reducing COR and increasing IgG. Moreover, Bacillus subtilis and Lactobacillus acidophilus, as a symbiotic bacterial system, combined with gamma-aminobutyric acid, achieve rapid colonization of rumen bacteria, which is beneficial for reducing oxidative stress and inflammatory responses in ruminants, thereby achieving the effect of rapidly relieving stress and promoting growth in ruminants.

[0010] Preferably, the preparation containing Bacillus subtilis and Lactobacillus acidophilus is in powder form and consists of Bacillus subtilis and Lactobacillus acidophilus, and contains Bacillus subtilis ≥ 1.0×10 6 CFU / g, Lactobacillus acidophilus ≥1.0×10 6 CFU / g, the carrier is one or more of vermiculite and calcium carbonate; the gamma-aminobutyric acid is in a coated form, and the coating material is one or more of silicon dioxide, sodium carboxymethyl cellulose, and palm oil.

[0011] A use of the composition of the present invention in preparing a product for rapidly relieving stress and promoting growth of ruminants.

[0012] Preferably, the application is specifically the application in resisting stress and promoting growth during the fattening stage of Hu sheep.

[0013] Preferably, the dosage of γ-aminobutyric acid is 0.2±0.05 g / day per ruminant, the dosage of the preparation containing Bacillus subtilis and Lactobacillus acidophilus is 0.1 to 0.5 g / day per ruminant, and the preparation containing Bacillus subtilis and Lactobacillus acidophilus contains Bacillus subtilis ≥1.0×10 6 CFU / g, Lactobacillus acidophilus ≥1.0×10 6 CFU / g.

[0014] Preferably, the ruminants in the above application include cattle and sheep, especially Hu sheep.

[0015] A functional feed containing the composition of the present invention. Preferably, the feed further comprises a basic diet component, wherein the basic diet component, calculated as 100% by mass, comprises: 31.6% corn, 16.0% soybean meal, 6.0% peanut meal, 6.0% wheat bran, 15.0% bean dregs, 9.0% peanut vines, 6.0% waste flour, 5.0% rice bran, 0.60% calcium hydrogen phosphate, 0.60% baking soda, 0.20% salt, and 4.00% premix;

[0016] Each kilogram of premix includes 1500kIU vitamin A, 350kIU vitamin D, 4000mg vitamin E, 300-420mg sodium selenite, 200mg biotin, 13000mg zinc, 35000mg copper, 5500mg manganese, 64mg selenium, 160-240mg iodine, and 48-72mg cobalt.

[0017] The present invention selects targeted probiotics according to the digestive physiological characteristics of Hu sheep, optimizes the formula, and combines it with the special component gamma-aminobutyric acid. Experiments have proven its palatability and health function for the diet, thereby improving the feeding and growth conditions of Hu sheep.

[0018] The beneficial effects of the present invention are:

[0019] The present invention constructs a control group (basal diet), a GABA group (basal diet + 0.1% GABA), a FAM group (basal diet + 0.1% FAM) and a combined group (basal diet + 0.1% GABA + 0.1% FAM). This project intends to adopt four different experimental models. By testing indicators such as growth, slaughter performance, meat quality, jejunum, ileum and blood biochemistry, immunity, etc., the volatile substances in the rumen, microbial flora, and metabolites in the rumen and blood are detected and evaluated.

[0020] The results showed that: (1) in terms of heat stress regulation, GABA and FAM alone could effectively reduce respiratory rate and rectal temperature, and combined treatment showed synergistic effect in the short term, but the long-term application effect fluctuated due to animal adaptability; (2) in terms of growth performance, single use of GABA had the best effect, which could significantly increase tail weight and average weight gain; while FAM could relieve stress, but its direct promotion to weight gain was weak, and the combined treatment did not show synergistic advantages; (3) slaughter performance analysis showed that GABA significantly optimized carcass morphology and increased spleen weight, indicating that it has both Growth promotion and immune regulation potential; (4) In terms of muscle quality, GABA and FAM improved pH and water retention in the early post-slaughter period, but had no significant effect on shear force and meat color stability; (5) Serum biochemical and immune indicators further revealed that GABA reduced cortisol (COR) and heat shock protein 70 (HSP70) levels, maintained metabolic homeostasis by enhancing total antioxidant capacity, inhibiting proinflammatory factors, reducing lipid peroxidation and regulating lipid metabolism; FAM increased immunoglobulins by regulating intestinal microecological balance and enhancing antioxidant capacity, and alleviated oxidative stress and inflammatory response. GABA and FAM treatment alone had significant effects in reducing stress markers (HSP70, COR), inhibiting proinflammatory factors (IL-1β, IL-6) and increasing some immunoglobulins (IgM), while combined treatment was more effective in reducing COR and increasing IgG. (6) In terms of rumen volatilization, microbial flora, and rumen and plasma metabolism, the concentration of GABA ammonia nitrogen was significantly reduced. GABA Bacillus (Bacillota) is a group with a high content in plants. The interaction between neuroactive ligands and receptors involves neurotransmitters such as GABA and glutamate. Heat stress may regulate the stress response of the central nervous system to high temperature by activating or inhibiting specific neurotransmitter receptors. As an inhibitory neurotransmitter, GABA may reduce nerve excitability through this pathway and alleviate the excessive stress response caused by heat stress. The intervention of FAM and GABA may maintain neural homeostasis by regulating such signaling pathways. FAM and GABA may regulate D-amino acid metabolism, ABC transporters and protein digestion and absorption pathways to synergistically optimize energy supply, enhance antioxidant capacity and maintain rumen microbial homeostasis, thereby alleviating heat stress in Hu sheep.

[0021] Studies have confirmed that GABA relieves stress response by regulating the neuroendocrine system, and FAM enhances the body's adaptability through the gut-immune axis. The combined use of the two can improve metabolic disorders, oxidative damage and immunosuppression caused by heat stress at multiple targets. The combined use of GABA and FAM can quickly relieve stress response. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the temperature and humidity index change curve of the sheep house during the experiment. DETAILED DESCRIPTION

[0023] The technical solution of the present invention is further described in detail below through specific examples. It should be understood that the implementation of the present invention is not limited to the following examples, and any form of modification and / or change made to the present invention will fall within the scope of protection of the present invention.

[0024] In the present invention, unless otherwise specified, all parts and percentages are by weight, and the equipment and raw materials used are commercially available or commonly used in the art. The methods in the following examples are all conventional methods in the art unless otherwise specified.

[0025] Unless otherwise specified, the reagents used in the following examples can be purchased from conventional biochemical reagent stores.

[0026] Example Application of an Anti-Ruminant Heat Stress Composition in Anti-Heat Stress of Hu Sheep in Fattening Stage 1.1 Test Materials

[0027] Experimental animals: 64 3-4 month old fattened Hu sheep lambs of similar weight and good health were purchased from Tongxiang Farm in Zhejiang Province. The feeding experiment was conducted at Tongxiang Farm in Zhejiang Province.

[0028] Gamma-aminobutyric acid (GABA), product name Yingjixiao 50, was purchased from Hangzhou Kangdequan Feed Co., Ltd. The GABA was in a coated form, and the carrier and coating materials were silicon dioxide, sodium carboxymethyl cellulose, and palm oil. The GABA content was ≥50.0%, and the moisture content was ≤10.0%.

[0029] The mixed preparation of Bacillus subtilis and Lactobacillus acidophilus was in powder form and the product name was Fu Ai Mei (FAM), which was purchased from Zhejiang Kangwan Dechuan Technology Co., Ltd. The carriers were vermiculite and calcium carbonate. The concentration of Bacillus subtilis was ≥1.0×10 6 CFU / g, Lactobacillus acidophilus ≥1.0×10 6 CFU / g, moisture ≤12.0%.

[0030] The composition and nutritional levels of the basal diet are shown in Table 1.

[0031] Table 1 Basic diet composition and nutritional levels

[0032]

[0033]

[0034] Note: Each kilogram of premix includes 1500kIU vitamin A, 350kIU vitamin D, 4000mg vitamin E, 300-420mg sodium selenite, 200mg biotin, 13000mg zinc, 35000mg copper, 5500mg manganese, 64mg selenium, 160-240mg iodine, and 48-72mg cobalt.

[0035] 1.2 Experimental design and feeding management

[0036] The study selected 64 fattening Hu sheep lambs aged 3-4 months of age, of similar weight and in good health. They were randomly divided into four groups: a control group (Con), a GABA group, a FAM group, and a GABA and FAM combination group. Each group consisted of four replicates, with four lambs per replicate. The experimental groups were given different feed treatments: the control group received only a standard basal diet; the GABA group received a daily supplement of 0.4g of GABA per lamb; the FAM group received a daily supplement of 0.5g of FAM per lamb; and the combined GABA and FAM group received both 0.4g of GABA and 0.5g of FAM per lamb. The entire trial lasted 42 days.

[0037] 1.3 Measurement indicators and methods

[0038] 1.3.1 Determination of heat stress indicators

[0039] (1) Temperature and Humidity Index (THI)

[0040] During the experiment, a standardized environmental monitoring plan was adopted. At three fixed time points every day at 8:00h, 14:00h and 20:00h, environmental parameters were collected using temperature and humidity recorders, and the temperature and humidity index was calculated according to the formula.

[0041] THI=(1.8×T+32)-(0.55-0.0055×RH)×(1.8×T-26)

[0042] Where: T represents temperature and RH represents humidity.

[0043] (2) Respiratory rate

[0044] A systematic random sampling method was used to select 8 test sheep that met the criteria from each group during the three time periods mentioned above every day. The number of fluctuations in the left thoracic area of ​​the test sheep was continuously collected and recorded within 3 minutes, and the average value was taken as the respiratory rate parameter of each group.

[0045] (3) Rectal temperature

[0046] Every Monday between 2:00 PM and 3:00 PM, a veterinary digital rectal thermometer was used. Following the WHO Animal Laboratory Procedure Guidelines, the probe was inserted to a uniform depth of 5 cm and the reading was recorded after 30 seconds of stable contact. Each Hu sheep was measured twice, with an interval of ≥5 minutes. If the difference in the readings exceeded 0.3°C, a third measurement was performed, and the average of the two closest valid values ​​was used for data recording.

[0047] 1.3.2 Determination of growth performance

[0048] At the beginning of the experiment, the initial weight of the Hu sheep was recorded by weighing them; at the end of the experiment, the sheep were weighed before morning feeding as the endpoint weight. Throughout the experiment, the feed amount and remaining feed amount (remaining feed amount was approximately 5% of the feed intake) were accurately recorded for each group. Based on this statistical data, the Hu sheep's dry matter intake (DMI), average daily gain (ADG), and feed-to-weight ratio (FCR) were calculated.

[0049] The dry matter intake (DMI) of experimental animals is calculated by dividing the difference between the daily diet and residual food intake of each group by their total amount. The average daily gain (ADG) is calculated by dividing the total experimental weight (final weight minus initial weight) by the number of experimental days. The feed-to-weight ratio (FCR) is calculated by dividing the dry matter intake by the average daily gain.

[0050] 1.3.3 Serum biochemical indicators

[0051] After the feeding experiment, 8 Hu sheep were randomly selected from each group, and 10 mL of blood was collected from the jugular vein. The serum was separated and dispensed into centrifuge tubes and stored at -20°C. The total protein (TP), albumin (ALB), urea nitrogen (BUN), triglyceride (TG), total cholesterol (TC), glucose (GLU), free fatty acids (NEFA), high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde (MDA), total antioxidant capacity (TAOC), β-hydroxybutyrate (BHB), and lactate dehydrogenase (LDH) in the serum of Hu sheep were determined using a full-automatic biochemical analyzer (Nanjing Detection Biological Co., Ltd.).

[0052] 1.3.4 Serum immune indicators

[0053] Serum was collected from Hu sheep and sent to Hangzhou Jinjinuo Co., Ltd. for determination of haptoglobin (HP), interleukin-6, lactic acid (D-LA), interleukin-1β (IL-1β), creatine kinase (CK), cortisol (COR), C-reactive protein (CRP), growth hormone (GH), heat shock protein 70 (HSP70), immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM), interleukin-10, insulin (Insulin), lipopolysaccharide binding protein (LBP) and tumor necrosis factor-a (TNF-a).

[0054] 1.4 Sample collection and indicator determination

[0055] 1.4.1 Feed sample collection and composition analysis

[0056] During the experiment, 500 grams of feed were collected every 14 days and stored in a refrigerator at -80°C. At the end of the experiment, all collected feed samples were thoroughly mixed for nutrient analysis. Prior to analysis, the feed was thawed at room temperature and then oven-dried at 65°C for 48 hours to completely remove moisture. The dried feed was pulverized and passed through 2 mm and 1 mm sieves. The screened feed samples were analyzed for dry matter, crude protein, crude ash, crude fat, calcium, and phosphorus according to AOAC standard methods. Acid detergent fiber and neutral detergent fiber were analyzed according to the methods of Vansoest et al. Acid-insoluble ash was determined according to the national standard GB / T 23742-2009.

[0057] 1.4.2 Stool collection and index determination

[0058] During the experiment, fecal samples were collected from Hu sheep using the rectal fecal collection method. After all feces were collected from each sheep, they were mixed evenly and stored in a -20°C refrigerator. The dry matter, crude protein, crude fat, acid detergent fiber, neutral detergent fiber, calcium, phosphorus, and acid-insoluble ash contents of the feces were subsequently measured. The nutrient composition of the feces was determined. The apparent digestibility of nutrients in fattened Hu sheep was calculated using the above measurement results: feed nutrient composition (%) = (% of a certain nutrient in feed / % of acid-insoluble ash in feed - % of a certain nutrient in feces / % of acid-insoluble ash in feces) / (% of a certain nutrient in feed / % of acid-insoluble ash in feed) × 100.

[0059] 1.4.3 Blood collection and index determination

[0060] On day 42 of the formal experimental period, before feeding in the morning, 10 ml of jugular venous blood was collected using a sodium heparin vacuum tube and stored at 4°C. The blood was then centrifuged at 3000 rpm for 15 minutes, and the plasma supernatant was collected. The supernatant was divided into several aliquots and stored frozen at -20°C until further use. Serum albumin, total protein, triglycerides, creatinine, glucose, free fatty acids, and total cholesterol were measured using kits (Nanjing Jiancheng Biotechnology Co., Ltd.).

[0061] 1.5 Data Analysis

[0062] The experimental data were preliminarily sorted using Excel 2024 and analyzed using SPSS27.0 software. One-way ANOVA and Duncan's multiple comparison method were used to test the significance of differences between groups. The results were expressed as "mean + standard deviation". P>0.05 indicated that the difference was not significant, P<0.05 indicated that the difference was significant, and P<0.01 indicated that the difference was extremely significant.

[0063] 2 Results and Analysis

[0064] 2.1 Effects of Yingjixiao and Fuaimei on heat stress indicators in fattening Hu sheep

[0065] 2.1.1 Temperature and humidity index change curve during the test

[0066] The temperature and humidity index change curve during the test is as follows: Figure 1 The three curves shown in the figure, including the maximum, average, and minimum daily temperature values, comprehensively reflect the dynamic changes in the temperature and humidity index during the experiment. The results show that the maximum temperature and humidity index during the experiment was around 75, the minimum was around 73, and the average was around 74, indicating that the Huyang were in a state of mild heat stress. The temperature dropped in the last week, and the heat stress was alleviated.

[0067] 2.1.2 Effects of Yingjixiao and Fuaimei on the respiratory rate of fattening Hu sheep

[0068] The effects of stress elimination, Fu Aimi and their combined treatment on the respiratory rate of fattening Hu sheep at different time periods during the experiment are shown in Table 2.

[0069] During the first week of the experiment, the respiratory rates of the three treatment groups were significantly lower than those of the control group (P<0.05). The respiratory rate of the Aimee group was significantly lower than that of the control group (P<0.05), and the respiratory rate of the stress elimination group was even lower (P<0.05). The respiratory rate of the combined group was the lowest, significantly lower than that of the other treatment groups (P<0.05).

[0070] In the second week of the experiment, the respiratory rate of the control group was significantly higher than that of the other three treatment groups (P<0.05), followed by the Fu Ai Mei group (P<0.05), and the combined group was significantly higher than the stress elimination group (all P<0.05). In the third and fourth weeks of the experiment, the respiratory rate of the three treatment groups was significantly lower than that of the control group (P<0.05), while there was no significant difference between the stress elimination group and the Fu Ai Mei group (P<0.05), but it was significantly higher than that of the combined group (P<0.05).

[0071] At the 5th week of the experiment, the respiratory rates of the three treatment groups were significantly lower than those of the control group (P<0.05), while there was no statistically significant difference in respiratory rates among the three treatment groups (P>0.05); at the 6th week of the experiment, the respiratory rates of the control group were significantly higher than those of the three treatment groups (P<0.05), and there was no significant difference in respiratory rate between the Fu Aimi group and the stress elimination group and the combined group (P>0.05).

[0072] The respiratory rate of the control group was the highest throughout the entire experimental period and was significantly higher than that of the other treatment groups at each time period (P < 0.05). The respiratory rate of the treatment groups using GABA or FAM alone was significantly lower than that of the control group (P < 0.05), but the difference between the two groups was small. The respiratory rate of the combined treatment group was further reduced (P < 0.05) and was the lowest among the multiple weeks.

[0073] Table 2 Effects of stress elimination and Fu Aimi on respiratory rate of fattening Hu sheep

[0074]

[0075] Note: Data are expressed as mean + standard error of the mean. No letters or the same letters in the same row indicate no significant difference (P>0.05), and different lowercase letters indicate significant difference (P<0.05). The same applies to the following tables.

[0076] 2.1.3 Effects of Yingjixiao and Fuaimei on rectal temperature of fattening Hu sheep

[0077] Table 3 shows the effects of Yingjixiao, Fu'aimei, and their combination on rectal temperature in fattening Hu sheep at different time points during the experimental period. From the first to the fifth week of the experiment, respiratory rates were significantly lower in all three treatment groups compared with the control group (P < 0.05), with no significant differences among the three treatment groups (P > 0.05). During the sixth week, respiratory rates were significantly lower in the Fu'aimei and combined treatment groups than in the control group (P < 0.05), but no significant differences were found between the two groups (P > 0.05). There were no significant differences between the Yingjixiao and other three treatment groups (P > 0.05). Rectal temperature was highest in the control group throughout the experimental period and remained significantly higher than that of the other treatment groups at all time points (P < 0.05). Temperatures in the groups treated with GABA or FAM alone, as well as in the combined treatment group, were significantly lower than those in the control group (P < 0.05).

[0078] Table 3 Effects of Yingjixiao and Fu Aimi on rectal temperature of fattening Hu sheep

[0079]

[0080] 2.2 Effects of Yingjixiao and Fuaimei on the growth performance of fattening Hu sheep

[0081] During the experimental phase, the effects of the stress-eliminating, fuaimei, and their combined treatments on the growth performance of fattening Hu sheep were evaluated. The results are detailed in Table 4. Initial body weights did not differ significantly among all experimental groups (P>0.05), demonstrating a sound experimental design and balanced baseline conditions. Final body weights were significantly increased in the stress-eliminating and combined treatment groups compared with the control group (P<0.05), while no significant differences were found between the two groups (P>0.05). The fuaimei group weighed significantly less than the stress-eliminating group (P<0.05), but did not differ significantly from the control group or the combined treatment group (P>0.05). Dry matter intake (DMI) was significantly higher in all three treatment groups than in the control group (P<0.05), with the stress-eliminating group achieving the highest DMI and significantly exceeding the combined treatment group (P<0.05). There was no significant difference between the fuaimei and combined treatment groups (P>0.05). In terms of average daily gain (ADG), all three treatment groups were significantly better than the control group (P < 0.05). The ADG of the stress-elimination group was significantly higher than that of the Fu Aimei group and the combined treatment group (P < 0.05), while the difference between the latter two groups was not significant (P > 0.05). In terms of feed conversion ratio, all three treatment groups were significantly lower than the control group (P < 0.05). The stress-elimination group had the lowest feed conversion ratio, which was significantly lower than the Fu Aimei group and the combined treatment group (P < 0.05), while there was no significant difference between the Fu Aimei group and the combined treatment group (P > 0.05).

[0082] Table 4 Effects of stress elimination and Fu Aimi on the growth performance of fattening Hu sheep

[0083]

[0084] 2.3 Effects of Yingjixiao and Fuaimei on the apparent digestibility of fattening Hu sheep

[0085] Table 5 reveals significant differences in nutritional indicators between treatments (P < 0.05). The dry matter content of the GABA-treated stress relief group was significantly lower than that of the control group, while the combined use of Foliumerin (Foliumerin) achieved the highest digestibility. Notably, crude protein content in the stress relief group was significantly lower than in the other groups (P < 0.05). These findings suggest that Foliumerin (Foliumerin) has a positive effect on nutrient digestion, while using stress relief alone may negatively impact nutrient absorption.

[0086] Table 5 Effects of Yingjixiao and Fu Aimi on the apparent digestibility of fattening Hu sheep

[0087]

[0088]

[0089] 2.4 Effects of Yingjixiao and Fuaimei on serum biochemical parameters of fattening Hu sheep

[0090] The effects of Yingjixiao, Fu Aimi and their combined treatment on serum biochemical indices of fattening Hu sheep during the experimental period are shown in Table 6.

[0091] Among the basal metabolic and nutritional indicators, total protein (TP) and albumin (ALB) showed significant differences between the groups (P < 0.05). The results showed that the protein content of the three treatment groups was significantly increased compared with the control group (P < 0.05), with the protein content of the stress elimination group being the highest, significantly higher than that of the Fu Ai Mei group (P < 0.05), while the difference between the two groups was not significant (P > 0.05). The serum albumin level was significantly increased in the stress elimination group (P < 0.05), while there was no statistically significant difference between the Mei Mei group and the combined group and the control group (P > 0.05). There was no statistically significant difference in urea nitrogen (BUN) between the two groups (P > 0.05). The antioxidant capacity (T-AOC) of the three treatment groups was significantly higher than that of the control group (P<0.05), but there was no significant difference in antioxidant capacity among the three treatment groups (P>0.05); the SOD activity of the Fu Aimi group was significantly increased compared with the control group (P<0.05), while there was no significant difference between the two groups (P>0.05); the glutathione peroxidase (GSH-PH) activity was significantly higher than that of the control group (P<0.05), while there was no significant difference between the stress elimination group and the control group (P<0.05); the malondialdehyde (MDA) content was significantly reduced under stress stimulation compared with the control group (P<0.05), while there was no significant difference between the Aimi group and the combined group and the control group (P>0.05).

[0092] In terms of tissue damage and cellular metabolism, lactate dehydrogenase (LDH) activity, which can well reflect the extent of tissue damage, was significantly lower in all three treatment groups compared to the control group (P < 0.05). The stress-elimination group had the lowest level, significantly lower than the Fu Aimi group and the combined treatment group (P < 0.05). Regarding glucose metabolism and energy metabolism, neither glucose (GLU) nor β-hydroxybutyrate (BHB) showed significant differences between the groups (P > 0.05).

[0093] Among the lipid metabolism indicators, non-esterified fatty acids (NEFA), as a marker of energy metabolism, were significantly higher in the stress elimination group and the combined group than in the control group (P < 0.05), while there was no significant difference between the Fu Aimi group and the control group (P > 0.05); triglycerides (TG) were significantly reduced in all three treatment groups (P < 0.05), and there was no significant difference among the three groups (P > 0.05); total cholesterol (TC) and high-density lipoprotein cholesterol (HDLC) were significantly increased in the stress elimination group (P < 0.05), and low-density lipoprotein cholesterol (HDLC) showed no significant difference among the groups (P > 0.05).

[0094] The use of Yingjixiao alone has significant effects on enhancing antioxidant capacity, improving lipid metabolism and reducing tissue damage, while Fuaimi has weaker effects on some indicators. Combined use may produce synergistic or antagonistic effects.

[0095] Table 6 Effects of stress elimination and Fu Aimi on serum biochemical parameters of fattening Hu sheep

[0096]

[0097] 2.5 Effects of Yingjixiao and Fuaimei on serum immune indicators of fattening Hu sheep

[0098] The effects of Yingjixiao, Fu Aimi and their combined treatment on the serum immune indicators of fattening Hu sheep during the experimental period are shown in Table 7.

[0099] Among stress-related indicators, decreases in HSP70 and COR were particularly prominent. Heat shock protein (HSP70) expression was significantly decreased in all three treatment groups (P < 0.05). There was no significant difference in the stress elimination group compared with the stress elimination and fulvestrant groups (P < 0.05), while there was no significant difference in the combined group compared with the stress elimination and fulvestrant groups (P > 0.05). Cortisol (COR) levels were significantly decreased in all three treatment groups compared with the control group (P < 0.05), while those in the combined group were significantly decreased (P < 0.05). Regarding tissue damage, creatine kinase (CK) levels were significantly lower in the stress elimination and combined treatment groups than in the control group (P < 0.05), while there was no significant difference in the fulvestrant group compared with the control group (P > 0.05). Lipopolysaccharide binding protein (LBP) levels did not show significant differences between the groups (P > 0.05).

[0100] In terms of immune regulation, there were no significant differences in C-reactive protein (CRP) and conjugated hemoglobin (HP) among the groups (P>0.05); the changes in cytokines IL-1β and IL-6 were significant (P<0.05). Compared with the control group, the three treatment groups were significantly lower than the control group (P<0.05), and the reduction in IL-1β in the stress-elimination group was more significant (P<0.05). However, there were no significant differences in cytokines TNF-α and IL-10 among the groups (P>0.05).

[0101] Among immunoglobulins, the IgG content of the three treatment groups was significantly higher than that of the control group (P<0.05), and the IgG content of the combined treatment group was significantly higher than that of the stress elimination group and the Fu Aimei group (P<0.05); there was no statistical significance in the IgM content between the Fu Aimei group and the control group (P>0.05); there was no statistical significance in the IgA content between the two groups (P>0.05).

[0102] Growth hormone (GH) levels were significantly elevated in both the stress elimination and combined groups (P < 0.05), with the stress elimination group showing significantly higher levels than the combined group (P < 0.05). Regarding metabolism and energy, insulin (INS) levels were significantly elevated in both the Fu Ai Mei and combined groups compared to the control group (P < 0.05), while no significant difference was observed in the stress elimination group (P > 0.05). D-lactic acid levels did not differ significantly between the groups (P > 0.05).

[0103] Treatment with Yingjixiao and Fuaimei alone was effective in reducing stress markers (HSP70, COR), inhibiting pro-inflammatory factors (IL-1β, IL-6) and increasing some immunoglobulins (IgM), while combined treatment was more effective in reducing COR and increasing IgG.

[0104] Table 7 Effects of stress elimination and Fu Aimi on serum immune indicators of fattening Hu sheep

[0105]

[0106]

[0107] 3 Discussions

[0108] 3.1 Effects of Yingjixiao and Fuaimei on heat stress indicators in fattening Hu sheep

[0109] In order to study the effects of GABA and FAM on heat stress in fattening Hu sheep, the present invention established a Hu sheep heat stress model, observed the effects of GABA, FAM and their combined treatment on the respiratory rate and rectal temperature of fattening Hu sheep, and analyzed the effects of the two additives in alleviating heat stress and their dynamic characteristics in regulating the physiological stress state of animals. The temperature and humidity index (THI) is a key indicator for measuring heat stress. The dynamic changes of THI during the experiment were observed and found that its value fluctuated between 73-75, with an average of about 74, indicating that Hu sheep were in a mild heat stress state for a long time. However, the degree of heat stress was alleviated in the sixth week of the experiment due to the drop in temperature. The overall heat stress level was stable and only slightly alleviated in the last week due to the drop in temperature. This environmental condition is consistent with the high temperature and high humidity climate commonly seen in summer. The synchronous reduction of respiratory rate and rectal temperature, as key indicators reflecting the metabolic intensity and stress level of animals, indicates that GABA and FAM may reduce the body's energy consumption and stress response by regulating the central nervous system or peripheral metabolic pathways. The respiratory rate and rectal temperature of the control group were significantly higher than those of the treatment group, indicating that the Hu sheep were in a mild stress state under conventional feeding conditions, and the intervention of GABA and FAM effectively alleviated this state. From the mechanism of action, GABA, as an inhibitory neurotransmitter, may reduce the release of stress hormones (such as cortisol) by inhibiting the activity of the hypothalamic-pituitary-adrenal (HPA) axis, thereby reducing the respiratory rate and heat production.

[106] ; FAM may indirectly reduce metabolic heat production by enhancing antioxidant capacity or improving energy metabolism efficiency

[107] The synergistic effect between the two was particularly significant in respiratory rate, but no synergistic effect was found in rectal temperature, which may reflect the different response pathways of the two indicators to intervention measures: respiratory rate is more directly regulated by neural regulation.

[108] , while body temperature regulation involves a more complex balance between heat production and heat dissipation

[109] , which may be affected by multiple factors such as ambient temperature and subcutaneous fat thickness.

[0110] GABA and FAM, used alone or in combination, can effectively alleviate the heat stress response of fattening Hu sheep, which is manifested by a significant decrease in respiratory rate and rectal temperature, helping to reduce energy loss and improve feed conversion efficiency, which is especially valuable in hot seasons or high-density breeding environments. The combined treatment showed synergistic advantages in most experimental stages, especially in the early and middle stages when heat stress is greater. However, the advantages of combined treatment are limited to short-term regulation of respiratory rate, while FAM has better stability in long-term applications. For example, in acute stress scenarios such as short-term transportation or vaccination, the combined use of GABA and FAM can quickly alleviate stress responses; while in the long-term fattening stage, the use of FAM alone may be more economical and effective.

[0111] 3.2 Effects of Yingjixiao and Fuaimei on the growth performance of fattening Hu sheep

[0112] Growth performance is a core indicator of animal husbandry. In the experimental design of the present invention, the initial body weights of the groups were similar, with no significant differences, and the basic conditions were consistent.

[0113] First, in terms of terminal body weight and average daily gain (ADG), the effect of GABA alone was significantly better than that of the control group and other treatment groups. This result may be closely related to its neuromodulatory function. The feed intake (DMI) of the GABA group in the present invention was significantly increased, which may be the result of its regulation of the feeding center or improvement of digestive function. However, the increase in ADG far exceeds the increase in DMI, indicating that the role of GABA is not limited to increasing feed intake, but may also promote growth by optimizing energy distribution (such as reducing energy loss under stress) or improving the metabolic efficiency of nutrients.

[0114] In contrast, although the improvement effect of FAM alone on terminal body weight and ADG was significantly better than that of the control group, it was weaker than that of the GABA group. This may be related to the different targets and metabolic pathways of FAM. FAM is composed of Bacillus subtilis and Lactobacillus acidophilus, and may play a role by regulating intestinal microbial communities, enhancing immune function, or providing specific metabolic precursors. However, in the present invention, the DMI of the FAM group was only slightly higher than that of the control group, and the increase in ADG was significantly lower than that of the GABA group, suggesting that its growth-promoting mechanism may be more dependent on metabolic regulation rather than simply increasing feed intake. This result is consistent with the previous study of heat stress indicators, that is, FAM is effective in alleviating physiological stress, but has limitations in directly promoting growth.

[0115] It is worth noting that the final body weight and ADG of the GABA and FAM combined group were significantly lower than those of the GABA alone group. This phenomenon suggests that the two additives may have antagonistic effects or competition in metabolic pathways. Possible explanations include: (1) Dose-dependent inhibition: When GABA and FAM are used simultaneously, an excess of one component may interfere with the absorption or signal transduction of the other component. For example, GABA receptor binding may be competitively inhibited by certain components of FAM; (2) Metabolic pathway conflict: GABA mainly regulates energy distribution through the neuroendocrine system, while FAM may act on intestinal microorganisms or liver metabolism. The synergistic effect of the two did not meet expectations and even led to an increase in metabolic burden. When GABA is used in combination with probiotics, low-dose combinations may enhance intestinal function, while high doses may cause bacterial imbalance.

[0116] GABA alone significantly improved the growth performance of fattening Hu sheep, as evidenced by increases in terminal body weight, ADMI, and ADG. While FAM alone or in combination alleviated heat stress, its direct effect on weight gain was limited. The ADG performance of the combined treatment group was weaker than that of GABA alone, suggesting the need for careful design of additive combinations.

[0117] 3.3 Effects of Yingjixiao and Fuaimei on serum biochemical parameters of fattening Hu sheep

[0118] Serum biochemical indicators are important for assessing an animal's metabolic state, antioxidant capacity, and tissue health. Their dynamic changes can intuitively reflect the comprehensive impact of heat stress on the body and the regulatory effects of intervention measures. This study analyzed the effects of GABA, FAM, and their combined treatment on serum biochemical indicators in fattening Hu sheep, revealing the unique mechanism of action of these two additives in alleviating heat stress, improving metabolic balance, and protecting tissue function.

[0119] Serum albumin (TP) and serum protein (ALB) are important indicators for assessing animal protein metabolism and liver function. Under high temperature stress conditions, animals' feeding capacity is reduced, and their energy metabolism is abnormal, which can lead to an accelerated rate of protein hydrolysis, resulting in a significant decrease in serum protein content.

[0120] In the present invention, total protein content in the stress-eliminating group was significantly higher than in the control group, and significant increases were also observed in the foamilide and combination groups, suggesting that GABA and FAM may promote protein synthesis or reduce protein degradation through different pathways. Notably, while TP levels in the combination group were higher than in the control group, they were lower than in the GABA alone group, suggesting that the synergistic effect of the two additives was not fully reflected in protein metabolism, possibly due to dosage matching or competition for target sites.

[0121] Albumin (ALB) is the main protein synthesized by the liver, and changes in its levels directly reflect the state of liver function. ALB levels in the stress-elimination group were significantly higher than in the control group, while no significant differences were observed in the fo-ami group or the combined group, suggesting that GABA may play a role by protecting liver cell function or enhancing liver synthesis capacity. In contrast, the fo-ami group had a limited effect on improving ALB, possibly because its effects focused more on regulating the intestinal microecology rather than directly regulating liver metabolic pathways.

[0122] Heat stress-induced oxidative damage is one of the core mechanisms of impaired animal health. In the present invention, the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were significantly improved in the stress elimination group and the combined group, but the malondialdehyde (MDA) content was significantly decreased, suggesting that GABA and FAM may improve the body's antioxidant defense system in various ways. The significant increase in T-AOC in the FuAimi group may be related to the metabolites of probiotics (such as short-chain fatty acids). Butyrate produced by probiotic metabolism can activate peroxisome proliferator-activated receptor γ (PPAR-γ), inhibit the NF-κB pathway, reduce the release of proinflammatory factors (such as TNF-α), and thus reduce the vicious cycle of oxidation and inflammation. However, the improvement effect of the FuAimi group on SOD and GSH-Px activity was weaker than that of the stress elimination group, suggesting that its antioxidant effect is more dependent on the regulation of the intestinal microenvironment rather than the direct activation of systemic antioxidant enzymes.

[0123] In addition, the increase in NEFA may reflect that GABA enhances the activity of hormone-sensitive lipase (HSL) and promotes the decomposition of triglycerides (TG). Moreover, the TG content in the stress-elimination group was significantly reduced, further confirming the effectiveness of its lipolytic effect. The regulation of lipid metabolism in the Fuaimi group showed different characteristics. Although the TG level decreased significantly, NEFA did not increase significantly, suggesting that its effect may be more focused on inhibiting fat synthesis rather than promoting decomposition. It is worth noting that the HDLC (high-density lipoprotein cholesterol) in the stress-elimination group was significantly increased, indicating that GABA may reduce the risk of atherosclerosis by improving lipid transport efficiency, which is of great significance to improving meat safety and long-term animal health.

[0124] Lactate dehydrogenase (LDH) is an important marker for evaluating tissue damage; elevated LDH activity reflects disruption of cell membrane integrity or exacerbated hypoxic metabolism. The stress-elimination group had the lowest LDH activity, significantly superior to the fo-Aimi group and the combined group, suggesting that GABA possesses unique advantages in protecting cell membrane structure and maintaining mitochondrial function. The decreased LDH activity in the fo-Aimi group may be related to its anti-inflammatory effects. Probiotics inhibit intestinal inflammation, reducing endotoxin entry into the bloodstream and indirectly reducing systemic tissue damage.

[0125] In terms of glucose metabolism, no significant differences were found between the groups in blood glucose (GLU) and beta-hydroxybutyrate (BHB), indicating that heat stress did not induce severe energy depletion and that the intervention primarily maintained energy homeostasis by optimizing lipid metabolism rather than glucose metabolism. This result is consistent with the physiological characteristics of Hu sheep as ruminants, whose energy supply primarily depends on volatile fatty acids rather than glucose.

[0126] Although the combined treatment showed an additive effect in some indicators (such as T-AOC and HDLC), its effect on total protein, SOD activity, and LDH activity was weaker than that of the GABA alone group. The two additives may be dose-dependent or pathway-competitive. GABA and FAM alleviated the negative effects of heat stress on serum biochemical indicators of Hu sheep through differentiated pathways. The core advantage of GABA is that it regulates the neuroendocrine system, inhibits oxidative stress, and optimizes lipid metabolism; while FAM enhances antioxidant and anti-inflammatory capabilities through probiotic-host interactions.

[0127] 3.4 Effects of Yingjixiao and Fuaimei on serum immune indicators of fattening Hu sheep

[0128] The inhibitory effect of heat stress on the animal immune system has been widely confirmed. It promotes the secretion of glucocorticoids by activating the hypothalamic-pituitary-adrenal axis (HPA axis), thereby inhibiting lymphocyte proliferation, antibody production and the release of pro-inflammatory factors, ultimately leading to a decline in immune function.

[0129] Heat shock protein 70 (HSP70) and cortisol (COR) are core biomarkers for evaluating the degree of heat stress. As a cytoprotective protein, HSP70 expression increases in the early stages of heat stress to maintain protein stability, but long-term high expression will inhibit normal immune function; COR, as the end product of the HPA axis, its increased level can directly inhibit the activity of immune cells. In the present invention, the HSP70 and COR contents of the control group were significantly higher than those of the treatment groups, indicating that the Hu sheep were in a state of chronic heat stress under conventional feeding conditions. The HSP70 and COR levels in the stress elimination group were the lowest, and were significantly better than those in the Fuaimei group and the combined group, suggesting that GABA indirectly reduces the expression pressure of HSP70 by inhibiting excessive activation of the HPA axis and reducing COR secretion. This mechanism is closely related to the central inhibitory effect of GABA: GABA inhibits the synthesis and release of corticotropin-releasing hormone (CRH) by activating the GABA_A receptor in the paraventricular nucleus of the hypothalamus, thereby blocking the HPA axis signal transduction. In addition, the peripheral immune regulatory effect of GABA cannot be ignored. It can regulate vagal nerve activity through 5-hydroxytryptamine (5-HT) secreted by intestinal chromaffin cells, reduce systemic inflammatory responses, and further reduce the release of COR. Although the HSP70 and COR levels in the Fuaimei group were significantly lower than those in the control group, the improvement effect was weaker than that in the stress elimination group, which may be related to the fact that the target of probiotics is concentrated on the regulation of intestinal microecology. Bacillus subtilis reduces the entry of endotoxins into the blood by competitively inhibiting the colonization of pathogens, thereby reducing the stimulation of the HPA axis by systemic inflammation; Lactobacillus acidophilus indirectly regulates the neuroendocrine system through metabolites (such as γ-aminobutyric acid precursors). However, the direct regulatory ability of probiotics on the central HPA axis is limited, resulting in its effect on alleviating stress markers being inferior to GABA. The levels of HSP70 and COR in the combined group did not show a synergistic effect. The significant decrease in COR may be due to the strong inhibition of GABA on the HPA axis, which masked the effect of probiotics, while the regulation of HSP70 was still dominated by GABA, suggesting that the two are insufficiently complementary in the stress pathway.

[0130] Heat stress-induced systemic inflammation is a key cause of immunosuppression. The significant reduction in IL-1β and IL-6 in the stress relief group confirms that GABA inhibits inflammatory responses through multiple pathways: (1) Central pathway: GABA reduces COR release by inhibiting the HPA axis, reducing COR's activation of monocytes; (2) Peripheral pathway: GABA directly acts on the GABA_B receptor on the surface of immune cells, inhibiting the activation of the NF-κB pathway and reducing the synthesis of IL-1β and IL-6. In addition, GABA can also reduce free radical-induced inflammatory damage by enhancing antioxidant capacity (such as increasing SOD activity), forming an "antioxidant-anti-inflammatory" synergistic effect. The IL-1β and IL-6 levels in the Fu Aimei group were also significantly lower than those in the control group, but its mechanism of action is more focused on the regulation of the intestinal-immune axis. However, the inhibitory effect of the Fu Aimei group on IL-1β was weaker than that of the stress relief group, which may be due to its limited ability to regulate the central inflammatory pathway, while the reduction in IL-6 was similar, suggesting that probiotics have a specific targeting effect on the regulation of systemic inflammation. The combined group did not perform better than the single treatment group in IL-1β and IL-6, and some indicators even increased, which may be related to the competitive regulation of GABA and probiotics in the inflammatory pathway.

[0131] Immunoglobulins (IgG, IgM, IgA) are the core effector molecules of humoral immunity, and their levels directly reflect the animal's ability to resist infection. In the present invention, the IgG and IgM levels in the stress elimination group were significantly higher than those in the control group, while the IgG level in the combined group (17.00 ng / mL) reached a peak, indicating that GABA and FAM can enhance antibody production through different pathways. The immune-enhancing effect of GABA may be related to its regulation of the Th1 / Th2 balance: GABA activates the GABA_A receptor on the surface of splenic B cells, promotes IL-4 secretion, and drives the class switch of IgG1 and IgE. In addition, GABA indirectly increases antibody levels by inhibiting COR release and relieving the inhibition of glucocorticoids on plasma cell differentiation. The significant increase in IgG in the Fuaimei group can be attributed to the mucosal immunomodulatory effect of probiotics. It is worth noting that the synergistic increase in IgG in the combined group may be the result of the complementary effects of GABA and probiotics in the Th2 pathway: GABA promotes IL-4 secretion, while probiotics enhance antigen presentation efficiency, jointly optimizing the B cell activation environment.

[0132] Growth hormone (GH) is not only involved in the regulation of growth and development, but also affects immune function by regulating macrophage activity and cytokine secretion. The GH level in the stress elimination group was significantly higher than that in the control group and the combined group, indicating that GABA promotes GH secretion. The increase in GH can further stimulate the liver to synthesize insulin-like growth factor-1 (IGF-1), which enhances immune surveillance function by enhancing T cell proliferation and NK cell activity. This mechanism is linked to the increase in IgG and IgM in the stress elimination group, and together they enhance the disease resistance of Hu sheep. In contrast, the limited increase in GH levels in the Fuaimei group may be related to its target being concentrated in the intestine rather than the neuroendocrine system, but it indirectly supports the metabolic-immune balance by improving intestinal health.

[0133] The combined treatment group showed synergistic advantages in some indicators (such as IgG, COR), but its effect on HSP70, IL-1β and GH was weaker than that of the GABA group alone.

[0134] GABA and FAM improve the immunosuppressive state of Hu sheep exposed to heat stress through differentiated mechanisms. GABA's core advantage lies in its central inhibition of HPA axis activity, reducing levels of COR and pro-inflammatory factors while directly enhancing antibody production. FAM, on the other hand, indirectly alleviates systemic inflammation by optimizing the intestinal immune microenvironment through probiotic-host interactions. Combined use of the two exhibits a synergistic effect in increasing IgG and reducing COR.

[0135] 4 Conclusion

[0136] (1) In terms of heat stress regulation, GABA and FAM can effectively reduce respiratory rate and rectal temperature when used alone. Combined treatment shows a synergistic effect in the short term and can achieve rapid relief of heat stress in ruminants.

[0137] (2) In terms of growth performance, GABA alone had the best effect, significantly increasing terminal body weight and average daily weight gain; while FAM could relieve stress, its direct promotion of weight gain was weak, and the combined treatment did not show a synergistic advantage, suggesting that there may be dose-dependency or metabolic pathway competition between the two.

[0138] (3) Serum biochemical and immune indicators further revealed that GABA reduced cortisol (COR) and heat shock protein 70 (HSP70) levels, maintained metabolic homeostasis by enhancing total antioxidant capacity, inhibiting proinflammatory factors, reducing lipid peroxidation, and regulating lipid metabolism; while FAM increased immunoglobulins by regulating intestinal microecological balance and enhancing antioxidant capacity, thereby alleviating oxidative stress and inflammatory response. GABA and FAM treatment alone were significantly effective in reducing stress markers (HSP70, COR), inhibiting proinflammatory factors (IL-1β, IL-6), and increasing some immunoglobulins (IgM), while combined treatment was particularly effective in reducing COR and increasing IgG.

Claims

1. Use of gamma-aminobutyric acid in the preparation of products for resisting stress and promoting growth in ruminants.

2. Use of gamma-aminobutyric acid in the preparation of products for regulating the neuroendocrine system of ruminants, inhibiting oxidative stress and / or optimizing lipid metabolism.

3. A composition for rapidly relieving stress and promoting growth in ruminants, characterized in that: The composition comprises gamma-aminobutyric acid and a preparation containing bacillus subtilis and lactobacillus acidophilus.

4. The composition according to claim 3, wherein: The preparation containing Bacillus subtilis and Lactobacillus acidophilus is in powder form and consists of Bacillus subtilis and Lactobacillus acidophilus, containing Bacillus subtilis ≥1.0×10 6 CFU / g, Lactobacillus acidophilus ≥1.0×10 6 CFU / g, the carrier is one or more of vermiculite and calcium carbonate; the gamma-aminobutyric acid is in a coated form, and the coating material is one or more of silicon dioxide, sodium carboxymethyl cellulose, and palm oil.

5. Use of the composition according to claim 3 in preparing a product for rapidly relieving stress and promoting growth in ruminants.

6. The use according to claim 5, characterized in that: The application is specifically for the anti-stress and growth promotion of Hu sheep during the fattening stage.

7. The use according to claim 5, characterized in that: The dosage of γ-aminobutyric acid is 0.2±0.05 g / day per ruminant. The dosage of preparations containing Bacillus subtilis and Lactobacillus acidophilus is 0.1 to 0.5 g / day per ruminant. The dosage of preparations containing Bacillus subtilis and Lactobacillus acidophilus is ≥1.0×10 6 CFU / g, Lactobacillus acidophilus ≥1.0×10 6 CFU / g.

8. The use according to claim 1, 2 or 6, characterized in that: The ruminants include cattle and sheep.

9. A functional feed comprising the composition according to claim 3.

10. The functional feed according to claim 9, characterized in that The feed also includes a basic diet component, which, based on mass, includes: 31.6% corn, 16.0% soybean meal, 6.0% peanut meal, 6.0% wheat bran, 15.0% bean dregs, 9.0% peanut seedlings, 6.0% off-starch, 5.0% rice bran, 0.60% calcium hydrogen phosphate, 0.60% baking soda, 0.20% salt, and 4.00% premix; Each kilogram of premix includes 1500 kIU of vitamin A, 350 kIU of vitamin D, 4000 mg of vitamin E, 300-420 mg of sodium selenite, 200 mg of biotin, 13000 mg of zinc, 35000 mg of copper, 5500 mg of manganese, 64 mg of selenium, 160-240 mg of iodine, and 48-72 mg of cobalt.

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