Composition for relieving beef cattle transportation stress syndrome and application thereof

Abstract

The invention provides a composition for relieving beef cattle transportation stress syndrome and application thereof, the composition is prepared by premixing dexmedetomidine hydrochloride, gastrodin and normal saline for injection according to the mass concentration ratio, the concentration of dexmedetomidine hydrochloride is 60-65 g / mL, and the concentration of gastrodin is 20-30 mg / mL. The composition is applied to beef cattle before transportation, the composition is intramuscularly injected, and the total injection volume is 10-20 mL. After the composition is injected before beef cattle transportation, immune dysfunction can be effectively improved, excessive HPA axis activation is inhibited, cortisol secretion is reduced, oxidative damage is relieved, and the stability of intestinal flora is maintained, so that inflammation is relieved, nervous excitability is reduced, calming and anxiety resisting effects are achieved, beef cattle transportation stress response is reduced, and the beef cattle transportation effect is improved. And the method is simple to operate and low in cost, and provides theoretical and practical support for preventing the beef cattle transportation stress syndrome.

Description

Technical Field

[0001] This invention belongs to the field of biological agent technology and relates to a composition for alleviating transport stress syndrome in beef cattle and its application. Background Art

[0002] Beef cattle transportation often faces various stressors, including drastic environmental changes, poor transportation conditions, and improper management. These stressors can lead to physiological, endocrine, and immune dysfunction, resulting in problems such as intestinal flora imbalance and bovine respiratory disease syndrome. These health issues not only reduce feed conversion ratio and growth performance but can also cause death in severe cases, seriously hindering the development of animal husbandry. In recent years, with increasing awareness of animal welfare and the expansion of transportation scale, significant progress has been made in alleviating the pressure of beef cattle transportation. Currently, methods such as optimizing transportation models and using feed additives or medications to effectively alleviate stress-induced physiological responses have been explored. However, these methods are often complex and costly to implement. Summary of the Invention

[0003] To address the problems of complex operation and high cost of the methods for alleviating the stress of beef cattle transportation described in the background art, the present invention provides a composition for alleviating beef cattle transportation stress syndrome and its application.

[0004] The composition of the present invention is prepared by premixing dexmedetomidine hydrochloride, gastrodin and physiological saline for injection in a certain mass concentration ratio, wherein the concentration of dexmedetomidine hydrochloride is 60-65µg / mL and the concentration of gastrodin is 20-30mg / mL.

[0005] Preferably, the total dose of dexmedetomidine hydrochloride is 2.5 µg / kg, the total dose of gastrodin is 1 mg / kg, and both dexmedetomidine hydrochloride and gastrodin are dissolved in physiological saline for injection to prepare an injection solution with a total volume of 10–20 mL.

[0006] Preferably, the dexmedetomidine hydrochloride and gastrodin are premixed in physiological saline for injection at a volume ratio of 1:400.

[0007] Preferably, the composition contains a phosphate buffer, and the pH of the composition is 4.5-7.0 to enhance stability and reduce tissue irritation.

[0008] Preferably, the concentration of dexmedetomidine hydrochloride is 62.5 µg / mL.

[0009] Preferably, the concentration of gastrodin is 25 mg / mL.

[0010] The present invention also proposes the application of a composition for alleviating transport stress syndrome in beef cattle, wherein the composition is administered intramuscularly before transport of beef cattle, with a total injection volume of 10–20 mL.

[0011] Preferably, the composition is packaged in a 10–20 mL sterile disposable injection vial for a single intramuscular injection.

[0012] Preferably, before transporting the beef cattle, an intramuscular injection is administered through the deep tendons of the neck or the muscles of the buttocks of the beef cattle, divided into 2–4 acupoints, with each acupoint containing no more than 5 mL, and the total injection volume being 10–20 mL.

[0013] Preferably, the composition is administered via intramuscular injection 15–30 minutes before transporting the beef cattle.

[0014] Compared with existing technologies, the composition of this invention for alleviating transport stress syndrome in beef cattle contains dexmedetomidine, a highly selective α2-adrenergic receptor agonist, which shows unique advantages in relieving transport stress in beef cattle. Its mechanism of action mainly involves activating the locus coeruleus and inhibiting norepinephrine release, thereby reducing inflammation, decreasing nerve excitability, producing sedative and anti-anxiety effects, and ultimately alleviating transport stress syndrome in beef cattle, demonstrating unique advantages in relieving transport stress in beef cattle. Gastrodin, as a traditional Chinese medicine, has neuroprotective effects and inhibits oxidative stress and inflammation. Verification shows that injecting the composition of this invention into beef cattle before transport can effectively improve immune dysfunction, inhibit excessive HPA axis activation, reduce cortisol secretion, reduce oxidative damage, and maintain the stability of intestinal flora, thereby reducing inflammation, decreasing nerve excitability, exerting sedative and anti-anxiety effects, reducing transport stress response in beef cattle, and is simple and low-cost to operate, providing theoretical and practical support for the prevention of transport stress syndrome in beef cattle. Attached Figure Description

[0015] Figure 1 The figure shows the effect of the composition provided by the present invention on glucose in transport-stressed beef cattle.

[0016] Figure 2 The figure shows the effect of the composition provided by this invention on ALT levels in transport-stressed beef cattle.

[0017] Figure 3 The figure shows the effect of the composition provided by the present invention on the AST of transport-stressed beef cattle.

[0018] Figure 4 The figure shows the effect of the composition provided by the present invention on ALB in transport-stressed beef cattle.

[0019] Figure 5 The figure shows the effect of the composition provided by the present invention on the total tidal volume (TC) of transport-stressed beef cattle.

[0020] Figure 6 The figure shows the effect of the composition provided by this invention on Mg in transport-stressed beef cattle.

[0021] Figure 7 The figure shows the effect of the composition provided by the present invention on calcium in transport-stressed beef cattle.

[0022] Figure 8 The figure shows the effect of the composition provided by the present invention on the Cor of transport stress in beef cattle.

[0023] Figure 9 The figure shows the effect of the composition provided by the present invention on NEFA in transport-stressed beef cattle.

[0024] Figure 10 The figure shows the effect of the composition provided by the present invention on SOD in transport-stressed beef cattle.

[0025] Figure 11 The figure shows the effect of the composition provided by the present invention on the CAT (causing stress) of transport-stressed beef cattle.

[0026] Figure 12-14 The figure shows the analysis results of the composition provided by the present invention on the α-diversity of gut microbiota in transport-stressed beef cattle.

[0027] Figure 15-16 The figure shows the analysis results of the composition provided by the present invention on the β diversity of gut microbiota in transport-stressed beef cattle. Detailed Implementation

[0028] To make the technical problems, technical solutions, and beneficial effects to be solved by this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and are not intended to limit the scope of this application.

[0029] A composition for alleviating transport stress syndrome in beef cattle is prepared by premixing dexmedetomidine hydrochloride, gastrodin, and physiological saline for injection in a certain mass concentration ratio, wherein the concentration of dexmedetomidine hydrochloride is 60-65 µg / mL and the concentration of gastrodin is 20-30 mg / mL.

[0030] Preferably, the total dose of dexmedetomidine hydrochloride is 2.5 µg / kg, the total dose of gastrodin is 1 mg / kg, and both dexmedetomidine hydrochloride and gastrodin are dissolved in physiological saline for injection to prepare an injection solution with a total volume of 10–20 mL.

[0031] Preferably, the dexmedetomidine hydrochloride and gastrodin are premixed in physiological saline for injection at a volume ratio of 1:400.

[0032] Preferably, the composition contains a phosphate buffer, and the pH of the composition is 4.5-7.0 to enhance stability and reduce tissue irritation.

[0033] Preferably, the concentration of dexmedetomidine hydrochloride is 62.5 µg / mL.

[0034] Preferably, the concentration of gastrodin is 25 mg / mL.

[0035] Application of a composition for alleviating transport stress syndrome in beef cattle, wherein the composition is administered intramuscularly prior to transport of the beef cattle, with a total injection volume of 10–20 mL.

[0036] Preferably, the composition is packaged in a 10–20 mL sterile disposable injection vial for a single intramuscular injection.

[0037] Preferably, before transporting the beef cattle, an intramuscular injection is administered through the deep tendons of the neck or the muscles of the buttocks of the beef cattle, divided into 2–4 acupoints, with each acupoint containing no more than 5 mL, and the total injection volume being 10–20 mL.

[0038] Preferably, the composition is administered via intramuscular injection 15–30 minutes before transporting the beef cattle.

[0039] Example Preparation of the composition: The total dose of dexmedetomidine hydrochloride is 2.5 µg / kg, and the total dose of gastrodin is 1 mg / kg. Dexmedetomidine hydrochloride and gastrodin are premixed in physiological saline for injection at a volume ratio of 1:400. The concentration of dexmedetomidine hydrochloride is 62.5 µg / mL, and the concentration of gastrodin is 25 mg / mL. Phosphate buffer is added to the composition to make the pH 6.5. The composition is packaged in sterile disposable injection bottles of 10–20 mL.

[0040] Simmental cattle around 12 months of age, in good health, and of similar weight were selected to analyze their effects on immune function and gut microbiota, providing a basis for the prevention of transport stress syndrome in beef cattle. The specific implementation steps are as follows.

[0041] Animal Selection: In this experiment, 10 healthy Simmental cattle of approximately 12 months age and similar weight were randomly selected from the same farm as experimental subjects. The animals were randomly divided into two groups (n=5 in each group). The DEX+Gas group was injected with a combination of dexmedetomidine and gastrodin; the control group was injected with an equal volume of physiological saline. Two transport vehicles were arranged for transportation. On October 15, 2024, the experimental cattle were transported from Zhumadian City, Henan Province to Huanggang City, Hubei Province, a journey of approximately 500 kilometers and 6 hours. During transportation, they were fed the same feed and water.

[0042] Sample Collection and Storage: To evaluate the effects of the composition, blood and rectal fecal samples were collected from cattle at four time points: before transport (D0), day 1 after arrival (D1), day 4 (D4), and day 7 (D7). Blood samples were collected from the tail vein of each group of cattle and transferred to anticoagulant tubes containing EDTA or heparin sodium. The samples in the anticoagulant tubes were centrifuged at 3,500 rpm for 15 minutes to separate the plasma, and then stored at −80 °C for analysis of biochemical parameters, hormones, and oxidative stress markers. Blood samples in EDTA tubes were stored at 4 °C for hematological examination. In addition, rectal fecal samples from all experimental cattle were collected into 50 mL centrifuge tubes and stored at -80 °C for 16S rRNA sequencing analysis.

[0043] The determination method is as follows: I. Serum Biochemical Indicators Determination: At the end of the experiment, the serum biochemical indicators of the two groups of beef cattle were detected using a fully automated biochemical analyzer, including: albumin (ALB), creatine kinase (CK), glucose (Glu), total cholesterol (TC), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), calcium (Ca), and magnesium (Mg).

[0044] II. Hormone and Oxidizing Factor Measurement: The following indicators in serum were detected by enzyme-linked immunosorbent assay (ELISA): cortisol (Cor), total cholesterol (TC), catalase (CAT) activity, superoxide dismutase (SOD) activity, and non-esterified fatty acid (NEFA) concentration.

[0045] III. Microbial diversity assessment: 16S rRNA sequencing analysis was performed on rectal fecal samples collected at four time points.

[0046] The data processing and analysis methods were as follows: GraphPad Prism 10.1 was used to plot the blood test results, which were expressed as mean ± standard error (SEM). Two-way ANOVA was used for group comparisons. All microbial diversity data were analyzed on the MajorBio cloud platform (https: / / cloud.majorbio.com).

[0047] The results are as follows: I. Effects of the Composition on Serum Biochemical Indicators: Before transportation, all biochemical indicators were within the normal physiological range. At day 1 after transportation, compared to day 0, serum glucose concentration decreased by 2.17% in the DEX+Gas group and increased by 7.17% in the CON group, with a significant difference between the groups (P<0.05). Figure 1There were no significant differences in ALT and AST levels between the two groups at any time point (P>0.05). Figure 2 , Figure 3 At day 1 (D1), albumin levels in the DEX+Gas group and the CON group increased by 16.36% and 21.67% respectively compared to day 0, but the differences between the groups were not statistically significant (P>0.05). Figure 4 Total cholesterol levels in all groups fluctuated within the normal range, with no significant differences between groups (P>0.05). Figure 5 Due to muscle damage caused by transportation stress, CK and LDH levels were significantly elevated and exceeded the instrument's detection limit; therefore, no statistical analysis was performed.

[0048] All ion concentrations measured in all groups fluctuated within the normal reference range. Although Mg levels showed a decreasing trend in all groups, the changes were not statistically significant (P>0.05). Figure 6 On Day 1, compared to pre-transport levels, the DEX+Gas group increased by 5.98%, and the CON group increased by 4.09%. The Ca level in the DEX+Gas group remained unchanged, while the CON group increased by 3.65% (P>0.05). Figure 7 ).

[0049] II. Effects of the composition on hormones and oxidative factors: At day 1, compared with pre-transport levels, cortisol increased by 93.46% in the DEX+Gas group and by 110.95% in the CON group, with no significant difference between groups (P>0.05). Figure 8 At day 1, the NEFA level decreased by 14.92% in the DEX+Gas group and by 15.85% in the CON group; the difference between the groups was not significant (P>0.05). Figure 9 Regarding SOD levels, compared to pre-transport levels, the DEX+Gas group saw an 8.51% decrease, and the CON group saw an 18.05% decrease; the difference between the groups was not significant (P>0.05). Figure 10 No significant differences were observed in CAT activity among the groups, and it remained relatively stable throughout the observation period (P>0.05). Figure 11 ).

[0050] III. Microbial Diversity Analysis: Microbial α-diversity results showed that after transportation, compared with the CON group, the DEX+Gas group exhibited higher Shannon and Pielou_e indices and lower Simpson indices, indicating higher diversity and evenness of the gut microbiota in the DEX+Gas group. At day 7, the Shannon and Pielou_e indices of the DEX+Gas group were significantly higher than those of the CON group (P<0.05). Figure 12 , Figure 14The Simpson index decreased significantly (P<0.05), while the Simpson index decreased significantly. Figure 13 The results in summary indicate that transport stress leads to a decrease in the α-diversity of the gut microbiota in Simmental cattle; while intervention with the combination of dexmedetomidine and gastrodin can effectively alleviate this downward trend and help maintain gut microbiota homeostasis.

[0051] Beta diversity was assessed using the unweighted UniFrac algorithm based on OTU sequences. Figure 15-16 The results showed a significant difference in OTU-level gut microbiota structure between the two groups (P=0.001). The microbiota structure in the DEX+Gas group showed slow changes after day 1, exhibiting a gradual recovery trend. The structural changes in the CON group were relatively smaller. Overall, the combined intervention of dexmedetomidine and gastrodin significantly altered the gut microbiota structure. This change was pronounced and persistent in the DEX+Gas group, remaining different from the CON group until the later stages of the experiment; while the structural changes in the CON group were smaller. These findings indicate that the combined intervention of dexmedetomidine and gastrodin has a significant regulatory effect on gut microbiota composition.

[0052] The results in summary demonstrate that the application of the composition of this invention can, to a certain extent, alleviate immune dysfunction caused by transportation stress in beef cattle, inhibit excessive HPA axis activation, reduce cortisol secretion, and mitigate oxidative damage. Furthermore, the combination of dexmedetomidine and gastrodin helps maintain intestinal homeostasis. These results provide theoretical and practical support for the feasibility of using the dexmedetomidine and gastrodin combination to alleviate transport stress syndrome in beef cattle during transportation.

[0053] The preferred embodiments of the present invention have been described in detail above with reference to the accompanying drawings and specific examples. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. A composition for alleviating transport stress syndrome in beef cattle, characterized in that: The composition is prepared by premixing dexmedetomidine hydrochloride, gastrodin, and physiological saline for injection in a certain mass concentration ratio, wherein the concentration of dexmedetomidine hydrochloride is 60-65 µg / mL and the concentration of gastrodin is 20-30 mg / mL.

2. The composition for alleviating transport stress syndrome in beef cattle according to claim 1, characterized in that: The total dose of dexmedetomidine hydrochloride is 2.5 µg / kg, and the total dose of gastrodin is 1 mg / kg. Both dexmedetomidine hydrochloride and gastrodin are dissolved in physiological saline for injection to prepare an injection solution with a total volume of 10–20 mL.

3. The composition for alleviating transport stress syndrome in beef cattle according to claim 2, characterized in that: The dexmedetomidine hydrochloride and gastrodin were premixed in physiological saline for injection at a volume ratio of 1:

400.

4. The composition for alleviating transport stress syndrome in beef cattle according to claim 1, characterized in that: The composition contains a phosphate buffer and has a pH of 4.5-7.

0.

5. The composition for alleviating transport stress syndrome in beef cattle according to claim 1, characterized in that: The concentration of dexmedetomidine hydrochloride was 62.5 µg / mL.

6. The composition for alleviating transport stress syndrome in beef cattle according to claim 5, characterized in that: The concentration of gastrodin is 25 mg / mL.

7. The application of the composition for alleviating transport stress syndrome in beef cattle according to any one of claims 1-6, characterized in that: Before transporting beef cattle, the composition is administered intramuscularly, with a total injection volume of 10–20 mL.

8. The application of the composition for alleviating transport stress syndrome in beef cattle according to claim 7, characterized in that: The composition is packaged in 10–20 mL sterile disposable injection vials for single intramuscular injection.

9. The application of the composition for alleviating transport stress syndrome in beef cattle according to claim 8, characterized in that: Before transporting beef cattle, administer intramuscular injections through the deep muscles of the neck or hip muscles of the cattle, divided into 2–4 acupoints, with each acupoint containing no more than 5 mL, and a total injection volume of 10–20 mL.

10. The application of the composition for alleviating transport stress syndrome in beef cattle according to claim 9, characterized in that: The composition is administered intramuscularly 15–30 minutes before transporting the beef cattle.

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