A composition for reducing stress in cats and uses thereof

Abstract

The application provides a composition for reducing stress reaction of cats and application thereof, and the composition is composed of Lactiplantibacillus plantarum ZYhyy-018, casein peptide and taurine; the Lactiplantibacillus plantarum ZYhyy-018 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 37293, and the preservation date is January 6, 2026. Experiments prove that the composition can significantly reduce the stress score of cats after stress, reduce the content of serum cortisol, improve the level of serum serotonin and gamma-aminobutyric acid, and maintain the number of granulocytes at a normal physiological level. The composition of the application synergistically acts on three target points of nerve, intestinal tract and nutrient metabolism, is natural and safe, is superior to existing products on the market, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of pet pharmaceutical formulation technology, and specifically relates to a composition for reducing stress response in cats and its application. Background Technology

[0002] In recent years, cats, as important companion animals, have provided significant emotional comfort to people, and their number as pets has continued to grow. However, with the increase in stocking density and the growing complexity of the environment, cat health problems have received increasing attention. Among these, stress reactions, due to their high frequency, ability to damage the human-cat relationship, and negative impact on the treatment process, have become a prominent problem troubling pet owners and veterinarians.

[0003] Cats are highly sensitive animals, with an extremely keen sense of smell. Environmental changes, such as new places, unfamiliar smells, or sounds, can easily trigger anxiety and fear in them. When cats are stressed, physiological signs often include ruffled fur, dilated pupils, rapid breathing, tachycardia, drooling, and stunted growth in kittens. Behavioral signs include stiffness, hiding, hissing, excessive grooming, persistent howling, and even prolonged hiding in secluded places such as under beds or sofas. Severe stress can further lead to vomiting, diarrhea, inappropriate defecation, urinary tract obstruction, refusal to eat, and even generalized convulsions, bulging eyes, and death.

[0004] Currently, stress-relieving products for cats on the market mainly fall into three categories: the first is pheromone products, which mimic the natural pheromones secreted by cats to deliver safety signals and are odorless to humans; the second is nutritional supplements, which relieve anxiety by regulating neurotransmitter balance and are usually oral preparations; and the third is functional foods, which incorporate soothing ingredients into daily diets and are suitable for long-term stress prevention. Although these products have met market demand to some extent, they still face multiple challenges, including inconsistent effectiveness, questionable safety, insufficient ease of use, and inconsistent market standards.

[0005] Therefore, there is an urgent need to develop a natural, safe, and effective composition that can systematically improve cats' emotional problems from multiple dimensions, including neuroregulation, gut health, and nutrient metabolism. Summary of the Invention

[0006] To address the problems existing in the prior art, the present invention aims to provide a composition for reducing stress response in cats and its application. The composition targets three core stress points in pet cats: the nervous system, the gut, and nutritional metabolism, thereby achieving a mood-relieving effect.

[0007] The objective of this invention is achieved through the following technical solution: The first aspect of the present invention provides a composition for reducing stress response in cats, the composition comprising Lactiplantibacillus plantarum ZYhyy-018, casein peptides and taurine; The *Lactobacillus plantarum* ZYhyy-018 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 37293 and deposit date of January 6, 2026.

[0008] Furthermore, in the composition, the effective dose of casein peptide is 70-80 mg / kg body weight / day.

[0009] Furthermore, in the composition, the effective dose of taurine is 10-20 mg / kg body weight / day.

[0010] Furthermore, in the composition, the effective dose of *Lactobacillus plantarum* ZYhyy-018 is: viable count ≥ CFU / kg body weight / day

[0011] Furthermore, when the composition is used on cats, the viable count of *Lactobacillus plantarum* ZYhyy-018 per kilogram of cat body weight is [missing information]. CFU, casein peptide 75mg, taurine 15mg.

[0012] The second aspect of the present invention provides the use of the composition described in the first aspect in the preparation of products that improve cat stress and alleviate cat mood.

[0013] Furthermore, the product in question is a pet medication.

[0014] Furthermore, the product is one or more of the following: (1) The product can reduce stress scores in cats after stress. (2) The product can increase the number of granulocytes in the serum of cats after stress; (3) The product can reduce the level of cortisol in the serum of cats after stress; (4) The product can increase the level of serotonin in the serum of cats after stress; (5) The product can increase the content of γ-aminobutyric acid in the serum of cats after stress.

[0015] Information on strain preservation: Lactiplantibacillus plantarum ZYhyy-018, classified as Lactiplantibacillus plantarum, is deposited at the China General Microbiological Culture Collection Center (CGMCC); address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, 100101, China; accession number: CGMCC No. 37293; deposit date: January 6, 2026.

[0016] The advantages of this invention compared to the prior art are as follows: 1. This invention combines *Lactobacillus plantarum* ZYhyy-018, casein peptides, and taurine to create a composition that synergistically targets three core aspects of cat stress response: the nervous system, gut microbiota, and nutritional metabolism, thereby comprehensively improving the stress state of cats from multiple dimensions. Specifically, the composition can reduce post-stress stress scores in cats, maintain the number of granulocytes in the serum of cats after stress at normal physiological levels, reduce the content of cortisol in the serum of cats after stress, increase the content of serotonin in the serum of cats after stress, and increase the content of γ-aminobutyric acid in the serum of cats after stress. 2. The composition for reducing stress response in cats described in this invention uses natural and safe substances with no adverse side effects. It can be added to cat food for a long time and has good prospects for industrial application. Attached Figure Description

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments: Figure 1 The stress scores of each group of cats are shown on day 0 and day 42. Figure 2 The number of granulocytes in the serum of each group of cats is shown. Figure 3 The table shows the cortisol levels in the serum of each group of cats. Figure 4 The levels of serotonin in the serum of each group of cats are shown. Figure 5 The table shows the levels of γ-aminobutyric acid (GABA) in the serum of each group of cats. Detailed Implementation

[0018] The embodiments described are provided to better illustrate the present invention, but are not intended to limit the scope of the invention to the embodiments described. Therefore, non-essential improvements and adjustments made to the embodiments by those skilled in the art based on the above description are still within the scope of protection of the present invention.

[0019] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0020] The present invention will be described in detail below through embodiments. It should be understood that the following embodiments are only used to exemplify and further explain and illustrate the content of the present invention, and are not intended to limit the present invention.

[0021] In the following examples, *Lactobacillus plantarum* ZYhyy-018 was provided by Zhongyuan Food Laboratory, and the strain was deposited at the China General Microbiological Culture Collection Center. Casein peptides were purchased from Guangdong Huatai Biotechnology Co., Ltd.; taurine was packaged by Jiahe Xuri and produced by Qianjiang Yong'an Pharmaceutical Co., Ltd.; and Shuxinbao was purchased from Jiangsu Yabo Animal Health Technology Co., Ltd.

[0022] Example 1: Preparation of a composition for reducing stress response in cats This embodiment provides a composition for reducing stress response in cats, consisting of Lactiplantibacillus plantarum ZYhyy-018 (CGMCC No. 37293), casein peptides, and taurine.

[0023] The method for preparing the composition includes the following steps: Step 1, Preparation of Lactobacillus plantarum ZYhyy-018 bacterial suspension (1) Take the frozen Lactobacillus plantarum ZYhyy-018 out of the -80℃ freezer and thaw it quickly in a 37℃ water bath. Inoculate the strain into 5 mL of liquid MRS medium at an inoculation rate of 0.02%~0.04% and culture it anaerobically at 37°C for 24-48 hours until the bacterial solution becomes turbid.

[0024] The MRS liquid culture medium was prepared as follows: 20.00 g glucose, 10.00 g peptone, 5.00 g yeast extract, 10.00 g beef extract, 1.00 g Tween 80, 5.00 g sodium acetate, 0.58 g magnesium sulfate, 0.25 g manganese sulfate, and 2.00 g dipotassium hydrogen phosphate were dissolved in 1 L of deionized water and sterilized at 121°C for 15 min.

[0025] (2) Take the above turbid bacterial solution and inoculate it on MRS solid medium. Activate it by anaerobic culture at 37°C for 24-48 hours. Inoculate the activated bacteria into primary medium and anaerobic culture at 37°C for 24-48 hours to obtain seed culture. Inoculate the seed culture into secondary medium at an inoculation rate of 1% to 2% and anaerobic culture at 37°C for 24-48 hours to obtain bacterial solution.

[0026] Both the primary and secondary culture media are MRS liquid culture media.

[0027] (3) Centrifuge the obtained bacterial culture at 4°C and 3000-5000 rpm for 15 min, collect the bacterial cells, wash with sterile physiological saline and resuspend to obtain a suspension; adjust the concentration of the suspension to be greater than CFU / mL.

[0028] Step 2, Preparation of the composition Weigh each cat and calculate the number of viable bacteria per kilogram of Lactobacillus plantarum ZYhyy-018 suspension (converted to live bacteria count). The proportions of CFU, casein peptide 75 mg, and taurine 15 mg were used to calculate the daily requirement of each ingredient for each cat. The three ingredients were then mixed thoroughly to obtain the composition for reducing stress in cats.

[0029] The casein peptides used in this embodiment were prepared from bovine casein using a biocontrolled enzymatic hydrolysis technique. The content of the hexapeptide YPVEPF (on a dry basis) was ≥0.8%, and the peptide content (on a dry basis) was ≥15.0%. The taurine content (on a dry basis) was 99.3%, which meets the standard range of 98.5%~101.5%.

[0030] To ensure the activity of Lactobacillus plantarum ZYhyy-018, the composition is preferably prepared fresh daily and added directly to the cat's daily diet.

[0031] Example 2: Evaluation of the effect of the composition on improving stress response in cats To evaluate the effect of the composition described in Example 1 on improving stress response in pet cats, the following animal experiments were conducted in this example: 1. Laboratory animals and grouping Twelve pet cats from the Zhongyuan Food Laboratory were randomly divided into three groups of four cats each: a control group, an experimental group, and a competitor group. The specific treatments were as follows: Control group: fed normal cat food, without any added stress relief products; Experimental group: The composition prepared in Example 1 was added to normal cat food, and the daily amount was calculated according to the actual weight of each pet cat; Competitor group: Add the commercially available product "Shuxinbao" to regular cat food, adding one packet daily according to the product instructions.

[0032] All experimental cats had completed their vaccinations and deworming before the experiment. The experimental environment operated on a 12-hour diurnal cycle, with the temperature controlled between 18-26℃ and the relative humidity between 40%-70%. The cattery was cleaned and disinfected twice daily. Each cat was fed twice daily, with 40 grams of basal food each time. The food was a baked staple food that met the nutritional standards of the Association of American Feed Control Officials (AAFCO, 2023) and the National Research Council (NRC, 2006). Free access to water was provided. The first week was an acclimatization period, followed by 42 days of the formal experimental period. After the experiment, blood samples were collected from the cats for evaluation.

[0033] All data were statistically analyzed using GraphPad Prism 10.1.2. The significance level was: This indicates that p < 0.05. This indicates that p < 0.01. This indicates that p < 0.001. This means p < 0.0001.

[0034] This study strictly followed international guidelines for the use of animals and the "Regulations on the Management of Laboratory Animals in China," and was approved by the Animal Ethics Committee of China Agricultural University (Approval No.: AW80215202-5-15). All experiments were conducted with the aim of minimizing animal suffering.

[0035] 2. Sample collection and indicator detection After the formal experimental period ended (day 42) and before the start of the experiment (day 0, as the baseline control), the following tests were performed on the cats in each group.

[0036] (1) Stress score Experimental cats were placed in airline crates and transported outdoors to an unfamiliar environment for 2 hours along a predetermined route to induce acute stress. Stress scores were assessed for each cat at 0, 15, 30, 60, 90, and 120 minutes after the start of transport, using a stress behavior assessment scale (see Table 1). Scores were completed independently by two trained researchers, and the average score was used.

[0037]

[0038] (2) Serum sample collection and index determination On day 0 and day 42, 2 mL of blood was collected from the forelimb vein of each cat and placed in a sterile centrifuge tube without anticoagulant. After standing at room temperature for 30 min, the blood was centrifuged at 3500 r / min for 10 min at 4℃ to separate the serum, which was then stored at -80℃ for later testing.

[0039] The testing indicators include: ① Granulocyte count: Blood routine tests were performed using a fully automated blood cell analyzer.

[0040] ② Cortisol content: determined using a cat-specific enzyme-linked immunosorbent assay (ELISA) kit.

[0041] ③ Serotonin (5-HT) content: determined using an ELISA kit.

[0042] ④ GABA content: determined using an ELISA kit.

[0043] 3. Statistical Analysis All data were statistically analyzed using GraphPad Prism 10.1.2 software. Paired t-tests were used for within-group comparisons, and one-way ANOVA was used for between-group comparisons. The significance level was set at: This indicates that p < 0.05. This indicates that p < 0.01. This indicates that p < 0.001. This means p < 0.0001.

[0044] 4. Experimental Results (1) Stress score Experimental results are as follows Figure 1 As shown, at the initial stress test on day 0, the stress scores of all three groups of cats showed a decreasing trend over time, reflecting the natural adaptation process of stress response. After 42 days of continuous feeding, the stress responses of each group showed significant differentiation.

[0045] Control group: Stress score decreased from 5.6 at 0 min to 3.5 at 120 min, which was completely consistent with the performance on day 0, indicating that there was no long-term improvement in stress response without intervention.

[0046] Experimental group: The stress score decreased from 4.4 points at 0 min to 1.7 points at 120 min, a decrease of 61.36%, and the endpoint score (1.7 points) was much lower than the performance at 0 days (3.3 points), indicating that long-term feeding of the composition for reducing cat stress response described in Example 1 significantly enhanced the cat's ability to alleviate acute stress.

[0047] Competitor group: The stress score dropped from 5.0 at 0 min to 2.1 at 120 min, a decrease of 58.0%, and the endpoint score (2.1) was lower than the performance on day 0 (3.8), indicating that long-term feeding of the competitor can alleviate the performance of cats under acute stress.

[0048] (2) Serum granulocyte count Experimental results are as follows Figure 2 As shown in the figure. On day 0, the median total granulocyte count of the three groups of cats was similar, with no significant difference between groups, indicating that the baseline immune status of each group was consistent before the experiment. On day 42, the total granulocyte count in the control group decreased significantly, with large fluctuations within the group; the total granulocyte count in the experimental group only decreased slightly, with overall stable values ​​and small data dispersion; the granulocyte count in the competing product group decreased, with the decrease falling between that of the control and experimental groups, and the fluctuations within the group were greater than those in the experimental group.

[0049] The above results indicate that the composition for reducing stress response in cats described in Example 1 can significantly alleviate the decrease in granulocyte count caused by stress and effectively maintain the stability of granulocyte count in cats under stress, with better results than the blank control group and the competing product group.

[0050] (3) Serum cortisol levels Experimental results are as follows Figure 3 As shown in the figure, there was no significant difference in cortisol concentration between day 0 and day 42 in the control group, indicating that stress-induced HPA axis activation persisted without effective intervention, and cortisol levels remained high. In the experimental group, the cortisol concentration was 16.67 ng / mL on day 0 and decreased to 9.87 ng / mL on day 42, with a highly significant difference within the group (p < 0.01), representing a decrease of 40.80%. This indicates that the composition described in Example 1 for reducing cat stress response can effectively inhibit excessive activation of the HPA axis and reduce stress-induced cortisol elevation. In the competing product group, the cortisol concentration was 16.72 ng / mL on day 0 and decreased to 12.10 ng / mL on day 42, with a significant difference within the group (p < 0.05), representing a decrease of 27.63%, which was weaker than the experimental group.

[0051] (4) Serum serotonin content Experimental results are as follows Figure 4 As shown in the figure, there was no significant difference in 5-HT concentration between the control group on day 0 and day 42. In the experimental group, the 5-HT concentration was 82.38 ng / mL on day 0, rising to 97.84 ng / mL on day 42, a significant difference within the group (p < 0.05), representing an increase of 15.13%. This indicates that the composition described in Example 1 for reducing cat stress response can promote 5-HT synthesis and improve emotional stability. The 5-HT concentration in the competing product group increased slightly on day 42 compared to day 0, but the increase was less than that in the experimental group.

[0052] (5) Serum γ-aminobutyric acid content Experimental results are as follows Figure 5 As shown in the figure, there was no significant difference in GABA concentration between the control group and day 42. In the experimental group, the GABA concentration was 4.19 μg / mL on day 0 and increased to 5.21 μg / mL on day 42, with a significant difference within the group (p < 0.05), representing an increase of 24.34%. This indicates that the composition of the present invention can increase GABA levels and enhance the inhibitory function of the central nervous system. In the competing product group, the GABA concentration was 4.02 μg / mL on day 0 and increased to 4.65 μg / mL on day 42, an increase of 15.67%, showing a weaker effect than the experimental group.

[0053] Finally, it should be noted that the above description is only used to illustrate the technical solutions of the present invention and is not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention.

Claims

1. A composition for reducing stress response in cats, characterized in that, The composition comprises Lactiplantibacillus plantarum ZYhyy-018, casein peptides, and taurine. The *Lactobacillus plantarum* ZYhyy-018 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 37293 and deposit date of January 6, 2026.

2. The composition for reducing stress response in cats according to claim 1, characterized in that, In the composition, the effective dose of casein peptide is 70-80 mg / kg body weight / day.

3. The composition for reducing stress response in cats according to claim 1, characterized in that, In the composition, the effective dose of taurine is 10-20 mg / kg body weight / day.

4. The composition for reducing stress response in cats according to claim 1, characterized in that, In the composition, the effective dose of *Lactobacillus plantarum* ZYhyy-018 is: viable count ≥ CFU / kg body weight / day 5. The composition for reducing stress response in cats according to any one of claims 1 to 4, characterized in that, When the composition is used on cats, the viable count of *Lactobacillus plantarum* ZYhyy-018 per kilogram of cat body weight is [missing information]. CFU, casein peptide 75mg, taurine 15mg.

6. The use of the composition according to any one of claims 1 to 5 in the preparation of products for improving cat stress and relieving cat mood, characterized in that, The product in question is a pet medication.

7. The application according to claim 6, characterized in that, The product is one or more of the following: (1) The product can reduce stress scores in cats after stress. (2) The product can increase the number of granulocytes in the serum of cats after stress; (3) The product can reduce the level of cortisol in the serum of cats after stress; (4) The product can increase the level of serotonin in the serum of cats after stress; (5) The product can increase the content of γ-aminobutyric acid in the serum of cats after stress.

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