Feed additive and application
By adding anthocyanins and methyl caffeate with a molar ratio of 1-3:1-3 to canine feed, the preventive problem of heat stress in canines was solved, and the temperature and stress indicators were significantly reduced, which had good economic benefits.
Patent Information
- Application Number
- CN202510719450.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-30
- Publication Date
- 2025-08-12
AI Technical Summary
The existing technology lacks effective preventive methods to improve the dog's resistance to heat stress. Traditional response measures are mostly post-processing and there are problems such as difficulty in identifying early symptoms and difficulty in treating them.
Anthocyanins and methyl caffeate with molar ratio of 1-3:1-3 are used as feed additives to improve the basal metabolic tolerance of dogs through daily diet adjustments and reduce the probability of heat stress.
It significantly reduces the increase in canine body temperature caused by heat stress, reduces stress indicators such as C-reactive protein, creatine kinase, malondialdehyde, etc., has low raw material costs and good economic benefits.
Smart Images
Figure CN120458192A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to feed, in particular to a feed additive and application thereof. Background Art
[0002] Canine heat stress is a pathological condition in which a dog's core body temperature is abnormally elevated due to an imbalance in body temperature regulation, which is common in high temperature, high humidity environments, or after strenuous exercise. Dogs mainly dissipate heat through panting, but their heat dissipation efficiency is limited, especially short-nosed dog breeds, which are more susceptible to the disease due to problems with the structure of their respiratory tract. Symptoms range from early shortness of breath, drooling, and restlessness, gradually developing into vomiting, collapse, confusion, and even organ failure, which can be fatal in severe cases. With global warming and accelerated urbanization, the risk of heat stress has increased significantly. Factors such as the urban heat island effect, confined spaces (such as high temperatures in cars), and frequent extreme weather have increased the probability of dogs being exposed to high temperature environments, and the imbalance between indoor and outdoor activities of pet dogs in modern lifestyles has further weakened their natural adaptability.
[0003] Traditional methods for managing heat stress in dogs rely primarily on physical cooling and emergency medical intervention, such as relocation to a cool area, cooling with cold water, and intravenous fluids. However, these traditional methods focus on post-event management, leaving many owners unable to recognize early symptoms. By the time dogs develop mid- to late-stage heat stress, they often suffer from multi-organ damage, significantly increasing the difficulty of treatment. Therefore, systematic methods to enhance long-term stress resistance are urgently needed.
[0004] In recent years, research on enhancing dogs' resistance to heat stress through feed conditioning has gradually received attention. The core of this research is to regulate physiological state through nutritional intervention. Compared with drugs or forced cooling, the advantage of feed conditioning is its preventive and low-invasiveness. By adjusting daily diet, the basal metabolic tolerance of dogs can be improved, the probability of heat stress can be reduced, and the side effects of drugs can be avoided. Summary of the Invention
[0005] Purpose of the invention: The purpose of the present invention is to provide a feed additive that can effectively alleviate heat stress in animals, especially canines. The second purpose is to provide applications of the feed additive.
[0006] Technical solution: The feed additive described in the present invention contains anthocyanidin and caffeic acid methyl ester in a molar ratio of 1-3:1-3.
[0007] Preferably, the feed additive contains anthocyanidin and caffeic acid methyl ester in a molar ratio of 1:1-3.
[0008] Preferably, the anthocyanidin is blueberry anthocyanidin, with the molecular formula C 15 H 11 ClO6, CAS No. 528-58-5; the molecular formula of the caffeic acid methyl ester is C 10 H 10O4, CAS number is 3843-74-1.
[0009] The invention relates to an application of the feed additive in preparing feed for alleviating animal stress.
[0010] Preferably, the feed is a feed for relieving stress in dogs; further, the feed is a feed for relieving heat stress in dogs.
[0011] Preferably, the feed preparation step comprises: adding feed additives to the feed according to the weight of the animal, and fully mixing to obtain feed that relieves animal stress, wherein the dosage of the feed additive is 10-20 mg per kilogram of animal weight.
[0012] Preferably, the dosage of the feed additive in the feed is 8-16 mg per kilogram of body weight.
[0013] Preferably, in the added feed additive, the dosage of blueberry anthocyanidins is 5-9 mg per kilogram of body weight, and the dosage of caffeic acid methyl ester is 3-7 mg per kilogram of body weight.
[0014] Beneficial effects: Compared with the existing technology, the present invention has the following significant advantages: 1. The feed additive can significantly reduce the increase in body temperature of dogs caused by heat stress, and at the same time significantly reduce stress indicators such as C-reactive protein, creatine kinase, and malondialdehyde, which can effectively alleviate the heat stress of dogs; 2. The raw material cost of the feed additive is low, it is easy to prepare and use, and has good economic benefit prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] Figure 1 This is a statistical chart of the biocompatibility results of blueberry anthocyanins and caffeic acid methyl ester;
[0016] Figure 2 This is a statistical chart showing the results of blueberry anthocyanidins and caffeic acid methyl ester used alone or in combination to alleviate heat damage to canine ileal epithelial cells;
[0017] Figure 3 This is the statistical result of digestibility of dogs after 30 days of different feeding methods;
[0018] Figure 4 This is the statistical result of dog weight after 30 days of different feeding methods;
[0019] Figure 5 The following is the statistical result of body temperature of dogs treated with normal room temperature of 25℃ or heat treatment of 35℃ for 30 days under different feeding methods;
[0020] Figure 6 The statistical results of inflammation, injury, and oxidative stress indicators in dogs treated with different feeding methods for 30 days after being treated at normal room temperature of 25℃ or heat-treated at 35℃. DETAILED DESCRIPTION
[0021] The technical solution of the present invention is further described below.
[0022] Example 1: Combination of blueberry anthocyanins and caffeic acid methyl ester alleviates heat stress damage to intestinal cells
[0023] Using the molecular formula C 15 H 11 ClO6, CAS number 528-58-5 blueberry anthocyanin; molecular formula C 10 H 10 O4, CAS number 3843-74-1 caffeic acid methyl ester was used for the experiment.
[0024] 1. Biocompatibility of blueberry anthocyanins and caffeic acid methyl ester
[0025] (1) Canine ileal epithelial cells were cultured at a rate of 5×10 3 The cells were seeded at a density of 100 cells / well in a 96-well cell culture plate and incubated for 16 h to allow the cells to adhere to the wall and grow further;
[0026] (2) A control group (0 μM), a blueberry anthocyanidin treatment group, and a caffeic acid methyl ester treatment group were set up. In the control group, DMEM complete medium was used to replace the original culture medium; in the blueberry anthocyanidin treatment group, DMEM complete medium containing blueberry anthocyanidin was added to replace the original culture medium, and the final concentrations of blueberry anthocyanidin were 2.5, 5, and 7.5 μM, respectively; in the caffeic acid methyl ester treatment group, DMEM complete medium containing caffeic acid methyl ester was added to replace the original culture medium, and the final concentrations of caffeic acid methyl ester were 2.5, 5, and 7.5 μM, respectively.
[0027] After 48 h of treatment, the cells were replaced with 90 μL of fresh DMEM complete medium and 10 μL of CCK8 was added to each well. The cells were incubated in a dark place in a cell culture incubator for 15 min. The absorbance was measured at 450 nm and the relative cell viability was calculated and normalized using formula (1):
[0028] Relative cell viability = (A s -A b ) / (A c -A b )×100% (1).
[0029] A s 、A b and A c are the absorbance values of sample, blank control and negative control, respectively.
[0030] The results are as follows Figure 1 As shown in the results, blueberry anthocyanins and caffeic acid methyl ester at a concentration of 2.5 μM did not cause damage to canine ileal epithelial cells.
[0031] 2. Combination of blueberry anthocyanins and caffeic acid methyl ester alleviates heat stress damage to intestinal cells
[0032] (1) Canine ileal epithelial cells were cultured at a rate of 5×10 3 The cells were seeded at a density of 100 cells / well in a 96-well cell culture plate and incubated for 16 h to allow the cells to adhere to the wall and grow further;
[0033] (2) The control group (CON), blueberry anthocyanidin treatment group, caffeic acid methyl ester treatment group, blueberry anthocyanidin and caffeic acid methyl ester combined treatment group (1:1), heat stress group (HS), heat stress + blueberry anthocyanidin treatment group (HS + blueberry anthocyanidin treatment group), heat stress + caffeic acid methyl ester treatment group (HS + caffeic acid methyl ester treatment group), heat stress + blueberry anthocyanidin and caffeic acid methyl ester combined treatment group (HS + 1:1) were set up.
[0034] Among them, the control group used DMEM complete medium to replace the original culture medium; the blueberry anthocyanin treatment group added DMEM complete medium containing 2.5 μM blueberry anthocyanin to replace the original culture medium; the caffeic acid methyl ester treatment group added DMEM complete medium containing 2.5 μM caffeic acid methyl ester to replace the original culture medium; the blueberry anthocyanin and caffeic acid methyl ester combined treatment group added DMEM complete medium containing 2.5 μM blueberry anthocyanin and 2.5 μM caffeic acid methyl ester to replace the original culture medium; the heat stress group, heat stress + blueberry anthocyanin treatment group, heat stress + caffeic acid methyl ester treatment group, and heat stress + blueberry anthocyanin and caffeic acid methyl ester combined treatment group were based on the above 4 groups. After the culture medium was replaced, they were subjected to simulated heat stress treatment at 39°C for 2 hours.
[0035] (3) After 48 h of treatment, the cells were replaced with 90 μL of fresh DMEM complete medium and 10 μL of CCK8 was added to each well. The cells were incubated in a dark place in a cell culture incubator for 15 min. The absorbance was measured at 450 nm and the relative cell viability was calculated and normalized using the aforementioned formula (1).
[0036] The results are as follows Figure 2 As shown, the combination of blueberry anthocyanins and caffeic acid methyl ester at a concentration of 2.5 μM can effectively alleviate heat stress damage to intestinal cells.
[0037] Example 2: Blueberry anthocyanin-caffeic acid methyl ester feed additive alleviates heat stress in dogs
[0038] Using the molecular formula C 15 H 11 ClO6, CAS number 528-58-5 blueberry anthocyanin; molecular formula C 10 H 10 O4, CAS number 3843-74-1 caffeic acid methyl ester was used for the experiment.
[0039] Equimolar amounts of blueberry anthocyanidins and caffeic acid methyl ester are mixed to obtain a feed additive.
[0040] (1) Twenty-four experimental beagle dogs were purchased from Jiangsu Leica Biotechnology Co., Ltd. and raised at the Lihong Livestock and Poultry Breeding Center in Jiangbei New District, Nanjing. During the experiment, the indoor temperature was maintained at 25±2℃. All experimental dogs had free access to food and water. All experimental dogs were randomly divided into normal wet food group (CON), normal wet food + blueberry anthocyanin group (blueberry anthocyanin), normal wet food + caffeic acid methyl ester group (caffeic acid methyl ester), and normal wet food + 1:1 blueberry anthocyanin and caffeic acid methyl ester combination group (1:1), with 6 dogs in each group.
[0041] (2) The normal wet food group was provided with normal wet food during daily feeding; the normal wet food + blueberry anthocyanidin group was provided with wet food containing 7.2 mg / kg of blueberry anthocyanidin; the normal wet food + caffeic acid methyl ester group was provided with wet food containing 4.9 mg / kg of caffeic acid methyl ester; and the normal wet food + 1:1 blueberry anthocyanidin and caffeic acid methyl ester combination group was provided with wet food containing feed additives, with a blueberry anthocyanidin dose of 7.2 mg / kg and a caffeic acid methyl ester dose of 4.9 mg / kg. This experiment lasted for 30 days, and the dogs' total intestinal digestibility and body weight were recorded at the end of the experiment.
[0042] The statistical results of total intestinal digestibility are as follows Figure 3 The weight statistics are shown in Figure 4 As shown, the use of blueberry anthocyanidins and caffeic acid methyl ester alone or in combination did not produce abnormal effects.
[0043] (3) The above four groups were then divided into two groups within each group, for a total of eight groups, including: normal room temperature and normal wet food group (CON), normal room temperature and normal wet food + blueberry anthocyanin group, normal room temperature and normal wet food + caffeic acid methyl ester group, normal room temperature and normal wet food + feed additive group (1:1), heat treatment and normal wet food group (HS), heat treatment and normal wet food + blueberry anthocyanin group (HS+blueberry anthocyanin group), heat treatment and normal wet food + caffeic acid methyl ester group (HS+caffeic acid methyl ester), and heat treatment and normal wet food + feed additive group (HS+1:1).
[0044] The normal room temperature group was treated at 25°C, while the heat treatment group was treated at 35°C for 3 hours. The normal wet food group showed increased water intake, decreased activity, and mild vomiting after 3 hours of treatment at 35°C. Body temperature and related blood parameters were measured in all eight experimental groups before and after treatment.
[0045] The body temperature statistics results are as follows Figure 5 As shown in the figure, the use of feed additives can effectively reduce the increase in body temperature of experimental dogs under heat treatment conditions; the statistical results of relevant blood indicators are shown in the figure. Figure 6As shown in the results, the use of feed additives can significantly reduce stress indicators including C-reactive protein, creatine kinase, and malondialdehyde, and the reduction rate is significantly higher than the use of blueberry anthocyanins or caffeic acid methyl ester alone. At the same time, feed additives can effectively increase the level of glutathione peroxide in the dog's blood, further indicating that the combined use of blueberry anthocyanins and caffeic acid methyl ester in feed additives can produce a synergistic effect, which can achieve more effective heat stress relief for dogs compared with the treatment of blueberry anthocyanins or caffeic acid methyl ester alone.
Claims
1. A feed additive, characterized in that The feed additive contains anthocyanidin and caffeic acid methyl ester in a molar ratio of 1-3:1-3.
2. The feed additive according to claim 1, characterized in that The feed additive contains anthocyanidin and caffeic acid methyl ester in a molar ratio of 1:1-3.
3. The feed additive according to claim 1 or 2, characterized in that The anthocyanin is blueberry anthocyanin, and its molecular formula is C 15 H 11 ClO6, CAS number is 528-58-5.
4. The feed additive according to claim 1 or 2, characterized in that The molecular formula of the caffeic acid methyl ester is C 10 H 10 O4, CAS number is 3843-74-1.
5. Use of the feed additive according to claim 1 or 2 in preparing feed for alleviating animal stress.
6. The use according to claim 5, characterized in that The feed is a feed for alleviating stress of dogs.
7. The use according to claim 6, characterized in that The feed is a feed for alleviating heat stress of dogs.
8. The use according to claim 5, characterized in that The feed preparation step comprises: adding a feed additive to the feed according to the weight of the animal, and fully mixing the feed to obtain a feed that relieves animal stress, wherein the dosage of the feed additive is 10-20 mg per kilogram of animal weight.
9. The use according to claim 8, characterized in that The dosage of the feed additive in the feed is 8-16 mg per kilogram of body weight.
10. The use according to claim 9, characterized in that In the feed additives, the dosage of blueberry anthocyanin is 5-9 mg per kilogram of body weight, and the dosage of caffeic acid methyl ester is 3-7 mg per kilogram of body weight.