Application of high-purity fish oil containing DHA and EPA to improvement of health and beauty of dog hair

Abstract

The invention discloses application of high-purity fish oil containing DHA (docosahexaenoic acid) and EPA (eicosapentaenoic acid) to improvement of health and beauty of dog hair, and belongs to the field of pet food. The invention relates to a feed additive, which is fish oil and contains Omega-3 fatty acid, the Omega-3 fatty acid comprises DHA (docosahexaenoic acid) and EPA (eicosapentaenoic acid), and the purity of the Omega-3 fatty acid is 95% or above based on the proportion of the Omega-3 fatty acid (comprising EPA and DHA) in total oil; fish oil is added into food to be quantitatively and continuously fed to dogs, and better dog hair conditions can be obtained.

Description

Technical Field

[0001] This application pertains to the field of pet food, and in particular relates to the application of a fish oil containing DHA and EPA to improve the health and appearance of a dog's coat. Background Technology

[0002] Humans have been keeping pets for at least two or three thousand years. The most common pets are cats and dogs, and there are many records of cat and dog keeping in ancient China.

[0003] In this application, fish oil refers to the oil extracted from fish. It is naturally rich in various polyunsaturated fatty acids such as EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid). It can be extracted from fatty deep-sea fish, such as herring, mackerel, sardines, and cod.

[0004] Fish oil is known to be used as food or medicine. In the 18th century, Germans and British people used cod liver oil to treat diseases such as rickets, rheumatism, gout, and tuberculosis.

[0005] Catalase (CAT), an important antioxidant enzyme in the body, efficiently catalyzes the decomposition of hydrogen peroxide (H2O2) produced by intracellular metabolism into water and oxygen, thereby protecting cells from oxidative damage caused by reactive oxygen species (ROS). However, insufficient catalase or excessive hydrogen peroxide in the body may pose a risk of oxidative damage. Therefore, it is necessary to provide targeted prevention and resistance solutions. Patents CN202311772631.0 and CN201280031582.9 also mention feeding pets substances containing DHA, but they do not discuss pet coat problems; the technical solutions they seek to address are for the prevention of serious diseases such as cancer, which is fundamentally different from this application.

[0006] Dog owners want their dogs to be healthy, live long, and have beautiful fur. The solution proposed in this application can effectively help dogs obtain healthier fibroblasts and skin, prevent and resist skin diseases, improve their health, and grow beautiful fur. Summary of the Invention

[0007] This application provides an application of high-purity fish oil containing DHA and EPA to improve the health and appearance of dog coats, including: An application of high-purity fish oil containing DHA and EPA to improve the health and appearance of dog coat, wherein the high-purity fish oil contains Omega-3 fatty acids, including DHA and EPA, and the purity is above 95% based on the proportion of Omega-3 fatty acids (including EPA and DHA) in the total oil, characterized in that the high-purity fish oil containing DHA and EPA is added to the food of dogs for feeding.

[0008] Furthermore, the high-purity fish oil containing DHA and EPA is extracted from deep-sea fish including herring, mackerel, sardines and cod.

[0009] Furthermore, feed at a rate of 50-80 mg / kg body weight / day.

[0010] Furthermore, feed continuously for more than 30 days without interruption.

[0011] Furthermore, feed continuously for more than 45 days without interruption.

[0012] Furthermore, feeding the dog before health problems with its coat can serve as a preventative and protective measure.

[0013] Furthermore, add it to the dog's wet food for a more even mix.

[0014] Furthermore, feed dogs liquid fish oil or capsules directly.

[0015] This application provides an application of high-purity fish oil containing DHA and EPA to improve the health and appearance of dog coats, taking into account factors such as scientific dosage, scientific feeding methods, and the dog's willingness to eat. Detailed Implementation

[0016] The technical solutions of this application will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0017] In the description of this application, it should be understood that the terms "upper," "lower," "left," "right," etc., are used only for the convenience of describing this application and for simplifying the description, and do not indicate or imply that any structure referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on this application. In particular, the understanding of the term "upper" following a noun in the claims should be understood as meaning that the entire inner and outer surfaces of the structure referred to by the noun conform to the definition of "upper."

[0018] It should be further clarified that when the terms "liquid" or "solution" appear in this application, their meaning is broad. In this application, they should not only be understood as clear and pure "liquid substances" and "solutions," but also as mixtures of insoluble solid particles suspended in a liquid, such as "suspensions," mixtures of small droplets dispersed in another immiscible liquid, such as "emulsions," "colloids," and "liquid crystals," and all substances that are essentially mobile phases and can flow freely. For ease of writing and expression, they are abstractly summarized as "liquid," which differs somewhat from the common meaning of the physics term "liquid."

[0019] It should be further clarified that when the term "organism" appears in this application, its meaning is broad. In this application, it should not only be understood as the cellular structure between cells and organs, but also, depending on the specific circumstances, as smaller cells, cytoplasm, or larger organs, or even the entire organism—anything composed of biological material within a certain range. For ease of writing and expression, it is abstractly summarized as "organism," which differs somewhat from the meaning of "organism" in the common biological or medical terminology.

[0020] It should be noted that "dog" and "canine" in this application are synonyms, referring to the same well-known animal, and can be used interchangeably.

[0021] The following detailed description, in conjunction with preferred embodiments, describes the specific implementation methods, structures, features, and effects provided in this application.

[0022] Example 1 The applicant initially considered cats and therefore commissioned the Institute of Animal Husbandry and Veterinary Medicine of the Zhejiang Academy of Agricultural Sciences to conduct experimental verification of the applicant's approach. The Institute purchased feline fibroblasts from the Kunming Cell Bank of the Chinese Academy of Sciences and conducted in vitro verification experiments. Using DHA and EPA as test targets and primary feline fibroblasts as a model, the pre-protective effects of DHA and EPA were systematically evaluated. The results showed that the negative control group cells maintained typical spindle or polygonal shapes, adhered firmly to the cell wall, and had a tight intercellular arrangement, with no obvious detachment or debris. The positive damage group, after 3 hours of treatment with 100 μM H2O2 alone, exhibited obvious characteristics of oxidative damage: cell volume decreased, cell bodies became rounded, intercellular spaces significantly widened, adhesion rate decreased, a large number of cells detached, cell membranes ruptured, and cell debris diffused. However, pretreatment with either 100 μM or 500 μM DHA or EPA failed to effectively alleviate H2O2-mediated cell damage. Even after pretreatment with DHA or EPA, the cells still exhibited significant damage characteristics: blurred cell outlines, shrunken cell bodies, and a significant reduction in cell number. High-concentration DHA or EPA (500 μM) treatment also failed to improve cell condition, with a noticeable loss of intact cells in the field of view. CCK-8 quantitative results further confirmed this phenomenon: the cell viability in the negative control group was set at 100%, while the cell viability in the positive damage group (treated with 100 μM H2O2 alone) decreased to approximately 45%. After pretreatment with 100 μM or 500 μM DHA and EPA, the cell viability remained around 50%, with no statistically significant difference compared to the positive damage group. In conclusion, under the current experimental conditions, neither 100 μM nor 500 μM DHA or EPA pretreatment provided effective antioxidant protection for cat fibroblasts, and the cell viability was essentially the same as the untreated oxidative damage group. These results suggest that DHA and EPA may lack direct antioxidant protection in cat fibroblasts, or require higher concentrations, longer pretreatment times, or synergistic effects with other antioxidants to exert their effects. However, detection of MDA levels in cells revealed that after 24 hours of pretreatment with DHA and EPA, stimulation with H2O2 reduced intracellular MDA levels by an average of approximately 37% and 25%, respectively, suggesting that DHA and EPA still exert in vitro antioxidant effects by inhibiting lipid peroxidation.

[0023] The above experimental results indicate that the effects of feeding DHA and EPA may not be immediate. The conventional approach only knows that DHA and EPA have antioxidant properties, and that applying them to cats can improve the appearance of their fur. However, it doesn't discuss the specific mechanisms or how to use them, which constitutes a technical bias. For example, Chinese invention patent CN202311772631.0 discusses that compositions containing DHA and EPA can have antioxidant and anti-inflammatory effects, but its conclusion stops there; it doesn't further analyze the specific mechanisms of their antioxidant effects, nor does it develop a refined feeding plan for a specific problem (such as coat condition) in a particular animal (e.g., a cat).

[0024] Example 2 Continuing with the above experiments, the applicant believes that gene expression regulation may be an important factor affecting cell growth and development and the activity of antioxidant enzymes. Pretreatment with DHA and EPA for 24 hours directly impacts cellular gene expression, and this effect requires more than 24 hours to manifest. The SOD gene, manganese superoxide dismutase, is a major antioxidant enzyme gene in mitochondria. The CAT gene encodes catalase, which is distributed in both the cytoplasm and mitochondria, protecting cells from oxidative damage by scavenging hydrogen peroxide produced by mitochondria; the GPX gene has multiple types. The applicant requested the Zhejiang Academy of Agricultural Sciences' Institute of Animal Husbandry and Veterinary Medicine to further determine the expression levels of genes exhibiting transcriptional changes using quantitative real-time PCR. The results showed that after 24 hours of incubation with DHA and EPA, the expression levels of antioxidant enzyme genes SOD, CAT, and GPX were all upregulated, and directly proportional to the EPA concentration. Therefore, DHA and EPA can promote the expression of antioxidant enzymes and enhance their activity. Simultaneously, the transcription of vascular endothelial growth factor (VEGFA) and insulin-like growth factor (IGF-1) in cells is upregulated, enhancing perifollicular vascularization, increasing blood supply to hair follicles, providing sufficient nutrition for hair growth, promoting the hair follicle growth cycle, increasing hair density, and reducing abnormal hair loss. EPA and DHA can also regulate fibroblast growth factor (FGF) to regulate the activation of hair follicle stem cells. Furthermore, DHA and EPA also alleviate folliculitis and reduce inflammatory damage by inhibiting inflammatory factors such as IL-4, thus reducing hair loss caused by skin inflammation.

[0025] The above experimental results reveal that the mechanism by which EPA and DHA improve cat hair is gene expression regulation. This mechanism of action requires a certain amount of time to take effect, suggesting that a better approach is to allow sufficient time for long-term maintenance of EPA and DHA use to improve cat hair. It is observed that temporary supplementation when cat hair problems occur is less effective than long-term maintenance.

[0026] Example 3 To confirm the effects of feeding live cats with fish oil containing DHA and EPA and to conduct quantitative analysis, the applicant commissioned the Institute of Animal Husbandry and Veterinary Medicine of the Zhejiang Academy of Agricultural Sciences and the Brand Pet Feeding Research and Development Center to conduct live cat experiments. It should be noted that this experiment was humane and in accordance with animal welfare ethics, and its content only included feeding, hair collection (brushing or trimming), physical examination, hair analysis, and comprehensive observation, without causing any pain or injury to the animals. Feeding and hair collection were conducted at the Brand Pet Feeding Research and Development Center's cat breeding base (located in Xianghu Scenic Area, Hangzhou), and hair analysis was conducted by the Institute of Animal Husbandry and Veterinary Medicine of the Zhejiang Academy of Agricultural Sciences. Fifteen healthy cats were selected, excluding those with skin problems, weighing between 4.25±1.38 kg, randomly assigned by sex, and housed individually. Before the experiment, the cats underwent adequate vaccination and deworming. The feeding period consisted of a 7-day pre-feeding period and a 45-day regular feeding period. The litter box was changed daily, the enclosures were cleaned, and the cats were ensured sufficient rest and playtime. The cats were divided into three groups of five each. The control group was given 40g of regular dry cat food and 40g of regular wet cat food. Control group 1 consisted of long-haired cats, who were given 200mg / cat / day of 95% high-purity fish oil in addition to the regular cat food. The fish oil was added to their wet food. Control group 2 consisted of short-haired cats, who were also given 200mg / cat / day of 95% high-purity fish oil in addition to the regular cat food. All cats were given 300ml of drinking water daily.

[0027] During the trial period, it was observed that the weight changes of the cats were not significant, and the average daily food intake did not differ significantly. Therefore, adding high-purity fish oil containing DHA and EPA to cat food does not cause obesity in cats; adding high-purity fish oil containing DHA and EPA to wet cat food does not cause anorexia in cats.

[0028] During the trial period, double-blind manual assessment revealed no significant difference in coat luster between the experimental and control groups. The difference in coat smoothness between the long-haired group (Experimental Group 1) and the control group was extremely significant at D30 and significant at D45; the difference in coat smoothness between the short-haired group (Experimental Group 2) and the control group was not significant. There was no significant difference in skin elasticity between the experimental and control groups. However, there was a significant difference in smoothness between the experimental group and the control group. Therefore, high-purity fish oil containing DHA and EPA did not significantly improve the feel and luster of the cat's coat; it significantly improved the feel and smoothness of the long-haired cat's coat from day 30 onwards; it did not significantly improve the feel and smoothness of the short-haired cat's coat; and it did not significantly improve the feel and elasticity of the cat's skin.

[0029] During the experiment, the following findings were observed: There were no significant differences in hair loss weight (D15, D30, D45, and D0) between the short-haired group (experimental group 2) and the control group. In the long-haired group (experimental group 1), hair loss weight (D5, D30, and D45) was significantly lower than D0. There were no significant differences in skin moisture, skin oil content, or skin pH between the experimental and control groups. At the end of the experiment, hair density (D45) in both experimental groups 1 and 2 was significantly higher than at the beginning (D0). In the control group, hair density did not differ significantly between the beginning and end of the experiment. Therefore, high-purity fish oil containing DHA and EPA has no significant effect on hair loss weight in short-haired cats; high-purity fish oil containing DHA and EPA significantly reduces hair loss weight in long-haired cats (i.e., reduces shedding); and high-purity fish oil containing DHA and EPA has no significant effect on skin moisture, oil content, or pH in cats. It should be noted that the skin moisture, oil content, and pH value were measured using an RBX-916 capacitive skin analyzer. The measurement site was the exposed skin around the eyebrows. The skin analyzer probe only needed to touch the skin surface to take the measurement. Three measurements were taken and the average value was taken. No inhumane methods such as damaging or removing the cat's skin were used.

[0030] During the trial period, it was found that there was no significant difference in the length-to-width ratio of the hair scales between the experimental and control groups under electron microscopy. Under electron microscopy, regardless of whether it was the long-haired or short-haired group, there were significant differences in the morphology and smoothness of the hair scales before and after fish oil feeding in the experimental group, with the outer periphery being smoother and neater.

[0031] In conclusion, feeding cats high-purity fish oil containing DHA and EPA results in significant improvements in their coats, regardless of whether they are long-haired or short-haired. The improvements are more pronounced in long-haired cats, with an immediate and noticeable reduction in shedding and a significant increase in coat softness starting after 30 days of continuous feeding. Short-haired cats also show improvement, although this improvement may not be immediately apparent to the naked eye; however, electron microscopy reveals healthier coats. Therefore, the applicant believes that to improve a cat's coat, feeding them high-purity (95%) fish oil containing DHA and EPA should be 40-60 mg per kilogram of body weight daily for at least 30 consecutive days.

[0032] Example 4 Based on the successful results of the aforementioned experiments, the applicant also considered another common pet, namely, the dog. The applicant hopes to obtain similar or identical experimental results with dogs. Therefore, the applicant commissioned the Institute of Animal Husbandry and Veterinary Medicine of the Zhejiang Academy of Agricultural Sciences and the Brand Pet Feeding Research and Development Center to conduct live dog experiments. It should be noted that this experiment is humane and in accordance with animal protection ethics, and its content includes only feeding, hair collection (brushing or trimming), physical examination, hair testing, and comprehensive observation, without causing any pain or injury to the animals. Feeding and hair collection were conducted at the Cui Jie Feeding Base in Huzhou, Zhejiang Province (located in Huzhou City) and volunteer families of the experimental dogs (located at various locations). Hair sample processing and data collection and analysis were carried out by the Brand Healthy Feeding Research and Development Center, and hair testing was conducted by the Institute of Animal Husbandry and Veterinary Medicine of the Zhejiang Academy of Agricultural Sciences. Twelve healthy dogs, excluding those with skin problems and weighing between 9.22±2.69 kg, were randomly selected by sex and were centrally housed. They underwent adequate vaccination and deworming before the experiment. Feeding was conducted over a 7-day pre-feeding period and a 45-day regular feeding period. Their enclosures were cleaned daily to maintain cleanliness, and the dogs were guaranteed sufficient rest and playtime. They were divided into three groups: a control group of 5 dogs fed regular dog food, and control group 1 of 7 dogs fed regular dog food supplemented with 75 mg / kg / day of 95% high-purity fish oil. Both the control group and control group 1 were housed at the Cui Jie Feeding Base in Huzhou. Thirteen healthy dogs or dogs with mild underlying skin conditions were selected for control group 2, excluding those with severe skin problems and weighing between 12.14±6.35 kg, randomly selected by sex and were centrally housed by volunteer families. They underwent adequate vaccination and deworming before the experiment and were fed over a 7-day pre-feeding period and a 45-day regular feeding period, supplemented with 75 mg / kg / day of 95% high-purity fish oil. All dogs had free access to food and water.

[0033] During the trial period, it was observed that the weight changes of the dogs were not significant, and the average daily food intake did not differ significantly. Therefore, adding high-purity fish oil containing DHA and EPA to dog food does not cause obesity in dogs; adding high-purity fish oil containing DHA and EPA to wet dog food does not cause anorexia in dogs.

[0034] During the trial period, double-blind sensory evaluation revealed the following: In the control group, hair shine did not differ significantly across sampling days. However, in experimental group 1, D30 hair shine was significantly higher than D0; in experimental group 2, D30 and D45 shine were significantly higher than D0. Smoothness was significantly improved in experimental group 1, with significant differences between D45 and D0, and also significantly different from the control group. In experimental group 2, D15 and D30 smoothness were significantly higher than D0, and the difference at D45 was extremely significant. There were no significant differences in skin elasticity among the control group, experimental group 1, and experimental group 2. Therefore, high-purity fish oil containing DHA and EPA significantly improves the sensory shine of dog coats; it significantly improves the sensory smoothness of dog coats; but it does not significantly improve the sensory skin elasticity of dog coats.

[0035] During the experiment, the following findings were observed: There was no significant difference in the proportion of hair loss between experimental group 1 and the control group as a percentage of initial hair loss. In experimental group 2, the proportion of hair loss at D45 was significantly lower than at D0. There was no significant difference in skin moisture between experimental group 1 and the control group, but experimental group 1 showed an increase at D15, with a highly significant difference from the initial value at D0, which subsequently returned to a non-significant level. There was no significant difference in skin oil content between experimental group 1 and the control group at D0, but experimental group 1 showed a significant difference at D45. Within each group, there were no significant differences in skin oil content. There were no significant differences in hair density between experimental group 1 and the control group, or between the beginning and end of the experiment. It is known that high-purity fish oil containing DHA and EPA may reduce hair loss in dogs, but the effect is not obvious for all dogs. There are unexplained variations in skin moisture data from high-purity fish oil containing DHA and EPA, which may be due to measurement errors or unforeseen factors; the exact cause is unclear. High-purity fish oil containing DHA and EPA has little effect on skin oil content in dogs. High-purity fish oil containing DHA and EPA has no significant effect on hair density in dogs. It should be noted that the skin moisture, oil content, and pH values ​​were measured using an RBX-916 capacitive skin analyzer. The measurement site was the shaved skin on both sides of the mid-back. The probe of the skin analyzer simply touched the skin surface for measurement. Three measurements were taken and the average value was calculated. No inhumane methods such as damaging or removing the dog's skin were used.

[0036] During the trial period, it was found that there was no significant difference in the length-to-width ratio of the hair scales between the experimental group and the control group under electron microscopy. Under electron microscopy, there were significant differences in the morphology and smoothness of the hair scales before and after fish oil feeding in the experimental group, with the outer periphery being smoother and more uniform.

[0037] Furthermore, in the volunteer family trial, we observed an additional phenomenon: in experimental group 2, six dogs with mild underlying skin conditions showed significant improvement in their skin health during the 45-day fish oil feeding period. Specifically, the skin symptoms of four dogs completely disappeared, reaching the clinical cure standard; the symptoms of one dog were alleviated, showing signs of improvement, and hair regrowth occurred in one dog with a patchy bald area on its tail; only one dog showed no significant change in its skin condition. Although this observation was not included in the pre-designed quantitative research system, it further suggests the positive impact of No. 1 high-purity fish oil on canine skin health from a clinical perspective. The applicant speculates that the Omega-3 polyunsaturated fatty acids abundant in fish oil may promote the recovery process of skin diseases through mechanisms such as regulating skin inflammatory responses and improving skin barrier function.

[0038] In conclusion, feeding dogs high-purity fish oil containing DHA and EPA significantly improves their coat, with even stronger improvements in sensory shine and smoothness compared to previous experiments on cats. However, the reduction in shedding in dogs only begins to appear after 45 days of continuous feeding, while improvements in shine and smoothness begin to appear after approximately 30 days. Electron microscopy also confirms that dogs fed high-purity fish oil containing DHA and EPA have healthier coats. Therefore, the applicant believes that to improve a dog's coat, feeding them high-purity (95%) fish oil containing DHA and EPA should be 50-80 mg per kilogram of body weight daily for at least 45 days. The reason for the higher dosage per unit body weight for dogs compared to cats is that dogs are significantly larger and have a higher metabolic rate and digestive capacity.

[0039] Example 5 The applicant also tested adding high-purity fish oil containing DHA and EPA to dry food for cats and dogs, respectively. The results showed that the fish oil made the dry food sticky, significantly altering its texture and palatability, thus noticeably reducing the cats' appetite. Furthermore, due to the small amount of fish oil and the large amount of dry food, cats would avoid the wet portion, making it difficult for them to prioritize the fish oil. However, this was not the case for dogs. Dogs did not dislike eating fish oil and could eat the wet dry food normally. Even so, the applicant still recommends adding fish oil to wet food because it is not easily mixed evenly in dry food. In addition, dogs generally accept and readily accept direct feeding of liquid fish oil or capsules.

[0040] The embodiments described above are merely preferred embodiments of this application. It should be noted that those skilled in the art can make various improvements and substitutions without departing from the technical principles of this application, and these improvements and substitutions should also be considered within the scope of protection of this application. Furthermore, it should be stated that the research results concerning cats are not donated, and the applicant has simultaneously applied for other patents for protection.

Claims

1. An application of high-purity fish oil containing DHA and EPA to improve the health and appearance of dog coat, wherein the high-purity fish oil contains Omega-3 fatty acids, including DHA and EPA, and the purity, based on the percentage of Omega-3 fatty acids (including EPA and DHA) in the total oil, is above 95%, characterized in that... The high-purity fish oil containing DHA and EPA is added to the food of dogs for feeding.

2. The application of high-purity fish oil containing DHA and EPA as described in claim 1 for improving the health and appearance of dog coat, characterized in that, The high-purity fish oil containing DHA and EPA is extracted from deep-sea fish including herring, mackerel, sardines, and cod.

3. The application according to claim 1, characterized in that, Feed at a dose of 50-80 mg / kg body weight / day.

4. The application according to claim 1, characterized in that, Feed continuously for more than 30 days without interruption.

5. The application according to claim 1, characterized in that, Feed continuously for more than 45 days without interruption.

6. The application according to claim 1, characterized in that, Feed your dog before it develops health problems with its coat, as a preventative and protective measure.

7. The application according to claim 1, characterized in that, Add it to the dog's wet food for a more even mix.

8. The application according to claim 1, characterized in that, Feed your dog liquid fish oil or capsules directly.