Composition and application thereof in preparation of product for relieving diarrhea

Abstract

The invention relates to the technical field of feed products, in particular to a composition and application thereof in preparation of a product for relieving diarrhea. The composition of the present invention comprises butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, or a precursor substance capable of releasing the aforementioned fatty acids in the digestive tract. The composition provided by the invention can effectively relieve diarrhea and inflammation, regulate intestinal flora and improve intestinal barrier integrity and intestinal health, and is used for preventing, relieving or treating intestinal diseases; the composition has an obvious relieving effect on diarrhea and enteritis of animals, compared with drugs such as antibiotics, the composition has higher safety and is not prone to drug resistance, and the composition can be used for treating intestinal diseases and can also be added into daily ration to improve intestinal health.

Description

Technical Field

[0001] The invention relates to the technical field of feeding products, and in particular to a composition and application thereof in preparing a product for alleviating diarrhea. Background Art

[0002] Gastrointestinal diseases are common diseases in animal husbandry, among which diarrhea is the most common clinical symptom. Microbial infection is one of the main causes of diarrhea. The infected pathogenic bacteria secrete bacterial toxins that cause intestinal epithelial homeostasis disorders, leading to symptoms such as diarrhea. Diarrhea can cause loss of appetite, sudden weight loss, dehydration, etc. in animals, and in severe cases, it can lead to death.

[0003] Currently, the main treatment for diarrhea is antibiotics, but due to the irrational use of antibiotics, drug-resistant bacteria gradually emerge, resulting in poor therapeutic effects of antibiotics. Therefore, it is urgent to develop non-antibiotic compositions that can effectively relieve diarrhea. Summary of the invention

[0004] The invention provides a composition and application of the composition in preparing a product for relieving diarrhea.

[0005] In the research and development of products for relieving diarrhea and improving intestinal health, the present invention unexpectedly discovered that butyric acid, lauric acid, arachidonic acid (ARA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA) can cooperate with each other in a certain ratio in repairing the integrity of the intestinal barrier, inhibiting harmful intestinal bacteria, regulating intestinal flora, and relieving inflammation, producing a synergistic effect, and significantly improving the effect of the composition on improving intestinal flora and intestinal health, and relieving intestinal inflammation and diarrhea; adding precursor substances that can release the above-mentioned fatty acids in the digestive tract through digestion and decomposition can have a considerable technical effect.

[0006] Specifically, the present invention provides the following technical solutions:

[0007] The present invention provides a composition comprising:

[0008] (1) butyric acid and / or a precursor substance capable of releasing butyric acid in the digestive tract; and

[0009] (2) lauric acid and / or a precursor substance capable of releasing lauric acid in the digestive tract; and

[0010] (3) arachidonic acid and / or a precursor substance capable of releasing arachidonic acid in the digestive tract; and

[0011] (4) docosahexaenoic acid and / or a precursor substance capable of releasing docosahexaenoic acid in the digestive tract; and

[0012] (5) eicosapentaenoic acid and / or a precursor substance capable of releasing eicosapentaenoic acid in the digestive tract;

[0013] Calculated by the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the weight proportions of the components are as follows: 2-5 parts of butyric acid, 1-3 parts of lauric acid, 7-10 parts of arachidonic acid, 15-30 parts of docosahexaenoic acid and 15-30 parts of eicosapentaenoic acid.

[0014] In the present invention, the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid refers to the sum of the mass of the above fatty acids contained in the composition and the mass of the above fatty acids that the precursor substances of the above fatty acids contained can theoretically produce. For example, if a butyric acid precursor is added, the amount of butyric acid added is calculated based on the amount of the butyric acid precursor added and its butyric acid content.

[0015] Preferably, based on the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the weight parts of each component are as follows: 2-4 parts of butyric acid, 1-2 parts of lauric acid, 7-9 parts of arachidonic acid, 18-25 parts of docosahexaenoic acid and 18-25 parts of eicosapentaenoic acid.

[0016] Preferably, based on the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the ratio of the total mass of butyric acid and lauric acid to the total mass of arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid is (3-6):(40-60).

[0017] The present invention finds that by controlling the ratio of the total mass of butyric acid and lauric acid to the total mass of arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid within the above range, the above saturated fatty acids and unsaturated fatty acids can produce a synergistic effect in alleviating diarrhea and intestinal inflammation and improving intestinal flora and its metabolism, thereby significantly improving the effect of the composition on diarrhea and intestinal inflammation.

[0018] Further preferably, based on the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the ratio of the total mass of butyric acid and lauric acid to the total mass of arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid is (3-6):(44-58).

[0019] In the present invention, the precursor substance refers to a substance that can provide target fatty acids (butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid, eicosapentaenoic acid) through biotransformation methods such as hydrolysis and esterification in the digestive system.

[0020] In the above composition, the precursor substance capable of releasing butyric acid in the digestive tract is one or more selected from butyrate, butyrate ester, and butyrate derivatives.

[0021] The precursor substance capable of releasing lauric acid in the digestive tract is one or more selected from laurate, lauric acid ester, and lauric acid ester derivatives.

[0022] The precursor substance capable of releasing arachidonic acid in the digestive tract is one or more substances selected from arachidonic acid salts, arachidonic acid esters, and arachidonic acid ester derivatives.

[0023] The precursor substance capable of releasing docosahexaenoic acid in the digestive tract is one or more selected from docosahexaenoic acid salts, docosahexaenoic acid esters, and docosahexaenoic acid ester derivatives.

[0024] The precursor substance capable of releasing eicosapentaenoic acid in the digestive tract is one or more substances selected from eicosapentaenoic acid salts, eicosapentaenoic acid esters, and eicosapentaenoic acid ester derivatives.

[0025] In the above-mentioned precursor substances, the salt includes one or more of sodium salt, potassium salt, calcium salt and magnesium salt. The ester and ester derivative include ethyl ester, glyceride, phospholipid type glyceride or its salt.

[0026] In some embodiments of the present invention, the ester is a glyceride. Among them, the butyric acid glyceride can be one or more of tributyrin, monobutyric acid glyceride, and dibutyric acid glyceride. The butyric acid glyceride derivative can be one or more of butyric acid glyceride salts, butyric acid lauric acid glyceride, butyric acid caprylic acid glyceride, butyric acid capric acid glyceride, or. Lauric acid glyceride can be one or more selected from polyglycerol laurate or its salt, lauric acid monoglyceride or its salt, dilauric acid glyceride or its salt, trilauric acid glyceride or its salt. Lauric acid glyceride derivatives can be one or more selected from lauric acid glyceride salts, esters of lauric acid and other short-chain or medium-chain fatty acids and glycerol or their salts.

[0027] In some embodiments of the present invention, the precursor of butyric acid is tributyrin.

[0028] In some embodiments of the present invention, the precursor of lauric acid is lauric acid monoglyceride.

[0029] Among them, arachidonic acid or its precursor can be obtained from plants or microorganisms (such as Mortierella alpina) through fermentation.

[0030] Docosahexaenoic acid, eicosapentaenoic acid or their precursors can be obtained by fermentation from plants, deep-sea fish or microorganisms (algae). The types of the above polyunsaturated fatty acids are relatively concentrated in the oils and fats obtained from microbial fermentation and deep-sea fish, and the content is relatively high, which can be used as the raw materials of the above fatty acids or their precursors. For example, the main components of fish oil are docosahexaenoic acid and eicosapentaenoic acid. In some embodiments of the present invention, fish oil with high EPA content and fish oil with high DHA content are used as raw materials to provide EPA and DHA, respectively.

[0031] In order to reduce the difficulty of application, the above-mentioned oils and fats are usually prepared into powders in the art, and the powders, in addition to the oils and fats containing the target fatty acids, mainly contain starch, casein, antioxidants and other substances as ingredients, which are only used as auxiliary materials for the powders and do not have the effects of relieving diarrhea and improving intestinal health in the body. Therefore, it can be understood by those skilled in the art that the use of powders as raw materials (for example, in some embodiments of the present invention, arachidonic acid powder is used as a raw material to provide arachidonic acid, and fish oil powder is used as a raw material to provide EPA and DHA) will not affect the efficacy of the composition of the present invention.

[0032] In some embodiments of the present invention, the composition comprises the following components: tributyrin, monolaurin, arachidonic acid, fish oil with a high docosahexaenoic acid content, and fish oil with a high eicosapentaenoic acid content. Based on the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid, and eicosapentaenoic acid contained in the above components, the weight parts of each component are as follows: 2-4 parts of butyric acid, 1-2 parts of lauric acid, 7-9 parts of arachidonic acid, 18-25 parts of docosahexaenoic acid, and 18-25 parts of eicosapentaenoic acid.

[0033] Optionally, the docosahexaenoic acid content of the fish oil with high docosahexaenoic acid content is 10-20%, and the eicosapentaenoic acid content is 1-4%, and the eicosapentaenoic acid content of the fish oil with high icosapentaenoic acid content is 9-18%, and the docosahexaenoic acid content is 4-8%.

[0034] The weight of each precursor substance in the composition can be calculated based on the content of the corresponding fatty acid in the precursor substance and the weight ratio of the above fatty acids.

[0035] The animals targeted by the above composition include mammals, preferably cats, and more preferably adult cats. The adult cats are cats over 6 months old. The above composition has an effective alleviating effect on diarrhea and intestinal inflammation in adult cats, can significantly reduce the diarrhea rate, and relieve intestinal inflammation and systemic inflammation. It has an excellent alleviating effect on bacterial diarrhea in adult cats.

[0036] The composition provided by the present invention is preferably a composition for cats. More preferably, it is a composition for adult cats. More preferably, it is a composition for alleviating diarrhea in adult cats.

[0037] The present invention also provides any of the following applications of the above-mentioned composition:

[0038] (1) Use in the preparation of products for preventing, alleviating or treating diarrhea in animals;

[0039] (2) Use in the preparation of products for preventing, alleviating or treating intestinal inflammation in animals;

[0040] (3) Application in the preparation of products for improving intestinal health of animals;

[0041] (4) Application in the preparation of products for improving intestinal flora of animals;

[0042] (5) Application in the preparation of products for increasing the content of short-chain fatty acids in the intestines of animals;

[0043] (6) Application in animal husbandry;

[0044] (7) Use in the preparation of feed, feed additives, medicines or foods.

[0045] In the above application, the animal is preferably a mammal.

[0046] More preferably, the animal is a feline.

[0047] In some embodiments of the invention, the animal is a cat.

[0048] In some embodiments of the invention, the animal is an adult cat.

[0049] In the above application, the product is one selected from feed, feed additives, medicine, and food, wherein the food includes ordinary food and health food.

[0050] In the above application, the dosage of the product when administered to animals is: 0.04-0.09 g / kg body weight / day, more preferably 0.06-0.09 g / kg body weight / day, based on the total amount of arachidonic acid, EPA, DHA, butyric acid and lauric acid.

[0051] The present invention provides a medicine comprising the above composition.

[0052] In addition to the above-mentioned composition, the drug may also contain excipients permitted in the pharmaceutical field. The drug may be in any dosage form, including but not limited to solid preparations such as powders and liquid preparations such as emulsions.

[0053] Preferably, the content of arachidonic acid in the drug is 2-10% based on the mass of arachidonic acid.

[0054] The above-mentioned medicine can be a medicine for preventing, alleviating or treating diarrhea or enteritis, and is preferably a medicine for cats.

[0055] The present invention provides a feed, which comprises the above composition.

[0056] Preferably, the content of arachidonic acid in the feed is 0.01-0.05% based on the mass of arachidonic acid.

[0057] In some embodiments of the present invention, the feed is cat feed (eg, cat canned food).

[0058] The present invention provides a method for alleviating or curing diarrhea in adult cats, comprising: administering the above composition to a subject, in a total amount of arachidonic acid, EPA, DHA, butyric acid and lauric acid, at 0.04-0.09 g / kg body weight / day, more preferably 0.06-0.09 g / kg body weight / day.

[0059] The present invention is verified through animal experiments that administering the composition of the present invention to adult cats in the above dosage can effectively reduce the diarrhea rate, relieve inflammation, increase the content of intestinal short-chain fatty acids and improve intestinal flora.

[0060] The beneficial effects of the present invention are as follows: the composition provided by the present invention can effectively relieve diarrhea and inflammation, regulate intestinal flora, improve intestinal barrier integrity and intestinal health, and is used to prevent, relieve or treat intestinal diseases; the composition has a significant alleviating effect on diarrhea and enteritis in animals (especially adult cats), and uses a nutrient substance, fatty acid, which has higher safety and is less likely to produce drug resistance than drugs such as antibiotics, and can be used for the treatment of intestinal diseases and can also be added to the diet to improve intestinal health. DETAILED DESCRIPTION

[0061] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0062] In the compositions of the following examples and comparative examples, butyric acid is provided as a raw material using tributyrin (butyric acid content is 60%), lauric acid is provided as a raw material using lauric acid monoglyceride (lauric acid content is 90%), ARA is provided as a raw material using arachidonic acid powder (ARA content is 20%), and DHA and EPA are provided as raw materials using fish oil powder (high EPA content, 7.4% DHA, 9.4% EPA) and fish oil powder (high DHA content, 12.81% DHA, 2.78% EPA).

[0063] The dosage of the above raw materials can be calculated based on the content of the target fatty acid and the dosage ratio of the target fatty acid.

[0064] Specifically, the composition of Example 1 includes the following raw material components in a mass ratio: the mass ratio of tributyrin powder, monolaurin powder, arachidonic acid powder, fish oil powder (high EPA content), and fish oil powder (high DHA content) is 1.73: 0.77: 13.88: 69.71: 13.90. The composition of Example 2 includes the following raw material components in a mass ratio: the mass ratio of tributyrin powder, monolaurin powder, arachidonic acid powder, fish oil powder (high EPA content), and fish oil powder (high DHA content) is 1.28: 0.43: 15.31: 69.19: 13.80. The composition of Example 3 includes the following raw material components in mass ratio: the mass ratio of tributyrin powder, lauric acid monoglyceride powder, arachidonic acid powder, fish oil powder (high EPA content), and fish oil powder (high DHA content) is 1.90: 0.63: 11.42: 71.73: 14.30. The composition of Comparative Example 1 includes the following raw material components in mass ratio: the mass ratio of tributyrin powder and lauric acid monoglyceride powder is 1.73: 0.77. The composition of Comparative Example 2 includes the following raw material components in mass ratio: the mass ratio of arachidonic acid powder, fish oil powder (high EPA content), and fish oil powder (high DHA content) is 13.88: 69.71: 13.90. The raw materials of the composition of Comparative Example 3 are to replace the fish oil powder (high EPA content) in Example 1 with 10% algae oil powder (the oil source is Nannochloropsis, which contains almost no DHA), and replace the fish oil powder (high DHA content) with 60% linolenic acid powder, and the amounts of the above two raw materials are calculated according to the amounts of EPA and linolenic acid. The raw materials of the composition of Comparative Example 4 are to replace the fish oil powder (high DHA content) and fish oil powder (high EPA content) in Example 1 with DHA algae oil powder derived from dinoflagellate (the content is 15%, and the EPA content in dinoflagellate is very small and negligible).

[0065] In the above raw materials, except for the target fatty acid, the content of other relevant functional long-chain polyunsaturated fatty acids is very low. For example, in arachidonic acid powder, except for arachidonic acid, most of it is stearic acid and palmitic acid, and the content of other polyunsaturated fatty acids is extremely low; similarly, in fish oil powder, except for high content of EPA and DHA, it is mainly stearic acid. The above raw materials are all raw materials commonly used to provide the target fatty acids of the present invention. Those skilled in the art can understand that trace amounts of other unsaturated fatty acids will not have a significant effect on the efficacy of the composition.

[0066] Example 1

[0067] This embodiment provides a composition comprising the following components in parts by weight: 8 parts of arachidonic acid, 20 parts of EPA, 20 parts of DHA, 3 parts of butyric acid, and 2 parts of lauric acid.

[0068] Example 2

[0069] This embodiment provides a composition comprising the following components in parts by weight: 8 parts of arachidonic acid, 18 parts of EPA, 18 parts of DHA, 2 parts of butyric acid, and 1 part of lauric acid.

[0070] Example 3

[0071] This embodiment provides a composition comprising the following components in parts by weight: 9 parts of arachidonic acid, 25 parts of EPA, 25 parts of DHA, 4 parts of butyric acid, and 2 parts of lauric acid.

[0072] Comparative Example 1

[0073] This comparative example provides a composition comprising the following components in parts by weight: 3 parts of butyric acid and 2 parts of lauric acid.

[0074] Comparative Example 2

[0075] This comparative example provides a composition comprising the following components in parts by weight: 8 parts of arachidonic acid, 20 parts of EPA, and 20 parts of DHA.

[0076] Comparative Example 3

[0077] This comparative example provides a composition, which is different from the composition of Example 1 only in that DHA is replaced by an equal amount of linolenic acid.

[0078] Comparative Example 4

[0079] This comparative example provides a composition, which differs from the composition of Example 1 only in that EPA is replaced by DHA, that is, it comprises the following components in parts by weight: 8 parts of arachidonic acid, 40 parts of DHA, 3 parts of butyric acid, and 2 parts of lauric acid.

[0080] Experimental example

[0081] Animal experiments were conducted on the compositions of the above embodiments and comparative examples to evaluate their efficacy in relieving diarrhea, inflammation, improving intestinal health, etc., as follows:

[0082] 1. Experimental animals and groups

[0083] Adult cats aged 1-2 years old, with an error of less than 4 months, half male and half female, weighing 2.6-3.5 kg, were selected for a pet breeding experiment, namely, an adult cat control group (Group A), an adult cat infection group (Group B), and Examples 1-3 and Comparative Examples 1-4.

[0084] 2. Experimental diet

[0085] The diet was a purchased regular basal adult cat diet.

[0086] 3. Test methods

[0087] After 2 days of adaptive feeding, the adult cat control group (Group A) was fed with a basic diet and PBS buffer; the adult cat infection group (Group B) and the various examples and comparative groups were fed with a basic diet and 5 mL of pathogenic Escherichia coli (10 8 CFU / mL), it is estimated that this stage is 7 days. When all soft stools or more than half of diarrhea (lasting more than two days, watery stools or porridge-like stools, daily diarrhea frequency is greater than three times, without obvious pus and blood, can be accompanied by abdominal pain, abdominal distension, tenesmus, fever, etc.) occur in the negative control group, this stage can be terminated in advance. After this, Group A and Group B are all only fed with basal diets, and each embodiment and comparative example group are fed with the compositions of corresponding embodiments and comparative examples respectively, and the compositions of Embodiments 1-3 and Comparative Examples 1-4 are all given according to the feeding amount of 0.07g fatty acid (i.e. the total amount of arachidonic acid, EPA, DHA, butyric acid and lauric acid, lacking components and calculating as 0 according to the dosage) / kg body weight / day.

[0088] During the experiment, each cat had an individual cage, and the food intake was recorded daily and the cats were weighed once a week.

[0089] 4. Feeding and management

[0090] All adult cats are raised at room temperature of 20-25℃, with light and dark alternating every 12 hours and relative humidity of about 40%-60%. The daily diet is purchased regular adult cat food, fed according to the recommended dosage, and the drinking water is uniformly matched. The indoor environment is kept well ventilated and the cage is maintained in a good sanitary environment.

[0091] During the entire experimental process, the adult cats were raised scientifically in accordance with the relevant regulations of the Experimental Animal Ethics Committee.

[0092] 5. Test indicators

[0093] Before the start of the trial, the adult cats were physically examined and blood samples were collected to obtain complete blood counts and serum biochemical profiles to ensure that all adult cats were healthy before the trial and laboratory tests were within normal ranges.

[0094] The detection indicators before and during the experiment include diarrhea conditions (feces morphology, diarrhea incidence), growth indicators (feed intake, daily weight gain, feed conversion rate), blood indicators (blood biochemical indicators, serum antibodies and inflammatory indicators), and feces indicators (volatile fatty acids in feces, flora in feces).

[0095] (1) Sample collection: Collect animal feces, hair and blood.

[0096] (2) The detection indicators and methods are as follows:

[0097] a. Diarrhea

[0098] After feeding pathogenic E. coli, pay close attention to the feces of each cat, take photos to record the feces morphology, and record the incidence of diarrhea every day. When all the negative groups have soft stools, in order to reduce the pain of adult cats and reduce the harm of pathogenic bacteria to animals, soft stools are assumed to be diarrhea.

[0099] The formula for calculating the diarrhea rate is as follows:

[0100] b. Growth indicators

[0101] Based on the daily recorded feed intake and weekly body weight, feed intake, daily weight gain and feed conversion rate of each group were calculated.

[0102] Feed intake: Record the amount of feed given daily, weigh the remaining amount of each group before feeding every evening, and calculate and record the daily feed intake.

[0103] Daily weight gain: Each cat was weighed once a week, and weekly and daily weight gain were calculated.

[0104]

[0105] c. Blood indicators

[0106] After the experiment, blood was collected from the animals for routine blood tests and detection of serum inflammatory indicators.

[0107] After collecting blood, put it into EDTA anticoagulant tube and use IDEXX ProCyte one blood cell analyzer for routine blood test. The blood in the blank tube was centrifuged at 3000rpm / min for 10min, and the upper serum was separated. The serum SAA concentration was detected by enzyme-linked immunosorbent assay (ELISA). For specific operation steps, please refer to the kit instructions.

[0108] d. Fecal indicators

[0109] The volatile fatty acids in the feces were detected by gas chromatography, and the bacterial flora in the feces were sent to the company for 16SrDNA determination.

[0110] The method for detecting volatile fatty acids in feces is as follows:

[0111] Weigh 0.4g of feces into a 1.5mL centrifuge tube, add ultrapure water at a ratio of 1:3, vortex mix and let stand for 10min, then centrifuge at 10000rpm for 10min; separate the supernatant, centrifuge at 10000rpm for 10min; take 1mL of supernatant and mix with 20μL of chromatographic grade phosphoric acid; pass the mixture through a 0.22μm disposable syringe filter and inject the filtrate directly into the chromatographic bottle. Use gas chromatograph to determine the content of SCFAs (mainly acetic acid, propionic acid, and butyric acid) in the cecal contents.

[0112] The fecal flora detection method is as follows:

[0113] Microbial DNA from adult cat feces was extracted using a DNA SPIN kit. Primers 341F (5'-CCTAYGGGRBGCASCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3') were used to amplify the V3-V4 region of the 16S rDNA gene in the samples. The pooled DNA products were used to construct an Illumina paired-end library according to the Illumina genomic DNA library preparation procedure. The amplicon library was then paired-end sequenced (2×250) on the Illumina Novaseq platform according to the standard protocol. The raw image data files obtained from high-throughput sequencing were converted into sequencing reads by base calling analysis.

[0114] Data optimization: Raw data were cleaned through three steps: raw read filtering, high-quality read assembly, and chimera removal.

[0115] OTU clustering: Using Usearch software, non-repetitive sequences (excluding single sequences) with similarity > 97% were assigned to the same OTU, and the representative sequence of each OTU was annotated with a threshold of 0.8.

[0116] Taxonomic analysis: uclust software was used to perform taxonomic analysis on OTU representative sequences, and the community composition of each sample was statistically analyzed at each classification level: domain, kingdom, plylum, class, order, family, genus, and species. The comparison database was RDP.

[0117] Sample diversity analysis (Alpha-diversity): Based on the obtained OTUs data, the sample flora abundance and flora diversity were analyzed through multiple indexes, and the R language tool was used to draw graphs.

[0118] Species composition: Based on the OTU sample table with a similarity level of 97%, the number of common and unique OTUs in multiple samples was counted, the similarity and overlap of each sample were compared, and the Veen diagram was produced using the R language tool.

[0119] Differential analysis: First, the nonparametric factor Kruskal-Wallis sum rank test was used to detect the characteristics with significant abundance differences, and the groups with significant differences in abundance were found. The difference was considered significant when P < 0.05. Finally, the linear discriminant analysis (LDA) was used to estimate the magnitude of the influence of each species abundance on the differential effect using LEfSE software.

[0120] 6. Experimental data processing

[0121] The experimental results were expressed as mean ± standard deviation; the data were statistically analyzed by SPSS20.0 software. Independent sample t test was used for comparison between two groups, and one-way analysis of variance (ANOVA) was used for comparison between multiple groups. Compared with the minimum value in the group, P < 0.05 was considered statistically significant, * indicated P less than 0.05, ** indicated P less than 0.01, and *** indicated P less than 0.001.

[0122] 7. Test results

[0123] (1) Effect of the composition on diarrhea rate in adult cats

[0124] Calculations showed that there were no significant differences in feed intake, daily weight gain and feed conversion rate among the groups of adult cats during the trial period.

[0125] Except for the adult cat negative control group (Group A), the adult cat infection group (Group B) and each embodiment and comparative example group were infected with E. coli. On the third day after infection, soft stools began to appear in each group, and all groups had soft stools within eight days. The diarrhea rate is shown in Table 1. Some adult cats had broken feces and no shape, and very few adult cats had bloody stools, which proved that the model was successful.

[0126] After bacterial infection, the infected adult cats (Group B) were fed with a basic diet and allowed to recover on their own; each Example and Comparative Example group was fed with the composition of the corresponding Example and Comparative Example, and the diarrhea rate was counted and the cat feces were observed daily.

[0127] After observation, the adult cats in the infected group (Group B) began to improve their fecal morphology three days after the infection was stopped. On the seventh day, half of the adult cats had normal feces and half had soft feces. For the adult cats fed with the composition of Example 1, most of the adult cats had partially soft feces after two days, and their feces returned to normal after seven days.

[0128] The results show (Table 2) that after adult cats have soft stools, supplementing with the composition of each embodiment can accelerate the recovery of adult cats' stools to normal and significantly shorten the time required for the stools to return to normal.

[0129] Table 1 Diarrhea rate on the eighth day after infection

[0130]

[0131] Table 2 The number of days it takes for the diarrhea rate to return to zero

[0132]

[0133]

[0134] (2) Effect of the composition on the growth performance of adult cats

[0135] Calculations showed that there were no significant differences in feed intake, daily weight gain and feed conversion rate among the groups of adult cats during the trial period.

[0136] (3) Effects of the composition on blood inflammation-related indicators in adult cats

[0137] The results showed that after the invasion of E. coli, the total number of white blood cells, the total number of neutrophils and the total number of monocytes in Group B and each embodiment and comparative group increased significantly, indicating that inflammation occurred after the body was infected with bacteria, the white blood cells in the body increased pathologically, and the neutrophils increased and decomposed the bacteria.

[0138] The test results of the blood inflammation-related indicators of adult cats after the test are shown in Table 3. The total white blood cell count, total neutrophil count and total monocyte count of Group B remain at a high level, which exceeds the normal reference range of adult cats, while the total white blood cell count, total neutrophil count and total monocyte count of each composition cat of the composition of Example 1-3 have been reduced to within the normal range. This shows that the composition of the embodiment of the present invention can significantly reduce the inflammatory cells in adult cats (P < 0.05) and effectively relieve inflammation. However, the total white blood cell count, total neutrophil count and total monocyte count of the blood of adult cats given the composition of Comparative Examples 1-4 remain at a high level, significantly exceeding the normal reference range of adult cats.

[0139] Table 3 Effects of the composition on blood inflammation-related indicators in adult cats

[0140]

[0141] (4) Effects of the composition on SCFAs and flora in adult cat feces

[0142] The results of the SCFAs content test in adult cat feces are shown in Table 4. It can be seen from Table 4 that among all adult cat groups, the SCFAs concentration in the feces of adult cats given the composition of Example 1 is the highest, which indicates that the SCFAs concentration in the feces of adult cats can be significantly increased after supplementing with the composition of Example 1, and its effect of promoting SCFAs production is significantly better than that of the compositions of the comparative examples. Similarly, the compositions of Examples 2-3 can also significantly increase the SCFAs concentration in feces.

[0143] Table 4 Effect of the composition on SCFAs in adult cat feces

[0144]

[0145] The results of the detection of bacterial abundance in adult cat feces are shown in Table 5, where the Chaos index represents the abundance of bacterial flora in feces.

[0146] Table 5 Effect of the composition on the abundance of bacterial flora in adult cat feces

[0147]

[0148] The results showed that the coverage index of each group was equal to 0.99, indicating that the probability of the sequence being detected in the samples of each group was equally high. After the cats in group B were invaded by pathogenic bacteria, the abundance of the flora decreased and the diversity of the flora decreased. After being invaded by pathogenic bacteria, supplementing the diet with the composition of Example 1 can increase the abundance and diversity of the flora, and its effect is better than that of each comparative example. Similarly, supplementing the diet with the composition of Examples 2-3 can also increase the abundance and diversity of the flora and improve the chaos index.

[0149] The relative abundance of adult cat fecal microorganisms at the genus level is shown in Table 6.

[0150] Table 6 Effect of the composition on the level of bacterial flora in adult cat feces

[0151]

[0152]

[0153] The results show that at the genus level, the compositions of the examples and comparative examples can increase the relative abundance of Bacteroides and Bifidobacterium to a certain extent, but there are significant differences in the relative abundance of Allisonella and Escherichia-Shigella. The composition of Example 1 can significantly increase the abundance of Allisonella in feces (P < 0.01) and reduce the relative abundance of Escherichia-Shigella, and its effect is significantly better than that of each comparative example. Allisonella belongs to the Firmicutes phylum, and the histamine produced by its metabolism is an important intestinal epithelial growth factor, which can prevent intestinal mucosal damage, promote intestinal mucosal repair, and inhibit intestinal bacterial translocation. Escherichia-Shigella belongs to the Proteobacteria phylum, which can penetrate epithelial cells to cause macrophage apoptosis and release IL-1β to induce intestinal inflammation. After supplementing with the composition of Example 1, the relative abundance of the bacteria is significantly reduced, which is beneficial to repair the barrier function damage of the intestinal epithelium. Similarly, after supplementation with the composition of Example 2-3, the abundance of Allisonella in feces also increased significantly, and the relative abundance of Escherichia-Shigella decreased.

[0154] From the above results, it can be seen that for adult cats attacked by bacteria, supplementing with the composition of the present invention can accelerate the recovery of feces to normal, reduce the diarrhea rate, and significantly reduce systemic inflammation. At the same time, it can increase the beneficial intestinal bacteria Allisonella, Bacteroides, and Bifidobacterium, reduce the harmful bacteria Escherichia-Shigella, and increase the concentration of SCFAs such as acetic acid and butyric acid in feces. It has a good effect on relieving diarrhea and inflammation, and improving intestinal health and intestinal flora.

[0155] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition, characterized in that It contains: (1) butyric acid and / or a precursor substance capable of releasing butyric acid in the digestive tract; and (2) lauric acid and / or a precursor substance capable of releasing lauric acid in the digestive tract; as well as (3) arachidonic acid and / or a precursor substance capable of releasing arachidonic acid in the digestive tract; as well as (4) docosahexaenoic acid and / or a precursor substance capable of releasing docosahexaenoic acid in the digestive tract; as well as (5) eicosapentaenoic acid and / or a precursor substance capable of releasing eicosapentaenoic acid in the digestive tract; Calculated by the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the weight proportions of the components are as follows: 2-5 parts of butyric acid, 1-3 parts of lauric acid, 7-10 parts of arachidonic acid, 15-30 parts of docosahexaenoic acid and 15-30 parts of eicosapentaenoic acid.

2. The composition according to claim 1, characterized in that Calculated by the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the weight proportions of the components are as follows: 2-4 parts of butyric acid, 1-2 parts of lauric acid, 7-9 parts of arachidonic acid, 18-25 parts of docosahexaenoic acid and 18-25 parts of eicosapentaenoic acid.

3. The composition according to claim 1 or 2, characterized in that Calculated by the mass of butyric acid, lauric acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid, the ratio of the total mass of butyric acid and lauric acid to the total mass of arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid is (3-6):(40-60).

4. The composition according to any one of claims 1 to 3, characterized in that The precursor substance capable of releasing butyric acid in the digestive tract is one or more selected from butyrate, butyrate ester, and butyrate derivatives; And / or, the precursor substance capable of releasing lauric acid in the digestive tract is one or more selected from laurate, lauric acid ester, and lauric acid ester derivative; And / or, the precursor substance capable of releasing arachidonic acid in the digestive tract is one or more selected from arachidonic acid salts, arachidonic acid esters, and arachidonic acid ester derivatives; And / or, the precursor substance capable of releasing docosahexaenoic acid in the digestive tract is one or more selected from docosahexaenoic acid salt, docosahexaenoic acid ester, and docosahexaenoic acid ester derivative; And / or, the precursor substance capable of releasing eicosapentaenoic acid in the digestive tract is one or more selected from eicosapentaenoic acid salts, eicosapentaenoic acid esters, and eicosapentaenoic acid ester derivatives.

5. Any of the following uses of the composition according to any one of claims 1 to 4: (1) Use in the preparation of products for preventing, alleviating or treating diarrhea in animals; (2) Use in the preparation of products for preventing, alleviating or treating intestinal inflammation in animals; (3) Application in the preparation of products for improving intestinal health of animals; (4) Application in the preparation of products for improving intestinal flora of animals; (5) Application in the preparation of products for increasing the content of short-chain fatty acids in the intestines of animals; (6) Application in animal husbandry; (7) Use in the preparation of feed, feed additives, medicines or foods.

6. The use according to claim 5, characterized in that: The animal is a mammal; Preferably, the animal is a feline; more preferably a cat.

7. The use according to claim 5 or 6, characterized in that: The product is one selected from feed, feed additives, medicine and food.

8. The use according to any one of claims 5 to 7, characterized in that: The dosage of the product when administered to animals is 0.04-0.09 g / kg body weight / day, calculated based on the total amount of arachidonic acid, EPA, DHA, butyric acid and lauric acid.

9. A drug, characterized in that The medicine comprises the composition according to any one of claims 1 to 4.

10. A feed, characterized in that: The feed comprises the composition according to any one of claims 1 to 4.

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