Compound seaweed source composition for assisting in improving oral health of dogs and cats and application of compound seaweed source composition

Abstract

The invention relates to the technical field of pet oral health care, in particular to a compound seaweed source composition for assisting in improving oral health of dogs and cats and application of the compound seaweed source composition. The invention discloses a compound seaweed source composition for assisting in improving oral health of dogs and cats. The compound seaweed source composition comprises sodium alginate and fucoidan, the value of mannuronic acid (M) / guluronic acid (G) in the sodium alginate is 0.96-1.18; the fucoidan is prepared from mannose, glucuronic acid, galactose, xylose and fucose; the weight ratio of the sodium alginate to the fucoidan is 9: 2. The compound seaweed source composition disclosed by the invention has the effects of inhibiting the proliferation of pathogenic bacteria in the oral cavity of a pet, slowing down the rising rate of health indexes of the oral cavity of the pet, inhibiting the generation of volatile organic compounds and ammonia gas in the oral cavity, improving saliva components, slowing down the generation of dental plaque and the like, and can effectively improve the health condition of the oral cavity of the pet.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pet oral health care, in particular to a compound seaweed source composition for assisting in improving the oral health of dogs and cats and application thereof. BACKGROUND

[0002] With the vigorous development of the pet industry, more and more people keep pets, and the oral health of pets has a crucial influence on the life quality and overall health of pets. The oral health status of pets has become a concern of many pet owners and is also highly valued in pet clinical treatment. The common oral problems of pets include dental plaque, dental calculus, gingivitis, periodontitis and other oral diseases, which may cause pets to have symptoms such as halitosis, drooling, pain and anorexia, and seriously affect the life quality of pets. According to the global research data of the World Small Animal Veterinary Association (WSAVA) in 2024, the incidence of oral diseases in dogs and cats over 3 years old is as high as 82%, of which periodontal disease accounts for 68%, and has become the second largest pet health killer after obesity.

[0003] Pet oral odor is the most important problem in pet oral care. Among them, hydrogen sulfide (H2S) and methyl mercaptan (CH3SH) play a major role in oral odor. These two compounds account for more than 90% of VSCs, and they are the most important two components of volatile sulfides (VSCs). They are produced by volatile sulfides through the metabolism of sulfur-containing amino acids by anaerobic bacteria in the oral cavity, and volatile sulfides are released into the oral cavity, thereby causing halitosis. Hydrogen sulfide has a rotten egg smell, and even at very low concentrations, it can produce a noticeable odor; while methyl mercaptan is also an important component of halitosis, and its odor index is much greater than that of hydrogen sulfide.

[0004] Dental plaque is a biofilm formed by bacteria and their byproducts, saliva components, oral impurities and a small number of epithelial cells and inflammatory cells. Within a few hours after cleaning the teeth, a biofilm will begin to form on the clean tooth surface. When oral hygiene management is lacking, dental plaque will gradually accumulate and thicken. With the accumulation of dental plaque, periodontitis-causing bacteria begin to multiply in large numbers, and this process only takes less than 24 hours. The accumulation of dental plaque can lead to the formation of dental calculus, causing gingivitis in pets.

[0005] The common solutions to pet oral odor, dental plaque and gingivitis and other oral problems at present include regular tooth brushing and tooth cleaning for pets, using pet-specific toothbrushes and toothpaste, and providing appropriate food and chew toys, etc. The above methods have their applicable conditions and potential drawbacks for alleviating pet oral health problems.

[0006] Marine algae are rich in bioactive substances, such as anti-inflammatory, immune enhancement, antibacterial, and antitumor activities, and are important raw materials for developing pet oral care products with oral protection. The composition of algal polysaccharides is a complex trait influenced by species genetics, growth and development (season / age), environmental conditions (light, temperature, salinity, nutrition), and key extraction and purification processes. This high variability is a challenge in research and application, but also an opportunity to find specific active components. SUMMARY

[0007] The present application aims to provide a complex seaweed source composition for assisting in improving the oral health of dogs and cats and its application, so as to solve the problems in the prior art.

[0008] The technical scheme adopted by the present application to solve its technical problems is: In a first aspect, the present application provides a complex seaweed source composition for assisting in improving the oral health of dogs and cats, which comprises: sodium alginate and fucoidan; wherein the mannuronic acid (M) / guluronic acid (G) value of sodium alginate is 1.08-1.18; and the fucoidan is composed of mannose, glucuronic acid, galactose, xylose and fucose.

[0009] Further, the weight ratio of sodium alginate to fucoidan is 9:2.

[0010] Further, the fucoidan is composed of the following components in terms of mass percentage: 2-4% mannose, 4-6% glucuronic acid, 10-12% galactose, 7-10% xylose, and 60-75% fucose.

[0011] The complex seaweed source composition for assisting in improving the oral health of dogs and cats provided by the present application can inhibit the production of volatile organic compounds and ammonia in the oral cavity of dogs and cats, inhibit the proliferation of pathogenic bacteria such as Streptococcus mutans and Porphyromonas gingivalis in the oral cavity of dogs and cats, effectively slow down the rate of increase of the oral health index of pets, and improve the oral health status of pets.

[0012] In a second aspect, the present application further provides a complex seaweed source composition for assisting in improving the oral health of dogs and cats, which is used for preparing an oral care product for preventing oral diseases of pets.

[0013] Further, the oral care product comprises tooth powder, tooth gel, tooth cleaning spray, toothpaste, and additives for oral health related products; and the oral diseases of pets include dental caries, periodontitis, gingivitis, periodontal disease, and halitosis.

[0014] In a third aspect, the present application further provides a pet food or health product, which comprises the above-mentioned complex seaweed source composition.

[0015] The present application has the beneficial effects of: (1) The complex seaweed source composition for assisting in improving the oral health of dogs and cats provided by the present application has the effects of inhibiting the proliferation of pathogenic bacteria in the oral cavity of pets, slowing the rate of increase of the oral health index of pets, inhibiting the generation of volatile organic compounds and ammonia gas in the oral cavity, improving saliva composition, and slowing the generation of dental plaque, and can effectively improve the oral health of pets.

[0016] (2) The complex seaweed source composition for assisting in improving the oral health of dogs and cats provided by the present application has the effects of inhibiting the proliferation of pathogenic bacteria in the oral cavity of pets, slowing the rate of increase of the oral health index of pets, inhibiting the generation of volatile organic compounds and ammonia gas in the oral cavity, improving saliva composition, and slowing the generation of dental plaque, and can effectively improve the oral health of pets.

[0017] (3) The composition provided by the present application can be applied to the preparation of oral care products for preventing oral diseases of pets, and has a broad market prospect in the application of health care products for protecting the oral cavity of pets. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the drawings needed in the specific embodiments or prior art description will be briefly introduced below. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0019] Figure 1 It is the analysis chart of the influence of different Ascophyllum nodosum on the oral health index of pets in Example 1.

[0020] Figure 2 It is the HPLC analysis chart for determining the monosaccharide composition of fucan in Example 3.

[0021] Figure 3 It is the local amplification spectrum chart of NMR detection of sodium alginate in Example 3.

[0022] Figure 4 It is the detection result chart of dog mouth odor value in Example 5.

[0023] Figure 5 It is the detection result chart of dog ammonia value in Example 5.

[0024] Figure 6 It is the detection result chart of cat mouth odor value in Example 5.

[0025] Figure 7 It is the detection result chart of cat ammonia value in Example 5.

[0026] Figure 8 It is the analysis chart of the species in the dental plaque of dogs that have undergone significant changes.

[0027] Figure 9Analysis chart for species significantly changed in cat dental plaque Figure 10 Analysis chart for Porphyromonas abundance in dog and cat dental plaque. DETAILED DESCRIPTION

[0028] The following examples facilitate a better understanding of the present application, but do not limit the present application. In the following examples, the experimental methods are conventional methods unless otherwise specified. In the following examples, the test materials used are purchased from conventional biochemical reagent companies unless otherwise specified. In the following examples, the quantitative tests are set up with three repeated experiments, and the results are averaged.

[0029] Example 1: Effect of different Fucus vesiculosus polysaccharides on pet oral health index Selection and treatment of Fucus vesiculosus: three Fucus vesiculosus samples from different origins and different batches were selected, and equal amounts of samples were oven-dried at 40°C and pulverized. 0.1 mol / L HC1 and 0.15% volume of HCHO were added at a solid-liquid ratio of 1:25 (m / v), and the mixture was stirred at room temperature for 1 h. The extraction was repeated twice to obtain the corresponding Fucus vesiculosus total polysaccharides, which were numbered as Fucus vesiculosus 1 polysaccharide, Fucus vesiculosus 2 polysaccharide, and Fucus vesiculosus 3 polysaccharide, respectively.

[0030] Experimental method: 20 pet dogs aged 3-4 years were randomly divided into four groups, one of which was a control group, and the other three were experimental groups, namely Fucus vesiculosus 1 polysaccharide group, Fucus vesiculosus 2 polysaccharide group, and Fucus vesiculosus 3 polysaccharide group. Under the same feeding conditions, the three groups of pet dogs were fed with Fucus vesiculosus 1 polysaccharide, Fucus vesiculosus 2 polysaccharide, and Fucus vesiculosus 3 polysaccharide, respectively, once a day, 70 mg / kg body weight each time, for 28 days. Then the oral health index of the four groups of pet dogs was detected every week.

[0031] Oral health index detection environment: the detection was carried out in a well-ventilated environment to reduce the influence of odor on the detection results. The experimental environment should be as quiet as possible to avoid the influence of noise on the mood and behavior of the pets.

[0032] Pet status: the test dogs should be in a relaxed state to avoid excessive tension or excitement during the detection process. The test dogs should be fasted before the detection to avoid the influence of food residues on the test results.

[0033] The dental plaque detector was aimed at the pet teeth and kept vertical and close, and attention should be paid to stability to avoid shaking to ensure accuracy. The detection direction was unified, starting from the right side of the oral cavity, and the same angle and position were maintained throughout the process. The red color of the dental plaque attachment under specific light was captured by high-definition photographic equipment to ensure sufficient light and clear pictures as the basis for subsequent analysis. The following scoring criteria were used for scoring at 1w, 2w, 3w, and 4w after tooth cleaning.

[0034] The pet oral health index OHI score criteria are shown in the following table: Table 1 Pet oral health index OHI score criteria The results are shown in Figure 1 The control group and the test group pet dogs' oral health index showed a slow upward trend after self-cleaning teeth. Compared with the control group, the pet dogs' oral health index of the Chondrus crispus 3 polysaccharide group was the lowest. Therefore, Chondrus crispus 3 polysaccharide has obvious improvement effect on the oral health of pet dogs, and the effect is the best.

[0035] Example 2: Screening of antibacterial activity of different Chondrus crispus polysaccharides (1) Preparation of culture medium: The culture medium is prepared according to the BHI medium formula. The volume of each component is accurately weighed according to the properties of the culture medium. A small amount of distilled water is first added to a triangular flask, then each component is added to prevent adhesion to the bottom of the flask, and then the remaining water is used to rinse the bottle wall. After the culture medium is completely dissolved, it is sealed and placed in a sterilization pot, and sterilized at 121°C for 15 min. After the culture medium is cooled to about 50°C, it is poured into a sterile plate in a clean bench with aseptic operation. Pour about 25 mL of culture medium into a 9 cm diameter plate, shake the bottom of the plate gently to make the culture medium flat on the bottom of the plate. After complete solidification, the plate culture medium is inverted and placed in a 37°C incubator for about 30 min. After the plate is dried, it is used.

[0036] (2) Bacterial activation and preparation of bacterial suspension: Prepare a test tube containing liquid medium and two plate culture media; open in a safety cabinet, burn the top of the test tube with an alcohol lamp, then quickly drip sterile water to break it, then knock it with tweezers; 0.5 mL of liquid medium is sucked into the bacterial freeze-dried tube, fully dissolved and then injected into the liquid medium test tube, mixed well; 0.2 mL of bacterial suspension is sucked into the plate culture medium and evenly coated, and two plate culture media are obtained by repeating; the liquid medium test tube and the plate culture medium are placed in the corresponding culture conditions for culture, and the bacterial strain is used when it grows. Streptococcus mutans and Porphyromonas gingivalis are activated and cultured according to the above steps, and all bacterial strains are subcultured for 2 generations before use.

[0037] Preparation of bacterial suspension: The activated bacteria are cultured under the corresponding conditions until turbidity, the absorbance OD600 is measured to be 0.6-0.8, 1 mL of bacterial suspension is placed in a sterile centrifuge tube, and centrifuged at 5 000 r / min for 1 min.

[0038] After removing the supernatant, resuspend with 1 mL of PBS buffer, centrifuge again, remove the supernatant to remove residual medium, and resuspend with 1 mL of PBS buffer for use. 10 1 ~ 10 10Gradient dilution preparation of bacterial suspension, 4 areas in each plate, 0.1 mL of different concentrations of bacterial suspension was dropped in each area, after the liquid was completely absorbed, it was placed in the corresponding culture condition, and the viable cell count was counted after culture to determine the dilution factor, and the bacterial concentration was adjusted to 10 7 CFU / mL for the determination of minimum inhibitory concentration.

[0039] (3) Determination of minimum inhibitory concentration Determination of minimum inhibitory concentration: the inhibitory concentration was determined by two-fold dilution method, and the concentration gradient of three kinds of Ascophyllum nodosum polysaccharide solution was 10.0 mg / mL, 5.0 mg / mL, 2.5 mg / mL, 1.25 mg / mL, 0.625 mg / ml, 0.3125 mg / ml, 0.156 mg / ml, 0.078 mg / ml, 0.039 mg / mL, 0.02 mg / mL, and sterile RO water was set as control, with 3 repeats for each. After 18 h of culture in an anaerobic incubator at 37℃, the OD600 value was determined by using an enzyme-labeled instrument, and the minimum concentration with significant difference between the experimental group and the blank control group after treatment and the negative control group was the minimum inhibitory concentration.

[0040] Table 2 Minimum inhibitory concentration of different Ascophyllum nodosum polysaccharides on oral pathogenic bacteria As shown in Table 2, based on the in vitro antibacterial experiment, the minimum inhibitory concentration of Ascophyllum nodosum 3 polysaccharide against Streptococcus mutans and Porphyromonas gingivalis was 0.3125 mg / ml, and compared with Ascophyllum nodosum 1 polysaccharide and Ascophyllum nodosum 2 polysaccharide, Ascophyllum nodosum 3 polysaccharide had the best antibacterial effect.

[0041] Example 3: Qualitative and quantitative analysis of the composition of Ascophyllum nodosum 3 polysaccharide Based on the excellent effect of Ascophyllum nodosum 3 polysaccharide on pet oral health index and in vitro antibacterial experiment, Ascophyllum nodosum 3 polysaccharide sample was selected for further study, and the main components and content of Ascophyllum nodosum 3 polysaccharide were analyzed, and the specific method and steps were as follows: (1) Select Ascophyllum nodosum 3 sample, dry at 40℃, crush, add 0.1 mol / L HCl and 0.15% volume of HCHO according to the ratio of 1:25 (m / v), extract for 1 h under stirring at room temperature; extract twice, and the combined 2 times of filter residue is the Ascophyllum nodosum 3 extract.

[0042] The residue was added with 1% NaCO3 solution at a ratio of 1:15 (m / v) and extracted at 60°C for 1 h; the extraction was performed twice. Centrifugation was performed, and the supernatant was adjusted to pH = 2 with HCl; standing, filtration. The gel was dissolved with 2% NaOH, bleached with 10% NaClO, and added with 2 times the volume of ethanol for alcohol precipitation overnight. Centrifugation was performed to obtain the precipitate, which was redissolved with double-distilled water, dialyzed in a 3500 Da dialysis bag, and freeze-dried to obtain product A (sodium alginate). The filtrate was neutralized with 0.5 mol / L NaOH, centrifuged, and the supernatant was concentrated, added with ethanol to a total volume of 60%, and then stood and centrifuged. The precipitate was redissolved with double-distilled water, dialyzed in a 3500 Da dialysis bag, and freeze-dried to obtain product B (fucoidan) crude product. The yield of product A and product B was analyzed, and the results are shown in Table 3.

[0043] Table 3 Analysis of main components of sample 3 of Ascophyllum nodosum Sample Product A Sodium alginate Product B Fucoidan Yield % 9 2 (2) The monosaccharide composition of fucoidan was determined by pre-column derivatization high performance liquid chromatography, and the chromatographic conditions were as follows: chromatographic column: Agilent Eclipse XDB-C18 (150 mm x 4.6 mm, 5 μm); mobile phase: 0.1 mol phosphate buffer (p H 6.8) and acetonitrile, with a ratio of 84:16 (v / v, %); detector: ultraviolet detector; detection wavelength: 245 nm; flow rate: 1.0 mL / min; column temperature: 30°C; injection volume: 10 μL; collection time: 45 min.

[0044] The results are shown in Table 3. Figure 2 The fucoidan was composed of the following components: 3.11% mannose, 5.40% glucuronic acid, 10.63% galactose, 7.86% xylose, and 73.00% fucose.

[0045] (3) The total sugar content of each component was determined by the sulfuric acid-phenol method.

[0046] According to the determination results of the monosaccharide composition of the sample, a standard solution of 0.5 mg / mL was prepared in proportion, and the specific method was as follows: working solution: take 0, 10, 20, 30, 40, 50, 60 μL of the standard solution, and add water to 100 μL, with concentrations of 0, 0.05, 0.10, 0.15, 0.20, 0.25, and 0.30 mg / mL, respectively; add 200 μL of 6% phenol, quickly add 1.5 mL of concentrated sulfuric acid, and shake uniformly; react at 100°C for 10 min; after cooling, use a UV-spectrophotometer to measure the absorbance at 490 nm wavelength, and fit the standard curve.

[0047] Sample solution: take 5 mg of the sample, add 5 mL of ultrapure water to prepare a polysaccharide solution of 1 mg / mL, take 30 μL, and add ultrapure water to 100 μL. Other operations are the same as the working solution treatment.

[0048] The results are shown in Table 4: Table 4 Analysis of the sugar content of the main components of sample 3 of Ascophyllum nodosum Sample Sodium alginate Fucoidan Total sugar content (mg / mL) 77.06 73.90 (4) The relative molecular weight of fucoidan was determined by high performance gel permeation chromatography combined with multi-angle laser light scattering (HPGPC-MALLS).

[0049] The Ascophyllum nodosum 3 polysaccharide solution (5 mg / mL) was centrifuged at 10,000 rpm for 10 min, filtered through a 0.22 μm filter membrane, and then injected.

[0050] The chromatographic conditions were as follows: Shodex Ohpak SB-802.5 HQ (8.0 μm x 300 mm) and Shodex Ohpak SB-803 HQ (8.0 μm x 300 mm) chromatographic columns in series; flow rate: 0.6 mL / min; column temperature: 35 °C; injection volume: 100 μL; collection time: 45 min; mobile phase: 0.1 mol / L sodium nitrate solution; detector: RI differential detector and multi-angle laser light scattering (MALLS) detector.

[0051] The molecular weight of fucoidan was determined to be 4.806 x 10 5 -6.165 x 10 5 Daltons.

[0052] (5) M / G value of sodium alginate 20 mg of pure sodium alginate in sample 3 of Ascophyllum nodosum was taken and dissolved in water to form a 10 mg / mL aqueous solution, and then adjusted to pH = 1 using HCl and degraded at 100 °C for 1 h; neutralized using NaOH, dialyzed, and lyophilized. After three heavy water exchanges, 600 μL of heavy water was used for dissolution, and the supernatant was taken after centrifugation and subjected to NMR (400 MHz) detection to determine the M / G value.

[0053] The results are shown in Table 4: Figure 3 After analysis, the M / G ratio of sodium alginate was determined to be 0.96-1.18.

[0054] Example 4: In vitro antibacterial properties and synergistic effects of different seaweed extracts (1) Minimum inhibitory concentration determination: The minimum inhibitory concentration was determined by the double dilution method. The solution concentration gradient of sodium alginate 1, sodium alginate 2, and fucoidan was 10.0 mg / mL, 5.0 mg / mL, 2.5 mg / mL, 1.25 mg / mL, 0.625 mg / mL, 0.3125 mg / mL, 0.156 mg / mL, 0.078 mg / mL, 0.039 mg / mL, and 0.02 mg / mL. Sterile RO water was set as a control, with 3 replicates for each. After incubation in an anaerobic incubator at 37°C for 18 h, the OD600 value was determined using a microplate reader. The minimum concentration with a significant difference in OD600 value between the experimental group and the blank control group and the negative control group was the minimum inhibitory concentration (MIC).

[0055] Table 5 Minimum inhibitory concentration of three active ingredients on oral pathogenic bacteria (2) Synergistic antibacterial activity of different seaweed extracts Synergistic inhibition of Streptococcus mutans and Porphyromonas gingivalis: The solution concentration gradient of sodium alginate 1 and sodium alginate 2 was set as 0.1565 mg / mL, 0.3125 mg / mL, and 0.625 mg / mL, respectively. The solution concentration gradient of fucoidan was 0.039 mg / mL, 0.078 mg / mL, and 0.156 mg / mL. Sterile RO water was set as a control, with 3 replicates for each. After incubation in an anaerobic incubator at 37°C for 18 h, the OD600 value was determined using a microplate reader. The experimental number corresponding to the minimum OD600 value was the optimal synergistic combination.

[0056] Table 6 Orthogonal test scheme table Table 7 Orthogonal test results The results are shown in Tables 6 and 7. Compared with the control group, different proportions of the combination showed certain inhibition effect on Streptococcus mutans and Porphyromonas gingivalis. When the concentration of sodium alginate 1 and 2 was 0.3125 mg / mL and the concentration of fucoidan was 0.078 mg / mL, the inhibition effect on Streptococcus mutans and Porphyromonas gingivalis was the strongest.

[0057] Example 5: Effect of compounded seaweed source composition on oral odor value and ammonia gas of dogs and cats Preparation of the compound seaweed source composition: a compound seaweed source composition was prepared, which consisted of sodium alginate and fucoidan, and the weight ratio of sodium alginate to fucoidan was 9:2; the M / G value of mannuronic acid (M) in sodium alginate was 1.08; the fucoidan was composed of 3.11% mannose, 5.40% glucuronic acid, 10.63% galactose, 7.86% xylose, and 73.00% fucose.

[0058] The dogs and cats were randomly divided into two groups, one group was the control group, and the other group was the experimental group. The experimental group was fed with a certain amount of compound seaweed source composition, and the other feeding conditions were the same.

[0059] The dogs and cats were measured after tooth cleaning, 1w, 2w, 3w, and 4w after the start of the experiment, using a portable gas chromatograph (Oral Chroma) to measure the breath value of the dogs and cats.

[0060] The sampling tube was inserted into the dog's mouth to a sufficient depth to collect the breath sample, avoiding damage to the oral cavity. Close the mouth for 1 minute, during which time slowly push and pull the piston to mix the breath components evenly. After 1 minute, remove the sampling tube, wipe the top with a paper towel, and inject the Oral Chroma inlet with 1 ml of gas left. The detection starts automatically, and the instrument is kept stable during the waiting period. Record the value after the detection is completed. During this process, pay attention to keep the detection instrument stable to ensure the accuracy of the detection results. Use your hand to help the pet close their lips, and start timing after observing the instrument value rising. Take it out for 15 seconds, read the highest value and record it, and repeat the measurement three times to take the average value.

[0061] When measuring ammonia, make sure that the ammonia detector is working properly and calibrated accurately. Align the gas inlet with the dog's mouth and keep it stable to avoid gaps. Observe the instrument value rising, start timing, take it out for 15 seconds, read the highest value and record it. To improve accuracy, measure three times and take the average value. Measure after tooth cleaning, 1w, 2w, 3w, and 4w after the start of the experiment.

[0062] The results are shown in Figure 4 - Figure 7 After 2 weeks of eating the compound seaweed source composition, the breath value and ammonia concentration of the dogs and cats were significantly reduced, and the improvement effect was further enhanced after four weeks of intervention.

[0063] Example 6: Effect of compound seaweed source composition on pet oral plaque microecology Plaque collection method: 2-4 hours before sampling, the test dogs and cats were fasted and gently fixed. Use a sterile spatula to scrape the plaque along the tooth surface (from the crown to the neck of the tooth), scrape 3 times at different positions, and pay attention to only touch the area covered with plaque. After scraping, use a sterile cotton swab to wipe off the plaque and put it into a sterile EP tube, while recording the pet number and sampling time. Finally, the sample was stored at -80°C.

[0064] The plaque samples were extracted according to the kit instructions to obtain total genomic DNA samples, and the samples were stored below -20°C before further analysis. The total DNA of each plaque sample was extracted using magnetic bead method. The qualified DNA samples were subjected to library preparation. The library was detected using Agilent 2100 Bioanalyzer and QPCR. After passing the library detection, the sample was sequenced. The raw data was quality controlled and dehosted using KneadData software, and the number of sequences of the species contained in the sample was calculated using Kraken2 and a self-built microbial nucleic acid database (screening the sequences of bacteria, fungi, archaea and viruses in NCBI NT nucleic acid database and RefSeq whole genome database), and the actual abundance of the species in the sample was estimated using Bracken.

[0065] SPSS 26.0 was used for statistical analysis of the data. The measurement data conforming to the normal distribution were expressed as x ± s, and the t-test was used for comparison between groups. R software (Version 4.1.2) was used for Alpha diversity index and relative abundance difference analysis between groups, and P<0.05 was considered statistically significant.

[0066] Alpha diversity index is a comprehensive index used to measure the species richness and evenness of a specific ecological sample. The main measurement indexes include chao, ace, shannon, simpson, etc. Among them, the indexes reflecting the richness of the community are chao and ace, and the indexes reflecting the diversity of the community are shannon and simpson.

[0067] Table 8 Analysis of plaque Alpha diversity index The results are shown in Table 8. The Coverage of each group was higher than 0.99, indicating that the species in the sample were basically detected. There was no significant difference in chao, ace, shannon and simpson between groups, indicating that the complex seaweed source composition had no significant effect on the Alpha diversity of the dog and cat plaque.

[0068] As Figure 8 - Figure 10Compared with the control group, the abundance of Fusobacterium, Christensenella R-7 group, Campylobacter, and Peptoniphilus in the dental plaque of the experimental group dogs was significantly reduced. Compared with the control group, the abundance of Odoribacter, Peptoniphilus sp. strain. Strain.W5053, Prevotella 195, Porphyromonas, and Eubacterium ATCC_35896 in the dental plaque of the experimental group cats was significantly reduced, indicating that the compound seaweed source composition can effectively improve oral health.

[0069] Finally, it should be noted that: the above only for the preferred embodiments of the present application, and not for the purpose of limiting the present application, although the foregoing detailed description of the present application is made with reference to the foregoing embodiments, for those skilled in the art, it still can be modified, or part of the technical features of the equivalent replacement of the technical solutions described in the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A compound seaweed-derived composition for assisting in improving oral health in dogs and cats, characterized in that, Its composition includes: sodium alginate and fucoidan; the mannuronic acid (M) / guluronic acid (G) ratio in the sodium alginate is 0.96-1.18; the fucoidan is composed of mannose, glucuronic acid, galactose, xylose and fucose.

2. The compound seaweed-derived composition for assisting in improving oral health in dogs and cats according to claim 1, characterized in that, The weight ratio of sodium alginate to fucoidan is 9:

2.

3. The compound seaweed-derived composition for assisting in improving oral health in dogs and cats according to claim 1, characterized in that, The fucoidan, calculated by weight percentage, consists of the following components: 2-4% mannose, 4-6% glucuronic acid, 10-12% galactose, 7-10% xylose, and 60-75% fucose.

4. The compound seaweed-derived composition for assisting in improving oral health in dogs and cats according to claim 1, characterized in that, The composition can inhibit the production of volatile organic compounds and ammonia in the oral cavity of dogs and cats.

5. The compound seaweed-derived composition for assisting in improving oral health in dogs and cats according to claim 1, characterized in that, The composition can inhibit the proliferation of oral pathogens in dogs and cats, such as Streptococcus mutans and Porphyromonas gingivalis.

6. The application of a compound seaweed-derived composition according to claim 1 for assisting in improving oral health in dogs and cats, characterized in that, Used to develop oral care products for the prevention of oral diseases in pets.

7. The application according to claim 6, characterized in that, The oral care products include tooth powder, teething gel, teething spray, toothpaste, and additives for oral health-related products.

8. The application according to claim 6, characterized in that, The oral diseases in pets include tooth decay, periodontitis, gingivitis, periodontal disease, and halitosis.

9. A pet food or health product, characterized in that, The pet food and health products contain the compound seaweed source composition as described in claim 1.

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