Cat food containing high-flax pickled Chinese cabbage seeds

Abstract

The invention discloses cat food containing high-linolenic-acid Chinese cabbage seeds, and belongs to the technical field of pet food. The cat food containing high-flax pickled Chinese cabbage seeds comprises the following components in parts by mass: 30-32 parts of chicken powder; 9 to 12 parts of flour; 9-12 parts of fish meal; 5-7 parts of beet pulp; 9-12 parts of poultry fat; 1.8 to 2.5 parts of chicken liver powder; 0.18 to 0.23 part of taurine; 0.05 to 0.07 part of a premix; 25 to 28 parts of puffed corn; and 3.8-4.2 parts of pickled Chinese cabbage seeds with high linseed content. The cat food containing the high-flax pickled Chinese cabbage seeds can improve the excrement quality of pet cats, improve the body health by reducing the concentration of glucose and inflammatory factors in serum, improve the hair quality by increasing the height of hair scales and reducing the thickness of the scales, and has no adverse effects on blood routine indexes, serum biochemical indexes and excrement microorganisms.

Description

Technical Field

[0001] The invention belongs to the technical field of pet feed, and in particular relates to cat food containing high-linolenic acid rapeseed. Background Art

[0002] At present, the pet industry is developing in the direction of high quality, high-end, technological and segmented. Pet cats are popular domestic pets. Improving the hair quality of pet cats and maintaining the health of pet cats are of great significance to the lives of pet owners and the development of the pet industry.

[0003] Polyunsaturated fatty acids (PUFAs) can improve the skin and hair quality of animals. Common pet hair care products on the market are rich in n-3PUFAs, such as fish oil supplements or functional food rich in deep-sea fish oil. Fish oil is rich in n-3PUFAs, but it may contain harmful substances such as dioxins and polychlorinated biphenyls. At the same time, fish oil is easily oxidized during storage, resulting in nutrient loss. In addition, the production and quality of marine fish are easily affected by factors such as season, climate and overfishing, which seriously limits the application of animal n-3PUFAs.

[0004] As vegetable oil has the advantages of stable supply, low price and less harmful substances, vegetable oil rich in α-linolenic acid is now more preferred as a source of n-3PUFAs. Rapeseed is a major oil crop, but the linolenic acid content of rapeseed is only about 8%, which is relatively low. Studies have shown that adding high-linolenic rapeseed oil to feed can effectively improve the health, antioxidant properties and nutritional content of yellow catfish.

[0005] At present, due to the low content of α-linolenic acid in rapeseed, rapeseed and other ingredients are rarely added to pet feeds such as pet cat food to provide the effective ingredient of α-linolenic acid, resulting in the high price of pet feed currently sold with fish oil ingredients rich in n-3 PUFAs, which is not conducive to the promotion and application of high-quality pet feed. Summary of the invention

[0006] The embodiment of the present invention provides a cat food containing high-linolenic acid rapeseed, aiming to solve the problem that the existing n-3 PUFAs rich in fish oil are easily oxidized and have high cost; and the α-linolenic acid content of ordinary rapeseed is low and the addition of it does not improve the skin and hair quality of animals.

[0007] The technical solution of the present invention is achieved in this way:

[0008] A cat food containing high-linolenic acid rapeseed, comprising the following components in parts by weight:

[0009]

[0010] Further preferably, the cat food containing high-linolenic acid rapeseed comprises the following components in parts by weight:

[0011]

[0012] Preferably, the high-linolenic acid rapeseed has an α-linolenic acid content of 35% or more. Further preferably, the high-linolenic acid rapeseed has an α-linolenic acid content of 36.82%.

[0013] Further preferably, the high linolenic acid rapeseed is high linolenic acid rapeseed overexpressing CsFAD2 and CsFAD3 genes.

[0014] Preferably, the premix consists of: CuCl2, FeSO4, ZnSO4, MnSO4, KIO3, Na2SeO3, CoSO4, vitamin A, vitamin B1, vitamin B2, vitamin E, vitamin C, vitamin B12, and vitamin B6.

[0015] Further preferably, each kilogram of the premix contains CuCl2 1-5 mg, FeSO4 2-5 mg, ZnSO4 2-4 mg, MnSO4 1-2 mg, KIO3 0.5-2 mg, Na2SeO3 0.5-1 mg, CoSO4 0.5-1 mg, vitamin A 4000-5000 IU, vitamin B1 5-15 mg, vitamin B2 3-5 mg, vitamin E 50-100 IU, vitamin C 200-400 mg, vitamin B12 0.05-0.10 mg, and vitamin B6 10-20 mg.

[0016] The percentage content of the nutritional level of the cat food containing high linolenic acid rapeseed is as follows:

[0017]

[0018] Furthermore, the percentage content of the nutritional level of the cat food containing high-linolenic acid rapeseed is as follows:

[0019]

[0020] The present invention selected 30 Chinese rural cats in good health, similar age and weight (3.53±1.16kg) through experiments, and randomly divided them into a control group (CON group), a wild-type rapeseed group (WT group) and a high-linolenic rapeseed group (TR group), with 10 replicates in each group. The control group was fed a basic diet, and the wild-type rapeseed group and the high-linolenic rapeseed group were fed a test diet supplemented with 4% wild-type rapeseed and 4% high-linolenic rapeseed, respectively. The experiment was divided into a pre-test period of 4d and a formal period of 40d. The test results showed that:

[0021] (1) Growth performance results showed that there were no significant differences in initial body weight, final body weight, and daily feed intake between the high-linolenic rapeseed group and the wild-type rapeseed group compared with the control group. The fecal scores of cats in the high-linolenic rapeseed group were significantly higher than those in the control group.

[0022] (2) Routine blood tests showed that there were no significant differences in the routine blood tests of the cats in each group, but the RDWc of the cats in the wild-type rapeseed group showed a downward trend compared with the control group. This indicates that the overall health status of the cats in the control group and the high-linolenic acid rapeseed group was relatively good, and the addition of high-linolenic acid rapeseed did not have a negative impact on the health status of the cats, while the wild-type rapeseed could lead to a decrease in the RDWc index, thus affecting the overall health status of the cats.

[0023] (3) Serum indicators showed that compared with the control group, the serum GLU concentration of cats in the high-linolenic acid rapeseed group was significantly reduced, and the serum IL-4 content was significantly increased. Compared with the control group, the serum T-AOC of cats in the wild-type rapeseed group was significantly reduced, while there was no significant difference in T-AOC of cats in the high-linolenic acid rapeseed group.

[0024] (4) Hair quality indicators showed that on the 40th day, there was no significant difference in hair score, hair length and hair diameter among the cats in each group. The hair scale height of the cats in the high-linolenic rapeseed group was significantly higher than that in the wild-type rapeseed group, and the hair scale thickness was significantly lower than that in the control group. The hair scale thickness of the cats in the high-linolenic rapeseed group decreased significantly on the 40th day compared with the 0th day.

[0025] (5) The results of fecal microbial diversity analysis showed that there were no significant differences in Chao1, ACE, Shannon, and Simpson indexes among the three groups. The results of PERMANOVA group detection showed that there were no significant differences in the composition of microbial flora among the three groups. There were no significant differences in the phylum, family, and genus levels of fecal microorganisms among the three groups. The dominant microbial flora at the phylum level were Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria; at the family level, the relatively high abundances were Rhodotorulaceae, Veillonellaceae, Lachnospiraceae, and Erysipelothrixaceae; at the genus level, Collinsella and Megamonas were the two main genera at the genus level of cat fecal microorganisms.

[0026] In summary, the technical solution of the present invention has the following beneficial effects compared with the prior art:

[0027] 1. Cat food containing high-linolenic acid rapeseed (adding 4% high-linolenic acid rapeseed) can improve the quality of pet cat feces, promote body health by reducing the concentration of glucose and inflammatory factors in serum, improve hair quality by increasing the height of hair scales and reducing the thickness of scales, and has no adverse effects on routine blood indicators, serum biochemical indicators and fecal microorganisms.

[0028] 2. The linolenic acid content of ordinary rapeseed is only about 8%. The linolenic acid content of ordinary rapeseed is low. A small amount of ordinary rapeseed cannot fully exert the effect of n-3 polyunsaturated fatty acids, and a large amount of ordinary rapeseed will affect the taste of pet feed and the overall health of pets. The present invention uses high-linolenic rapeseed that overexpresses CsFAD2 and CsFAD3 genes to increase the linolenic acid content in rapeseed as the source of high-linolenic rapeseed (the α-linolenic acid content of high-linolenic rapeseed is more than 35%), and through feeding experiments, the effects of cat food containing high-linolenic rapeseed on the growth performance, blood routine and serum biochemical indicators, hair quality and fecal microbiome of pet cats are explored. The results show that cat food containing high-linolenic rapeseed can improve the fecal quality of pet cats, improve the health of the body by reducing the concentration of glucose and inflammatory factors in serum, and improve the hair quality by increasing the height of hair scales and reducing the thickness of scales, and has no adverse effects on blood routine indicators, serum biochemical indicators and fecal microorganisms; its effect is significantly better than that of adding the same amount of wild-type rapeseed. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 A reference diagram of the Weihao stool scoring system used in the examples;

[0030] Figure 2 The results of the effect of high linolenic acid rapeseed on cat hair diameter in the examples are shown in Figures A, B, and C. Figures A, B, and C are the hair diameters of the CON, WT, and TR groups at 40 days, Figure D is the hair diameters of the three groups at 40 days, Figures E and F are the hair diameters of the TR group at 0 days and 40 days, and Figure G is the hair diameters of the TR group at 0 days and 40 days. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

[0031] Figure 3 The results of the effect of high-linolenic rapeseed on the height of cat hair scales in the examples are shown in Figures A, B, and C, which are the heights of hair scales in the CON, WT, and TR groups at 40 days, Figure D is the height of hair scales among the three groups at 40 days, Figures E and F are the heights of hair scales in the TR group at 0 days and 40 days, and Figure G is the height of hair scales in the TR group at 0 days and 40 days. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

[0032] Figure 4The results of the effect of high linolenic acid rapeseed on the thickness of cat hair scales in the examples are shown in Figures A, B, and C, which are the thickness of hair scales in the CON, WT, and TR groups at 40 days, Figure D is the thickness of hair scales among the three groups at 40 days, Figures E and F are the thickness of hair scales in the TR group at 0 days and 40 days, and Figure G is the thickness of hair scales in the TR group at 0 days and 40 days. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

[0033] Figure 5 This is a result diagram of the fecal microbiome dilution curve based on the Alpha diversity index in the example.

[0034] Figure 6 It is a Venn diagram based on the OTU level in the embodiment.

[0035] Figure 7 This is a PCoA analysis result diagram based on the OTU level and Bray-Curtis algorithm in the embodiment.

[0036] Figure 8 Graph showing the abundance of fecal microorganisms in the examples.

[0037] Fig. 9 Graph showing the abundance of fecal microorganism families in the examples.

[0038] Fig.10 Graph showing the abundance of fecal microorganisms at the genus level in the examples.

[0039] Fig.11 This is a graph showing the results of the correlation analysis between fecal microbial flora and inflammatory factors in the examples.

[0040] Fig.12 This is a graph showing the correlation analysis results between fecal microbial flora and antioxidant indicators in the examples. DETAILED DESCRIPTION

[0041] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only 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.

[0042] In addition, the technical solutions between the various embodiments can be combined with each other, but it must be based on the fact that ordinary technicians in the field can implement it. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0043] At present, existing n-3PUFAs are mostly provided by fish oil, which is easily oxidized and has high cost; and the α-linolenic acid content of ordinary rapeseed is low and does not improve the skin and hair quality of animals after adding it. In order to solve the above technical problems, the present invention proposes a cat food containing high-linolenic rapeseed.

[0044] Example 1

[0045] The present invention selects 30 Chinese rural cats with good health, similar age and weight (3.53±1.16kg) through experiments, and randomly divides them into a control group (CON group), a wild-type rapeseed group (WT group) and a high-linolenic rapeseed group (TR group), with 10 replicates in each group. The control group is fed with a basic diet, and the wild-type rapeseed group and the high-linolenic rapeseed group are fed with a test diet supplemented with 4% wild-type rapeseed and 4% high-linolenic rapeseed, respectively. The experiment is divided into a pre-test period of 4d and a formal period of 40d.

[0046] 1. Test materials and test methods

[0047] 1.1 Determination of fatty acid content in wild-type rapeseed and high-linolenic rapeseed

[0048] Wild-type rapeseed (wild-type rapeseed is ordinary rapeseed without overexpression of CsFAD2 and CsFAD3 genes) and high-linolenic rapeseed (the high-linolenic rapeseed is high-linolenic rapeseed overexpressing CsFAD2 and CsFAD3 genes. The high-linolenic rapeseed overexpressing CsFAD2 and CsFAD3 genes comes from the College of Plant Science and Technology of Huazhong Agricultural University, and is published in MolBreed.2024Jan 29, Improving linolenic acid content in rapeseed oil by overexpression of CsFAD2 and CsFAD3 genes.), the fatty acid compositions of wild-type rapeseed and high-linolenic rapeseed are shown in Table 1,

[0049] Table 1 Fatty acid content in wild rapeseed and high linolenic acid rapeseed

[0050]

[0051]

[0052] Determination showed that the content of α-linolenic acid (C18:3n3) in wild-type rapeseed is 8.37%, while the content of α-linolenic acid (C18:3n3) in high-linolenic acid rapeseed reaches 36.82%. The content of α-linolenic acid (C18:3n3) in high-linolenic acid rapeseed is significantly higher than that in conventional wild-type rapeseed.

[0053] 1.2 Experimental animals, groups and experimental time

[0054] Methods Thirty Chinese rural cats in good health, with similar age (2-4 years old) and weight (3.53±1.16kg) were selected and randomly divided into three groups: control group (CON group), wild-type rapeseed group (WT group), and high-linolenic rapeseed group (TR group), with 10 cats in each group; the pre-test period was 4 days and the formal test period was 40 days.

[0055] 1.3 Experimental diet and feeding management

[0056] As shown in Table 2, the composition and nutritional level of the three diets for the experimental cats meet the requirements of the Chinese national standard GB / T 31217-2014 Complete Pet Food Cat Diet. The control group was fed with a basal diet, and the wild-type rapeseed group and the high-linolenic rapeseed group were fed with the experimental diets supplemented with 4% wild-type rapeseed and 4% high-linolenic rapeseed, respectively. Each cat was fed 70-130 g of cat food at 08:30 every day. The experimental cats were kept in single cages with free access to water. The indoor temperature was 26°C and the relative humidity was 46%. The cat cages were fully cleaned and disinfected before the start of the experiment, and the experimental cats were given epidemic prevention and deworming according to routine procedures. The experimental site was cleaned and disinfected regularly during the experiment.

[0057] Table 2 Daily diet formula and nutritional level of pet cats (dry matter basis)

[0058]

[0059] Note: ① The composition of premix is: CuCl2, FeSO4, ZnSO4, MnSO4, KIO3, Na2SeO3, CoSO4, VA, VB1, VB2, VE, VC, VB12, VB6. ② The nutrient composition of each feed is calculated value.

[0060] Each kilogram of the premix contains CuCl2 1-5mg, FeSO4 2-5mg, ZnSO4 2-4mg, MnSO4 1-2mg, KIO3 0.5-2mg, Na2SeO3 0.5-1mg, CoSO4 0.5-1mg, vitamin A 4000-5000IU, vitamin B1 5-15mg, vitamin B2 3-5mg, vitamin E 50-100IU, vitamin C 200-400mg, vitamin B12 0.05-0.10mg, and vitamin B6 10-20mg. 2. Test items and methods

[0061] 2.1 Growth performance determination

[0062] All experimental cats were weighed individually on the 0th day of the formal period, and the initial body weight (IBW) was recorded. They were weighed on the 40th day of the experiment to obtain the final body weight (FBW). The amount of residual feed was recorded at 8:30 am every day, and the feed intake and average daily feed intake (ADFI) were calculated.

[0063] Average daily feed intake (ADFI) = total feed intake (g) / days (d);

[0064] During the experiment, cats were scored for feces (FS) every day according to the following criteria: Figure 1 and Table 3, referring to the Waltham stool scoring system (The The Faeces Scoring System (WFS) divides stool characteristics into 9 levels from 1 to 5:

[0065] Table 3. Waltham feces scoring system

[0066]

[0067] 2.2 Determination of hair quality indexes

[0068] Hair samples were collected on the 0th and 40th days of the experiment. On the 0th day, a pet-specific electric hair clipper was used to shave the hair of about 4cm×4cm on both sides of the middle of the lower back, close to the skin. The hair sample was sealed and dried for storage. On the 40th day, new hair was shaved from the same position to measure the hair length and hair scale structure index. The hair length was measured by using a ruler to select the guard hair.

[0069] The scale structure of the hair samples was observed using a field emission scanning electron microscope (Tianmei Scientific Instrument Co., Ltd., SU8010). The dried samples were fixed on a short aluminum plate with double-sided tape, and a metal film was coated on the samples using a vacuum coater to make them conductive. Sample images were captured at 700, 1500, and 15,000 times magnification under a voltage of 15 kV. The diameter, scale height, and scale thickness of the hair samples were measured using image J.

[0070] On the 40th day, the hair of the test cats was scored under the same lighting conditions. The hair scoring criteria are shown in Table 4, referring to the method of Rees et al. (2001).

[0071] Table 4: Pet dog and cat hair scoring standards

[0072]

[0073] 2.3 Blood routine and serum index determination

[0074] On the 40th day of the experiment, 5 ml of blood was collected from the jugular vein and divided into EDTA anticoagulant tubes and ordinary blood collection tubes. The whole blood in the EDTA anticoagulant tubes was used to measure the blood routine indicators using a routine blood test automatic analyzer (Zoetis, HM5 five-category routine blood test).

[0075] The ordinary blood collection tube was left at room temperature for 30 minutes, and then centrifuged at 3000 rmin for 15 minutes. The supernatant was placed in a 1.5 mL cryotube and stored in a -80°C refrigerator for later use. It was used to measure serum biochemical indices, serum immunoglobulins, inflammatory factors, and antioxidant indices.

[0076] Alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), total protein (TP), creatinine (CR), triglyceride (TG), cholesterol (TC), alkaline phosphatase (ALP), glucose (GLU), urea (UREA), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) in serum were detected by automatic biochemical analyzer (Shandong Bo Ke Biological Industry Co., Ltd., BK-280).

[0077] Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total cholesterol (T-AOC) and malondialdehyde (MDA) in serum were detected by using an enzyme label detector (USCNK, SMR60047) according to the method provided by the kit of Nanjing Jiancheng Company. Catalase (CAT) was detected by using a spectrophotometer (721G, a subsidiary of Shanghai Yidian Holding Company) according to the method provided by the kit of Nanjing Jiancheng Company.

[0078] The levels of immunoglobulin G, immunoglobulin M, interleukin 6, interleukin 4 and tumor necrosis factor α in serum were detected by ELISA kit (ELISA Biotechnology, MM-6752502).

[0079] 2.4 Fecal microbial sequencing

[0080] Fresh fecal samples were collected on day 40 of the experiment and transferred to 5 mL sterile fecal collection tubes for fecal microbial sequencing. The specific sequencing process is as follows:

[0081] (1) Sample DNA extraction

[0082] Total genomic DNA of fecal microorganisms was extracted according to the instructions of TIANamp Stool DNA Kit (DP328). The concentration and quality of the extracted DNA were detected using NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and 1% agarose gel electrophoresis, respectively.

[0083] (2) PCR amplification

[0084] The forward primer 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and the reverse primer 806R (5'-GGACTACHVGGGTWTCTAAT-3') were used to generate the PCR product. Amp9700) was used to amplify the V3-V4 region of the bacterial 16S rRNA gene. The relative concentration and fragment length of the PCR amplicon in each sample were determined using 2% agarose gel. The amplicon was then purified, quantified, and normalized using AgencourtAMPure Beads (Beckman Coulter, Indianapolis, IN) to form a sequencing library. The constructed library was first subjected to library quality inspection, and the library that passed the quality inspection was sequenced using Numina Hiseq 2500.

[0085] (3) Sequencing data analysis

[0086] UPARSE (version v.7.10) software was used to cluster the sequences at a similarity level of 97% to obtain operational taxonomic units (OTUs), and the OTUs were annotated taxonomically (at the phylum, family, and genus levels) based on the bacterial taxonomy database. The authenticity of the sequencing results and the feasibility of the sequencing depth were reflected by OTU coverage and the Shannon index based on the OTU level. At a similarity level of 97%, the Venn diagram can be used to display the number of common or unique OTUs between different groups. MOTHUR software (version v.1.30) was used to evaluate the sample Alpha diversity index by changes in Shannon, Simpson, ACE, Chao1, and PD_whole_tree indices. QIME software (QIIME, v1.8.0) was used to evaluate the sample Beta diversity using principal component analysis (PCoA) based on the Bray-Curtis algorithm.

[0087] 2.5 Data processing and analysis

[0088] The experimental data were preliminarily processed in Excel 2019 and then analyzed by single-factor AVONA in SPSS 25.0. The Duncan method was used for multiple comparisons and significance analysis. Welch's ANOVA was used in the case of unequal variance, and then Dunnett's T3 test was used for comparative analysis. The hair scale structure data were processed and analyzed using GraphPad Prism 9 software and plotted. The statistical results were expressed in the form of "mean ± standard deviation" (Mean ± SD), with P < 0.05 as the criterion for judging the significance of the difference, P < 0.01 as the criterion for judging the extremely significant difference, and 0.05 ≤ P < 0.10 indicating a trend of significant difference.

[0089] 3 Results and analysis

[0090] 3.1 Effects of high-linolenic acid rapeseed on growth performance in cats

[0091] The effect of high-linolenic acid rapeseed on the growth performance of cats is shown in Table 4. There was no significant difference in the initial body weight, final body weight and daily feed intake of cats in each group (P>0.05). The fecal score of cats in the high-linolenic acid rapeseed group was significantly higher than that in the control group (P<0.05), and the fecal score was maintained at a relatively healthy score (2.66 points).

[0092] Table 4 Effects of high-linolenic acid rapeseed on body weight, food intake and fecal scores of cats

[0093]

[0094] 3.2 Effects of high-linolenic acid rapeseed on routine blood indicators in cats

[0095] The effects of high-linolenic rapeseed on the blood routine indices of cats are shown in Table 5. There was no significant difference in the blood routine indices of cats in each group (P>0.05), but the RDWc of cats in the wild-type rapeseed group showed a downward trend compared with the control group (P<0.1). That is, adding 4% high-linolenic rapeseed and wild-type rapeseed had no significant effect on the blood routine indices of cats.

[0096] Table 5 Effects of high-linolenic acid rapeseed on routine blood indicators in cats

[0097]

[0098]

[0099]

[0100] 3.3 Effects of high-linolenic acid rapeseed on cat serum parameters

[0101] 3.3.1 Effects of high-linolenic acid rapeseed on cat serum biochemical parameters

[0102] The effects of high-linolenic rapeseed on cat serum biochemical indices are shown in Table 6. The serum GLU concentration of cats in the high-linolenic rapeseed group was significantly lower than that in the control group (P < 0.05), and there was no significant difference in the serum GLU concentration of cats in the wild-type rapeseed group and the control group (P > 0.05). There was no significant difference in the contents of TP, TG, ALT, AST, ALP, ALB, TC, LDL-C, HDL-C, UREA and CR among the groups (P > 0.05).

[0103] Table 6 Effects of high linolenic acid rapeseed on cat serum biochemical parameters

[0104]

[0105]

[0106] 3.3.2 Effects of high-linolenic acid rapeseed on cat serum inflammatory factors and immunoglobulins

[0107] The effects of high-linolenic rapeseed on cat serum immunoglobulin and inflammatory factors are shown in Table 7. The IL-4 content in the serum of cats in the high-linolenic rapeseed group was significantly higher than that in the control group (P < 0.05). There were no significant differences in the contents of IgG, IgM, IL-6 and TNF-α among the groups (P > 0.05).

[0108] Table 7 Effects of high linolenic acid rapeseed on cat serum inflammatory factors and immunoglobulins

[0109]

[0110]

[0111] 3.3.3 Effect of high-linolenic acid rapeseed on the antioxidant capacity of cat serum

[0112] The effect of high-linolenic acid rapeseed on cat serum antioxidant indexes is shown in Table 8. The wild-type rapeseed group cat serum T-AOC was significantly lower than the control group and high-linolenic acid rapeseed group (P < 0.05). There was no significant difference in the MDA content, SOD activity, GSH-px activity, and CAT content of the cat serum among the groups (P > 0.05).

[0113] Table 8 Effect of high linolenic acid rapeseed on the antioxidant capacity of cat serum

[0114]

[0115] 3.4 Effect of high-linolenic acid rapeseed on cat hair quality

[0116] 3.4.1 Effects of high-linolenic acid rapeseed on hair score and hair length in cats

[0117] The effects of high-linolenic acid rapeseed on cat hair score and hair length are shown in Table 9. On the 40th day, the hair score of cats in the high-linolenic acid rapeseed group was 10.86% higher than that in the control group and the wild-type rapeseed group, but the difference was not significant (P>0.05). There was no significant difference in the hair length of cats in each group (P>0.05).

[0118] Table 9 Effects of high linolenic acid rapeseed on hair score and length of cats on day 40

[0119]

[0120] 3.4.2 Effect of high-linolenic acid rapeseed on cat hair diameter

[0121] Effects of high-linolenic acid rapeseed on cat hair diameter Figure 2 As shown, compared with the control group and wild-type rapeseed group, the hair diameter of cats in the high-linolenic rapeseed group decreased by 12.86% and 6.26%, respectively, but there was no significant difference (P>0.05). There was no significant difference in hair diameter between the 40th day and the 0th day in the high-linolenic rapeseed group (P>0.05).

[0122] 3.4.3 Effect of high-linolenic acid rapeseed on cat hair scale height

[0123] Effects of high-linolenic acid rapeseed on cat hair scale height Figure 3As shown, on the 40th day, the hair scale height of the cats in the high-linolenic rapeseed group was significantly higher than that in the wild-type rapeseed group (P < 0.0001). There was no significant difference in the hair scale height of the cats in the high-linolenic rapeseed group on the 40th day compared with the 0th day (P > 0.05).

[0124] 3.4.4 Effect of high-linolenic acid rapeseed on cat hair scale thickness

[0125] Effects of high-linolenic acid rapeseed on cat hair scale thickness Figure 4 As shown, on the 40th day, the hair scale thickness of the cats in the high-linolenic rapeseed group was significantly lower than that in the control group (P < 0.05). The hair scale thickness of the cats in the high-linolenic rapeseed group decreased significantly on the 40th day compared with the 0th day (P < 0.05).

[0126] 3.5 Effects of high-linolenic acid rapeseed on cat fecal microorganisms

[0127] 3.5.1 Sample OTU analysis and dilution curve

[0128] The dilution curve results are as follows Figure 5 The curve shows the rate of new species emergence under continuous sampling. As the number of sequencing increases, the curve is steeper and in an upward stage, indicating that a large number of species have been discovered. When the curve tends to be flat, it means that the number of species in this environment will not increase significantly with the increase in the number of sequencing, which can be used for later experimental data analysis. The Venn diagram results show that ( Figure 6 ), a total of 2005 OTUs were obtained from the fecal samples of the three groups, and the number of OTUs shared by the three groups was 185, of which the numbers of OTUs unique to the control group, wild-type rapeseed group, and high-linolenic rapeseed group were 590, 616, and 450, respectively.

[0129] 3.5.2 Effect of high-linolenic acid rapeseed on the alpha diversity of cat fecal microorganisms

[0130] As shown in Table 10, there were no significant differences in Chao1, ACE, Shannon, Simpson index and PD_whole_tree index among the three groups (P>0.05).

[0131] Table 10 Effect of high linolenic acid rapeseed on cat Alpha diversity index

[0132]

[0133]

[0134] 3.5.3 Effect of high-linolenic acid rapeseed on cat fecal microbial Beta diversity

[0135] By principal coordinates analysis ( Figure 7) analysis found that the distribution of samples within the three groups was relatively concentrated, indicating that the differences in the microbial flora of the samples within the groups were small; the results of PERMANOVA between groups showed that there was no significant difference in the composition of the microbial flora among the three groups (P>0.05).

[0136] 3.5.4 Effect of high-linolenic acid rapeseed on cat fecal flora structure

[0137] Depend on Figure 8 It can be seen that the relative abundance of each group of microorganisms at the phylum level. The figure selects the top ten species in each group. The relatively high abundance at the phylum level includes Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria, which together account for more than 97% of the fecal flora. Firmicutes has the highest relative abundance in the three groups, followed by Actinobacteria. These two bacteria account for more than 84.5% and are dominant phyla. There was no significant difference in the fecal microbial phylum level among the three groups (P>0.05).

[0138] Depend on Fig. 9 It can be seen that the relative abundance of microorganisms in each group at the family level, Coriobacteriaceae, Veillonellaceae, Lachnospiraceae, Erysipelotrichaceae and Prevotellaceae were relatively high at the family level. There was no significant difference in the family level of fecal microorganisms among the three groups (P>0.05).

[0139] Depend on Fig.10 It can be seen that the relative abundance of microorganisms in each group at the family level, the highest relative abundance of each group of bacteria is Collinsella, followed by Megasphaera, Solobacterium, Blautia, Prevotella_9, etc. There was no significant difference in the genus level of fecal microorganisms among the three groups (P>0.05).

[0140] 3.5.5 Correlation analysis between fecal microorganisms and inflammatory factors

[0141] In order to further explore the relationship between changes in cat fecal microbial flora and inflammatory factors, the correlation analysis between the more abundant bacterial genera and serum inflammatory factors was performed. Fig.11 It was found that Solobacterium was negatively correlated with TNF-α (P<0.05).

[0142] 4.5.6 Correlation analysis between fecal microorganisms and antioxidant indicators

[0143] In order to further explore the relationship between changes in cat fecal microbial flora and antioxidant properties, the correlation analysis between the more abundant bacterial genera and serum antioxidant indicators was performed. Fig.12 It was found that Collinsella and Solobacterium were positively correlated with T-AOC (P<0.05), Peptoclostridium and Blautia were positively correlated with CAT (P<0.05), Megasphaera was negatively correlated with CAT (P<0.01), and Prevotella_9 was positively correlated with SOD (P<0.05).

[0144] 5. Discussion of results

[0145] 5.1 Effects of high-linolenic acid rapeseed on growth performance in cats

[0146] In this study, compared with the control group, adding high-linolenic acid rapeseed to the diet significantly improved the fecal score of cats, maintaining their fecal score at a healthier score (2.66 points), indicating that feeding high-linolenic acid rapeseed rich in n-3 PUFAs can help improve the fecal score of cats.

[0147] 5.2 Effects of high-linolenic acid rapeseed on cat blood routine

[0148] Routine blood tests can be used to assess the overall health of animals. Red blood cells (RBC) and hemoglobin (HGB) can be used to assess the nutritional status of animals and whether they are anemic. Hematocrit (HCT) refers to the percentage of red blood cells in the whole blood. Mean corpuscular volume (MCV) refers to the average volume of a single red blood cell in the body. Mean corpuscular hemoglobin content (MCH) refers to the average amount of hemoglobin contained in each red blood cell. Mean corpuscular hemoglobin concentration (MCHC) is the ratio of hemoglobin concentration in the blood to hematocrit in the blood. Red blood cell volume distribution width (RDWC) is an indicator of red blood cell heterogeneity. The total white blood cell count (WBC), lymphocyte count (LYM) and neutrophil (NEU) count reflect the body's immune function level and inflammatory state. Platelet indicators reflect the body's coagulation function. The results of this study found that adding high-linolenic acid rapeseed and wild-type rapeseed to the diet had no significant effect on the routine blood indicators of cats, but the RBC, HGB, and HCT of cats in the high-linolenic acid rapeseed group and the control group were higher than those in the wild-type rapeseed group, and the RDWc of cats in the wild-type rapeseed group showed a downward trend compared with the control group, indicating that the overall health status of cats in the control group and the high-linolenic acid rapeseed group was relatively good, and the addition of high-linolenic acid rapeseed would not have a negative impact on the health status of cats.

[0149] 5.3 Effects of high-linolenic acid rapeseed on cat serum parameters

[0150] 5.3.1 Effects of high-linolenic acid rapeseed on cat serum biochemical parameters

[0151] Serum biochemical indicators can directly reflect the health status of the animal body. GLU content reflects the body's ability to metabolize glucose, and normal blood sugar levels help maintain physiological balance. High blood sugar levels may be one of the symptoms of diabetes, while hypoglycemia may be the result of excessive insulin or insufficient diet. In this study, compared with the control group, the addition of high-linolenic acid rapeseed to the diet significantly reduced the serum glucose concentration of cats, while wild-type rapeseed had no significant effect on the blood glucose concentration of cats, indicating that ALA in high-linolenic acid rapeseed can effectively reduce the blood glucose concentration of cats and improve the sugar metabolism of cats.

[0152] The concentrations of TC and TG in serum reflect the lipid metabolism status of the animal body. The main function of LDL-C is to transport cholesterol from the liver to extrahepatic tissues, and HDL transports cholesterol from extrahepatic tissues back to the liver. The results of this study found that feeding high-linolenic acid rapeseed in the diet had no significant effect on the concentrations of serum TC, TG, LDL-C and HDL-C in cats.

[0153] ALT and AST are important indicators of liver function. Increased concentrations of ALT and AST mean that liver function and liver cells are damaged. ALP can usually reflect the condition and severity of liver cell damage. TP concentration reflects changes in the body's protein anabolism and immune status. GLB is synthesized by immune organs and has immune function. ALB is synthesized by liver cytoplasm and can maintain plasma osmotic pressure and transport metabolites in the blood. Creatinine (CR) and UREA concentrations are important indicators of renal function and can also measure the body's nitrogen utilization capacity. The results of this study found that feeding high-linolenic acid rapeseed or wild-type rapeseed in the diet had no significant effect on the concentrations of serum TP, ALT, AST, GLB, ALP, ALB, UREA and CR in cats, indicating that 4% addition of high-linolenic acid rapeseed and wild-type rapeseed will not cause adverse effects on cat blood.

[0154] 5.3.2 Effects of high-linolenic acid rapeseed on cat serum inflammatory factors and immunoglobulins

[0155] IL-4 and IL-6 are mainly produced by Th2 cells in T lymphocytes, and TNF-α mainly comes from Th1 cells in lymphocytes. IL-6 and TNF-α are pro-inflammatory factors, and IL-4 is an anti-inflammatory factor. Linolenic acid has an anti-inflammatory effect by affecting fat metabolism in body tissues and reducing the inflammatory response of inflammatory mediators in peptides. In this study, high-linolenic acid rapeseed can significantly increase the concentration of IL-4 in cat serum, but has no significant effect on the concentrations of IL-6 and TNF-α, indicating that high-linolenic acid rapeseed can have an anti-inflammatory effect to a certain extent.

[0156] Immunoglobulin is a globulin that can be used as an antibody and has a chemical structure similar to that of an antibody. Immunoglobulins play a role in antigen-specific binding and regulating immune responses, and measuring serum immunoglobulin concentrations is one of the most commonly used methods to evaluate immune function. Serum immunoglobulins mainly include IgG, IgM, and IgA, among which IgG has the highest content and plays a major role in the body's immune regulation. In this study, the IgG concentrations in the high-linolenic rapeseed group and the wild-type rapeseed group were 17.98% and 20.35% higher than those in the control group, respectively, and the IgM concentrations were 11.8% and 9.54% higher, respectively, but there were no significant differences.

[0157] 5.3.3 Effect of high-linolenic acid rapeseed on the antioxidant capacity of cat serum

[0158] T-AOC is an overall indicator for evaluating the antioxidant capacity of an organism; the antioxidant enzyme system is an important defense system in the body to combat oxidative stress and cell damage, including a series of enzymes and other molecules that can remove free radicals and other harmful oxidizing substances to protect cells from oxidative damage. These enzymes include superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-px). SOD can dismutate oxygen free radicals to produce hydrogen peroxide, and CAT and GSH-px can decompose hydrogen peroxide into water. CAT can clean up oxygen free radicals in the body. MDA is the main active aldehyde that appears during the peroxidation reaction of polyunsaturated fatty acids in biological membranes and can be used as an indicator to judge the degree of peroxidation. Excessive intake of unsaturated fatty acids will lead to an increase in the oxidative stress index and myeloperoxidase activity in rats. The results of this study found that compared with the control group, the wild-type rapeseed group had significantly lower T-AOC in cat serum, while the high-linolenic rapeseed group had no significant difference in T-AOC. This may be due to the anti-nutritional factors contained in wild-type rapeseed, which led to a decrease in total antioxidant capacity, while the high AlA expression in high-linolenic rapeseed inhibited the decrease in antioxidant capacity caused by rapeseed addition. There were no significant differences in SOD, GSH-Px activity, CAT level, and MDA level in the serum of cats in the high-linolenic rapeseed and wild-type rapeseed groups compared with the control group.

[0159] 5.4 Effect of high-linolenic acid rapeseed on cat hair quality

[0160] 5.4.1 Effects of high-linolenic acid rapeseed on hair score and hair length in cats

[0161] Hair scoring is a subjective method for evaluating the hair quality of pets. It requires observing the glossiness, smoothness and greasiness of pet hair under the same lighting conditions to score the hair. The results of this study found that adding high-linolenic acid rapeseed to the diet can improve the hair score of cats, but there is no significant effect. Hair length is related to skin health and hair follicle development, and is regulated by multiple genes. The results of this study found that there was no significant effect on hair length in each group.

[0162] 5.4.2 Effects of high-linolenic acid rapeseed on cat hair scale structure

[0163] Scale structure is an important indicator for evaluating hair quality. The thickness, density, shape and arrangement structure of scales can directly affect the gloss, softness and feel of hair. The smaller the thickness of the scales, the smaller the friction coefficient of the hair and the smoother the hair surface. The larger the scale height, the fewer scales per unit length and the smaller the scale density, resulting in smoother hair. The larger the hair diameter, the worse its softness. The results of this study found that the addition of high-linolenic rapeseed to the diet had no significant effect on the diameter of cat hair. On the 40th day, the height of the hair scales of the cats in the high-linolenic rapeseed group was significantly higher than that of the wild-type rapeseed group, and the thickness of the hair scales was significantly lower than that of the control group, and the thickness of the hair scales of the cats in the high-linolenic rapeseed group was significantly lower than that of the 0th day. This shows that high-linolenic rapeseed can improve the hair scale structure of cats and make their hair softer and smoother.

[0164] 5.5 Effects of high-linolenic acid rapeseed on cat fecal microorganisms

[0165] The intestinal flora plays an important role in maintaining the homeostasis of the intestinal environment, improving intestinal health, promoting nutrient digestion and absorption, participating in host metabolism, and regulating the body's immune function. Diet, age, and diarrhea may affect the composition of the intestinal flora. At present, studies usually use Alpha diversity and Beta diversity to evaluate the diversity of intestinal microbial flora. The results of this experiment showed that there were no significant differences in the Chao1, ACE, Shannon, and Simpson indices among the three groups, indicating that the diversity of cat fecal flora was relatively small. When PCoA was used to analyze Beta diversity, it was found that the microbial flora of each sample within the three groups was slightly different, and the composition of the microbial flora between the three groups was not significantly different.

[0166] This study detected fecal microorganisms in the control group, wild-type rapeseed group, and high-linolenic acid rapeseed group and found that the dominant fecal microorganisms at the phylum level were Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria. The results of this experiment found that the relative abundance of Firmicutes in cat feces in the high-linolenic acid rapeseed group and wild-type rapeseed group was 4.16% and 4.49% higher than that in the control group, respectively, and the relative abundance of Actinobacteria in cat feces in the high-linolenic acid rapeseed group and wild-type rapeseed group was 7.14% and 5.97% lower than that in the control group, respectively, but neither reached a significant difference.

[0167] At the family level, the Red Bug Family can be negatively correlated with liver glucose and glycogen levels, and its increased abundance can be regarded as a sign of impaired glucose metabolism. In this study, the Red Bug Family in the high-linolenic rapeseed group was lower than that in the control group, which was consistent with the decrease in serum GLU concentration in cats in the high-linolenic rapeseed group. The Lachnospiraceae can produce short-chain fatty acids, which are beneficial for maintaining intestinal epithelial health. The results of this study showed that the Lachnospiraceae in the feces of cats in the high-linolenic rapeseed group and the wild-type rapeseed group were 30% and 32.66% higher than those in the control group, indicating that high-linolenic rapeseed and wild-type rapeseed can regulate the abundance of beneficial bacteria in the cat intestine to a certain extent and maintain intestinal homeostasis.

[0168] At the genus level, the bacteria with higher relative abundance in each group were Collinsella, Megamonas, Solobacterium, Blautia, Prevotella_9, Subdoligranulum, and unclassified_Clostridia_UCG_014. The results of this experiment showed that the Collinsella genus in the high-linolenic acid rapeseed group and the wild-type rapeseed group were 21.58% and 25% lower than those in the control group, respectively. Megasphaera is a beneficial bacterium in the intestine that can produce volatile fatty acids such as lactic acid, propionic acid, and butyric acid, which can promote nutrient absorption and inhibit the proliferation of harmful bacteria. Solobacterium belongs to the Erysipelothrix family, which plays an important role in obese patients and patients with inflammatory bowel disease. The results of this experiment showed that the abundance of Megasphaera and Solobacterium did not change significantly, indicating that the addition of 4% high-linolenic acid rapeseed and wild-type rapeseed to the diet had no significant effect on the fecal microbial flora of cats.

[0169] In summary, adding 4% high-linolenic acid rapeseed to the diet can improve the fecal quality of pet cats, promote body health by reducing the concentration of glucose and inflammatory factors in serum, improve hair quality by increasing the height of hair scales and reducing the thickness of scales, and has no adverse effects on routine blood indices, serum biochemical indices and fecal microorganisms.

[0170] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A cat food containing high-linolenic acid rapeseed, characterized in that: The composition comprises the following components in parts by weight:

2. The cat food containing high linolenic acid rapeseed according to claim 1, characterized in that: The composition comprises the following components in parts by weight:

3. The cat food containing high linolenic acid rapeseed according to claim 1 or 2, characterized in that: The high-linolenic acid rapeseed has an α-linolenic acid content of more than 35%.

4. The cat food containing high linolenic acid rapeseed according to claim 1 or 2, characterized in that: The high-linolenic acid rapeseed has an α-linolenic acid content of 36.82%.

5. The cat food containing high linolenic acid rapeseed according to claim 1 or 2, characterized in that: The high-linolenic acid rapeseed is a high-linolenic acid rapeseed that overexpresses CsFAD2 and CsFAD3 genes.

6. The cat food containing high linolenic acid rapeseed according to claim 1 or 2, characterized in that: The premix consists of: CuCl2, FeSO4, ZnSO4, MnSO4, KIO3, Na2SeO3, CoSO4, VA, VB1, VB2, VE, VC, VB12, and VB6.

7. The cat food containing high linolenic acid rapeseed according to claim 6, characterized in that: Each kilogram of the premix contains CuCl2 1-5mg, FeSO4 2-5mg, ZnSO4 2-4mg, MnSO4 1-2mg, KIO30.5-2mg, Na2SeO3 0.5-1mg, CoSO40.5-1mg, vitamin A 4000-5000IU, vitamin B1 5-15mg, vitamin B2 3-5mg, vitamin E 50-100IU, vitamin C 200-400mg, vitamin B12 0.05-0.10mg, and vitamin B610-20mg.

8. The cat food containing high linolenic acid rapeseed according to claim 1 or 2, characterized in that: The percentages of its nutritional levels are as follows:

9. The cat food containing high linolenic acid rapeseed according to claim 1 or 2, characterized in that: The percentages of its nutritional levels are as follows:

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