Application of pediococcus pentosaceus in preparation of products for improving pork quality and intestinal flora

Abstract

The invention provides application of pediococcus pentosaceus in preparation of a product for improving pork quality and intestinal flora, and relates to the technical field of animal feeding. The pediococcus pentosaceus disclosed by the invention is pediococcus pentosaceus LL2018368, is separated from the digestive tract of a Diqing Tibetan pig and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.18709. The pediococcus pentosaceus disclosed by the invention has the advantages that the pediococcus pentosaceus can be used for preparing the pediococcus pentosaceus; the pediococcus pentosaceus LL2018368 is added into drinking water of the Du-Si pigs, the influence of the pediococcus pentosaceus LL2018368 on archaea, meat quality, tissue fat-soluble vitamins and excrement metabolite composition in the excrement of the Du-Si pigs is studied, and the result shows that the pediococcus pentosaceus LL2018368 enables the species of the archaea in the excrement to tend to be consistent, so that the content of the archaea in the excrement of the Du-Si pigs in the excrement of the Du-Si pigs in the excrement of the Du-Si pigs is increased. The contents of vitamin A and all-trans-retinol in kidney tissues are increased, and the probiotic functions of amino acid and fat metabolism related bacteria and enzymes are improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of animal breeding, and specifically relates to the application of Pediococcus pentosaceus in the preparation of products for improving pork quality and intestinal flora. Background Art

[0002] The Dusa pig is a pig breed with specific genetic characteristics and production performance, mainly used for breeding and farming. It has a relatively fast growth rate and excellent muscle quality. However, currently, when raising this pig breed, there are still problems such as the abuse of feed additives, which leads to the imbalance of the pig intestinal flora and further affects the pork quality. Pediococcus pentosaceus belongs to the genus Pediococcus of the family Streptococcaceae, is a Gram-positive bacterium, and is a kind of lactic acid bacterium. Pediococcus pentosaceus has the effects of increasing the total free amino acids and antioxidant activity in the dough after fermentation, can promote the absorption of iron in the body, and reduce the content of nitrite in pickles. For a long time, Pediococcus pentosaceus has been used as a biological preservative for fermented foods. In recent years, it has shown the potential of probiotics, including anti-inflammatory, antioxidant, and antagonistic activities against pathogens. For example, after adding Pediococcus pentosaceus to the chicken diet, the level of chain fatty acids in the intestine was increased, the flavor of chicken meat was improved, and the intestinal flora structure was regulated. Regarding the probiotic characteristics of Pediococcus pentosaceus extracted from sow milk, in a pig feeding experiment, oral administration of Pediococcus pentosaceus SMM914 could reduce the potential liver damage during weaning by decreasing the levels of ALT and TP in the serum (Antioxidant potential of Pediococcus pentosaceus strains from the sow milk bacterial collection in weaned piglets, Microbiome, 2022). However, the effects of Pediococcus pentosaceus from specific sources on the growth performance, fecal flora structure, and metabolites of animals may be different. Therefore, seeking a strain that can replace feed additives is of great significance for the green breeding of Dusa pigs. Summary of the Invention

[0003] In view of this, the purpose of the present invention is to provide the application of Pediococcus pentosaceus in the preparation of products for improving pork quality and intestinal flora.

[0004] To achieve the above-mentioned invention purpose, the present invention provides the following technical solutions: The present invention provides the application of Pediococcus pentosaceus in the preparation of products for improving pork quality and intestinal flora, and the Pediococcus pentosaceus is Pediococcus pentosaceus LL2018368, which is preserved in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No. 18709.

[0005] Preferably, the product includes a feed additive.

[0006] More preferably, the addition amount of Pediococcus pentosaceus in the feed additive is 10 8 ~10 12 CFU / L.

[0007] Preferably, the improvement of pork quality includes increasing the total amount of amino acids in pork.

[0008] Preferably, the improvement of pork quality also includes increasing the contents of vitamin A and all-trans retinol in pig kidney tissues.

[0009] Preferably, the improvement of the intestinal flora includes promoting the growth of beneficial bacteria, inhibiting the growth of harmful bacteria, and improving the consistency of fecal archaea.

[0010] More preferably, the beneficial bacteria include one or more of Eubacterium, Selenomonas, Ruminococcaceae, Clostridium, and Lachnospira.

[0011] More preferably, the harmful bacteria include Methanobrevibacter.

[0012] Preferably, the breed of the pig is Dusa pig.

[0013] Compared with the prior art, the present invention has the following beneficial effects: After adding Pediococcus pentosaceus LL2018368 to the drinking water of Dusa finishing pigs in the present invention, the species of archaea in the feces of Dusa pigs tend to be consistent, the contents of isobutyric acid, butyric acid, and isovaleric acid in the feces are reduced, the contents of vitamin A and all-trans retinol in the kidneys are increased, the content of α-tocopherol in the serum is reduced, and the relative abundances of microbial functional genes related to signal transduction, cell movement, amino acid, and lipid metabolism in the feces are increased, which can improve the probiotic functions of amino acid- and lipid-metabolism-related bacteria and enzymes.

[0014] Biological preservation certificate Pediococcus pentosaceus LL2018368, classified and named as Pediococcus pentosaceus, preservation unit: China General Microbiological Culture Collection Center (abbreviated as CGMCC), address of the preservation unit: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, preservation number: CGMCC No. 18709, preservation date: October 21, 2019. Brief description of the drawings

[0015] Figure 1 It is the α-diversity analysis result of the fecal flora of Dusa pigs after adding Pediococcus pentosaceus. In the figure, A is the Shannon index based on the genus level, B is the Simpson index based on the genus level, FC is the control group sample, and FA is the experimental group sample; Figure 2Principal coordinate analysis results of the fecal microbiota of Dusa pigs after adding Pediococcus pentosaceus. In the figure, FA represents the experimental group samples, and FC represents the control group samples; Figure 3 Bar chart analysis of the species composition community of the fecal microbiota of Dusa pigs after adding Pediococcus pentosaceus. In the figure, A is at the phylum level, B is at the species level, and C is the analysis of the difference in community composition based on LDA; Firmicutes: Firmicutes, Bacteroidetes: Bacteroidetes, Proteobacteria: Proteobacteria, Spirochaetes: Spirochaetes, Fibrobacteres: Fibrobacteres, Actinobacteria: Actinobacteria, Euryarchaeota: Euryarchaeota, Verrucomicrobia: Verrucomicrobia, Lentisphaerae: Lentisphaerae, Tenericutes: Tenericutes, Fusobacteria: Fusobacteria, Planctomycetes: Planctomycetes, Chloroflexi: Chloroflexi, Chlamydiae: Chlamydiae, Cyanobacteria: Cyanobacteria, Synergistetes: Synergistetes, Chytridiomycota: Chytridiomycota, Ascomycota: Ascomycota, Mucoromycota: Mucoromycota, Others: Others; Bacteria_unclassified: Unclassified bacteria, Eubacterium: Eubacterium, Eubacteriaceae: Eubacteriaceae, Selenomonadaceae: Selenomonadaceae, Selenomonadales: Selenomonadales, Ruminococcaceae: Ruminococcaceae, Clostridium: Clostridium, Lachnospiraceae_unclassified: Unclassified Lachnospiraceae, Clostridiaceae: Clostridiaceae, Roseburia: Roseburia, Lachnospiraceae: Lachnospiraceae, Clostridia: Clostridia, Clostridiales: Clostridiales; Figure 4 Alpha diversity analysis results of the archaea in the feces of Dusa pigs after adding Pediococcus pentosaceus. A is the Simpson index based on the genus level, B is the Shannon index based on the genus level, and C is the Chao1 index based on the genus level; Figure 5 Principal coordinate analysis results of the archaea in the feces of Dusa pigs after adding Pediococcus pentosaceus. FC represents the control group samples, and FA represents the experimental group samples; Figure 6Bar chart analysis of the species composition of archaea in the feces of Dusa pigs after adding Pediococcus pentosaceus. In the figure, A is at the phylum level, B is at the genus level, and C is the analysis of the differences in community composition based on LDA; Euryarchaeota: Euryarchaeota, Candidatus_Thorarchaeota: Candidatus Thorarchaeota; Candidatus_Bathyarchaeota: Candidatus Bathyarchaeota; Crenarchaeota: Crenarchaeota; Candidatus_Lokiarchaeota: Candidatus Lokiarchaeota; Candidatus_Woesearchaeota: Candidatus Woesearchaeota; Thaumarchaeota: Thaumarchaeota; Candidatus_Altiarchaeota: Candidatus_; Candidatus_Heimdallarchaeota: Candidatus Heimdallarchaeota; Candidatus_Korarchaeota: Candidatus Korarchaeota; Candidatus_Aenigmarchaeota: Candidatus Aenigmarchaeota; Nanoarchaeota: Nanoarchaeota; Candidatus_Verstraetearchaeota: Candidatus Verstraetearchaeota; Candidatus_Diapherotrites: Candidatus Diapherotrites; Candidatus_Marsarchaeota: Candidatus Marsarchaeota; Methanobrevibacter: Methanobrevibacter; Methanosarcina: Methanosarcina; Methanoculleus: Methanoculleus; Methanosphaera: Methanosphaera; Methanocorpusculum: Methanocorpusculum; Thermococcus: Thermococcus; Methanohalophilus: Methanohalophilus; Methanobacterium: Methanobacterium; Methanothrix: Methanothrix; Methanomicrobium: Methanomicrobium; Methanococcoides: Methanococcoides; Nitrosopumilus: Nitrosopumilus; Methanococcus: Methanococcus; g__Thermoplasmata_unclassified: Unclassified Thermoplasmata; o__Thermoplasmata_unclassified: Unclassified Thermoplasmata; f__Thermoplasmata_unclassified: Unclassified Thermoplasmata; o__Methanosarcinales: Methanosarcinales; s_Thermoplasmata_archaeon:;f_Methanosarcinaceae: Methanosarcinaceae; c_Thermoprotei: Thermoprotei; c__Thermococci: Thermococci; g__Methanohalophilus: Methanohalophilus; Figure 7 To analyze the results of differential metabolites after adding Pediococcus pentosaceus using the partial least squares discriminant analysis (PLS-DA) method, FA in the figure is the sample of the experimental group, and FC is the sample of the control group; Figure 8 It is a volcano plot of differential metabolites. Specific implementation mode

[0016] The present invention provides an application of Pediococcus pentosaceus in the preparation of products for improving pork quality and intestinal flora. In the present invention, the Pediococcus pentosaceus is Pediococcus pentosaceus LL2018368, which is isolated from the digestive tract (cecum) of Diqing Tibetan pigs and preserved in the General Microbiological Center of the China Microbial Culture Collection Center. The preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; The preservation date: October 21, 2019; The preservation number is CGMCC No. 18709. Pediococcus pentosaceus LL2018368 is disclosed in the patent document ZL202110843467.2. In the present invention, the product preferably includes a feed additive. In the present invention, when the Pediococcus pentosaceus is used as a feed additive, the addition amount is preferably 10 8 ~10 12 CFU / L, more preferably 10 10 CFU / L.

[0017] After adding Pediococcus pentosaceus LL2018368 to the drinking water of the experimental group in the present invention, it can improve pork quality and the intestinal flora of pigs. In the present invention, the improvement of pork quality preferably includes increasing the total amount of amino acids in pork and increasing the contents of vitamin A and all-trans retinol in pig kidney tissues.

[0018] In the present invention, the improvement of intestinal flora includes promoting the growth of beneficial bacteria, inhibiting the growth of harmful bacteria, and improving the consistency of fecal archaea. In the present invention, the beneficial bacteria preferably include one or more of Eubacterium, Selenomonas, Ruminococcaceae, Clostridium, and Lachnospiraceae. The harmful bacteria preferably include Methanobrevibacter.

[0019] In the present invention, the breed of the pigs is preferably Dusa pigs. Experiments have shown that Pediococcus pentosaceus LL2018368 has an improvement effect on the archaea in the feces of Dusa pigs, meat quality, tissue fat-soluble vitamins, and the composition of fecal metabolites.

[0020] The technical solutions provided by the present invention will be described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0021] In the following experimental methods of the embodiments, unless otherwise specified, they are all conventional methods. The test materials used in the following embodiments are all commercially available products unless otherwise specified.

[0022] Example 1 1 Materials and Methods 1.1 Test Materials Pediococcus pentosaceus LL2018368 used in the test was isolated from the digestive tract (cecum) of Diqing Tibetan pigs by this laboratory, identified as Pediococcus pentosaceus LL2018368, and is currently preserved in the China General Microbiological Culture Collection Center CGMCC No. 18709. The composition and nutrient levels of the basal diet are shown in Tables 1 and 2. The animal feeding trial was carried out in Luquan Renhe Breeding Technology Co., Ltd. Before the start of the trial, the empty house was cleaned and disinfected, the pigs were weighed and numbered. During the feeding process, free access to food and water was adopted, and other management was carried out according to the management procedures.

[0023] Table 1 Composition of basal diet

[0024] Table 2 Nutrient levels of basal diet

[0025] 1.2 Experimental Design Twenty Dusa finishing pigs (with 50% Duroc and 50% Saba pig bloodlines) with a body weight of about 77 kg were selected as experimental animals and randomly divided into 2 groups (experimental group FA and control group FC), with 2 replicates in each group and 5 pigs in each replicate. The two groups were fed the same basal diet. Pediococcus pentosaceus LL2018368 (10 10 CFU / L) was added to the drinking water of the experimental group, and nothing was added to the drinking water of the control group. The experimental period was 28 days. After the end of the experiment, 3 pigs were randomly selected from each pen (each replicate), and fresh pig feces were collected and quickly placed in liquid nitrogen for subsequent metagenomic analysis. The animal experiment was approved by the Animal Ethics Committee of the Yunnan Academy of Animal Husbandry and Veterinary Sciences, approval number YNASVI01-2023011.

[0026] 1.3 Metagenomics Sequencing and Bioinformatics Analysis Total DNA extraction of the samples and metagenomic sequencing were completed by LC Bio Technology CO., Ltd. Hangzhou. The sequencing mode of Illumina Novaseq 6000 (LC Bio Technology CO., Ltd., Hangzhou, China) was PE150. The raw data was quality controlled and host-filtered to obtain valid data. Starting from the valid data after quality control of each sample, the reads of all samples were put together for mixed assembly in order to discover low-abundance species information in the samples. Starting from the Scaffold after mixed assembly, gene prediction was performed using Prodigal, and redundancy removal was carried out to construct a gene catalogue. Combining the valid data of each sample, the abundance information of the gene catalogue in each sample could be obtained.

[0027] 1.4 Sample collection After the experiment, 3 pigs were randomly selected from each pen (each replicate), and fresh pig feces were collected and quickly placed in liquid nitrogen for subsequent metabolomics analysis. After slaughter, the longissimus dorsi muscle was taken for meat quality analysis, and blood, liver, and kidney were taken for fat-soluble vitamin detection.

[0028] 1.5 Meat color detection Meat quality traits such as meat color were determined with reference to the agricultural industry standard "Technical Specification for Determination of Pig Muscle Quality NY / T 821-2004". The determination of muscle amino acids was processed by acid hydrolysis method and measured by liquid chromatography.

[0029] 1.6 Metabolomics analysis The detection of fecal short-chain fatty acid content and fat-soluble vitamin content in tissues was analyzed by targeted metabolomics. The sample detection was completed by LC Bio Technology CO., Ltd. Hangzhou. The analytical instrument was an ultra-high pressure liquid system LC-MS / MS (Vanquish Flex UPLC, Thermo Company, USA). The mass spectrometry condition parameters were as follows: the sampling mode was multiple reaction monitoring (MRM), the ion source condition was ESI source, and the positive ion scan mode (Positive).

[0030] 1.7 Data analysis One-way ANOVA was performed using SPSS 17.0 statistical software. P < 0.05 was considered significantly different, and P < 0.01 was considered extremely significantly different. The data was expressed as mean ± standard deviation. All metagenomic sequencing data were analyzed on the bio-cloud platform of LC Bio Technology CO., Ltd. Hangzhou.

[0031] 2 Results and analysis 2.1 Metagenomic sequencing information Twelve Dusa pig feces samples were sequenced using Illumina Novaseq 6000, resulting in a total of 1,169,302,598 raw sequences. On average, each sample had 97,441,883.17 raw sequences, and the average sequencing depth was 14.62 Gb. After quality control, 1,127,107,386 clean sequences were obtained, with an average of 93,925,615.50 clean sequences per sample. After optimizing the sequences and performing assembly and splicing, a total of 8,807,131 contigs were obtained, with an average of 7,339,277.58 per sample. After removing redundancy, 6,094,097 complete genes were obtained.

[0032] 2.2 Effects of Pediococcus pentosaceus on the fecal bacteria of Dusa pigs 2.2.1 Alpha diversity analysis The analysis of the alpha diversity index of species at the genus level is shown in Table 3 and Figure 1 . After adding Pediococcus pentosaceus LL2018368 to the drinking water, its Shannon index ( Figure 1 A in Figure 1 ) increased significantly (P < 0.05), and the Simpson index ( Figure 1 B in ) increased extremely significantly (P < 0.01). The alpha diversity of the species in the samples increased significantly, indicating that adding Pediococcus pentosaceus LL2018368 to the drinking water of Dusa pigs can significantly improve species diversity.

[0033] Table 3 Analysis of alpha diversity index

[0034] Note: Different lowercase letters in the superscripts of the same row of data indicate significant differences (P < 0.05), and different capital letters indicate extremely significant differences (P < 0.01). 2.2.2 Beta diversity analysis The principal coordinates analysis (PCoA) is shown in Figure 2 . The samples of the two groups could not be completely separated and partially overlapped, indicating that after adding Pediococcus pentosaceus LL2018368, there were both similar and different parts between the two groups of samples. Adding Pediococcus pentosaceus LL2018368 had little effect on the microorganisms with relatively low relative abundances in the fecal bacteria, and the species similarity tended to be consistent. In summary, after adding Pediococcus pentosaceus LL2018368 to the diet, some dominant components in the fecal microorganisms of pigs changed.

[0035] 2.2.3 Species composition analysis Sequencing results showed that among the annotated species, 73.40% of the genes were annotated to bacteria, 0.26% to archaea, 0.46% to viruses, 0.10% to eukaryotes, and 25.41% of the genes were not annotated to known species. A total of 186 phyla, 322 classes, 550 orders, 1112 families, 4031 genera, and 26232 species were obtained in the species annotation.

[0036] The ratio of Firmicutes to Bacteroidetes was positively correlated with host fat storage, body weight, visceral fat, and subcutaneous fat. As Figure 3 shown in A of [reference], at the phylum level, the dominant species with the highest relative abundance in the control group FC was Firmicutes (23.29%), followed by unclassified bacteria (Bacteria_unclassified, 23.04%), Bacteroidetes (19.76%), Proteobacteria (2.25%), and Spirochaetes (1.97%). Unclassified species accounted for 26.13%. The dominant species with the highest relative abundance in the experimental group FA was Firmicutes (25.45%), followed by unclassified bacteria (21.40%), Bacteroidetes (21.19%), Proteobacteria (2.02%), and Spirochaetes (1.65%). Unclassified species accounted for 25.41%. The relative abundances of Firmicutes and Bacteroidetes were the highest in both groups. After adding Pediococcus pentosaceus LL2018368, the ratio of Firmicutes to Bacteroidetes in the experimental group FA increased, but the difference was not significant. As Figure 3 shown in B of [reference], at the species level, the genus with the highest relative abundance in the control group was Prevotella_copri (1.29%), followed by unclassified Prevotella (Prevotella_unclassified, 1.17%). The genus with the highest relative abundance in the experimental group was Prevotella_copri (1.23%), followed by unclassified Prevotella (Prevotella_unclassified, 1.14%).

[0037] To explore the effect of adding Pediococcus pentosaceus LL2018368 on the microbial species in the feces of Dusa pigs, differential analysis of community composition based on LDA ( Figure 3In C), when the LDA value was greater than 3.0, a total of 20 biomarkers were obtained in the two groups. Among them, 14 biomarkers were obtained in the experimental group. The bacteria with the largest LDA value in the FC group were unclassified bacteria, and the bacteria with the largest LDA values in the FA group were Clostridiales and Clostridia. At the phylum level, the relative abundance of Firmicutes in the FA group was significantly higher than that in the FC group; at the class level, the relative abundances of Negativicutes and Clostridia in the FA group were higher than those in the FC group; at the order level, the relative abundances of Selenomonadales and Clostridiales in the FA group were higher than those in the FC group; at the family level, the relative abundances of Eubacteriaceae, Selenomonadaceae, Ruminococcaceae, Clostridiaceae, and Lachnospiraceae in the FA group were higher than those in the FC group.

[0038] 2.3 Archaeal diversity and composition 2.3.1 α-diversity analysis The analysis of species α-diversity index is shown in Figure 4 . After adding Pediococcus pentosaceus LL2018368 to the drinking water, there were no significant differences in its Simpson index ( Figure 4 in A), Shannon index ( Figure 4 in B), and Chao1 ( Figure 4 in C) indices.

[0039] 2.3.2 β-diversity analysis Principal Coordinates Analysis (PCoA) is shown in Figure 5 . The samples of the two groups overlapped, and the samples of the experimental group had higher consistency, indicating that after adding Pediococcus pentosaceus LL2018368, there were both similar and different parts in the samples of the two groups. After adding Pediococcus pentosaceus LL2018368, the similarity of archaeal species in Duthiea porci feces tended to be consistent. In summary, after adding Pediococcus pentosaceus LL2018368 to the diet, the consistency of archaea in pig feces was higher.

[0040] 2.3.3 Species composition analysis A total of 23 phyla, 40 classes, 57 orders, 87 families, 191 genera, and 770 species were obtained by species annotation.

[0041] As Figure 6 shown in A, at the phylum level, the dominant species with the highest relative abundances in both the control group FC and the experimental group FA were Candidatus_Thorarchaeota, followed by Candidatus_Bathyarchaeota and Crenarchaeota. As Figure 6As shown in B in [reference], at the genus level, the genus with the highest relative abundance in both the control group and the experimental group was Methanobrevibacter, followed by Candidatus_Methanomethylophilus and Methanosarcina. After adding Pediococcus pentosaceus LL2018368, the relative abundance of Methanobrevibacter in the experimental group decreased (P > 0.05).

[0042] To explore the effects of adding Pediococcus pentosaceus LL2018368 on the archaeal community in Dusa pig feces, a differential analysis of community composition based on LDA ( Figure 6 as shown in C in [reference]) was used. When the LDA value was greater than 3.0, a total of 14 biomarkers were obtained in both groups. Among them, 14 biomarkers were obtained in the experimental group. The unclassified Thermoplasmatales had the largest LDA value in the experimental FA group. At the class level, the relative abundances of Candidatus_Poseidoniia, Thermoprotei, and Thermococci in the FA group were higher than those in the FC group; at the order level, the relative abundances of unclassified Thermoplasmatales, Methanosarcinales, and Candidatus_Poseidoniales in the FA group were higher than those in the FC group; at the family level, the relative abundances of Thermoplasmata_unclassified, Methanosarcinaceae, and Candidatus_Poseidoniales_unclassified in the FA group were higher than those in the FC group; at the genus level, the relative abundances of unclassified Thermoplasmatales, Candidatus_Poseidoniales_unclassified, and Methanohalophilus in the FA group were higher than those in the FC group; at the species level, the relative abundances of Thermoplasmata_archaeon and archaeon_SCG_AAA382B04 in the FA group were higher than those in the FC group.

[0043] 2.4 Determination of meat quality and analysis of muscle chemical composition results Table 4 Results of meat color determination

[0044] The analysis of meat quality is shown in Table 4. After adding Pediococcus pentosaceus LL2018368, the lightness value L and the red-green value a of the experimental group samples increased, while the yellow-blue value b decreased.

[0045] Table 5 Chemical composition and amino acid analysis (calculated based on air-dried samples)

[0046] The amino acid and chemical composition analysis of the samples is shown in Table 5. After adding Pediococcus pentosaceus LL2018368, glycine, cystine, valine, isoleucine, leucine, phenylalanine, lysine, histidine, arginine, proline, and the total amount of amino acids showed an upward trend.

[0047] 2.5 Metabolomics analysis 2.5.1 Analysis of short-chain fatty acid components The analysis of short-chain fatty acid content in feces is shown in Table 6. After adding Pediococcus pentosaceus LL2018368, the contents of isobutyric acid, butyric acid and isovaleric acid in the experimental group samples decreased significantly (P < 0.05).

[0048] Table 6 Determination results of short-chain fatty acids in feces (μg / g)

[0049] Note: Different lowercase letters in the superscript of the same row of data indicate significant differences (P < 0.05). 2.5.2 Analysis of fat-soluble vitamin content The fat-soluble vitamin contents in the liver, kidney and serum are shown in Table 7. After adding Pediococcus pentosaceus LL2018368, the contents of vitamin A and all-trans retinol in the kidney increased significantly (P < 0.05), while the content of α-tocopherol in the serum decreased significantly (P < 0.05).

[0050] Table 7 Determination results of fat-soluble vitamins in tissues (μg / g)

[0051] Note: ND indicates not detected. Different lowercase letters in the superscript of the same item in the same row indicate significant differences (P < 0.05), different capital letters indicate extremely significant differences (P < 0.01), and no letter or the same letter in the superscript of the data indicates no significant difference (P > 0.05).

[0052] 2.5.3 Non-targeted metabolome characteristics Table 8 Number of metabolites

[0053] As can be seen from Table 8, a total of 23,993 primary metabolites were identified from the fecal samples of two groups of Dusa finishing pigs, including 9,048 positive ion metabolites and 14,945 negative ion metabolites; 18,784 metabolites were annotated to the KEGG database and 20,738 metabolites were annotated to the HMDB database.

[0054] 2.5.4 PLS-DA analysis The partial least squares discriminant analysis (PLS-DA) method was used to analyze the differential metabolites to visually display the classification effect by discriminant analysis. As Figure 7 shown, the two groups of samples were completely separated with a large separation degree, indicating that the composition of metabolites in pig feces changed after adding Pediococcus pentosaceus.

[0055] 2.5.5 Differential metabolites There were a total of 606 differential metabolites in the two groups ( Figure 8 ). Compared with the control group, 30 metabolites were significantly up-regulated, 25 were significantly down-regulated, and 551 showed no significant difference in the experimental group. KEGG pathway enrichment analysis of the differential metabolites found 20 enriched pathways, among which 13 were significantly different, namely: degradation of flavonoids, metabolism of linoleic acid, production of cutin, suberin and wax, arginine biosynthesis, biosynthesis of valine, leucine and isoleucine, cannabinoid signaling, biosynthesis of phenylpropanoids, longevity regulating pathway - worm, biosynthesis of unsaturated fatty acids, autophagy - animal, biosynthesis of fatty acids, herpesvirus infection; 7 metabolic pathways with no significant difference were: biosynthesis of plant secondary metabolites, biosynthesis of glycosylphosphatidylinositol, autophagy - others, autophagy - yeast, pathogenic Escherichia coli infection, efferocytosis, choline metabolism in cancer, involving degradation of flavonoids, production of fatty acids and amino acids, metabolism of linoleic acid, etc., and the most significant enriched pathway was degradation of flavonoids.

[0056] Table 9 Gene Enrichment Analysis

[0057] GSEA (Gene Set Enrichment Analysis) is applied to the effective information mining of minor substances, and can more comprehensively explain the regulatory role of a certain functional unit, and can complement traditional enrichment analysis. When |NES| > 1, NOM.pval < 0.05, FDR.qval < 0.25, 3 genes were screened out (Table 9): glycerophospholipid metabolism, glycine, serine and threonine metabolism, and biosynthesis of unsaturated fatty acids.

[0058] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Application of Pediococcus pentosaceus in the preparation of products for improving pork quality and intestinal flora, characterized in that: The Pediococcus pentosaceus is Pediococcus pentosaceus LL2018368, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with a deposit number of CGMCC No.18709; The breed of pigs is Dusa pigs; Improving pork quality includes increasing the total amount of amino acids in pork; The improving pork quality also includes increasing the content of vitamin A and all-trans retinol in pig kidney tissue; The improvement of intestinal flora includes promoting the growth of beneficial bacteria, inhibiting the growth of harmful bacteria and improving the consistency of fecal archaea; The beneficial bacteria include one or more of Eubacterium, Lunar Mononas, Ruminococcaceae, Clostridium and Lachnospiraceae; The harmful bacteria include Methanobacterium brevis.

2. The use according to claim 1, characterized in that: The products include feed additives.

3. The use according to claim 2, characterized in that: The amount of Pediococcus pentosaceus added in the feed additive is 10 8 ~10 12 CFU / L.

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