Allicin-e-containing pig feed

Abstract

Provided in the present invention is an allicin-E-containing pig feed. Specifically, provided in the present invention is the use of a compound of formula (I) for preparing a pig feed. The pig feed is a piglet feed, and can effectively improve the feed dry-matter digestibility of pigs and prevent or reverse inflammations during piglet breeding.

Description

A pig feed containing allicin E Technical Field

[0001] This invention relates to the field of livestock and poultry industry, specifically to the use of garlic E in pig feed, and the corresponding pig feed products. Background Technology

[0002] Pig farming is a vital component of global agriculture, with a profound impact on the food supply chain and the economy. Feed costs typically account for 60%-70% of total pig farming costs; therefore, reducing the feed conversion ratio (the amount of feed required per unit of weight gain) in piglets is a key factor in increasing economic efficiency.

[0003] Currently, methods to reduce the feed conversion ratio (FCR) in piglets mainly include improving feed formulation, optimizing feeding management, genetic selection, and using feed additives. Regarding feed formulation, researchers adjust the ratios of protein, amino acids, energy, and fiber to meet the nutritional needs of piglets at different growth stages. Feed management involves environmental control, disease prevention, and early weaning techniques, aiming to create conditions conducive to healthy piglet growth. In terms of genetic selection, breeding with fast-growing pigs and high feed conversion efficiency can gradually improve the performance of offspring. Furthermore, the use of various additives, such as enzyme preparations, probiotics, and acidifiers, has also been shown to improve the digestive and absorptive capacity of piglets, thereby reducing the FCR.

[0004] While the methods described above have reduced the feed conversion ratio in piglets to some extent, several limitations and challenges remain. For example, feed costs remain high, and the production of high-protein feeds can burden the environment. Furthermore, piglets need to resist various diseases during their growth, but the side effects and drug resistance of traditional antibiotics are becoming increasingly serious problems.

[0005] In view of the above problems, there is a need to develop a more efficient, environmentally friendly, sustainable piglet feed formula that does not rely on antibiotics, in order to further reduce the feed conversion ratio of piglets, while ensuring the health and welfare of piglets and reducing the impact on the environment.

[0006] Summary of the Invention

[0007] The purpose of this invention is to provide a high-efficiency, environmentally friendly, sustainable, and antibiotic-free piglet feed formula.

[0008] In a first aspect, the invention provides the use of a compound of formula (I) for the preparation of pig feed:

[0009] In another preferred embodiment, the pig feed is piglet feed.

[0010] In another preferred embodiment, the pig feed is used to improve the dry matter digestibility of pig feed.

[0011] In another preferred embodiment, the content of the compound represented by formula (I) in the pig feed is 10-500 mg / kg, more preferably 10-400 mg / kg, and even more preferably 10-300 mg / kg.

[0012] In a second aspect, the present invention provides a pig feed composition comprising a compound of formula (I):

[0013] In another preferred embodiment, the content of the compound represented by formula (I) in the pig feed composition is 10-500 mg / kg, more preferably 10-400 mg / kg, and even more preferably 10-300 mg / kg.

[0014] In another preferred embodiment, the pig feed composition comprises the following components:

[0015] 0.001 to 0.05 parts by weight of compound (I), 60 to 70 parts by weight of corn, 10 to 20 parts by weight of soybean meal, and 10 to 25 parts by weight of concentrate.

[0016] In another preferred embodiment, the pig feed composition further includes the following components: 0.1 to 5 parts by weight of oil powder, 0 to 10 parts by weight of wheat bran, and 0 to 5 parts by weight of fish meal.

[0017] In another preferred embodiment, the pig feed composition further includes the following components: 0.001 to 0.04 parts by weight of compound of formula (I), 61 to 68 parts by weight of corn, 12 to 19 parts by weight of soybean meal, 0.5 to 3 parts by weight of oil powder, 0 to 8 parts by weight of wheat bran, 0 to 3 parts by weight of fish meal, and 11 to 23 parts by weight of concentrate.

[0018] In another preferred embodiment, the pig feed composition further includes the following components: 0.001 to 0.03 parts by weight of compound of formula (I), 61 to 66 parts by weight of corn, 14 to 18 parts by weight of soybean meal, 0.5 to 2 parts by weight of oil powder, 0 to 5 parts by weight of wheat bran, 0 to 2 parts by weight of fish meal, and 12 to 20 parts by weight of concentrate.

[0019] In another preferred embodiment, the pig feed is also used to improve the liver function of piglets.

[0020] In another preferred embodiment, the pig feed is also used to downregulate serum triglyceride, urea, alanine aminotransferase and / or aspartate aminotransferase levels.

[0021] In another preferred embodiment, the pig feed is also used to upregulate cytokines in serum, preferably, the cytokines being selected from the group consisting of IgG, IgM, IgA, IL-6, or combinations thereof.

[0022] In another preferred embodiment, the pig feed is also used to reduce serum endotoxin levels.

[0023] In another preferred embodiment, the pig feed is also used to enhance the antioxidant capacity of piglets.

[0024] In another preferred embodiment, the pig feed is also used to increase the total short-chain fatty acid (SCFA) level in piglet feces.

[0025] In another preferred embodiment, the pig feed is also used to regulate the abundance of probiotics in the piglets' gut.

[0026] In another preferred embodiment, the pig feed is also used to reduce the abundance of probiotics in the intestines of piglets selected from the group consisting of: *Succinivibrio*, *Clostridium sensu*, *Roseburia*, *UCG-002* (family Oscillatoriaceae), and *Lachnospiraceae*.

[0027] In another preferred embodiment, the pig feed is also used to increase the abundance of probiotics in the intestines of piglets selected from the group consisting of: *Lactobacillus reuteri*, *Acinetobacter*, *Limnohabitans*, *Collinsella* and *Alistipes*, *Acinetobacter junii*, *Coprococcus catus*, *Bacteroides acidifaciens*, and *Lactobacillus mucosa*.

[0028] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0029] Figure 1 shows the flowchart of pig modeling.

[0030] Figure 2 shows the effect of allicin on feed intake in weaned piglets.

[0031] Figure 3 shows the effect of allicin on the weight of weaned piglets.

[0032] Figure 4 shows the effect of allicin on the feed conversion ratio of weaned piglets.

[0033] Figure 5 shows the total oxygen free radical uptake capacity (ORAC) of allicin E.

[0034] Figure 6 shows the scavenging ability of allicin E on DPPH free radicals.

[0035] Figure 7 shows the iron-reducing capacity of garlic E.

[0036] Figure 8 shows the effect of garlic E on the NMDS of piglet fecal microbiota.

[0037] Figure 9 shows the effect of garlic E on differences in gut microbiota genera in piglets.

[0038] Figure 10 shows the effect of garlic E on species-level differences in the gut microbiota of piglets. Detailed Implementation

[0039] Through extensive and in-depth research, and after numerous experiments and screenings, the inventors unexpectedly discovered a pig feed comprising the compound of formula (I). Experiments show that using the pig feed of this invention can improve the dry matter digestibility of pig feed, improve liver function in piglets, enhance the antioxidant capacity of piglets, regulate the abundance of beneficial bacteria in the piglet gut, and reduce the feed conversion ratio, thereby reducing pig farming costs. Based on this, the present invention was completed.

[0040] the term

[0041] To facilitate understanding of the invention, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains. Before describing the invention, it should be understood that the invention is not limited to the specific methods and experimental conditions described, as such methods and conditions can vary. It should also be understood that the terminology used herein is intended only to describe particular embodiments and is not intended to be restrictive; the scope of the invention will be limited only by the appended claims.

[0042] As used herein, the term “comprising” or its variations such as “including” or “comprising” are understood to include the said element or component without excluding other elements or other components.

[0043] The term “about” can refer to a value or composition within an acceptable margin of error for a particular value or composition as determined by a person skilled in the art, depending in part on how the value or composition is measured or determined. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0044] As used herein, unless otherwise stated, any concentration range, percentage range, proportion range, or integer range shall be understood to include any integer value within the range and, where appropriate, its fractional value (e.g., one-tenth and one-hundredth of an integer).

[0045] As used herein, the term “and / or” refers to and covers any and all possible combinations of one or more of the related listed items.

[0046] Garlic E

[0047] ALE (Allicin E) is a single compound synthesized biomimetically by modifying the structure outside the antibacterial active group of ordinary allicin. Its structure is shown in the following formula:

[0048] Garlic E has broad-spectrum bactericidal and green safety characteristics, and can effectively replace traditional antibiotics, supporting environmental disinfection, animal and plant protection, preservation, and upgrading of the biopharmaceutical industry.

[0049] Pig feed formulation

[0050] This invention provides a pig feed composition comprising a compound of formula (I):

[0051] In a preferred embodiment, the content of the compound represented by formula (I) in the pig feed composition is 10-500 mg / kg, more preferably 10-400 mg / kg, and even more preferably 10-300 mg / kg.

[0052] Since the addition of garlic E element in this invention can effectively reduce the feed conversion ratio and improve the digestibility of feed protein, it can effectively reduce the amount of crude protein in the feed composition. In a preferred pig feed composition formulation, the pig feed composition includes the following components:

[0053] 0.001 to 0.05 parts by weight of compound (I), 60 to 70 parts by weight of corn, 10 to 20 parts by weight of soybean meal, and 10 to 25 parts by weight of concentrate.

[0054] In a preferred embodiment, the pig feed composition further includes the following components: 0.1 to 5 parts by weight of oil powder, 0 to 10 parts by weight of wheat bran, and 0 to 5 parts by weight of fish meal.

[0055] In a preferred embodiment, the pig feed composition further includes the following components: 0.001 to 0.04 parts by weight of compound of formula (I), 61 to 68 parts by weight of corn, 12 to 19 parts by weight of soybean meal, 0.5 to 3 parts by weight of oil powder, 0 to 8 parts by weight of wheat bran, 0 to 3 parts by weight of fish meal, and 11 to 23 parts by weight of concentrate.

[0056] In a preferred embodiment, the pig feed composition further includes the following components: 0.001 to 0.03 parts by weight of compound of formula (I), 61 to 66 parts by weight of corn, 14 to 18 parts by weight of soybean meal, 0.5 to 2 parts by weight of oil powder, 0 to 5 parts by weight of wheat bran, 0 to 2 parts by weight of fish meal, and 12 to 20 parts by weight of concentrate.

[0057] The pig feed composition of the present invention has certain health benefits due to the addition of garlic E. In a preferred embodiment, the pig feed is also used to improve the liver function of piglets.

[0058] In another preferred embodiment, the pig feed is also used to downregulate serum triglyceride, urea, alanine aminotransferase and / or aspartate aminotransferase levels.

[0059] In another preferred embodiment, the pig feed is also used to upregulate cytokines in serum, preferably, the cytokines being selected from the group consisting of IgG, IgM, IgA, IL-6, or combinations thereof.

[0060] In another preferred embodiment, the pig feed is also used to enhance the antioxidant capacity of piglets.

[0061] In another preferred embodiment, the pig feed is also used to regulate the abundance of probiotics in the piglets' gut.

[0062] In another preferred embodiment, the pig feed is also used to reduce the abundance of probiotics in the intestines of piglets selected from the group consisting of: *Succinivibrio*, *Clostridium sensu*, *Roseburia*, *UCG-002* (family Oscillatoriaceae), and *Lachnospiraceae*.

[0063] In another preferred embodiment, the pig feed is also used to increase the abundance of probiotics in the intestines of piglets selected from the group consisting of: *Lactobacillus reuteri*, *Acinetobacter*, *Limnohabitans*, *Collinsella* and *Alistipes*, *Acinetobacter junii*, *Coprococcus catus*, *Bacteroides acidifaciens*, and *Lactobacillus mucosa*.

[0064] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0065] Material

[0066] The allicin used in the experiment was obtained from Shanghai Laishen Biotechnology Co., Ltd. (99%, 5 mg / ml dissolved in water), and allicin was purchased from Henan Munihei Biotechnology Co., Ltd. (25%).

[0067] The basal diet was customized by the Shanghai Academy of Agricultural Sciences. The main components and their contents are shown in Table 1.

[0068] Table 1. Basal Diet Formulation (%)

[0069] a The 29-35 day concentrate contains: crude protein ≥30%; crude fiber ≤4%; crude ash ≤30%; total P ≥1%; Ca ≥2%; vitamin A ≥15,000 IU / kg; vitamin D3 ≥2400 IU / kg; vitamin E ≥100 mg / kg; Cu ≥100 mg / kg; Fe ≥500 mg / kg; Zn ≥480 mg / kg; lysine ≥1.5%; NaCl 1.0-3.5%; and H2O ≤12%.

[0070] b The 29-35 day concentrate contains crude protein ≥17.0%, crude fiber ≤5.0%, crude ash ≤40.0%, total P ≥1.5%, Ca 4.0-9.0%, vitamin A 41600-58300 IU / kg, vitamin D3 ≥10,400-41,600 IU / kg, vitamin E ≥166 mg / kg, Cu 73-290 mg / kg, Fe 600-1600 mg / kg, Zn 500-1,250 mg / kg, lysine ≥1.0%, NaCl 2.0-6.0%, and H2O ≤12%.

[0071] General Method

[0072] 1. Animal modeling

[0073] Two hundred and ten 28-day-old weaned piglets were randomly divided into three groups, with seven pens per group and ten piglets per pen. One group was fed a basal diet as a control group (C group); another group was fed a basal diet supplemented with 50 mg / kg allicin (L group); and the third group was fed a basal diet supplemented with 50 mg / kg allicin (M group) (Table 2). Figure 1 shows the flowchart of pig modeling.

[0074] The experiment lasted for four weeks, with feces collected midway through the experiment to determine feed digestibility. On days 14 and 28 (i.e., 35 and 56 days of age), two animals were randomly selected from each pen to collect blood and feces.

[0075] Table 2

[0076] 2. Animal husbandry and sample collection and testing

[0077] The experiment lasted 28 days, with piglets having free access to feed and water. The experiment lasted 4 weeks, with piglet feed administered for the first week and piglet feed administered for the following 3 weeks. Growth performance parameters, including feed intake, live weight, average daily weight gain, and feed conversion ratio, were recorded weekly. Fresh fecal samples were collected consecutively at the midpoint of the experiment and 3 days before the end of the experiment (days 12-14 and 26-28, corresponding to days 40-42 and 54-56 based on age). One portion was divided into two sterile centrifuge tubes and stored at -20°C for the determination of short-chain fatty acids (SCFA) and gut microbiota composition; the other portion was fixed with 10% dilute hydrochloric acid for the determination of feed digestibility (dry matter, crude protein, and crude fat) in both periods. Serum samples were collected for the determination of complete blood count, blood biochemical indicators, antioxidant levels, and immune-related indicators.

[0078] 3. Serum-related index measurement

[0079] Complete blood count and basic blood biochemistry indicators were measured by Shanghai Qianmai Bomier Medical Laboratory Co., Ltd.

[0080] Serum endotoxin was detected using the Limulus amebocyte lysate (LAL) endotoxin detection kit (catalog number: EC80545S) manufactured by Xiamen Limulus amebocyte lysate Reagent Biotechnology Co., Ltd. Serum MDA and total SOD were measured using kits manufactured by Nanjing Jiancheng Biological Research Institute Co., Ltd. (MDA kit catalog number: A003-2-1, SOD kit catalog number: A001-3-2). The procedures were performed in accordance with the kit instructions.

[0081] Serum IgG and IgM were measured using kits manufactured by Nanjing Jiancheng Biological Research Institute Co., Ltd. (catalog numbers: E026-1-1 and E025-1-1, respectively). Serum IL-2, IL-6, and IFN-γ were measured using ELISA. The procedures were performed according to the kit instructions.

[0082] 4. In vitro antioxidant test of garlic E.

[0083] The total oxygen free radical scavenging capacity (ORAC), nitrogen free radical scavenging capacity (DPPH), and iron ion reducing capacity of garlic E were determined using reagent kits (catalog numbers A006, A001, and A003, respectively) manufactured by Shanghai Congyi Technology Co., Ltd., and the operation was performed in accordance with the kit instructions.

[0084] 5. Determination of apparent digestibility

[0085] The collected samples (feces and feed) were placed in an oven and dried at 65°C for 48 hours. After standing at room temperature for 24 hours, the samples were weighed and then pulverized to prepare air-dried samples. Crude protein was determined by the Kjeldahl method, and crude fat was determined by ether extraction. The dry matter was dried to constant weight at 105°C. The digestibility of crude protein (CP), crude fat (EE), and dry matter (DM) was calculated. The calculation formula is as follows: 1 - (acid-insoluble ash content in feed / acid-insoluble ash content in feces) × (nutrient content in feces / nutrient content in feed) × 100%.

[0086] 6. Determination of short-chain fatty acids in feces

[0087] Accurately weigh the sample and add 10 times its volume of double-distilled water, mixing thoroughly. Then, take 1.00 ml of the supernatant, add 0.20 ml of crotonic acid metaphosphate, and store overnight at -20°C. After thawing, centrifuge at 12000 rpm for 10 min, collect the supernatant for storage, and pass a small amount through a 0.22 μm syringe filter before analysis. Inject the filtrate directly into the sample.

[0088] Chromatographic conditions

[0089] Instrument Agilent 7890B

[0090] Capillary column 30m × 0.32mm × 0.25μm film thickness

[0091] Column temperature 130℃

[0092] vaporization temperature 180℃

[0093] Hydrogen ion flame detector, detection temperature 180℃

[0094] The carrier gas is nitrogen at a pressure of 60 kPa, hydrogen at a pressure of 50 kPa, and oxygen at a pressure of 50 kPa.

[0095] 7. Fecal 16S sequencing analysis

[0096] Fecal 16S sequencing analysis was performed by Beijing Novogene Technology Co., Ltd.

[0097] 8. Data Statistics

[0098] The means of each group were analyzed by one-way ANOVA using SPSS. The significance of the differences between the means was determined by ANOVA of repeated determinations and the least significant difference (LSD) method. P < 0.05 was considered significant.

[0099] Example 1: Effect of Garlic E Addition on Piglet Production Performance

[0100] As shown in Figure 2, there was no significant difference in feed intake among all groups in the early and late stages of the experiment (P>0.05). The original data can be found in Appendix Table 1.

[0101] As shown in Figure 3, there were no significant differences in the body weight and average daily weight gain of piglets in the groups at days 0, 14, and 28 (see Appendix Table 2) (P>0.05).

[0102] As shown in Figure 4, there was no significant difference in feed conversion ratio among the groups in the early stage of the experiment. In the later stage of the experiment, the feed conversion ratio of piglets fed with garlic E (L) was significantly lower than that of piglets fed with traditional garlic extract (M). Feed is a major cost factor in the livestock industry. A lower feed conversion ratio means that farmers can greatly reduce feed input and lower production costs. Therefore, the feed of this invention can effectively improve the feed conversion ratio in the piglet feeding process.

[0103] Example 2: Effect of Garlic E Addition on Apparent Digestibility of Major Nutrients in Feed

[0104] Apparent digestibility of feed reflects the rate at which animals utilize the nutrients in the feed.

[0105] Table 3. Effects of Garlic E on the Apparent Digestibility of Major Nutrients in Piglet Feed.

[0106] DM: Dry matter digestibility; CP: Crude protein digestibility; EE: Crude fat digestibility. C: Control group; L: Garlic E-added group; M: Traditional garlic extract-added group.

[0107] As shown in Table 3, the addition of allicin or allicin E to the diet significantly improved the digestibility of feed dry matter (P<0.05). The addition of allicin E to the feed improved the digestibility of feed protein, but had no significant effect on the digestibility of fat (P>0.05).

[0108] As described in Example 1, it was found that the addition of garlic E significantly reduced the feed conversion ratio compared to traditional allicin. This result explains to some extent why garlic E reduces the feed conversion ratio. However, even with a slight increase in dry matter digestibility, the feed conversion ratio of traditional allicin is still significantly lower than that of garlic E.

[0109] The results showed that the addition of conventional allicin slightly decreased the digestibility of crude fat, but the difference was not statistically significant. The apparent digestibility of nitrogen-free extract was not measured in this study. Protein, fat, and nitrogen-free extract were the main sources of animal weight gain and also the main factors affecting the feed conversion ratio.

[0110] Example 3: Effects of Garlic E Addition on Blood Routine and Blood Biochemical Indicators in Piglets

[0111] A complete blood count (CBC) is one of the simplest and most common methods for assessing the health status of an organism. Therefore, CBCs are tested in piglets during the feeding process to determine the effectiveness of garlic E supplementation.

[0112] Table 4. Results of routine blood tests on piglets on day 7 of the experiment.

[0113] C: Control group; L: Garlic E-added group; M: Traditional garlic E-added group.

[0114] Table 5. Results of routine blood tests on piglets on day 28 of the experiment.

[0115] C: Control group; L: Garlic E-added group; M: Traditional garlic E-added group.

[0116] Tables 4-5 show the effects of garlic E or allicin on routine blood tests in piglets. The results indicate that the addition of garlic E and allicin promoted hemoglobin production (at 7 and 28 days) and significantly increased the proportion of eosinophils. However, due to the small magnitude of these changes, their impact on health cannot be determined without further data.

[0117] Table 6. Results of blood biochemical tests on piglets on day 7 of the experiment.

[0118] ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; MDA: Malondialdehyde; SOD: Superoxide dismutase. C: Control group; L: Garlic E-added group; M: Traditional allicin-added group.

[0119] Table 7. Results of blood biochemical tests on piglets on day 28 of the experiment.

[0120] ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; MDA: Malondialdehyde; SOD: Superoxide dismutase. C: Control group; L: Garlic E-added group; M: Traditional allicin-added group.

[0121] Tables 6 and 7 show the effects of garlic E or allicin on common biochemical indicators in piglet blood. The results indicate that the addition of garlic E and allicin reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations in piglets and increased the albumin-globulin ratio, suggesting that garlic E has a certain protective effect on the piglet liver. Simultaneously, the addition of garlic E reduced serum urea levels.

[0122] Urea is a metabolic product of protein. Under normal physiological conditions, the catabolism of protein is maintained at a low level, suggesting that the addition of garlic E increases protein synthesis or reduces protein breakdown, which is consistent with the fact that garlic E improves the digestibility of crude protein.

[0123] Triglycerides and cholesterol are both important components of blood lipids. Serum biochemical analysis results showed that the addition of garlic E significantly increased the serum cholesterol content of piglets, but the test results at 28 days of age also showed that garlic E significantly reduced the serum triglyceride content of piglets.

[0124] Example 4: Effects of Garlic E Addition on Immune-Related Indicators in Piglets

[0125] The effects of garlic E on serum endotoxin, IgG, IgM, IL-2, IL-6, and IFN-γ in piglets are shown in Table 8.

[0126] Table 8. Effects of Garlic E on Immune-Related Indicators in Piglets

[0127] C: Control group; L: Garlic E-added group; M: Traditional garlic E-added group.

[0128] Table 8 shows that the addition of garlic E significantly reduced the serum endotoxin content in piglets (P<0.05). At day 28 of the experiment, the serum IgG and IL-6 concentrations in piglets fed a diet supplemented with garlic E were significantly increased (P<0.05). Endotoxins are phospholipid-polysaccharide-protein complexes produced by Gram-negative bacteria, with lipopolysaccharide as the main component, and they have toxic effects on animals. IgG is the main immunoglobulin for fighting infection, and IL-6 is a cytokine produced by monocytes / macrophages that can promote the secretion of IgG, IgM, and IgA.

[0129] The results showed that allicin E could enhance the body's immunity and reduce serum endotoxin levels by increasing IL-6 expression and promoting IgG secretion, and its enhancement level was further improved compared with traditional allicin.

[0130] Example 5: Antioxidant capacity analysis of allicin E

[0131] The in vitro antioxidant capacity of garlic E was evaluated by total oxygen free radical uptake capacity (ORAC), nitrogen free radical scavenging capacity (DPPH), and iron ion reducing capacity. The results are shown in Figures 5, 6, and 7, respectively.

[0132] As calculated from Figures 5-7, 1g of garlic E has a total oxygen free radical absorption capacity equivalent to 1.4mmol Trolox, a DPPH free radical scavenging capacity equivalent to 47.8μmol Trolox, and an iron ion reducing capacity equivalent to 1.2mmol Trolox, indicating that it has a certain antioxidant capacity.

[0133] The in vivo antioxidant capacity of garlic E was assessed by measuring serum MDA and SOD enzyme activities, and the data are shown in Tables 6-7. The results showed that on day 7 of the experiment, serum SOD enzyme activity in piglets fed with garlic E was significantly increased (P<0.05). On day 28, serum SOD enzyme activity in piglets fed with garlic E was also increased, but the difference was not statistically significant (P>0.05). There were no differences in serum MDA levels among the groups of piglets.

[0134] Example 6: Effect of Garlic E Addition on Short-Chain Fatty Acid Content in Piglet Feces

[0135] The vast majority of short-chain fatty acids (SCFAs) are produced by bacterial fermentation of undigested carbohydrates in the intestines. Small amounts of SCFAs are also produced from dietary intake and the metabolism of proteins. SCFAs are beneficial to the gut health of animals.

[0136] Table 9. SCFA content in feces after 14 days (mg / g)

[0137] Note: C: Control group; L: Garlic E-added group; M: Traditional garlic E-added group.

[0138] Table 10. SCFA content in feces after 28 days (mg / g)

[0139] Note: C: Control group; L: Garlic E-added group; M: Traditional garlic E-added group.

[0140] As shown in Tables 9 and 10, the addition of garlic E and allicin to the feed can increase the total acid level in piglet feces (P<0.05); at 28 days of the experiment, the content of acetic acid and propionic acid in the feed containing garlic E and allicin was higher than that in the control group (P<0.05).

[0141] This indicates that allicin E and allicin have the effect of regulating intestinal flora and promoting the production of SCFA, a conclusion consistent with the 16S sequencing results.

[0142] Example 7: Effects of Garlic E on the Fecal Microbiota Structure of Piglets

[0143] 7.1 Effects of Garlic E on the α-diversity of fecal microbiota in piglets

[0144] Table 11 Effects of allicin E on fecal microbiota α diversity in piglets

[0145] Note: C: Control group; L: Garlic E-added group; M: Traditional garlic E-added group.

[0146] As shown in Table 11, garlic E (group L) had a significant effect on the number of observed species and species richness (Chao1) of piglet fecal microbiota, increasing the variety of microbiota in piglet feces (P<0.05).

[0147] 7.2 Effects of Garlic E on NMDS in Piglet Fecal Microbiota

[0148] Non-Metric Multi-Dimensional Scaling (NMDS) is a statistical method suitable for ordination in ecological studies. NMDS analysis can reflect differences between and within groups of a sample by using the distances between points.

[0149] As shown in Figure 8, there were certain differences in the gut microbiota among the control group (C), the garlic E group (L), and the ordinary allicin group (M). The calculated stress value of 0.186 indicates that the NMDS analysis was effective (a stress value less than 0.2 indicates that NMDS can accurately reflect the degree of difference between samples).

[0150] 7.3 Effects of Garlic E on Differences in Intestinal Microbiota Genus in Piglets

[0151] Pairwise comparisons of the abundance at the genus level in the three groups (Figure 9) revealed that, compared with the control group C, the abundance of *Succinivibrio*, *Clostridium sensu*, *Roseburia*, *UCG-002*, and *lachnospiraceae* in the feces of piglets fed with garlic E (group L) was significantly reduced (P<0.05); while the abundance of *Limosilactobacillus*, *Acinetobacter*, *Limnohabitans*, *Collinsella*, and *Alistipes* was significantly increased (P<0.05). *Limosilactobacillus* exhibits numerous beneficial functions, including inhibiting pathogens, improving the intestinal environment, regulating intestinal flora, and reducing serum cholesterol levels.

[0152] Comparisons at the genus level have many limitations because a genus often includes many species, and the characteristics of different species can vary greatly. An analysis was then conducted at the species level, and the results are shown in Figure 14.

[0153] As shown in Figure 10, compared with the control group C, the abundance of Lactobacillus johnsonii, Lactobacillus reuteri, Acinetobacter junii, Coprococcus catus, Bacteroides acidifaciens, and Lactobacillus mucosa was significantly increased in piglets fed with garlic E (group L) (P<0.05), while the abundance of Clostridium butyricum was significantly decreased (P<0.05).

[0154] Lactobacillus johnsonii, Lactobacillus reuteri, and Lactobacillus mucosa are common probiotics of the Lactobacillus genus; Acinetobacter jonense has a degrading effect on protein in residues and can promote the digestion and utilization of protein; Strychnine foetida has the effect of promoting the production of SCFA, which can promote the increase of animal weight.

[0155] The results showed that garlic E has the effect of increasing the abundance of intestinal probiotics, which may be one of the mechanisms by which it promotes the increase of fecal SCFA and improves digestibility.

[0156] in conclusion

[0157] 1. Adding 50 mg / kg of garlic E to the feed of piglets did not cause abnormal mortality, nor did it cause significant changes in routine blood tests or common blood biochemical indicators exceeding the normal range, nor did it cause a large increase in pathogenic bacteria in piglet feces. This indicates that short-term feeding of diets containing a certain amount of garlic E will not have adverse effects on the health of piglets.

[0158] 2. Adding 50 mg / kg of garlic E to the feed of piglets had no significant effect on feed intake and daily weight gain (P>0.05); compared with the addition of traditional garlic, the addition of garlic E to the feed reduced the feed conversion ratio of piglets to a significant level (P=0.030), which theoretically can help reduce the cost of pig farming.

[0159] 3. Garlic E can reduce the feed conversion ratio, which is related to its ability to improve the digestibility of nutrients.

[0160] 4. Garlic E has a certain in vitro antioxidant capacity and can enhance the activity of SOD enzyme in piglet serum.

[0161] 5. Garlic E can regulate the immune function of piglets, increase IL-6 expression, increase serum IgG concentration, and reduce serum endotoxin content.

[0162] 6. Garlic E can regulate the intestinal flora of piglets, thereby promoting the production of SCFA and having a beneficial effect on the intestinal health of piglets.

[0163] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. Use of a compound of formula (I), characterized in that, Used in the preparation of pig feed:

2. The use as described in claim 1, characterized in that, The pig feed is piglet feed.

3. The use as described in claim 1, characterized in that, The pig feed contains a compound of formula (I) in a concentration of 10–500 mg / kg, preferably 10–400 mg / kg, and more preferably 10–300 mg / kg.

4. A pig feed composition, characterized in that, The pig feed composition comprises a compound of formula (I):

5. The pig feed composition according to claim 4, characterized in that, The content of the compound shown in formula (I) in the pig feed composition is 10-500 mg / kg.

6. The pig feed composition according to claim 4, characterized in that, The content of the compound shown in formula (I) in the pig feed composition is 10-400 mg / kg.

7. The pig feed composition according to claim 4, characterized in that, The content of the compound shown in formula (I) in the pig feed composition is 10-300 mg / kg.

8. The pig feed composition according to claim 4, characterized in that, The pig feed composition comprises the following components: 0.001 to 0.05 parts by weight of compound (I), 60 to 70 parts by weight of corn, 10 to 20 parts by weight of soybean meal, and 10 to 25 parts by weight of concentrate.

9. The pig feed composition according to claim 8, characterized in that, The pig feed composition further includes the following components: 0.1 to 5 parts by weight of oil powder, 0 to 10 parts by weight of wheat bran, and 0 to 5 parts by weight of fish meal.

10. The pig feed composition according to claim 8, characterized in that, The pig feed composition further includes the following components: 0.001 to 0.04 parts by weight of compound (I), 61 to 68 parts by weight of corn, 12 to 19 parts by weight of soybean meal, 0.5 to 3 parts by weight of oil powder, 0 to 8 parts by weight of wheat bran, 0 to 3 parts by weight of fish meal, and 11 to 23 parts by weight of concentrate.

11. The pig feed composition according to claim 8, characterized in that, The pig feed composition further includes the following components: 0.001 to 0.03 parts by weight of compound (I), 61 to 66 parts by weight of corn, 14 to 18 parts by weight of soybean meal, 0.5 to 2 parts by weight of oil powder, 0 to 5 parts by weight of wheat bran, 0 to 2 parts by weight of fish meal, and 12 to 20 parts by weight of concentrate.

Images