Use of fullerenes in the preparation of a product against intestinal infections in weaned piglets

Abstract

The application discloses application of fullerene in preparation of a product for resisting weaned piglet intestinal infection and belongs to the technical field of livestock and poultry breeding. The application first provides the role of fullerene in resisting weaned piglet intestinal Escherichia coli infection and uses the fullerene as a feed additive for weaned piglets. Experimental results show that, when 100 mg / kg of the fullerene is added into weaned piglet feed, the reduction of ileum and colon villus height caused by Escherichia coli K88 infection can be significantly relieved, the ileum villus crypt ratio is improved, the proliferation of a plurality of beneficial bacteria such as lactobacillus and Prevotellaceae in the colon is promoted, the intestinal morphological damage and the colon flora structure disorder of piglets caused by the Escherichia coli infection are improved, the intestinal health of the piglets is maintained, and the resistance to pathogenic microorganisms is improved. The application provides a non-toxic, efficient and novel functional feed additive for resisting weaned piglet Escherichia coli infection.

Description

Technical Field

[0001] This invention relates to the field of livestock and poultry farming, and in particular to the application of fullerenes in the preparation of products for treating intestinal infections in weaned piglets. Background Technology

[0002] In modern large-scale pig farming, weaned piglets are highly susceptible to pathogenic bacteria, such as Escherichia coli, due to their immature intestinal development and weak barrier function, leading to intestinal damage and barrier dysfunction. E. coli can adhere to intestinal epithelial cells and, through the secretion of enterotoxins, disrupt the intestinal mucosal barrier structure, causing villus atrophy, crypt hyperplasia, and a decrease in the villus height / crypt depth ratio, thereby reducing the intestinal villus absorption area and lowering nutrient utilization efficiency. Furthermore, intestinal microecological imbalance can also lead to a decrease in the abundance of beneficial bacteria, further impairing intestinal immunity and barrier function. Previously, production often relied on antibiotics and high-dose zinc oxide for intervention; however, long-term use of antibiotics easily induces drug resistance and carries food safety risks; high-dose zinc oxide also leads to environmental pollution. Therefore, there is an urgent need to develop a safe and environmentally friendly new functional feed additive to protect the intestinal barrier of piglets from damage caused by pathogenic bacteria such as E. coli.

[0003] Fullerene (C 60 Fullerene (C6N) is a carbon nanomaterial with a spherical conjugated structure. It possesses excellent free radical scavenging capabilities, effectively reducing reactive oxygen species (ROS) levels. It is not easily absorbed in the gastrointestinal tract, primarily exerting its effects locally within the intestinal lumen and being excreted with feces, exhibiting high biocompatibility. Therefore, the development of fullerenes (C6N) is a promising area of ​​research. 60 As a functional additive for piglet feed, it has extremely high production and application value. Summary of the Invention

[0004] The purpose of this invention is to provide the application of fullerene in the preparation of products for treating intestinal infections in weaned piglets, in order to solve the problems existing in the prior art. This invention provides a novel, non-toxic, and highly effective functional feed additive for treating Escherichia coli infection in weaned piglets.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides fullerene C 60 Application in the preparation of products for preventing intestinal infections in weaned piglets.

[0006] Furthermore, the intestinal infection is an intestinal infection caused by Escherichia coli.

[0007] Furthermore, the Escherichia coli is Escherichia coli K88.

[0008] Furthermore, the product is feed, feed additive, or medicine.

[0009] Furthermore, when the product is animal feed, the fullerene C 60 The content is not less than 100 mg / kg.

[0010] Furthermore, the fullerene C 60 The content is 100 mg / kg.

[0011] Furthermore, the anti-intestinal infection in weaned piglets includes improving intestinal morphological damage and improving colonic flora dysbiosis.

[0012] The present invention discloses the following technical effects: This invention provides, for the first time, the role of fullerene in combating Escherichia coli infection in the intestines of weaned piglets and its application as a feed additive. Experimental results show that adding 100 mg / kg fullerene to the feed of weaned piglets significantly alleviates the reduction in villus height in the ileum and colon caused by Escherichia coli K88 infection, increases the ileum-to-villus ratio, and promotes the proliferation of various beneficial bacteria in the colon, such as Lactobacillus and Prevotella. This improves the intestinal morphological damage and colonic flora imbalance caused by E. coli infection, thereby maintaining intestinal health and enhancing resistance to pathogenic microorganisms. This invention provides a novel, non-toxic, and highly effective functional feed additive for combating E. coli infection in weaned piglets. Attached Figure Description

[0013] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0014] Figure 1 Add C to the diet 60 Figure 1 shows the effect of Escherichia coli K88 infection on the intestinal morphology of piglets; where A is a photomicrograph of the ileum and colon after HE staining; B is the villus height, crypt depth, and villus-crypt ratio of the ileum; and C is the crypt depth of the colon. Figure 2 Add C to the diet 60 Figure showing the effects of Escherichia coli K88 infection on the colonic microbial community structure induced in piglets; where A is the PCoA diagram of colonic microbiota based on Bray-Curtis distance at the OUT level; and B is the α-diversity of the colonic microbial community (Sobs and Chao indices). Figure 3 Add C to the diet 60The results of the influence of Escherichia coli K88 infection on the colonic microbial composition of piglets are shown in the figure; where AC represents the relative abundance of the microbial community at the phylum (A), genus (B), and species (C) levels; D is the score plot of LEfSe analysis; and EF represents the relative abundance levels of key differentially expressed genera (E) and species (F). Detailed Implementation

[0015] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0016] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0017] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0018] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0019] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0020] Example 1 1. Experimental treatment The experimental groups and treatment methods are shown in Table 1.

[0021] Table 1 Experimental Groups and Treatments 2. Test Methods Forty-eight healthy 7-day-old Duroc × Landrace × Yorkshire piglets of similar parity and weight were selected. They were randomly divided into four treatment groups, with 12 piglets in each treatment group and one piglet in each replicate.

[0022] The trial period was from 7 to 28 days old (22 days in total) for piglets.

[0023] Throughout the trial, piglets had free access to feed with or without added vitamin C. 60 All piglets were weaned at 21 days of age using the provided feed.

[0024] Group K88 and C 60 Piglets in the +K88 group were challenged with E. coli K88 at 22, 25, and 28 days of age (intraperitoneal injection of E. coli K88 bacterial solution, bacterial solution concentration: 2×10⁻⁶). 9 (CFU / mL), the injection doses for different ages were 1, 2, and 3 mL respectively. CON group and C 60 Each group was injected with the same dose of physiological saline.

[0025] Four hours after piglets were challenged with the virus at 28 days of age, four piglets from each group were randomly selected for slaughter and sampling: ileum and colon tissue samples were collected for HE staining to observe intestinal morphology; colonic chyme was collected for 16S rRNA sequencing to analyze changes in intestinal flora.

[0026] 3. Test Results 3.1 Intestinal morphology results like Figure 1 As shown, E. coli K88 challenge significantly reduced ileal villus height and villus-to-crypto ratio. P <0.05); while dietary C supplementation 60 This trend has been significantly reversed. P <0.05), restoring it to a level comparable to the CON group.

[0027] There was no significant difference in ileal crypt depth among the groups. Regarding the colon, E. coli K88 challenge also significantly reduced colonic villus height. P <0.05); Add C 60 There is a trend towards improving fluff height. Two-factor analysis shows that C 60 Escherichia coli K88 had a significant main effect on ileal villus height, ileocecal-cryptotropic ratio, and colonic villus height. P <0.01), but the interaction between the two was not significant ( P >0.05). The results showed that dietary supplementation with C 60 It can effectively reduce the morphological damage to the ileum and colonic mucosa of piglets caused by Escherichia coli K88 infection and protect the integrity of the intestinal structure.

[0028] 3.2 Colonic microbiota structure Figure 2 Analysis of the diversity and community structure of the colonic microbiota in piglets showed that there were no significant differences in Alpha diversity indicators such as Sobs and Chao index among the treatment groups. Figure 2 B, P >0.05); but if Figure 2 As shown in Figure A, principal coordinate analysis (PCoA) based on OTU levels indicates that different treatments significantly altered the community structure of the colonic microbiota (R=0.18056, P =0.029).

[0029] The results showed that dietary supplementation with C 60 It can reshape the colonic microecological community structure disrupted by E. coli K88 challenge.

[0030] 3.3 Composition of Colonic Microbiota Figure 3 Further analysis was conducted on the compositional characteristics and key differentially expressed microbiota of the colon. Microbiota composition and LEfSe differential analysis ( Figure 3 (AD) shows that C 60 The group was significantly enriched with Lactobacillus reuteri ( s_Lactobacillus_reuteri ), Lactobacillus amyloliquefaciens ( s_Lactobacillus_amylovorus Beneficial bacteria such as C were added under challenge conditions; 60 Lactobacillus species were significantly enriched in the colon. g_Lactobacillus Lactobacillus johnsonii ( s_Lactobacillus_johnsonii It also contains beneficial bacteria such as Lactobacillus (f.Lactobacillaceae).

[0031] Quantitative analysis of key differentially expressed microbiota further confirmed ( Figure 3 (EF), C in the two-factor analysis 60 It has a significant main effect on Lactobacillus and Prevotellaceae. P <0.05); Compared to the K88 group and the CON group, the dietary supplementation with C 60 Significantly increased the relative abundance of Prevostidae, Lactobacillus, and Lactobacillus johnsonii. P <0.05).

[0032] The results showed that dietary supplementation with C 60 It can effectively inhibit the accumulation of intestinal pathogens caused by Escherichia coli K88 infection, and regulate and improve the balance of colonic flora in weaned piglets by promoting the proliferation of beneficial bacteria such as Lactobacillus and Prevotella.

[0033] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. Fullerene C 60 Application in the preparation of products for preventing intestinal infections in weaned piglets.

2. The application as described in claim 1, characterized in that, The intestinal infection mentioned is an intestinal infection caused by Escherichia coli.

3. The application as described in claim 2, characterized in that, The Escherichia coli mentioned is Escherichia coli K88.

4. The application as described in claim 1, characterized in that, The product is feed, feed additive, or medicine.

5. The application as described in claim 4, characterized in that, When the product is animal feed, the fullerene C 60 The content is not less than 100 mg / kg.

6. The application as described in claim 5, characterized in that, The fullerene C 60 The content is 100 mg / kg.

7. The application as described in claim 1, characterized in that, The anti-intestinal infection in weaned piglets includes improving intestinal morphological damage and improving colonic flora structure imbalance.

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