A synbiotic and uses thereof
By combining probiotics and prebiotics to prepare synbiotic microspheres, the problems of unstable treatment effects and drug resistance in poultry colitis are solved, achieving intestinal health repair and immune regulation, and making it suitable for simple treatment in poultry farming.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- YUNNAN AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-02-26
- Publication Date
- 2026-05-29
AI Technical Summary
Existing technologies for treating avian colitis have problems such as unstable efficacy, long-term use leading to drug resistance and intestinal flora imbalance, and existing prevention and control measures are difficult or costly to implement.
By using a synbiotic approach, probiotics (such as Lactobacillus plantarum) are combined with prebiotics (such as fructooligosaccharides), and microspheres are formed by encapsulating them with low-ester pectin and chitosan. The therapeutic effect is enhanced through a dual mechanism of live bacteria colonization and substrate supply.
It effectively repairs the intestinal mucosal barrier, reduces intestinal permeability, regulates immune balance, optimizes the microbial community structure, simplifies the usage method, and is easy to promote in farms.
Smart Images
Figure CN122097435A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of microbial technology, and particularly to a synbiotic and its uses. Background Technology
[0002] Colitis is one of the major health threats in poultry farming and a key scientific issue hindering the healthy development of the industry. Colitis is caused by bacterial, parasitic, or mixed infections. Current main solutions for poultry colitis include: Using probiotic preparations to regulate the intestinal microecology, a compound probiotic isolated from the duodenum of poultry ( Bacteroides caecicola , B. uniformis , B. salanitronis It can enhance the expression of immune-related genes in the intestinal mucosa, reduce inflammatory response, and inhibit the proliferation of harmful bacteria. However, long-term use of the preparation is required to maintain its effect, and the efficacy of probiotic preparations against complex infections caused by different strain combinations is unstable, making large-scale production difficult.
[0003] Traditional Chinese medicine formulas and extracts have significant preventive effects against coccidiosis and can reduce dependence on chemical drugs. For example, berberine and oxidized berberine can reduce inflammation by inhibiting the TLR4 / NF-κB signaling pathway and regulate the gut microbiota (e.g., increasing...). Akkermansia (Abundance); however, its complex composition leads to unclear mechanisms of action, some prescriptions have poor palatability affecting feed intake, and there is a lack of dosage optimization studies for specific poultry breeds.
[0004] Precision antibiotic therapy and resistance management, along with antibiotic use guided by drug susceptibility testing (e.g., for E. coli infections sensitive to neomycin), can rapidly control the disease. Targeted medication can control mixed infections, but long-term use can lead to the spread of resistance (e.g., tetracycline resistance in Salmonella enteritidis) and disrupt the intestinal flora balance, increasing the risk of secondary infections.
[0005] While environmental management and biosecurity measures, such as all-in-all-out systems, biological thermal fermentation of manure, and flame disinfection, can effectively kill coccidia oocysts and pathogens, they are difficult to implement in free-range poultry farming models, require continuous investment in facilities, and have excessively high labor costs.
[0006] In summary, the above prevention and control measures have significant shortcomings. Currently, research on how to treat poultry, especially chicken colitis, is scarce. Summary of the Invention
[0007] To address the technical problems existing in the prior art, this invention proposes a synbiotic, comprising: probiotics or their bacterial lysates and fermentation products, wherein the probiotics are selected from one or more of the following groups: Bifidobacterium, Streptococcus, Lactobacillus, Saccharomyces, and Escherichia coli; the prebiotics are selected from one or more of the following groups: human milk oligosaccharides, fructooligosaccharides, inulin, lactulose, galactooligosaccharides, soybean oligosaccharides, resistant dextrin, xylooligosaccharides, isomaltooligosaccharides, and chitosan oligosaccharides; and a wall material; wherein the probiotics or their bacterial lysates and fermentation products are coupled with the prebiotics, and the wall material encapsulates the coupling.
[0008] As described above, the probiotics in the synbiotic are selected from one or more of the following groups: Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, Yellow Flag Bacillus infantis, Bifidobacterium lactis, Bifidobacterium longum, Saccharomyces boulardii, Streptococcus salivarius, Bacillus coagulans, Clostridium butyricum, Enterococcus faecalis, and Bacillus subtilis.
[0009] As described above, the probiotics in the synbiotic are selected from Lactobacillus plantarum.
[0010] As described above, the prebiotic is selected from fructooligosaccharides.
[0011] As described above, the wall material of the synbiotic comprises oligopectin and / or chitosan.
[0012] A method for preparing a synbiotic includes: adding *Lactobacillus plantarum* cells to a fructooligosaccharide solution to obtain a fructooligosaccharide bacterial suspension; wherein the concentration of the fructooligosaccharide solution is (1-8) mg / mL; adding a low-ester pectin solution to the bacterial suspension to obtain a mixture; wherein the concentration of the low-ester pectin solution is (0.05-0.1) g / mL; preferably, the concentration of the low-ester pectin solution is 0.063 g / mL; wherein the volume ratio of the bacterial suspension to the low-ester pectin solution is 1:(1-3); preferably, the volume ratio of the bacterial suspension to the low-ester pectin solution is 1:2; dripping the mixture into a metal ion solution and stirring to obtain synbiotic microspheres; wherein the concentration of the metal ions is (0.1-0.5) M; preferably, the concentration of the metal ions is (0.1-0.3) M; more preferably, the concentration of the metal ions is 0.2 M. M; Add the synbiotic microspheres to a chitosan solution and stir for a certain period of time; wherein the concentration of the chitosan solution is (1-8) mg / mL; preferably, the concentration of the chitosan solution is (2-5) mg / mL; more preferably, the concentration of the chitosan solution is 3 mg / mL; add the obtained product to a metal ion solution; wherein the concentration of the metal ion is (0.02-0.5) M; preferably, the concentration of the metal ion is (0.5-0.3) M; more preferably, the concentration of the metal ion is 0.1 M; wash the product to obtain the synbiotic.
[0013] The preparation method described above further includes: placing the bacterial suspension in an environment of 28-37°C and shaking it for 20-60 min.
[0014] In the preparation method described above, the metal ion is selected from one or more of the following group: calcium ion, magnesium ion, zinc ion, iron ion, and ferrous ion.
[0015] Use of the synbiotic as described above or the synbiotic prepared by any of the methods described above in the preparation of a drug or food additive for treating avian colitis.
[0016] For the purposes described above, the poultry is selected from chickens; preferably, the poultry is selected from one or more of the following group: Yunnan Wuliangshan Black-boned Chicken, Wuding Chicken, Yanjin Black-boned Chicken, Zhenyuanpiao Chicken, Daweishan Miniature Chicken, Camellia Chicken, and commercial broiler chickens; more preferably, the poultry is selected from Yunnan Wuliangshan Black-boned Chicken.
[0017] The synbiotic formulation in this application is simple, containing no added chemical drugs or antibiotics. It can repair damaged intestinal mucosal barriers, reduce intestinal permeability, regulate immune balance, and optimize gut microbiota structure, thereby more effectively treating colitis. Furthermore, it is easy to use and readily applicable to farms. Attached Figure Description
[0018] The preferred embodiments of the present invention will now be described in further detail with reference to the accompanying drawings, wherein: Figure 1 According to an embodiment of the present invention L.plantarum Quantitative results of co-cultivation with FOS; among which, L.plantarum It is Lactobacillus plantarum, and FOS is fructooligosaccharide; Figure 2 According to an embodiment of the present invention L.plantarum The optimal gel-forming concentration of @LMP / CS gel microspheres was determined; among which... L.plantarum It is Lactobacillus plantarum, LMP is low-ester pectin, and CS is chitosan; Figure 3 According to an embodiment of the present invention L.plantarum SEM images of LMP / CS gel microspheres; L.plantarum Lactobacillus plantarum, LMP is low-ester pectin, and CS is chitosan; Scale bar: 500 nm / 1 µm; Figure 4 This refers to a control group, a 3% DSS challenge, and a 7% DSS challenge, according to one embodiment of the present invention. L.plantarum Colon HE staining image of the LMP / CS treatment group; Scale bar: 1 mm / 100 µm; Figure 5This refers to a control group, a 3% DSS challenge, and a 7% DSS challenge, according to one embodiment of the present invention. L.plantarum @Statistical results of gut microbiota abundance in the LMP / CS treatment group; Figure 6 This refers to a control group, a 3% DSS challenge, and a 7% DSS challenge, according to one embodiment of the present invention. L.plantarum The quantification results of the cecal ELISA kit in the LMP / CS treatment group were detected. Detailed Implementation
[0019] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0020] In the following detailed description, reference can be made to the accompanying drawings, which form part of this application and illustrate specific embodiments of the present application. In the drawings, similar reference numerals describe substantially similar components in different figures. Specific embodiments of the present application are described in sufficient detail below to enable those skilled in the art to implement the technical solutions of the present application. It should be understood that other embodiments or modifications to the embodiments of the present application may also be utilized.
[0021] This invention enhances the therapeutic effect of Lactobacillus plantarum and fructooligosaccharides through a dual mechanism of "live bacteria colonization and substrate supply," which can solve the colitis problem faced by chickens, especially Wuliangshan black-bone chickens, and has important practical value in production, ensuring the quality and yield of this local specialty product.
[0022] Existing research largely focuses on the gut health of common broiler chickens (such as white-feathered and yellow-feathered broilers). This invention, for the first time, establishes a synergistic model of the synergistic effect of fructooligosaccharides and Lactobacillus plantarum in a chicken colitis model. Based on the experimental results, a feed-compatible, microencapsulated synergistic formulation was developed, achieving a feed additive form that is resistant to gastric acid and heat and targets the gut. 16S sequencing, metagenomics, and mycology were combined to evaluate the mechanism of action of the synergistic, forming a comprehensive analytical framework. This invention is distinctive and innovative in its selection of research subjects, research focus, and research approach.
[0023] Unless otherwise specified, all experimental methods involved in this application are conventional methods existing in this technical field, and all ingredients or materials used, unless otherwise specified, are commercially available. The influence of different brands or batches of ingredients or materials on the experimental results of this application is negligible.
[0024] Some terms appearing in this application have the following meanings: The term "birds" as used in this application refers to poultry and birds of prey. In some embodiments, birds may be classified into pheasants, anatidae, land birds, waterfowl, wading birds, birds of prey, songbirds, climbing birds, etc.
[0025] The term "chicken" as used in this application refers to a type of poultry belonging to the Phasianidae family. There are many breeds of chickens, generally categorized as broiler chickens, laying hens, fighting chickens, ornamental chickens, medicinal chickens, and dual-purpose (meat and egg) chickens. In some embodiments, chickens include, but are not limited to, local Yunnan chicken breeds, such as the Wuliang Mountain Black-boned Chicken, Wuding Chicken, Yanjin Black-boned Chicken, Zhenyuan Piao Chicken, Daweishan Miniature Chicken, and Camellia Chicken; commercial broiler chickens, such as white-feathered broiler chickens and yellow-feathered broiler chickens; and various local chicken breeds from across China.
[0026] The "black-boned chicken" mentioned in this application, also known as the Silkie chicken, is a type of poultry whose entire body, including skin, meat, bones, and internal organs, is black. It belongs to the genus *Gallus* of the family Phasianidae and is known for its medicinal and tonic value. Common black-boned chickens include, but are not limited to, the Wuliang Mountain Black-boned Chicken of Yunnan, the Yanjin Black-boned Chicken, the Taihe Black-boned Chicken, the Black Phoenix Chicken, the Yugan Black-boned Chicken, and the Wumeng Black-boned Chicken.
[0027] The term "colitis" as used in this application refers to intestinal inflammation in poultry, including the colon and small intestine, and is a common problem in poultry farming.
[0028] The term "synbiotic" as used in this application refers to a mixture of probiotics or their metabolites and prebiotics. Metabolites include, but are not limited to, cell lysates and fermentation broths.
[0029] In some embodiments, probiotics or their metabolites are beneficial to the gut. These probiotics include, but are not limited to, Bifidobacterium, Streptococcus, Lactobacillus, Saccharomyces, and Escherichia coli. Furthermore, probiotics include, but are not limited to, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, Yellow Flag Bacillus infantis, Bifidobacterium lactis, Bifidobacterium longum, Saccharomyces boulardii, Streptococcus salivarius, Bacillus coagulans, Clostridium butyricum, Enterococcus faecalis, and Bacillus subtilis.
[0030] The "Lactobacillus plantarum" mentioned in this application is a type of lactic acid bacteria that can promote food breakdown and relieve constipation and diarrhea. The Lactobacillus plantarum mentioned in this application is a commercially available product purchased directly; its manufacturer or production batch has little impact on the technical effect of this application, and therefore is not limited thereto.
[0031] In some embodiments, prebiotics refer to substances that can selectively and minimally stimulate the activity or growth of one or more probiotics in the host's gut, without being digested or absorbed by the host, thereby promoting the colonization and growth of probiotics in the gut. In some embodiments, prebiotics include, but are not limited to: human milk oligosaccharides, fructooligosaccharides, inulin, lactulose, galactooligosaccharides, soybean oligosaccharides, resistant dextrin, xylooligosaccharides, isomaltooligosaccharides, and oligosaccharides such as chitosan oligosaccharides.
[0032] The term "fructooligosaccharide" as used in this application refers to a short-chain carbohydrate consisting of 2-9 fructose molecules linked by β-2,1 glycosidic bonds. Fructose oligosaccharides are commonly found in citrus fruits, onions, garlic, bananas, chicory roots, etc., and can promote the proliferation of bifidobacteria, inhibit harmful bacteria, and improve intestinal motility and calcium absorption.
[0033] In some embodiments, the present invention combines fructooligosaccharides with Lactobacillus plantarum. Fructooligosaccharides can not only act as a prebiotic to selectively promote the proliferation of Lactobacillus plantarum in the intestine, but can also be fermented by the strain to produce short-chain fatty acids (such as acetic acid, propionic acid, and butyric acid), which together reduce the intestinal pH value and inhibit the growth of pathogenic bacteria such as Escherichia coli and Salmonella.
[0034] The term "low-ester pectin" as used in this application refers to pectin with a degree of esterification of less than 50%. Low-ester pectin can form irreversible gels with the participation of metal ions such as calcium ions. In the food industry, low-ester pectin can be used as a gelling agent, a pharmaceutical sustained-release formulation, etc. In some embodiments, the metal ions include, but are not limited to, magnesium ions, zinc ions, iron ions, ferrous ions, and calcium ions.
[0035] The "chitosan" mentioned in this application refers to a natural polysaccharide extracted from the shells of crustaceans or insects, which is made from chitin through deacetylation and has biocompatibility, biodegradability and antibacterial properties.
[0036] The “DSS” mentioned in this application refers to sodium dextran sulfate, a chemically synthesized compound produced by sulfation modification, used to induce acute or chronic ulcerative colitis models.
[0037] The "PBS buffer" mentioned in this application is a phosphate buffer solution, a solution that simulates the in vivo environment of a living organism. PBS buffer has salt balance and an adjustable pH buffering effect, and is mainly used to maintain the stability of the liquid's acidity or alkalinity during experimental operations, creating a stable experimental environment.
[0038] The "HE staining" mentioned in this application refers to hematoxylin-eosin staining, which is the most commonly used staining technique in pathology. Hematoxylin stains the cell nucleus blue, while eosin stains the cytoplasm and collagen fibers pink, thus clearly revealing the microstructure of colon tissue.
[0039] This application provides a synbiotic whose active ingredient is a combination of fructooligosaccharides and Lactobacillus plantarum, which effectively relieves colon inflammation by promoting the growth of beneficial bacteria and inhibiting the growth of pathogenic bacteria.
[0040] The synbiotic in this application can repair the damaged intestinal mucosal barrier, reduce intestinal permeability, regulate immune balance, and optimize the microbiota structure, thereby more effectively treating colitis.
[0041] This application discloses a synbiotic comprising: probiotics or their bacterial lysates or fermentation products, wherein the probiotics are selected from one or more of the following groups: Bifidobacterium, Streptococcus, Lactobacillus, Saccharomyces, and Escherichia coli; prebiotics are selected from one or more of the following groups: human milk oligosaccharides, fructooligosaccharides, inulin, lactulose, galactooligosaccharides, soybean oligosaccharides, resistant dextrin, xylooligosaccharides, isomaltooligosaccharides, and chitosan oligosaccharides; and a wall material; wherein the probiotics or their bacterial lysates or fermentation products are coupled with the prebiotics, and the wall material encapsulates the coupling.
[0042] In some embodiments, the probiotics are selected from one or more of the following groups: Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, Yellow Flag Bacillus infantis, Bifidobacterium lactis, Bifidobacterium longum, Saccharomyces boulardii, Streptococcus salivarius, Bacillus coagulans, Clostridium butyricum, Enterococcus faecalis, and Bacillus subtilis.
[0043] In some embodiments, the probiotics are selected from Lactobacillus plantarum.
[0044] In some embodiments, the prebiotic is selected from fructooligosaccharides.
[0045] In some embodiments, the wall material includes oligopectin and / or chitosan.
[0046] The preparation method of the above-mentioned synbiotic includes: adding *Lactobacillus plantarum* cells to an oligofructose solution to obtain an oligofructose bacterial suspension; wherein the concentration of the oligofructose solution is (3-8) mg / mL; preferably, the concentration of the oligofructose solution is 5 mg / mL; adding a low-ester pectin solution to the bacterial suspension to obtain a mixture; wherein the concentration of the low-ester pectin solution is (0.05-0.1) g / mL; preferably, the concentration of the low-ester pectin solution is 0.063 g / mL. g / mL; wherein the volume ratio of bacterial suspension to low-ester pectin solution is (1-3):1; preferably, the volume ratio of bacterial suspension to low-ester pectin solution is 2:1; the mixture is dripped into a metal ion solution and stirred to obtain synbiotic microspheres; wherein the concentration of the metal ion is (0.1-0.5) M; preferably, the concentration of the metal ion is (0.1-0.3) M; more preferably, the concentration of the metal ion is 0.2 M; the synbiotic microspheres are added to a chitosan solution and stirred for a certain period of time; wherein the concentration of the chitosan solution is (1-8) mg / mL; preferably, the concentration of the chitosan solution is (2-5) mg / mL; more preferably, the concentration of the chitosan solution is 3 mg / mL. mg / mL; the obtained product is added to a metal ion solution; wherein the concentration of the metal ion is (0.02-0.5) M; preferably, the concentration of the metal ion is (0.5-0.3) M; more preferably, the concentration of the metal ion is 0.1 M; the product is washed to obtain synbiotic.
[0047] In some embodiments, the preparation method further includes: placing the bacterial suspension in an environment of 28-37°C and shaking it for 20-60 min.
[0048] In some embodiments, the metal ion is selected from one or more of the group consisting of calcium ions, magnesium ions, zinc ions, ferric ions, and ferrous ions.
[0049] In some embodiments, the drug or food additive is used to treat or alleviate symptoms in a subject associated with abnormal bacterial niche metabolism.
[0050] Preferably, in some embodiments, the drug or food additive is used to treat or alleviate conditions such as obesity, inflammatory bowel disease, Crohn's disease, colitis, asthma, cystic fibrosis, allergies, diabetes, psoriasis, eczema, atopic dermatitis, gastrointestinal reflux disease, gastrointestinal cancer, infection, autism spectrum disorder, bacterial vaginosis, rheumatoid arthritis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, necrotizing enterocolitis, bacterial infection, viral infection, and parasitic infection in the subject.
[0051] In some embodiments, the route of administration of the drug or food additive is selected from oral, rectal, vaginal, pulmonary, ocular, or parenteral administration. Preferably, in some embodiments, the drug or food additive may be incorporated into the subject's drinking water or feed.
[0052] In some embodiments, the subjects are poultry. In some embodiments, the poultry is selected from chickens; preferably, the poultry is selected from one or more of the following group: Yunnan Wuliangshan Black-boned Chicken, Wuding Chicken, Yanjin Black-boned Chicken, Zhenyuanpiao Chicken, Daweishan Miniature Chicken, Camellia Chicken, and commercial broiler chickens; more preferably, the poultry is selected from Yunnan Wuliangshan Black-boned Chicken.
[0053] The above-mentioned synbiotic or the synbiotic prepared by the method described herein may be used in the preparation of drugs or food additives for treating avian colitis.
[0054] The technical solution of this application will be described below through specific embodiments. Those skilled in the art should understand that the following embodiments are only for illustrating the specific implementation process of the technical solution of this application, and are not intended to limit the technical solution of this application.
[0055] Example 1: The process of constructing a synbiotic from fructooligosaccharides and Lactobacillus plantarum Preparation of Lactobacillus plantarum suspension: Activated Lactobacillus plantarum (… L. plantarum After culturing to the logarithmic growth phase, the bacterial cells were collected by centrifugation, washed, and resuspended in PBS to obtain a bacterial suspension with a concentration of 6.83 × 10⁻⁶. 9 CFU / mL.
[0056] Preparation of FOS (Fructooligosaccharide) solution: Weigh FOS powder and dissolve it in deionized water to prepare FOS solutions with a final concentration of 1-5 mg / mL. Stir magnetically (300 rpm) for 15 min until completely dissolved and without precipitation. Filter through a 0.22 μm filter membrane for sterilization and store at 4℃ for later use.
[0057] Low-ester pectin (LMP) solution: Weigh low-ester pectin (LMP) powder, dissolve it in deionized water to a final concentration of 0.063 g / mL, stir magnetically (300 rpm, 60℃) until completely dissolved, cool to room temperature, filter through a 0.45 μm filter membrane for sterilization, and store at 4℃ for later use.
[0058] Chitosan (CS) solution: Take chitosan powder and dissolve it in 1% (v / v) sterile acetic acid solution to a final concentration of 3 mg / mL. Stir magnetically (500 rpm, 50℃) until completely dissolved, cool to room temperature, filter through a 0.45 μm filter membrane for sterilization, and store at 4℃ for later use.
[0059] Take 4 ml of the *Lactobacillus plantarum* bacterial suspension obtained by the aforementioned method, centrifuge to collect the bacterial cells, wash them, and resuspend them in 1 mL of fructooligosaccharide (FOS) solutions with concentrations of 1 mg / mL, 2.5 mg / mL, and 5 mg / mL to obtain bacterial suspensions. Incubate at 37°C with shaking at 150 rpm for 30 min to enhance the adsorption of bacteria and FOS, obtaining mixed bacterial suspension A. Suspension A includes... L. plants / FOS complex.
[0060] Figure 1 An embodiment according to this application is shown. L.plantarum Quantitative results of co-culturing with FOS. The results showed that the bacterial cell growth was best at a concentration of 5 mg / mL FOS compared to the control group.
[0061] Mixed bacterial suspension A with 0.063 g / mL low-ester pectin (LMP) solution at a ratio of V(bacterial suspension A):V(LMP solution) of 1:2. The mixture was stirred with a magnetic stirrer (150 rpm) for 5 min to obtain mixed bacterial solution B.
[0062] Crosslinking by dripping: Mixed bacterial suspension B was dripped into 0.2 M CaCl2 solution at a rate of 1 drop / second through a 0.45 mm needle hole, and crosslinked for 10 min by magnetic stirring (100 rpm) to form gel microspheres. L.plantarum @LMP, rinse the microspheres three times with deionized water to remove surface CaCl2 residue.
[0063] Will get L.plantarum @LMP gel microspheres were immersed in a 3 mg / mL chitosan solution and magnetically stirred (100 rpm) for 15 minutes to form a dense coating. The microspheres were then rinsed three times with deionized water.
[0064] After rinsing, the microspheres were transferred to 0.1 M CaCl2 solution and allowed to stand for a second cross-linking process for 5 min. The microspheres were then rinsed with PBS buffer to remove residual acetic acid and Ca. 2+ ,form L.plantarum @LMP / CS Biostime
[0065] Figure 2 An embodiment according to this application is shown. L.plantarum The optimal gelation concentration of @LMP / CS gel microspheres was determined.
[0066] like Figure 2As shown in Figure A, after different concentrations of LMP (55 mg / ml, 60 mg / ml, 65 mg / ml, 70 mg / ml) were treated with different concentrations of CaCl2 (0.1 M, 0.2 M, 0.3 M), the 65 mg / ml LMP and 0.2 M CaCl2 showed the most obvious spherical formation, while the others formed fluffy or flake-like shapes.
[0067] like Figure 2 As shown in B, LMP at different concentration gradients (61 mg / ml, 63 mg / ml, 65 mg / ml, 67 mg / ml, 69 mg / ml) was crosslinked with 0.2 M CaCl2 and then treated with different concentrations of CS (0.1 M, 0.3 M). The results showed that 63 mg / ml LMP, 0.3 M CS, and 0.2 M CaCl2 produced obvious spheroids without significant tailing, and were ultimately determined to be the optimal gelation concentration.
[0068] Figure 3 An embodiment according to this application is shown. L.plantarum @LMP / CS Biostime SEM results image, SEM results display. L.plantarum It is rod-shaped, and the @LMP / CS surface is densely plate-like, while L.plantarum @LMP / CS display L.plantarum Inlaid within @LMP / CS, indicating that @LMP / CS was successfully modified. L.plantarum The surface, i.e. L.plantarum @LMP / CS assembly successful.
[0069] Example 2: Animal husbandry Animal selection. Forty-five 35-day-old Wuliangshan black-boned chickens with fully developed intestines were selected as the research subjects and randomly divided into three groups: control group, challenge group, and treatment group, with 15 chickens in each group. They were raised in cages in a three-dimensional manner.
[0070] Challenge and treatment. Both the challenge and treatment groups were fed drinking water containing 3% DSS for three consecutive days to establish a challenge model. During the experiment, the control and 3% DSS challenge groups were fed a basal diet, while the treatment group was fed a diet containing 7% DSS. L.plantarum Continue feeding with LMP / CS Biostime diet for 7 more days.
[0071] The basal feed was purchased from Shennong Group (2508130001). During the trial, the pens were regularly washed to maintain ventilation, cleanliness, dryness, and hygiene. Other feeding management and immunization programs were carried out in accordance with the regulations of normal production procedures.
[0072] Sample collection and preservation. After on-site slaughter and dissection, spleen, kidney, jejunum, ileum, colon, and cecum tissue segments were collected. Some were fixed with fixative and used for section analysis and immunohistochemistry, while others were frozen at -80℃ for molecular biological analysis.
[0073] Intestinal barrier function assessment. Fixed intestinal tissue samples were paraffin-embedded, sectioned, and stained with hematoxylin and eosin (HE) to observe the morphology and structure of the intestinal mucosa, assess the integrity and development of the intestinal mucosa, and evaluate the abundance of intestinal flora.
[0074] Figure 4 This refers to a control group, a 3% DSS challenge, and a 7% DSS challenge, according to one embodiment of the present invention. L.plantarum Colon HE staining image of the LMP / CS synbiotic treatment group, as shown Figure 4 As shown, compared with the control group, the intestinal villi epithelium in the challenge group was largely shed and necrotic, and the crypt structure was destroyed and expanded due to inflammatory cell infiltration. After ingesting synbiotic, the above-mentioned symptom areas began to return to normal, and the treatment effect was obvious.
[0075] Figure 5 This refers to a control group, a 3% DSS challenge, and a 7% DSS challenge, according to one embodiment of the present invention. L.plantarum The results of gut microbiota abundance statistics in the LMP / CS synbiotic treatment group are as follows: Figure 5 As shown, intestinimonas were less abundant in the control and 3% DSS groups, but significantly increased in the treatment group. Intestinimonas are associated with the production of short-chain fatty acids, and their increase indicates a regulatory effect of the treatment group on gut health; Desulfovibrio, through the production of... It exacerbates intestinal inflammation and damage, and is significantly elevated in the 3% DSS challenge group. After synbiotic intake, the abundance of Desulfovibrio decreases.
[0076] Figure 6 This refers to a control group, a 3% DSS challenge, and a 7% DSS challenge, according to one embodiment of the present invention. L.plantarum @LMP / CS treatment group cecal ELISA kit detection quantitative results, such as Figure 6 As shown, compared with the control group and the challenge group, the levels of pro-inflammatory factors IL-1β and TNF-α were significantly increased. After treatment with synbiotics, the levels of pro-inflammatory factors IL-1β and TNF-α were significantly reduced, and the inflammation was relieved.
[0077] The above embodiments are for illustrative purposes only and are not intended to limit the invention. Those skilled in the art can make various changes and modifications without departing from the scope of the invention. Therefore, all equivalent technical solutions should also fall within the scope of the invention.
Claims
1. A synbiotic, comprising: Probiotics or their bacterial lysates or fermentation products, wherein the probiotics are selected from one or more of the following groups: Bifidobacterium, Streptococcus, Lactobacillus, Saccharomyces, and Escherichia coli. Prebiotics are selected from one or more of the following group: human milk oligosaccharides, fructooligosaccharides, inulin, lactulose, galactooligosaccharides, soybean oligosaccharides, resistant dextrin, xylooligosaccharides, isomaltooligosaccharides, chitosan oligosaccharides; and wall material; The probiotics or their bacterial lysates, fermentation products, and prebiotics are coupled together, and the wall material encapsulates the coupling.
2. The synbiotic according to claim 1, wherein the probiotic is selected from one or more of the following groups: Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, Yellow Flag Bacillus infantis, Bifidobacterium lactis, Bifidobacterium longum, Saccharomyces boulardii, Streptococcus salivarius, Bacillus coagulans, Clostridium butyricum, Enterococcus faecalis, and Bacillus subtilis.
3. The synbiotic according to claim 1, wherein the probiotic is selected from Lactobacillus plantarum.
4. The prebiotic according to claim 1, wherein the prebiotic is selected from fructooligosaccharides.
5. The synbiotic according to claim 1, wherein the wall material comprises oligopectin and / or chitosan.
6. A method for preparing a synbiotic, comprising: Lactobacillus plantarum cells are added to an oligofructose solution to obtain an oligofructose bacterial suspension; wherein the concentration of the oligofructose solution is (1-8) mg / mL; preferably, the concentration of the oligofructose solution is 5 mg / mL; A low-ester pectin solution is added to the bacterial suspension to obtain a mixture; wherein the concentration of the low-ester pectin solution is (0.05-0.1) g / mL; preferably, the concentration of the low-ester pectin solution is 0.063 g / mL; wherein the volume ratio of the bacterial suspension to the low-ester pectin solution is 1:(1-3); preferably, the volume ratio of the bacterial suspension to the low-ester pectin solution is 1:
2. The mixture is dripped into a metal ion solution and stirred to obtain synbiotic microspheres; wherein the concentration of the metal ions is (0.1-0.5) M; preferably, the concentration of the metal ions is (0.1-0.3) M; more preferably, the concentration of the metal ions is 0.2 M; The synbiotic microspheres are added to a chitosan solution and stirred for a certain period of time; wherein the concentration of the chitosan solution is (1-8) mg / mL; preferably, the concentration of the chitosan solution is (2-5) mg / mL; more preferably, the concentration of the chitosan solution is 3 mg / mL; The obtained product is added to a metal ion solution; wherein the concentration of the metal ion is (0.02-0.5) M; preferably, the concentration of the metal ion is (0.05-0.3) M; more preferably, the concentration of the metal ion is 0.1 M. The product is cleaned to obtain synbiotic.
7. The preparation method according to claim 6, further comprising: The bacterial suspension was placed in an environment of 28-37℃ and shaken for 20-60 min.
8. The preparation method according to claim 6, wherein the metal ion is selected from one or more of the following group: calcium ion, magnesium ion, zinc ion, iron ion, ferrous ion.
9. Use of the synbiotic as described in any one of claims 1-5 or the synbiotic prepared by any one of claims 6-8 in the preparation of a medicament or food additive for treating avian colitis.
10. The use according to claim 9, wherein the poultry is selected from chickens; preferably, the poultry is selected from one or more of the following: Yunnan Wuliangshan Black-boned Chicken, Wuding Chicken, Yanjin Black-boned Chicken, Zhenyuanpiao Chicken, Daweishan Miniature Chicken, Camellia Chicken, and commercial broiler chickens; more preferably, the poultry is selected from Yunnan Wuliangshan Black-boned Chicken.