A kind of Megamonas monomorpha and its application in preventing or treating stress in livestock and poultry

Abstract

The present invention provides a Megamonas monomorpha and an application thereof in preventing or treating stress in livestock and poultry. The Megamonas monomorpha can improve the growth performance of Tianfu broilers and regulate stress response; the Megamonas monomorpha can improve the effects of high-density feeding on the production performance and behavior of Tianfu broilers; and improve the effects of high-density feeding on antioxidant indicators and hormone indicators of Tianfu broilers; the present invention explores the application of Megamonas monomorpha in high-density feeding stress, has good practical application value for alleviating high-density feeding stress in the breeding process, and has good prospects for promotion and application.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to a Megamonas monomorpha and application thereof in preventing or treating stress in livestock and poultry. Background Art

[0002] High-density feeding stress is a common situation in broiler production. It is mainly due to the fact that high-density feeding reduces the living space of broilers, increases the ammonia concentration, carbon dioxide concentration and harmful microorganisms in the environment, resulting in decreased broiler production performance; decreased immunity of broilers; disordered intestinal microbial structure of broilers; dysfunction of 5-hydroxytryptamine (5-HT) and decreased animal welfare level. Its typical characteristics include decreased broiler production performance, reduced market weight and feed conversion rate; changes in broiler behavior, more abnormal behaviors such as fighting and feather pecking, prone to group explosion, and severe death of chickens; disordered hormone secretion of broilers, increased serum cortisol (CORT) concentration, and decreased hippocampal 5-HT concentration. The broiler breeding mode of ground flat breeding, net flat breeding and cage breeding is often accompanied by the problem of high-density feeding stress. High-density feeding stress can directly or indirectly cause broiler illness or death, increase the use of drugs in the breeding process, reduce production efficiency and product treatment, and increase potential food safety risks.

[0003] In recent years, more and more scholars have paid attention to the research on the brain-gut axis, and the view that microorganisms are involved in host emotions and stress responses has been recognized by more and more researchers. Currently, the use of probiotics has performed well in experiments to improve the host's ability to resist stress.

[0004] Since high-density feeding stress is common in broiler farming, and broiler stress is difficult to identify and chickens are prone to death, the broiler farming industry is in urgent need of a new, non-toxic, green, safe and effective probiotic for alleviating and / or treating high-density feeding stress in broilers. Summary of the invention

[0005] In view of this, in order to solve the above-mentioned technical problems, the purpose of the present invention is to provide a Megamonas monomorpha and its application in preventing or treating livestock and poultry stress, wherein the Megamonas monomorpha can improve the growth performance of Tianfu broilers and regulate stress response; the Megamonas monomorpha can improve the effects of high stocking density on the production performance and behavior of Tianfu broilers; and improve the effects of high stocking density on the antioxidant indicators and hormone indicators of Tianfu broilers; the present invention explores the application of Megamonas monomorpha in high stocking density stress, which has great practical application value for alleviating high stocking density stress in the breeding process and has good prospects for promotion and application.

[0006] In order to achieve the above purpose, the technical solutions adopted are as follows:

[0007] A single-shaped giant monocytic bacterium, characterized in that the single-shaped giant monocytic bacterium is a genus of giant monocytic bacterium of the phylum Firmicutes, a Gram-negative anaerobic bacterium, and the optimal growth conditions can be 37°C and anaerobic; the colonies cultured on a blood plate medium for 48 hours can be light yellow, flat, with irregular wavy edges, and the bacteria are rod-shaped under a microscope. The single-shaped giant monocytic bacterium has the ability to produce acetic acid and propionic acid and a small amount of lactic acid and succinic acid. The blood plate medium is TSA+5% defibrinated sheep blood. The single-shaped giant monocytic bacterium is purchased from Wuhan Zhili Zhongte Biotechnology Co., Ltd., with a product number of Bio-107321 and an original number of JCM14723=DSM19343.

[0008] Furthermore, the nucleotide sequence of 16S rDNA of the Megamonas monomorpha is shown in SEQ ID NO.1.

[0009] The culture of Megamonas monomorpha also belongs to the protection scope of the present invention. The culture of Megamonas monomorpha is a substance obtained by culturing Megamonas monomorpha in a microbial culture medium (such as a fermentation broth containing Megamonas monomorpha and a substance secreted into a liquid culture medium, or a fermentation broth containing Megamonas monomorpha and a substance secreted into a solid culture medium).

[0010] A bacterial agent contains the Megamonas monomorpha and / or the culture.

[0011] The microbial agent may further include a carrier. The carrier may be a solid carrier or a liquid carrier. The solid carrier is a mineral material or a biological material; the mineral material may be at least one of peat, clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica and diatomaceous earth; the biological material may be at least one of various crop straws, pine husks, straw, peanut shells, corn flour, bean flour, starch, peat and animal feces; the liquid carrier may be water; the formulation of the microbial agent may be a variety of formulations, such as liquid, emulsion, suspension, powder, granule, wettable powder or water dispersible granule.

[0012] Surfactants (such as Tween 20, Tween 80, etc.), binders, stabilizers (such as antioxidants), pH regulators, etc. may also be added to the bacterial agent.

[0013] Furthermore, the bacterial agent has at least one of the following characteristics:

[0014] A1) Treat or alleviate stress-related diseases in poultry / livestock;

[0015] A2) Increase the slaughter weight of poultry and / or livestock;

[0016] A3) Improve feed conversion efficiency of poultry and / or livestock;

[0017] A4) Improving the antioxidant capacity of poultry and / or livestock serum;

[0018] A5) Reducing serum cortisol concentrations in poultry and / or livestock;

[0019] A6) Reducing serum norepinephrine concentrations in poultry and / or livestock;

[0020] A7) Improving the immunity of poultry and / or livestock;

[0021] A8) Improving brain 5-HT metabolism in poultry and / or livestock;

[0022] A9) Improving animal welfare of poultry and / or livestock.

[0023] Further, at least one of the following uses of Megamonas monomorpha:

[0024] B1) Use of the Megamonas monomorpha described above in the preparation of products for treating or alleviating stress-related diseases in poultry and / or livestock;

[0025] B2) Use of the above-mentioned Megamonas monomorpha in increasing the product weight of poultry and / or livestock;

[0026] B3) Use of the above-mentioned Megamonas monomorpha in improving feed conversion rate of poultry and / or livestock;

[0027] B4) Use of the Megamonas monomorpha described in improving the antioxidant capacity of poultry and / or livestock serum;

[0028] B5) Use of the Megamonas monomorpha described in a product for reducing serum cortisol concentration in poultry and / or livestock;

[0029] B6) Use of the Megamonas monomorpha described above for maintaining serum norepinephrine concentration in poultry and / or livestock;

[0030] B7) Use of the Megamonas monomorpha described above in improving the immunity of poultry and / or livestock;

[0031] B8) The role of the described Megamonas monomorpha in increasing 5-HT metabolism in the brain of poultry and / or livestock;

[0032] B9) Use of the Megamonas monomorpha described in improving the welfare of poultry and / or livestock animals.

[0033] Furthermore, at least one of the following applications of the bacterial agent:

[0034] C1) Use of the bacterial agent in the preparation of products for treating or alleviating stress-related diseases in poultry and / or livestock;

[0035] C2) Use of the bacterial agent in preparing products for increasing the slaughter weight of poultry and / or livestock;

[0036] C3) Application of the bacterial agent in improving feed conversion rate of poultry and / or livestock;

[0037] C4) Use of the bacterial agent in improving the antioxidant capacity of poultry and / or livestock serum;

[0038] C5) Use of the bacterial agent in products for reducing serum cortisol concentration in poultry and / or livestock;

[0039] C6) Use of the bacterial agent in maintaining serum norepinephrine concentration in poultry and / or livestock;

[0040] C7) Use of the bacterial agent in products for improving the immunity of poultry and / or livestock;

[0041] C8) Use of the bacterial agent in increasing 5-HT metabolites in the brain of poultry and / or livestock;

[0042] C9) Use of the bacterial agent in improving the welfare of poultry and / or livestock.

[0043] Furthermore, the poultry is broiler chickens, the stress-related disease is high stocking density stress; and the product is feed or medicine.

[0044] SEQ ID NO. 1 sequence listing:

[0045] 1 ctttgacata agcttccgct tgaagatgag cttgcgtctg attagctagt tggtgagggt

[0046] 61 aaaggcccac caaggcgacg atcagtagcc ggtctgagag gatgaacggc cacattggga

[0047] 121 ctgagacacg gcccagactc ctacgggagg cagcagtggg gaatcttccg caatgggcga

[0048] 181 aagcctgacg gagcaacgcc gcgtgaacga tgaaggtctt aggatcgtaa agttctgttg

[0049] 241 ttagggacga aggataagga ttataataca gtctttgttt gacggtacct aacgaggaag

[0050] 301 ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gcggcaagcg ttgtccggaa

[0051] 361 ttattgggcg taaagggagc gcaggcggga aactaagcgg atcttaaaag tgcggggctc

[0052] 421 aaccccgtga tggggtccga actggttttc ttgagtgcag gaggaaag cggaattccc

[0053] 481 agtgtagcgg tgaaatgcgt agatattggg aagaacacca gtggcgaagg cggctttctg

[0054] 541 gactgtaact gacgctgagg ctcgaaagct agggtagcga acgggattag ataccccggt

[0055] 601 agtcctagcc gtaaacgatg gatactaggt gtgggaggta tcgactcctt ccgtgccgga

[0056] 661 gttaacgcaa tagtatccc gcctggggag tacggccgca aggttgaaac tcaaaggaat

[0057] 721 tgacggggc ccgcacaagc ggtggagtat gtggtttaat tcgacgcaac gcgaagaacc

[0058] 781

[0059] 841 agaaaacagg tggtgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc

[0060] 901 gcaacgagcg caacccctat actatgttgc cagcatttcg gatgggaact catagtagac

[0061] 961 tgccgcggac aacgcggagg aaggcgggga tgacgtcaag tcatcatgcc ccttacgtct

[0062] 1021 tgggctacac acgtactaca atgggatgaa cagagggaag cgaaatcgcgaggtggagcg

[0063] 1081 gatccctaaa agcatctctc agttcggatt gtaggctgaa actcgcctacatgaagtcgg

[0064] 1141 aatcgctagt aatcgcaggt cagcatactg cggtgaatac gttcccgggccttgtacaca

[0065] 1201 ccgcccgtca caccacgaaa gtcattcaca cccgaagccg gctaagggcctatggtaccg

[0066] 1261 accgtctaag gtgggggcga tgattggggt gaagtcgtaa caaggtagccgtatcggaag

[0067] 1321 gtgcggctgg atcacctcct ta

[0068] Beneficial effects:

[0069] The beneficial effects of the present invention are embodied in:

[0070] The present invention provides a Megamonas monomorpha and an application thereof in preventing or treating stress in livestock and poultry. The Megamonas monomorpha can improve the growth performance of Tianfu broilers and regulate stress response; the Megamonas monomorpha can improve the effects of high-density feeding on the production performance and behavior of Tianfu broilers; and improve the effects of high-density feeding on antioxidant indicators and hormone indicators of Tianfu broilers; the present invention explores the application of Megamonas monomorpha in high-density feeding stress, has good practical application value for alleviating high-density feeding stress in the breeding process, and has good prospects for promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0072] Figure 1 This is a comparison diagram of the effect of gavage with Megamonas monomorpha on the body weight of broiler chickens in the embodiments of the present invention;

[0073] Figure 2 This is a comparison diagram of the effect of gavage of Megamonas monomorpha on the feed-to-meat ratio in an embodiment of the present invention;

[0074] Figure 3 This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on serum SOD in the embodiments of the present invention;

[0075] Figure 4 This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on serum IL-6 concentration in the embodiments of the present invention;

[0076] Figure 5 This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on serum INF-γ concentration in the embodiments of the present invention;

[0077] Figure 6 This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on serum IL-2 concentration in an embodiment of the present invention;

[0078] Figure 7 This is a comparison diagram of the effects of intragastric administration of Megamonas monomorpha on serum CORT concentration in the embodiments of the present invention;

[0079] Figure 8 This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on serum NE concentration in the embodiments of the present invention;

[0080] Fig. 9 This is a comparison diagram of the effects of intragastric administration of Megamonas monomorpha on the concentration of 5-HT in the hippocampus in an embodiment of the present invention;

[0081] Fig.10 This is a comparison diagram of the effect of gavage of Megamonas monomorpha on the relative expression of tph2 gene in hippocampus in the embodiment of the present invention;

[0082] Fig.11 This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on the relative expression of HTR1a gene in hippocampus in the embodiment of the present invention;

[0083] Fig.12This is a comparison diagram of the effect of intragastric administration of Megamonas monomorpha on the relative expression level of HTR2a gene in the hippocampus in the examples of the present invention. DETAILED DESCRIPTION

[0084] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0085] Embodiment 1:

[0086] 1.1 Experimental Animal Management

[0087] Tianfu broiler chickens are high-quality chickens with green feet and feathers that are fast-growing and bred by the poultry breeding team of Sichuan Agricultural University. They were approved by the National Livestock and Poultry Genetic Resources in 2018. In this experiment, commercial roosters of Tianfu broiler chickens were selected as experimental animals. The nutritional level of the feed is shown in Table 1, which meets the nutritional standards of commercial generation of Tianfu broiler chickens. During the breeding process, the chickens were free to eat and drink water. The immunization program is shown in Table 2, the brooding temperature and humidity are shown in Table 3, and the lighting system is shown in Table 4.

[0088] Table 1 Nutritional standards for commercial Tianfu broiler chickens

[0089]

[0090] Table 2 Immunization schedule

[0091]

[0092] Table 3 Brooding temperature and humidity

[0093]

[0094] Table 4 Lighting system of chicken house

[0095]

[0096] 1.2 Main Reagents

[0097] (1) Megamonas funiformis was purchased from Zhili Zhongte (Wuhan) Biotechnology Co., Ltd., product number Bio-107321, original number JCM14723=DSM19343.

[0098] (2) Biochemical kit: Total superoxide dismutase (SOD) assay kit, purchased from Nanjing Jiancheng Bioengineering Institute;

[0099] (3) Elisa kits: 5-hydroxytryptamine (5-HT), cortisol (CORT), norepinephrine (NE), interleukin-2 (IL-2), interleukin-6 (IL-6), and INF-γ (interferon γ) assay kits were purchased from Jiangsu Baolai Biotechnology Co., Ltd.

[0100] (4) Blood agar medium (TSA + 5% defibrinated sheep blood), Qingdao Haibo.

[0101] 2 Test methods

[0102] The experiment used one-day-old commercial broiler chicks as the experimental material, and the stocking density was set at 20 chickens / m 2 The mice were gavaged with 1 mL of sterile saline daily as a positive control group (P-CON) and the stocking density was set at 40 mice / m 2 The negative control group (N-CON) was gavaged with 1 mL of sterile saline daily, and the stocking density was 40 mice / m 2 3×10 8 cfu / mL (1mL) of Megamonas monomorpha treatment group (Bacterial), a total of 3 treatments, each treatment set 3 replicates, 35 chickens in each group, a total of 315 chickens, weighed at the beginning of the experiment to ensure that there was no significant difference in weight between groups. The specific grouping is shown in Table 5. The growth performance of broilers was recorded, the behavioral characteristics of broilers were analyzed through video recording, and samples were taken at 30 days of age to measure the differences in welfare indicators and stress indicators. The effect of high density on the stress state of broilers was analyzed.

[0103] Table 5 Experimental grouping and experimental treatment

[0104]

[0105] Biochemical analysis:

[0106] Antioxidant assay: The activity of superoxide dismutase (SOD) in serum was determined using a commercial biochemical kit according to the instructions provided by the manufacturer (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

[0107] Determination of immune indexes: The contents of IL-2 (interleukin-2), IL-6 (interleukin-6), and INF-γ (interferon gamma) in serum were determined using commercial biochemical kits according to the instructions provided by the manufacturer (Jiangsu Baolai Biotechnology Co., Ltd., Jiangsu).

[0108] Determination of hormone indicators: The contents of 5-HT (5-hydroxytryptamine) in the supernatant of hippocampal tissue homogenate, NE (norepinephrine) and CORT (corticosterone) in serum were determined using commercial biochemical kits according to the instructions provided by the manufacturer (Jiangsu Baolai Biotechnology Co., Ltd., Jiangsu).

[0109] Gene expression test:

[0110] RNA extraction, reverse transcription and quantitative PCR:

[0111] ① RNA extraction (Trizol method)

[0112] a. Add 200 μL of chloroform and shake vigorously for 30 s. When the solution becomes turbid, let it stand for 10 mm. Centrifuge at 12000 r / min for 15 min at 4 °C until it becomes clear.

[0113] b. Aspirate the supernatant as much as possible, add 200 μL of chloroform, shake vigorously for 30 s, let it stand for 10 min until it becomes turbid, and centrifuge it at 12000 r / min, 15 min, 4 ℃.

[0114] c. Pipette 400-500 μL of supernatant, add 600 μL of pre-cooled isopropanol, shake for 30 s, let stand for 10 min, and centrifuge at 12000 r / min, 15 min, 4 ℃.

[0115] d. Pour out the supernatant, add 1 mL of 75% ethanol diluted with DEPC water or ddH2O, shake, vortex to shake off the precipitate, and centrifuge at 12000 r / min, 5 min, 4 ℃.

[0116] e. Pour off the supernatant, add 1 mL of 75% ethanol diluted with DEPC water or ddH2O, shake, vortex to shake off the precipitate, and centrifuge at 12000 r / min, 5 min, 4 ℃.

[0117] f. Pour out the supernatant, invert the EP tube on clean filter paper, let it stand for 3-5 minutes, and add ddH2O to dissolve the precipitate after it is dry. The amount added depends on the volume of the supernatant.

[0118] g. Check the concentration and run the gel.

[0119] Note: A260 / A280 should be 1.8-2.0, A260 / A230 should be above 1.6, and best close to 2. The gel should have three bands, 5S light, and bright on the spot well indicates protein contamination, without dragging.

[0120] ②Reverse transcription

[0121] The extracted RNA was reverse transcribed according to the instructions of TAKARA reverse transcription kit Primer Script RTreagent Kit With gDNA Eraser (RR047A). The specific steps are as follows:

[0122] a. Removal of genomic DNA: Add the corresponding volume ratio of reagents to the RNase-free EP tube on ice to prepare the reaction mixture; dispense 9 μL of the prepared mixture into the pre-numbered EP tubes, and then add 1 μL RNA (the RNA sample quality is guaranteed to be 1 μg) in sequence. Store on the PCR instrument at 42 ℃, 2 min, 4 ℃ to remove genomic DNA. See Table 6 for the specific reaction system.

[0123] Table 6 Reaction system for removing residual genomic DNA from RNA

[0124]

[0125] b. Reverse transcription: Continue to add reagents to the reaction solution obtained in the previous step, operate on ice, centrifuge briefly to mix, and perform reverse transcription on a common PCR: 37 ℃, 15 min; 85 ℃, 5 s; 4 ℃, ∞, to obtain cDNA, and store it at -20 ℃. See Table 7 for the specific reaction system.

[0126] Table 7 Reverse transcription PCR reaction system

[0127]

[0128] ③ Fluorescence quantitative PCR

[0129] According to the system in Table 6, the samples were added to the 96-well fluorescence quantitative plate, sealed and centrifuged, and quantified using the Bio-Rad iQ5 fluorescence quantitative PCR instrument. Each sample was technically repeated 3 times. Two-step amplification: 95 °C, 2 min; 95 °C, 5 s, annealing temperature 30 s, repeated 40 cycles; 95 °C, 10 s; 65 °C, 2 s, 95 °C, 5 s. The reaction system is shown in Table 8.

[0130] Table 8 Fluorescence quantitative reaction system

[0131]

[0132] According to the above reaction system and conditions, GADPH was used as the internal reference gene, and RNA was subjected to quantitative PCR analysis on a fluorescent quantitative PCR instrument. The primer information is shown in Table 9.

[0133] Table 9 Fluorescence quantitative target gene primers

[0134]

[0135] Cultivation of Megamonas monomorpha:

[0136] For the first activation of freeze-dried products, all dry powder should be used up and no reserve should be made. Use a sterile pipette to absorb 0.3 mL-0.5 mL of culture solution or sterile water, drop it into the freeze-dried tube, and shake it gently until it dissolves. Absorb all bacterial suspension and inoculate it in blood plate culture medium (TSA + 5% defibrinated sheep blood). Place the culture dish inoculated with bacteria in an anaerobic bag and culture at 37°C for 24-48 hours. Then use a sterilized inoculation loop to scrape the cultured bacteria and store them in 10% sterilized glycerol. The bacterial plate count method was used to detect the viability of the prepared bacterial solution. The test solution was first made into multiple 10-fold incremental dilutions. 1 mL of each dilution was evenly spread on the surface of nutrient agar. The coated agar plate was incubated at 37°C for 24-48 h. After the incubation, the colonies formed on the plate were observed and counted. The average colony number closest to 30 CFU or 300 CFU was selected and multiplied by the dilution factor to calculate the bacterial concentration of the test solution. The prepared concentration was 3×10 8 cfu / mL bacterial solution was stored at -20℃ and slowly thawed before use.

[0137] Data Analysis:

[0138] The experimental data were statistically analyzed using Excel 2019, the difference significance analysis was performed using JMP10 software, and the graphics were drawn using GraphPad Prism 7.04 software. The results were expressed as mean ± standard error (SEM). All trait data were analyzed using one-way analysis of variance, and the difference was considered significant when P < 0.05.

[0139] Results and Analysis:

[0140] 1. Effects of Megamonas monomorpha on the production performance of Tianfu broiler chicks under high-density rearing

[0141] like Figure 1 As shown, the body weight of Tianfu broiler chicks in the positive control group and the Megamonas monomorpha treatment group was significantly increased compared with the negative control group (P < 0.05).

[0142] like Figure 2 As shown in the figure, feed-to-meat ratio refers to the amount of feed consumed to increase the weight of livestock and poultry by 1 kg. It is an important indicator for evaluating feed return. A high feed-to-meat ratio indicates high input and low output. Compared with the negative control group, the feed-to-meat ratio of the positive control group and the Megamonas monomorpha treatment group was significantly reduced (P < 0.05). This shows that Megamonas monomorpha can promote the absorption of nutrients and improve feed utilization efficiency.

[0143] 2. Effects of Megamonas monomorpha on the behavior of Tianfu broiler chicks under high-density rearing

[0144] Table 10 shows that compared with the negative control group, the frequency of movement of Tianfu broiler chicks in the positive control group treated with Megamonas monomorpha increased significantly (P<0.05).

[0145] Table 10 Effects of Megamonas monomorpha on the frequency of daily behaviors of Tianfu broiler chicks

[0146]

[0147] Note: Behavioral data are expressed as arithmetic mean ± standard error, and different letters indicate significant differences (P<0.05).

[0148] 3. Effects of Megamonas monomorpha on antioxidant indices of Tianfu broiler chicks under high-density rearing

[0149] like Figure 3 As shown in the figure, serum SOD is an antioxidant enzyme, which mainly helps to remove free radicals in the body, reduce the damage of oxidative stress to cells and tissues, and help protect the health of the body. Compared with the negative control group, the SOD activity in the serum of Tianfu broiler chicks in the positive control group and the group treated with Megamonas monomorpha was significantly increased (P < 0.05), indicating that oral administration of Megamonas monomorpha can alleviate the oxidative stress damage caused by high-density feeding.

[0150] 4. Effects of Megamonas monomorpha on immune parameters of Tianfu broiler chicks under high-density rearing

[0151] like Figure 4 As shown in the results, serum IL-6 is involved in the occurrence and development of inflammatory-related diseases. Infection, inflammation, trauma, and stress can lead to increased IL-6 concentrations. IL-6 may also increase abnormally due to chronic diseases or immune regulation imbalance. Compared with the negative control group, the positive control group and the Megamonas monomorpha treatment group significantly reduced the IL-6 concentration in the serum of Tianfu broiler chicks (P < 0.05).

[0152] like Figure 5 and Figure 6As shown in the results, interferon-γ (INF-γ) is an immunomodulatory factor that can enhance the activity of macrophages and natural killer cells, promote antigen presentation, enhance the activity of T cells, etc., thereby enhancing the body's immune response to pathogens. Interleukin-2 (IL-2) is a T cell growth factor. Its functions include promoting the proliferation and differentiation of T cells, enhancing the activity of natural killer cells, regulating immune responses, etc., thereby enhancing the body's ability to clear pathogens. Compared with the negative control group, the positive control group and the group treated with Megamonas monomorpha significantly increased the concentrations of INF-γ and IL-2 (P < 0.05). This shows that oral administration of Megamonas monomorpha can improve the immune ability of chicks and eliminate the adverse effects of high-density feeding on the immune system of chicks.

[0153] 5. Effects of Megamonas monomorpha on hormone levels of Tianfu broiler chicks under high-density rearing

[0154] like Figure 7 As shown in the figure, CORT is a corticosteroid hormone secreted by the adrenal cortex. It is an important regulator of the body's stress response. Under stress, the serum CORT concentration will increase significantly. Compared with the negative control group, the CORT concentration in the serum of chicks in the positive control group and the Megamonas monomorpha treatment group was significantly reduced (P < 0.05), indicating that high-density feeding causes stress and affects the CORT secretion of chicks, and Megamonas monomorpha can regulate the secretion of CORT and alleviate the stress response.

[0155] like Figure 8 As shown in the data, NE is an important neurotransmitter and endocrine substance, which participates in stress response by regulating the activity of the sympathetic nervous system and neuroendocrine system. The serum NE concentration increased significantly under stress state, and the serum NE concentration of the positive control group and the M. monocytogenes treatment group was significantly lower than that of the negative control group (P < 0.05), indicating that high-density feeding caused stress and affected the NE secretion of chicks, while M. monocytogenes could regulate the secretion of NE and alleviate the stress response.

[0156] like Fig. 9 As shown in the results, 5-HT in the hippocampus of the brain can affect emotional regulation, inhibit anxiety and depression, and the 5-HT concentration in the chicken brain decreases under stress, resulting in abnormal behavior. Compared with the negative control group, the 5-HT concentration in the hippocampus of the positive control group and the group treated with Megamonas monomorpha increased significantly (P < 0.05). This indicates that high-density feeding causes stress and affects the secretion of 5-HT in the central nervous system of chicks, and Megamonas monomorpha can regulate the secretion of 5-HT and relieve stress response.

[0157] 6. Effect of Megamonas monomorpha on 5-HT gene expression in Tianfu broiler chicks under high-density rearing

[0158] like Fig.10 , Fig.11 and Fig.12 As shown in the figure, tph2 is a 5-HT synthase gene in the central nervous system. Under stress, the expression of tph2 decreases, resulting in a decrease in the 5-HT concentration in the hippocampus. The HTR1a gene encodes the 5-HT receptor HTR1a. The HTR1a receptor can bind to 5-HT to inhibit the secretion of CORT by the hypothalamus-pituitary-adrenal axis to regulate stress response. Activation of HTR1a helps to alleviate anxiety and depression symptoms. The HTR2a gene encodes the 5-HT receptor HTR2a. HTR2a can bind to 5-HT to promote the continuous secretion of CORT by the hypothalamus-pituitary-adrenal axis. Activation of HTR2a may be related to the aggravation of anxiety and depression symptoms. Compared with the negative control group, the relative expression of tph2 and HTR1a genes in the hippocampus of the positive control group and the Megamonas monomorpha treatment group increased, while the relative expression of HTR2a gene decreased (P < 0.05). This indicates that high-density rearing causes stress and affects the expression of 5-HT synthase and receptor genes in the central nervous system of chicks, while Megamonas monomorpha can regulate the expression of 5-HT synthase and receptor genes and alleviate the stress response.

[0159] SEQ ID NO. 1 sequence listing:

[0160] 1 ctttgacata agcttccgct tgaagatgag cttgcgtctg attagctagt tggtgagggt

[0161] 61 aaaggcccac caaggcgacg atcagtagcc ggtctgagag gatgaacggc cacattggga

[0162] 121 ctgagacacg gcccagactc ctacgggagg cagcagtggg gaatcttccg caatgggcga

[0163] 181 aagcctgacg gagcaacgcc gcgtgaacga tgaaggtctt aggatcgtaa agttctgttg

[0164] 241 ttagggacga aggataagga ttataataca gtctttgttt gacggtacct aacgaggaag

[0165] 301 ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gcggcaagcg ttgtccggaa

[0166] 361 ttattgggcg taaagggagc gcaggcggga aactaagcgg atcttaaaag tgcggggctc

[0167] 421 aaccccgtga tggggtccga actggttttc ttgagtgcag gaggaaag cggaattccc

[0168] 481 agtgtagcgg tgaaatgcgt agatattggg aagaacacca gtggcgaagg cggctttctg

[0169] 541 gactgtaact gacgctgagg ctcgaaagct agggtagcga acgggattag ataccccggt

[0170] 601 agtcctagcc gtaaacgatg gatactaggt gtgggaggta tcgactcctt ccgtgccgga

[0171] 661 gttaacgcaa tagtatccc gcctggggag tacggccgca aggttgaaac tcaaaggaat

[0172] 721 tgacggggc ccgcacaagc ggtggagtat gtggtttaat tcgacgcaac gcgaagaacc

[0173] 781

[0174] 841 agaaaacagg tggtgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc

[0175] 901

[0176] 961 tgccgcggac aacgcggagg aaggcgggga tgacgtcaag tcatcatgcc ccttacgtct

[0177] 1021 tgggctacac acgtactaca atgggatgaa cagagggaag cgaaatcgcgaggtggagcg

[0178] 1081 gatccctaaa agcatctctc agttcggatt gtaggctgaa actcgcctacatgaagtcgg

[0179] 1141 aatcgctagt aatcgcaggt cagcatactg cggtgaatac gttcccgggccttgtacaca

[0180] 1201 ccgcccgtca caccacgaaa gtcattcaca cccgaagccg gctaagggcctatggtaccg

[0181] 1261 accgtctaag gtgggggcga tgattggggt gaagtcgtaa caaggtagccgtatcggaag

[0182] 1321 gtgcggctgg atcacctcct ta

[0183] In summary, the present invention provides a Megamonas monomorpha and its application in preventing or treating livestock and poultry stress, wherein the Megamonas monomorpha can improve the growth performance of Tianfu broilers and regulate stress response; the Megamonas monomorpha can improve the effects of high stocking density on the production performance and behavior of Tianfu broilers; and improve the effects of high stocking density on the antioxidant indicators and hormone indicators of Tianfu broilers; the present invention explores the application of Megamonas monomorpha in high stocking density stress, which has great practical application value for alleviating high stocking density stress in the breeding process and has good prospects for promotion and application.

[0184] The above description is not intended to impose any form of limitation on the present invention. Although the present invention has been disclosed through the above embodiments, it is not intended to limit the present invention. Any technician familiar with the profession can make some changes or modifications to equivalent embodiments of equivalent changes using the technical contents disclosed above without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention still falls within the scope of the technical solution of the present invention.

Claims

1. A use of Megamonas funiformis in preparing a product for treating or alleviating broiler stress; characterized in that: The original number of the Megamonas monomorpha is JCM14723=DSM19343; the stress is high-density breeding stress.

2. Use of Megamonas funiformis as claimed in claim 1 in the preparation of a product for treating or alleviating broiler stress, wherein the nucleotide sequence of 16S rDNA of the Megamonas funiformis is shown in SEQ ID NO.

1.

3. Use of Megamonas funiformis as claimed in claim 1 in preparing a product for treating or alleviating broiler stress, wherein the product is feed or medicine.

4. Use of Megamonas funiformis according to claim 1 in preparing a product for treating or alleviating broiler stress, wherein the product can improve feed conversion rate.

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