Application of farnesol in preparation of medicine or feed additive for treating or preventing chicken necrotic enteritis

Abstract

The invention discloses an application of farnesol in preparation of a medicine or a feed additive for treating or preventing necrotic enteritis of chicken, the medicine or the feed additive is an oral liquid, a tablet, a granule or a powder prepared from farnesol or farnesol as an effective component and a solvent or an auxiliary material, and the use dosage is 10-100mg / kg body weight; or farnesol is directly added into the feed, and the addition amount of farnesol is 0.05-0.1% of the total weight of the chicken feed. It is found that farnesol serving as a medicine or a feed additive is high in safety, free of toxic and side effects and capable of effectively relieving chicken production performance reduction, proinflammatory cytokine increase and intestinal absorption and barrier function damage caused by necrotic enteritis.

Description

Technical Field

[0001] The present application relates to the technical field of prevention and treatment of chicken enteritis, and specifically to the application of farnesol in the preparation of drugs or feed additives for treating or preventing necrotic enteritis in chickens. Background Art

[0002] my country is a major producer and consumer of chicken. Ensuring the health and efficiency of broiler farming to improve chicken production and quality is related to the national economy and people's livelihood. However, necrotic enteritis (NE) seriously restricts the health and efficiency of broiler farming. NE is one of the main gastrointestinal diseases that endanger the healthy farming of broilers worldwide. It is mainly caused by infection with Clostridium perfringens (CP) type A and G. In the broiler production process, some abnormal environmental factors (such as heat stress), pathogens (such as coccidia) infection, and dietary types (such as wheat-based diets) provide conditions for CP infection. NE mainly occurs in the small intestine, mainly in the ileum, which leads to impaired physical and immune barrier functions of the broiler intestine, imbalance of intestinal flora structure, and then causes intestinal inflammatory damage.

[0003] Previously, the use of antibiotics was the main measure to prevent and control NE in broiler chickens. However, excessive use of antibiotics has brought many hazards: (1) Increased drug resistance: Long-term use of antibiotic additives can cause microorganisms to develop drug resistance, which can cause some pathogenic microorganisms to develop drug resistance through mutation, leading to a sharp increase in the number of antibiotic-resistant bacteria worldwide; (2) Excessive veterinary drug residues in animal-derived foods: Excessive use of veterinary drugs can cause excessive veterinary drug residues in animal-derived foods, posing a potential threat to human health; (3) Damage to the ecological environment: The discharge of metabolites from poultry that eat antibiotic-containing feed can damage the ecological environment and pollute drinking water or other agricultural products; (4) Impact on human health: If antibiotics remaining in livestock and poultry products are ingested by the human body, they will increase the burden on the liver and kidneys during metabolism, and long-term intake may cause lesions; (5) Reduced immunity: The abuse of antibiotics may cause an imbalance in the normal microbial system in the chicken, resulting in a decrease in the chicken's immune function and resistance. In view of the serious consequences of antibiotics, antibiotics have been banned on a large scale. However, since the ban on antibiotics in feed, pathogens in the gastrointestinal tract of broiler chickens have taken the opportunity to proliferate rapidly, which is bound to destroy their intestinal immune barrier and aggravate the negative impact of NE on broiler chickens.

[0004] It can be seen that NE is a key issue that needs to be urgently addressed in the healthy and efficient breeding of broilers in the post-antibiotic era. The prevention and control of NE through nutritional measures has gradually become one of the key topics of concern in broiler breeding. Summary of the invention

[0005] In view of this, the purpose of the present application is to provide the use of farnesol in the preparation of a drug or feed additive for treating or preventing necrotic enteritis in chickens. In this application, farnesol and a composition with farnesol as the main component can effectively inhibit the pathogenic bacteria of chicken enteritis. The farnesol and the composition with farnesol as the main component have the advantages of high safety, stable efficacy, no toxic side effects, etc., and have a significant preventive and control effect on necrotic enteritis in chickens.

[0006] To achieve the above objectives, this application provides at least the following technical solutions:

[0007] Application of farnesol in preparing medicine or feed additive for treating or preventing necrotic enteritis in chickens.

[0008] Compared with the prior art, the technical solution provided by this application has at least the following beneficial effects:

[0009] 1. Farnesol in this application is derived from natural plants, has high safety, no toxic side effects, and can be used as a feed additive for a long time to prevent necrotic enteritis in chickens. Long-term use will not lead to the production of drug-resistant strains, which is an advantage that other anti-disease drugs do not have.

[0010] 2. The present application shows through in vitro tests that farnesol or a composition comprising farnesol can inhibit the proliferation of Clostridium perfringens.

[0011] 3. The present application shows through in vivo experiments that farnesol or a composition comprising farnesol can inhibit the expression of pro-inflammatory cytokines, promote the expression of antioxidant enzymes, enhance intestinal health and thus improve the feed conversion efficiency of broilers with necrotic enteritis. DETAILED DESCRIPTION

[0012] In order to make the purpose, technical solution and advantages of the present application more clear, the present application is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described here are only used to explain the present application and are not used to limit the present application.

[0013] Those skilled in the art will appreciate that, unless otherwise stated, the words "said", "the", and "the aforementioned" used in the text of the present application may also include plural forms. It should be further understood that the wording "including" used in the specification of the present application means the presence of the features, steps, and operations, but does not exclude the presence or addition of one or more other features, integers, and steps.

[0014] Those skilled in the art will appreciate that, for any specific experimental steps or conditions not specified in the examples, the experiments may be carried out according to the conventional experimental steps or conditions described in the literature in the art; any raw materials or instruments used without specifying the manufacturer are conventional products that can be purchased commercially.

[0015] Those skilled in the art will understand that, unless otherwise specified in the present application, when a numerical range is given in an embodiment, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this application are consistent with the prior art mastery of those skilled in the art and the records of this application, and any methods, equipment and materials of the prior art similar or equivalent to the methods, equipment and materials described in the embodiments of this application may also be used to implement this application.

[0016] Farnesol is an active ingredient extracted from the bark of the acacia tree. Its main components are flavonoids and triterpenoids. Farnesol can also be found in citronella, lemongrass, tuberose, cyclamen, rose, orange blossom, balsam and musk. Its chemical formula is C 15 H 25 OH, molecular weight is 223.72g / mol. Studies have found that farnesol has a variety of biological effects such as antibacterial, anti-inflammatory, antioxidant, immune regulation and lipid metabolism. In clinical practice, farnesol is used to treat allergic asthma, diabetes, atherosclerosis, obesity and hyperlipidemia. Due to its green and safe characteristics, farnesol is widely used in food, skin care products, medicine and other fields. In livestock and poultry production, there are few reports on farnesol, and there are no reports on the effect of farnesol on necrotic enteritis in broilers.

[0017] The examples of the present application have shown through in vitro tests that farnesol or a composition comprising farnesol can inhibit the proliferation of Clostridium perfringens. In vivo tests have shown that farnesol or a composition comprising farnesol can inhibit the expression of pro-inflammatory cytokines, promote the expression of antioxidant enzymes, enhance intestinal health and thus improve the feed conversion efficiency of broilers with necrotic enteritis. The above results can provide basic data for the development of farnesol in poultry health drugs and feed additives.

[0018] Based on this, the embodiments of the present application provide the use of farnesol in the preparation of a drug or feed additive for treating or preventing necrotic enteritis in chickens.

[0019] In certain embodiments, the necrotic enteritis in chickens is caused by infection with Clostridium perfringens alone or with coccidia.

[0020] In certain embodiments, the drug or feed additive is farnesol, that is, farnesol can be directly used as a drug or feed additive for treating or preventing necrotic enteritis in chickens.

[0021] In certain embodiments, the medicine or feed additive is a farnesol composition formed by adding excipients with farnesol as an active ingredient.

[0022] In certain embodiments, the auxiliary material is at least one of anhydrous ethanol, propylene glycol, dimethyl sulfoxide, edible oil, and water.

[0023] In certain embodiments, the auxiliary material is at least one of lactose, maltose, mannitol, and soluble starch.

[0024] In certain embodiments, the auxiliary material is at least one of montmorillonite, medical stone, vermiculite, zeolite powder, white carbon black, defatted rice bran, corn cob powder, starch, and bran.

[0025] In certain preferred embodiments, the farnesol composition is a solution of farnesol mixed with anhydrous ethanol and water, wherein the concentration of farnesol is 10 to 120 mg / mL.

[0026] In certain preferred embodiments, the farnesol composition is a powdery mixture of farnesol, wheat bran and soluble starch, wherein the mass fraction of farnesol in the powdery mixture is 50%.

[0027] In certain preferred embodiments, the farnesol composition is a flaky mixture of farnesol, lactose or maltose, and soluble starch, wherein the mass fraction of farnesol in the flaky mixture is 50%.

[0028] In certain preferred embodiments, the farnesol composition is a granular mixture of farnesol, lactose or maltose, and soluble starch, wherein the mass fraction of farnesol in the granular mixture is 50%.

[0029] In certain embodiments, the farnesol composition may be prepared in the form of at least one of an oral solution, a granule, a tablet, and a powder, and may be prepared using conventional pharmaceutical preparation methods.

[0030] In certain embodiments, the method of using the drug or feed additive comprises at least one of the following methods:

[0031] (1) Orally administering farnesol to chickens at a dosage of 10 to 100 mg / kg of chicken body weight;

[0032] (2) farnesol granules, powder or tablets are added to chicken feed as a feed additive according to the farnesol content of 0.05% to 0.1% by weight of chicken feed;

[0033] (3) When used for daily prevention of enteritis in chickens, the farnesol powder, granules or tablets are ground into powder and added to the feed or drinking water so that the farnesol content is 0.05% by weight of the chicken feed or drinking water;

[0034] (4) When used for clinical treatment of chicken enteritis: orally administer the drug to chickens at a farnesol content of 100 mg / kg chicken body weight; or grind the granules, powder or tablets into powder and feed directly to chickens at a farnesol content of 100 mg / kg chicken body weight; or grind the granules, powder or tablets into powder and add it to the feed at a farnesol content of 0.1% by weight of chicken feed; or grind the granules, powder or tablets into powder and add it to the drinking water at a farnesol content of 0.1% by weight, three times a day, morning, noon and evening, for one week.

[0035] The technical solution of the present application and the technical effects achieved are described in detail below through more specific embodiments.

[0036] Example 1

[0037] This embodiment provides a method for preparing farnesol oral liquid, the steps of which are:

[0038] Weigh 10 g of farnesol, 30 mL of anhydrous ethanol, and an appropriate amount of water for injection and set aside; first dissolve farnesol in anhydrous ethanol and mix evenly, then slowly add water for injection to make up to 100 mL to obtain a farnesol oral solution with a concentration of 100 mg / mL.

[0039] When the above-mentioned farnesol oral liquid is used for the prevention of chicken enteritis, it is added to feed or drinking water at a dosage of 10 mg / kg of chicken body weight, or it is added to feed at a dosage of 0.05% of the total weight of feed, and it is used during the entire feeding cycle; when the above-mentioned farnesol oral liquid is used for the clinical treatment of chicken enteritis, it is orally administered at a dosage of 100 mg / kg of chicken body weight, or the oral liquid is added to feed or drinking water at a dosage of 100 mg / kg of chicken body weight, or it is added to feed at a dosage of 0.1% of the total weight of feed, and it is added three times a day in the morning, noon and evening for one week.

[0040] Embodiment 2:

[0041] This embodiment provides a method for preparing farnesol powder, the steps of which are:

[0042] Weigh 50 g of farnesol, 20 g of wheat bran and 30 g of soluble starch for later use; mix the farnesol, wheat bran and soluble starch evenly, pass through a 60-mesh sieve, and package to obtain a farnesol powder, in which the mass fraction of farnesol is 50%.

[0043] When the above-mentioned farnesol powder is used for the prevention of chicken enteritis, it is added to feed or drinking water at a dosage of 10 mg / kg of chicken body weight, or it is added to feed at a dosage of 0.05% of the total weight of feed, and it is used during the entire feeding cycle; when the above-mentioned farnesol powder is used for the clinical treatment of chicken enteritis, it is orally administered at a dosage of 100 mg / kg of chicken body weight, or the powder is added to feed or drinking water at a dosage of 100 mg / kg of chicken body weight, or it is added to feed at a dosage of 0.1% of the total weight of feed, and it is added three times a day in the morning, noon and evening for one week.

[0044] Embodiment 3:

[0045] This embodiment provides a method for preparing farnesol tablets, the steps of which are:

[0046] Weigh 50 g of farnesol, 20 g of lactose or maltose, and 30 g of soluble starch for later use; mix farnesol, lactose or maltose, and soluble starch, press and dry with a tablet press; obtain a farnesol tablet, wherein the mass fraction of farnesol is 50%.

[0047] When the above-mentioned farnesol tablets are used for the prevention of chicken enteritis, they are added to feed or drinking water at a dosage of 10 mg / kg of chicken body weight, or added to feed at a dosage of 0.05% of the total weight of feed, and used throughout the entire breeding cycle; when the above-mentioned farnesol tablets are used for the clinical treatment of chicken enteritis, they are orally administered at a dosage of 100 mg / kg of chicken body weight, or the tablets are added to feed or drinking water at a dosage of 100 mg / kg of chicken body weight, or added to feed at a dosage of 0.1% of the total weight of feed, and added three times a day in the morning, noon and evening for one week.

[0048] Embodiment 4:

[0049] This embodiment provides a method for preparing farnesol tablets, the steps of which are:

[0050] Weigh 50 g of farnesol, 20 g of lactose or maltose, and 30 g of soluble starch for later use; mix farnesol, lactose or maltose, and soluble starch to granulate, dry, and then package to obtain a farnesol granule, in which the mass fraction of farnesol is 50%.

[0051] When the above-mentioned farnesol granules are used for the prevention of chicken enteritis, they are added to feed or drinking water at a dosage of 10 mg / kg of chicken body weight, or added to feed at a dosage of 0.05% of the total weight of feed, and used throughout the entire feeding cycle; when the above-mentioned farnesol granules are used for the clinical treatment of chicken enteritis, they are orally administered at a dosage of 100 mg / kg of chicken body weight, or the granules are added to feed or drinking water at a dosage of 100 mg / kg of chicken body weight, or added to feed at a dosage of 0.1% of the total weight of feed, and added three times a day in the morning, noon and evening for one week.

[0052] In vitro inhibitory effect of farnesol on Clostridium perfringens

[0053] The examples of the present application verify the in vitro inhibitory effect of farnesol on Clostridium perfringens, and the verification method is as follows:

[0054] 1. Materials and Methods

[0055] 1.1 Bacteria: Clostridium perfringens was isolated, preserved and donated by the National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0056] 1.2 Experimental drugs: Weigh farnesol (purchased from Sigma, purity > 99%), dissolve it with physiological saline to prepare a farnesol stock solution with a concentration of 0.1 g / mL, and then dilute it step by step with physiological saline to 0.05 g / mL, 0.025 g / mL, 0.0125 g / mL, and 0.0063 g / mL, respectively.

[0057] 1.3 Test process:

[0058] The Oxford cup method was used to evaluate the antibacterial effect of farnesol on Clostridium perfringens by evaluating the diameter of the inhibition zone of different concentrations of farnesol. Physiological saline was used as the blank control group; enramycin (purchased from Sinopharm Group, purity>99%) was used as the positive control. In the inhibition zone test, enramycin was dissolved in physiological saline and its dose was 0.1μg / mL; farnesol was set at 0.1g / mL, 0.05g / mL, 0.025g / mL, 0.0125g / mL, and 0.0063g / mL, with a total of 5 gradients; each treatment group was repeated 3 times.

[0059] In the specific test process, Clostridium perfringens was cultured in cooked meat medium, and the concentration was 1×10 5 CFU / mL of bacterial liquid was used to prepare cooked meat medium solid culture plates and cooked meat semi-solid culture medium. When the cooked meat semi-solid culture medium was cooled to about 37°C, 100 μL of 1×10 5CFU / mL of Clostridium perfringens solution was added to the cooked meat semi-solid culture medium and shaken well, and then quickly poured onto the cooked meat culture medium solid culture plate, and then Oxford cups were evenly placed on the culture plate. After the semi-solid culture medium solidified, the Oxford cups were taken out and 50 μL of the above-mentioned treatment solutions were added to the ring holes, and the plates were placed in a 37°C constant temperature incubator for 24 hours; then the diameter of the inhibition zone was measured using a ruler (accuracy of 0.01 cm).

[0060] 2. Test results

[0061] The in vitro inhibition results of farnesol on Clostridium perfringens from chicken are shown in Table 1 below.

[0062] Table 1

[0063] Group Diameter of inhibition zone (cm) Blank control group (normal saline) / Positive control group (0.1 μg / mL enramycin) 2.03±0.01 0.1 g / mL farnesol group 1.87±0.02 0.05 g / mL farnesol group 1.76±0.14 0.025 g / mL farnesol group 1.34±0.07 0.0125 g / mL Farnesol group 1.12±0.11 0.0063 g / mL Farnesol group /

[0064] From the results in Table 1, it can be seen that farnesol has an inhibitory effect on Clostridium perfringens from chicken, and the diameter of the inhibition zone increases with the increase of farnesol dosage.

[0065] Effects of farnesol on growth performance and intestinal health of broiler chickens infected with Clostridium perfringens

[0066] The present application example verifies the effect of farnesol on the growth performance and intestinal health of broilers infected with Clostridium perfringens, and the verification method is as follows:

[0067] 1.1 Materials and methods

[0068] The experiment involved 720 healthy AA + Broiler male chicks were evenly divided into 3 treatment groups, each treatment group was divided into 8 replicate groups, and each replicate group had 30 chickens; the 3 treatment groups were: control group, Clostridium perfringens (CP) infection group, and farnesol + CP infection group. The broilers in the control group and CP infection group were fed a "corn-soybean meal" type basal diet, and the diet formula was formulated in accordance with the nutritional requirements standard of Chinese broilers (NY / T33-2004); the broiler diet of the farnesol + CP infection group was supplemented with 0.05% of the feed weight of farnesol (purchased from Aladdin, purity> 80%) in the basal diet; on the 14th to 21st day of the experiment, 1 mL of CP (CVCC2030 strain, dose of 1.0×10 9CFU / mL), and the control group broilers were gavage-fed with an equal volume of CP medium; during the experiment, broilers were free to eat and drink, and a 24-h lighting system was adopted; on the 22nd day of the experiment, all broilers were fasted for 8 h, then weighed and feed consumption was counted, and growth performance-related indicators such as average daily gain (ADG), average daily feed intake (ADFI), and feed-to-weight ratio (FCR) were calculated; thereafter, one broiler with uniform weight was selected from each replicate group to collect subwing venous blood and separate serum for testing.

[0069] 2 Data analysis

[0070] The experimental data were analyzed by one-way ANOVA in SPSS23.0 statistical software, and Duncan's method was used for multiple comparisons between groups. P < 0.05 was regarded as the standard of significant difference, and P < 0.01 was regarded as the standard of extremely significant difference. The results were expressed as mean ± standard deviation.

[0071] 3. Test results

[0072] 3.1 Effects of farnesol on growth performance of broilers infected with Clostridium perfringens

[0073] Table 2 shows the effects of farnesol on the growth performance of broilers infected with Clostridium perfringens, where a, b, and c are significant differences. Data with different letters on their shoulders in the table indicate significant differences, P<0.05.

[0074] Table 2

[0075] Test items Control group CP infection group Farnesol + CP infection group P-value d21-average body weight (g) <![CDATA[786.11±43.02 a ]]> <![CDATA[672.21±32.19 c ]]> <![CDATA[725.22±21.34 b ]]> <0.001 ADFI(g) <![CDATA[53.04±3.12 a ]]> <![CDATA[48.91±3.19 b ]]> <![CDATA[51.11±3.01 ab ]]> 0.032 ADG(g) <![CDATA[39.3±1.34 a ]]> <![CDATA[33.5±1.98 c ]]> <![CDATA[36.25±0.78 b ]]> <0.001 FCR <![CDATA[1.36±0.03 b ]]> <![CDATA[1.46±0.09 a ]]> <![CDATA[1.41±0.06 ab ]]> 0.012

[0076] As shown in Table 2, Clostridium perfringens infection reduces the average body weight, average daily feed intake, and average daily weight gain of broilers, and improves the feed-to-weight ratio; adding farnesol to the diet can alleviate the decrease in the average body weight and average daily weight gain of broilers caused by Clostridium perfringens infection, and has a trend of improving the feed-to-weight ratio. This shows that farnesol has a protective effect on broilers infected with Clostridium perfringens.

[0077] 3.2 Effect of farnesol on serum D-xylose and diamine oxidase levels in broiler chickens infected with Clostridium perfringens

[0078] Table 3 shows the effects of farnesol on serum D-xylose and diamine oxidase (DAO) levels in broilers infected with Clostridium perfringens, where a, b, and c are significant differences. Data with different letters on the shoulders in the table indicate significant differences P<0.05.

[0079] Table 3

[0080] Test items Control group CP infection group Farnesol + CP infection group P-value D-Xylose, mmo / L <![CDATA[2.35±0.31 a ]]> <![CDATA[0.89±0.06 c ]]> <![CDATA[1.52±0.31 b ]]> 0.011 DAO,U / L <![CDATA[21.06±4.21 a ]]> <![CDATA[41.07±6.38 b ]]> <![CDATA[30.88±3.31 b ]]> <0.001

[0081] Xylose can be absorbed by the intestine but cannot be used by the body. Therefore, by evaluating the D-xylose content in the blood, the intestinal absorption function can be evaluated. DAO exists in the intestine in a healthy state and is released into the blood when the intestine is damaged. Therefore, the higher the DAO activity in the serum, the more obvious the intestinal damage. As shown in Table 3, the addition of farnesol to the diet can alleviate the decrease in D-xylose content in the blood of broilers caused by Clostridium perfringens infection and the increase in DAO levels, indicating that farnesol can protect broilers from the decrease in intestinal absorption function and barrier damage caused by Clostridium perfringens infection.

[0082] 3.3 Effect of farnesol on serum inflammatory cytokine levels in broiler chickens infected with Clostridium perfringens

[0083] Table 4 shows the effects of farnesol on the levels of serum inflammatory cytokines in broilers infected with Clostridium perfringens, where a, b, and c are significant differences. Data with different letters on the shoulders in the table indicate significant differences (P<0.05); IL-1β and IL-6 represent interleukin 1β and interleukin 6, respectively, TNF-α represents tumor necrosis factor-α, and IFN-γ represents interferon-γ. The detection of pro-inflammatory cytokines was carried out using the chicken ELISA kit of Beijing Solebow Company.

[0084] Table 4

[0085] Test items Control group CP infection group Farnesol + CP infection group P-value IL-1β, pg / mL <![CDATA[113.12±12.31 c ]]> <![CDATA[287.13±31.06 a ]]> <![CDATA[201.13±30.11 b ]]> 0.038 TNF-α, pg / mL <![CDATA[77.92±10.21 b ]]> <![CDATA[141.07±31.38 a ]]> <![CDATA[130.88±38.11 a ]]> <0.001 IL-6, pg / mL <![CDATA[132.89±29.12 c ]]> <![CDATA[254.19±42.21 a ]]> <![CDATA[170.53±27.65 b ]]> <0.001 IFN-γ, pg / mL <![CDATA[173.97±47.65 b ]]> <![CDATA[301.02±71.23 a ]]> <![CDATA[287.89±66.39 a ]]> 0.043

[0086] As shown in Table 4, Clostridium perfringens infection causes an increase in the levels of pro-inflammatory cytokines, such as IL-1β, TNF-α, IL-6, and IFN-γ; and the addition of farnesol to the diet can alleviate the increase in serum IL-1β and IL-6 in broiler chickens caused by Clostridium perfringens infection, indicating that farnesol has an alleviating effect on the inflammatory response of broiler chickens caused by Clostridium perfringens infection.

[0087] Effects of farnesol on growth performance and intestinal health of broiler chickens infected with coccidia and Clostridium perfringens

[0088] The present application example verifies the effect of farnesol on the growth performance and intestinal health of broilers infected with coccidia and Clostridium perfringens. The verification method is as follows:

[0089] 1. Materials and Methods

[0090] A 2×2 factorial design was used to test 480 healthy AA +Broiler male chicks were divided into 4 treatment groups, each of which was divided into 8 replicate groups, each replicate group included 15 chickens; the 4 treatment groups were: control group, coccidia and Clostridium perfringens combined infection group (CCP), farnesol group, farnesol + CCP group. The control group and CCP infection group broilers were fed a "corn-soybean meal" basal diet, and the diet formula was formulated in accordance with the Chinese nutrient requirement standard for broilers (NY / T33-2004). The broiler diets of the farnesol group and the farnesol + CCP group were supplemented with 0.05% of the feed weight of farnesol (purchased from Aladdin Company, purity > 80%) in the basal diet; on the 8th and 10th days of the experiment, the broiler chickens in the CCP group and the farnesol + CCP group were vaccinated with an ultra-high dose of coccidia vaccine (purchased from Foshan Zhengdian Company, the dose was 30 times the recommended dose) to establish intestinal coccidiosis infection. On the 14th to 21st days of the experiment, the broiler chickens in the CCP group and the farnesol + CCP group were gavaged with 1 mL of CP (CVCC2030 strain, dose of 1.0×10 9 CFU / mL), and the control group and the farnesol group broilers were gavage-fed with an equal volume of CP medium; during the experiment, the broilers were free to eat and drink, and a 24-h lighting system was adopted; on the 22nd day of the experiment, all broilers were fasted for 8 h, and then weighed and the feed consumption was counted, and the growth performance related indicators such as average daily gain (ADG), average daily feed intake (ADFI), and feed-to-weight ratio (FCR) were calculated; thereafter, one broiler with uniform weight was selected from each replicate group to collect subwing venous blood and separate serum for testing; further, sodium pentobarbital (at a dose of 50 mg / kg live weight) was injected into the subwing vein for anesthesia, and the broilers were slaughtered, and the ileum tissue was dissected to make slices for observation, and ileum samples were taken for subsequent analysis.

[0091] 2. Data Analysis

[0092] The experimental data were analyzed by two-way ANOVA using SPSS23.0 statistical software. The main effects of the model included farnesol, coccidia and Clostridium perfringens infection, and the interaction between the two. When there was an interaction between the two, Duncan's method was used for multiple comparisons between groups, with P<0.05 as the standard of significant difference and P<0.01 as the standard of extremely significant difference. The results were expressed as mean ± standard error.

[0093] 3. Test results

[0094] 3.1 Effects of farnesol on growth performance of broilers infected with coccidia and Clostridium perfringens

[0095] Table 5 shows the effects of farnesol on the growth performance of broilers co-infected with coccidia and Clostridium perfringens, where a, b, and c are significant difference markers, and data with different letters on the shoulders in the table indicate significant differences P<0.05; -CCP means no co-infection with coccidia and Clostridium perfringens, +CCP means co-infection with coccidia and Clostridium perfringens, -HH means no farnesol was added to the diet, and +HH means farnesol was added to the diet; SEM means standard error.

[0096] Table 5

[0097]

[0098] As shown in Table 5, the combined infection of coccidia and Clostridium perfringens increases the feed-to-weight ratio, and the addition of farnesol to the diet can alleviate the negative impact of the combined infection of coccidia and Clostridium perfringens on the feed-to-weight ratio of broilers. The results showed that farnesol improved the growth performance of broilers infected with coccidia and Clostridium perfringens.

[0099] 3.2 Effects of farnesol on antioxidant function in blood and intestine of broilers infected with coccidia and Clostridium perfringens

[0100] Table 6 shows the effects of farnesol on the antioxidant function of broiler blood infected with coccidia and Clostridium perfringens, where a and b are significant differences. Data with different letters on the shoulders in the table indicate significant differences (P<0.05); -CCP means no co-infection with coccidia and Clostridium perfringens, +CCP means co-infection with coccidia and Clostridium perfringens, -HH means no farnesol was added to the diet, +HH means farnesol was added to the diet; SEM means standard error; T-AOC means antioxidant capacity, CAT means catalase, H2O2 means hydrogen peroxide, and MDA means malondialdehyde. The indicators in the table were all measured using a kit (purchased from Nanjing Jiancheng Bioengineering Institute), and the final values ​​were calibrated with total protein as the denominator.

[0101] Table 6

[0102]

[0103] Table 7 shows the effects of farnesol on the antioxidant function of the ileum of broiler chickens co-infected with coccidia and Clostridium perfringens, where a, b, and c are significant difference markers, and different letters on the shoulders of the data in the table indicate significant differences P<0.05; -CCP means no co-infection with coccidia and Clostridium perfringens, +CCP means co-infection with coccidia and Clostridium perfringens, -HH means no farnesol was added to the diet, and +HH means farnesol was added to the diet; SEM means standard error, GSH-Px means glutathione peroxidase, MPO means myeloperoxidase, T-SOD means superoxide dismutase, and MDA means malondialdehyde. The indicators in the table were all measured using a kit (purchased from Nanjing Jiancheng Bioengineering Institute), and the final values ​​were calibrated with total protein as the denominator.

[0104] Table 7

[0105]

[0106] As shown in Tables 6 and 7, the combined infection of coccidia and Clostridium perfringens down-regulated the content of CAT in serum, up-regulated the content of MDA, and down-regulated the content of GSH-Px and T-SOD in the ileum. Adding farnesol to the diet can alleviate the abnormal changes of the above indicators caused by the combined infection of coccidia and Clostridium perfringens. The results show that farnesol can enhance the antioxidant function of broilers with combined infection of coccidia and Clostridium perfringens.

[0107] 3.3 Effects of farnesol on ileal gene expression in broiler chickens infected with coccidia and Clostridium perfringens

[0108] Table 8 shows the effects of farnesol on ileum gene expression in broiler chickens co-infected with coccidia and Clostridium perfringens, where a, b, and c are significant difference markers, and data with different letters on the shoulders in the table indicate significant differences P<0.05; -CCP means no co-infection with coccidia and Clostridium perfringens, +CCP means co-infection with coccidia and Clostridium perfringens, -HH means no farnesol was added to the diet, and +HH means farnesol was added to the diet; SEM means standard error; TNF-α means tumor necrosis factor-α, IFN-γ means interferon-γ, IL-18 means interleukin-18, IL-8 means interleukin-8, IL-1β means interleukin-1β, ZO-1 means occludens protein-1, Claudin-1 means tight junction protein 1, and occludin means claudin.

[0109] Table 8

[0110]

[0111] As shown in Table 8, the combined infection of coccidia and Clostridium perfringens down-regulated the transcription levels of ileal barrier-related genes, Occludin and ZO-1, and up-regulated the mRNA levels of pro-inflammatory cytokines TNF-α, IFN-γ, and IL-1β in broilers. Adding farnesol to the diet can alleviate the abnormal changes of the above genes caused by the combined infection of coccidia and Clostridium perfringens. The results showed that farnesol has a protective effect on intestinal inflammation and impaired barrier function in broilers caused by the combined infection of coccidia and Clostridium perfringens.

[0112] In summary, farnesol can be used to prepare drugs or feed additives for treating or preventing necrotic enteritis in chickens, and the prepared drugs or feed additives have the ability to resist various pathogens that cause necrotic enteritis in chickens, such as Clostridium perfringens. The dosage forms of the drugs and feed additives include oral liquid, granules, powders, tablets and the like.

[0113] The present application is described in detail above. Specific examples are used in this article to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the present application and its core ideas. It should be pointed out that for ordinary technicians in this technical field, without departing from the principles of the present application, several improvements and modifications can be made to the present application, and these improvements and modifications also fall within the scope of protection of the claims of the present application.

Claims

1. Application of farnesol in the preparation of medicines or feed additives for treating or preventing necrotic enteritis in chickens.

2. The use according to claim 1, characterized in that: The chicken necrotic enteritis is caused by infection of Clostridium perfringens alone or in combination with coccidia.

3. The use according to claim 1, characterized in that: The medicine or feed additive is farnesol.

4. The use according to claim 1, characterized in that: The medicine or feed additive is a farnesol composition formed by adding excipients to farnesol as an effective ingredient, wherein the excipients are at least one of (1) to (3): (1) at least one of anhydrous ethanol, propylene glycol, dimethyl sulfoxide, edible oil, and water; (2) at least one of lactose, maltose, mannitol, and soluble starch; (3) at least one of montmorillonite, medical stone, vermiculite, zeolite powder, white carbon black, defatted rice bran, corn cob powder, starch and bran.

5. The use according to claim 4, characterized in that: The farnesol composition is a solution formed by mixing farnesol with anhydrous ethanol and water, wherein the concentration of farnesol is 10-120 mg / mL.

6. The use according to claim 4, characterized in that: The farnesol composition is a powdery mixture of farnesol, wheat bran and soluble starch, wherein the mass fraction of farnesol in the powdery mixture is 50%.

7. The use according to claim 4, characterized in that: The farnesol composition is a flaky mixture of farnesol, lactose or maltose and soluble starch, wherein the mass fraction of farnesol in the flaky mixture is 50%.

8. The use according to claim 4, characterized in that: The farnesol composition is a granular mixture of farnesol, lactose or maltose and soluble starch, wherein the mass fraction of farnesol in the granular mixture is 50%.

9. The use according to claim 4, characterized in that: The farnesol composition may be prepared in the form of at least one of an oral solution, a granule, a tablet, and a powder, and may be prepared by conventional pharmaceutical preparation methods.

10. The use according to claim 1, characterized in that: The method for using the drug or feed additive includes at least one of (1) to (4): (1) Orally administering farnesol to chickens at a dosage of 10 to 100 mg / kg of chicken body weight; (2) farnesol granules, powder or tablets are added to chicken feed as a feed additive according to the farnesol content of 0.05% to 0.1% by weight of chicken feed; (3) When used for daily prevention of chicken enteritis, the farnesol powder, granules or tablets are ground into powder and added to the feed or drinking water so that the farnesol content is 0.05% by weight of the chicken feed or drinking water; (4) When used for clinical treatment of chicken enteritis: orally administer the drug to chickens at a farnesol content of 100 mg / kg chicken body weight; or grind the granules, powder or tablets into powder and feed directly to chickens at a farnesol content of 100 mg / kg chicken body weight; or grind the granules, powder or tablets into powder and add it to the feed at a farnesol content of 0.1% by weight of chicken feed; or grind the granules, powder or tablets into powder and add it to the drinking water at a farnesol content of 0.1% by weight, three times a day, morning, noon and evening, for one week.

Images