Application of farnesol in the preparation of drugs or feed additives for the treatment or prevention of necrotizing enteritis in chickens

Abstract

This application discloses the use of farnesol in the preparation of drugs or feed additives for the treatment or prevention of necrotic enteritis in chickens. The drugs or feed additives are farnesol or oral liquids, tablets, granules, or powders made with farnesol as the active ingredient and added solvents or excipients, at a dosage of 10–100 mg / kg body weight; or farnesol can be directly added to the feed at a dosage of 0.05%–0.1% of the total weight of the chicken feed. This application finds that farnesol, as a drug or feed additive, has high safety, no toxic side effects, and can effectively alleviate the reduced chicken production performance, increased pro-inflammatory cytokines, and impaired intestinal absorption and barrier function caused by necrotic enteritis.

Description

Technical Field

[0001] This application relates to the field of chicken enteritis prevention and control technology, specifically to the application of farnesol in the preparation of drugs or feed additives for the treatment or prevention of necrotizing enteritis in chickens. Background Technology

[0002] my country is a major producer and consumer of chicken meat, and ensuring the healthy and efficient breeding of broilers to improve chicken yield and quality is crucial to the national economy and people's livelihood. However, necrotic enteritis (NE) severely restricts the healthy and efficient breeding of broilers. NE is one of the major gastrointestinal diseases endangering the healthy breeding of broilers worldwide, mainly caused by infection with Clostridium perfringens types A and G. Abnormal environmental factors (such as heat stress), pathogenic factors (such as coccidiosis) infection, and dietary types (such as wheat-based diets) during broiler production provide conditions for CP infection. NE mainly occurs in the small intestine, primarily the ileum, leading to impaired physical and immune barrier function of the broiler intestine, dysbiosis of the intestinal flora, and consequently, inflammatory damage to the intestine.

[0003] Previously, the use of antibiotics was the main measure to control non-steroidal anti-inflammatory disease (NE) in broilers. However, excessive use of antibiotics has brought many harms: (1) Increased drug resistance: Long-term use of antibiotic additives can lead to drug resistance in microorganisms, causing some pathogenic microorganisms to develop drug resistance through mutation, resulting in 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 emissions of metabolites from poultry fed with antibiotics can damage the ecological environment and pollute drinking water or other agricultural products; (4) Impact on human health: Antibiotic residues in livestock and poultry products, if ingested by humans, can 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 lead to an imbalance in the normal microbial system in chickens, causing a decline in the immune function and resistance of chickens. 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 broilers have been proliferating rapidly, which inevitably damages their intestinal immune barrier and exacerbates the negative effects of norepinephrine (NE) on broilers.

[0004] It is evident that non-steroidal anti-inflammatory disease (NE) is a key issue that urgently needs to be addressed for the healthy and efficient breeding of broilers in the post-antibiotic era. Controlling NE through nutritional measures has gradually become one of the key topics of concern in broiler farming. Summary of the Invention

[0005] In view of this, the purpose of this application is to provide the use of farnesol in the preparation of drugs or feed additives for the treatment or prevention of necrotic enteritis in chickens. In this application, farnesol and compositions with farnesol as the main component can effectively inhibit pathogenic bacteria causing enteritis in chickens. Farnesol and compositions with farnesol as the main component have advantages such as high safety, stable efficacy, and no toxic side effects, 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 the preparation of drugs or feed additives for the treatment or prevention of necrotizing enteritis in chickens.

[0008] The technical solution provided in this application has at least the following advantages compared with the prior art:

[0009] 1. The farnesol in this application is derived from natural plants, has high safety and 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 development of drug-resistant strains, which is an advantage not possessed by other anti-disease drugs.

[0010] 2. This application demonstrates through in vitro experiments that farnesol or compositions including farnesol can inhibit the proliferation of Clostridium perfringens.

[0011] 3. This application demonstrates through in vivo experiments that farnesol or compositions including farnesol can inhibit the expression of pro-inflammatory cytokines, promote the expression of antioxidant enzymes, improve intestinal health, and thus improve the feed conversion efficiency of broilers with necrotizing enteritis. Detailed Implementation

[0012] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0013] Those skilled in the art will understand that, unless otherwise stated, the terms "the," "the," and "the foregoing" used in this application may also include plural forms. It should be further understood that the word "comprising" as used in the specification of this application means the presence of the stated features, steps, or operations, but does not exclude the presence or addition of one or more other features, integers, or steps.

[0014] Those skilled in the art will understand that, where specific experimental steps or conditions are not specified in the embodiments, they can be performed according to the conventional experimental steps or conditions described in the literature in this field; and where the manufacturers of the raw materials or instruments and equipment used are not specified, they are all conventional products that can be obtained commercially.

[0015] Those skilled in the art will understand that, unless otherwise stated in this application, when numerical ranges are given in the embodiments, 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, as well as the prior art known to those skilled in the art and the descriptions in this application, can be implemented using any prior art methods, devices, and materials similar to or equivalent to those described, used, or made by means of methods, devices, and materials in the embodiments of this application.

[0016] Farnesol is an active ingredient extracted from the bark of the acacia tree, and its main components are flavonoids and triterpenoids. Farnesol is also found in lemongrass, citronella, tuberose, cyclamen, rose, orange blossom, balsam, and musk; its chemical formula is C63-C62 ... 15 H 25 With a molecular weight of 223.72 g / mol, farnesol has been found to possess various biological functions, including antibacterial, anti-inflammatory, antioxidant, and immunomodulatory effects, as well as regulation of lipid metabolism. Clinically, 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, skincare products, and pharmaceuticals. However, there are few reports on the use of farnesol in livestock and poultry production, and no reports have been found regarding its effects on necrotic enteritis in broilers.

[0017] The embodiments of this application have shown in vitro experiments that farnesol or compositions including farnesol can inhibit the proliferation of Clostridium perfringens. In vivo experiments have shown that farnesol or compositions including farnesol can inhibit the expression of pro-inflammatory cytokines, promote the expression of antioxidant enzymes, improve intestinal health, and thus improve the feed conversion efficiency of broilers with necrotizing enteritis. The above results can provide basic data for the development of farnesol in poultry health care drugs and feed additives.

[0018] Based on this, embodiments of this application provide the use of farnesol in the preparation of drugs or feed additives for the treatment or prevention of necrotizing enteritis in chickens.

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

[0020] In some embodiments, the drug or feed additive is farnesol, meaning that farnesol can be directly used as a drug or feed additive to treat or prevent necrotic enteritis in chickens.

[0021] In some embodiments, the drug or feed additive is a farnesol composition formed by adding farnesol as an excipient and using farnesol as the active ingredient.

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

[0023] In some embodiments, the excipient is at least one selected from lactose, maltose, mannitol, and soluble starch.

[0024] In some embodiments, the excipient is at least one selected from montmorillonite, maifanite, vermiculite, zeolite powder, silica, defatted rice bran, corn cob powder, starch, and wheat bran.

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

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

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

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

[0029] In some embodiments, the farnesol composition is formulated as at least one of oral liquid, granules, tablets, and powder, and can be prepared using conventional pharmaceutical formulation methods.

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

[0031] (1) Administer the medication orally to chickens at a dose of 10-100 mg / kg of chicken body weight with farnesol content;

[0032] (2) The content of farnesol is 0.05% to 0.1% of the weight of chicken feed; farnesol granules, powders or tablets are added to chicken feed as feed additives.

[0033] (3) When used for daily prevention of chicken enteritis, the farnesol content is 0.05% of the weight of chicken feed or drinking water. The farnesol powder, granules or tablets are crushed and added to the feed or drinking water.

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

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

[0036] Example 1

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

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

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

[0040] Example 2:

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

[0042] Weigh out 50g of farnesol, 20g of wheat bran, and 30g of soluble starch, and set aside. Mix farnesol, wheat bran, and soluble starch evenly, pass through a 60-mesh sieve, and package to obtain a farnesol powder, wherein the mass fraction of farnesol is 50%.

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

[0044] Example 3:

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

[0046] Weigh out 50g of farnesol, 20g of lactose or maltose, and 30g of soluble starch, and set aside. Mix the farnesol, lactose or maltose, and soluble starch, then compress and dry them using a tablet press. This will yield a farnesol tablet with a farnesol mass fraction of 50%.

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

[0048] Example 4:

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

[0050] Weigh out 50g of farnesol, 20g of lactose or maltose, and 30g of soluble starch, and set aside. Mix the farnesol, lactose or maltose, and soluble starch, granulate, dry, and package to obtain a farnesol granule, wherein the mass fraction of farnesol is 50%.

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

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

[0053] This application's embodiments verified 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 State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0056] 1.2 Test drug: Weigh farnesol (purchased from Sigma, purity >99%), dissolve it in physiological saline to prepare a farnesol stock solution with a concentration of 0.1 g / mL, and then dilute it stepwise 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 Experimental Procedure:

[0058] The Oxford cup method was used to evaluate the inhibition zone diameter of different concentrations of farnesol against Clostridium perfringens to assess its antibacterial effect. Physiological saline served as the blank control group; enramycin (purchased from Sinopharm Group, purity >99%) served as the positive control, and in the inhibition zone test, enramycin was dissolved in physiological saline at a dose of 0.1 μg / mL; farnesol was administered at five concentrations: 0.1 g / mL, 0.05 g / mL, 0.025 g / mL, 0.0125 g / mL, and 0.0063 g / mL; each treatment group had three replicates.

[0059] In the specific experiment, Clostridium perfringens was cultured on a meat culture medium and a concentration of 1×10⁻⁶ was obtained. 5 Prepare solid and semi-solid culture plates for butcher's meat agar using CFU / mL bacterial suspension. After cooling the semi-solid culture medium to approximately 37°C, take 100 μL of each plate containing 1×10⁻⁶ CFU / mL bacterial suspension. 5CFU / mL Clostridium perfringens bacterial suspension was added to the semi-solid meat culture medium and shaken well. The mixture was then quickly poured onto a solid meat culture plate. Oxford cups were then evenly placed on the plate. After the semi-solid medium solidified, the Oxford cups were removed and 50 μL of each of the above treatment solutions was added to the ring wells. The plate was then incubated at 37°C for 24 hours. Subsequently, the diameter of the inhibition zone was measured using a ruler (accuracy 0.01 cm).

[0060] 2. Test Results

[0061] The in vitro inhibitory effects of farnesol on chicken-derived Clostridium perfringens are shown in Table 1 below.

[0062] Table 1

[0063] Group Diameter of the inhibition zone (cm) Blank control group (physiological 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] As shown in Table 1, farnesol has an inhibitory effect on Clostridium perfringens from chickens, and the diameter of the inhibition zone increases with the increase of farnesol dosage.

[0065] Effects of farnesol on growth performance and gut health in broilers infected with Clostridium perfringens

[0066] This application's embodiments verified the effects of farnesol on the growth performance and gut health of broilers infected with Clostridium perfringens. The verification method is as follows:

[0067] 1.1 Materials and Methods

[0068] The experiment used 720 healthy AA (Animal Humidity) 1-day-old birds with uniform weight. + Broiler chicks were randomly divided into three treatment groups, each with eight replicates, and each replicate group consisting of 30 chicks. The three treatment groups were: a control group, a Clostridium perfringens (CP) infection group, and a farnesol + CP infection group. The control and CP infection groups were fed a corn-soybean meal basal diet formulated according to the Chinese broiler nutritional requirements standard (NY / T33-2004). The farnesol + CP infection group received a diet supplemented with 0.05% farnesol (purchased from Aladdin Company, purity >80%) by feed weight added to their basal diet. From days 14 to 21 of the experiment, the CP infection group and the farnesol + CP infection group were administered 1 mL of CP (CVCC2030 strain, dosage 1.0 × 10⁻⁶) daily by gavage. 9The control group was fed an equal volume of CP medium (CFU / mL). During the experiment, the broilers had free access to feed and water and were under a 24-hour light regime. On day 22, all broilers were fasted for 8 hours, then weighed and their feed consumption was recorded. Growth performance-related indicators such as average daily gain (ADG), average daily feed intake (ADFI), and feed conversion ratio (FCR) were calculated. Subsequently, one broiler of uniform weight was selected from each replicate group to collect blood from the wing vein and separate the serum for testing.

[0069] 2 Data Analysis

[0070] The experimental data were analyzed by one-way ANOVA in SPSS 23.0 statistical software. Duncan's method was used to perform multiple comparisons between groups. P < 0.05 was used as the significance criterion, and P < 0.01 was used as the extremely significant criterion. The results are 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 effect of farnesol on the growth performance of broilers infected with Clostridium perfringens. In the table, a, b, and c are indicators of significant differences. Different letters on the data indicate significant differences (P < 0.05).

[0074] Table 2

[0075] Testing items control group CP infection group Acacia alcohol + 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] Table 2 shows that *Clostridium perfringens* infection reduces the average body weight, average daily feed intake, and average daily weight gain of broilers, while increasing the feed conversion ratio. Adding farnesol to the diet can alleviate the reduction in average body weight and average daily weight gain caused by *Clostridium perfringens* infection in broilers, and shows a trend towards improving the feed conversion ratio. This indicates that farnesol has a protective effect against *Clostridium perfringens* infection in broilers.

[0077] 3.2 Effects 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 broiler chickens infected with Clostridium perfringens. In the table, a, b, and c are indicators of significant differences, and different letters on the data indicate significant differences (P < 0.05).

[0079] Table 3

[0080] Testing items control group CP infection group Acacia alcohol + CP infection group P-value D-xylose, mmol / 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 intestines but cannot be utilized by the body. Therefore, evaluating the D-xylose content in the blood can assess intestinal absorption function. In a healthy state, DAO exists in the intestines; when the intestines are damaged, it is released into the bloodstream. Therefore, the higher the DAO activity in the serum, the more significant the intestinal damage. Table 3 shows that adding farnesol to the diet can alleviate the decrease in D-xylose content and the increase in DAO levels in the blood of broilers caused by Clostridium perfringens infection, indicating that farnesol can protect broilers from the decrease in intestinal absorption function and barrier damage caused by Clostridium perfringens infection.

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

[0083] Table 4 shows the effect of farnesol on the serum inflammatory cytokine levels in broiler chickens infected with Clostridium perfringens. a, b, and c are statistical markers, and different letters under the data indicate statistically 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. Pro-inflammatory cytokines were detected using a chicken ELISA kit from Beijing Solarbio Co., Ltd.

[0084] Table 4

[0085] Testing items control group CP infection group Acacia alcohol + 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 caused an upregulation of pro-inflammatory cytokines such as IL-1β, TNF-α, IL-6, and IFN-γ. However, the addition of farnesol to the diet could alleviate the increase in serum IL-1β and IL-6 in broilers caused by Clostridium perfringens infection, indicating that farnesol has a mitigating effect on the inflammatory response in broilers caused by Clostridium perfringens infection.

[0087] Effects of farnesol on growth performance and gut health in broilers co-infected with coccidia and Clostridium perfringens

[0088] This application's embodiments verified the effects of farnesol on the growth performance and gut health of broilers co-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 in the experiment, which included 480 one-day-old healthy AA birds with uniform body weight. +Broiler male chicks were randomly assigned to four treatment groups, each with eight replicates, and each replicate consisting of 15 chicks. The four treatment groups were: control group, coccidia and Clostridium perfringens co-infection group (CCP), farnesol group, and farnesol + CCP group. Broilers in the control and CCP infection groups were fed a corn-soybean meal basal diet formulated according to the Chinese broiler nutritional requirements standard (NY / T33-2004). The diets of broilers in the farnesol group and the farnesol + CCP group were supplemented with 0.05% farnesol (purchased from Aladdin Company, purity >80%) by feed weight in their basal diet. On days 8 and 10 of the experiment, broilers in the CCP group and the farnesol + CCP group were vaccinated with an ultra-high dose of coccidiosis vaccine (purchased from Foshan Zhengdian Company, dose 30 times the recommended dose) to establish an intestinal coccidiosis infection state. From days 14 to 21 of the experiment, broilers in the CCP group and the farnesol + CCP group were gavaged daily with 1 mL of CP (CVCC2030 strain, dose 1.0 × 10⁻⁶). 9 Broilers in the control and farnesol groups were fed the same volume of CP medium (CFU / mL). During the experiment, broilers had free access to feed and water and were under a 24-hour light regime. On day 22, all broilers were fasted for 8 hours, then weighed and their feed consumption was recorded. Growth performance indicators such as average daily gain (ADG), average daily feed intake (ADFI), and feed conversion ratio (FCR) were calculated. Subsequently, one broiler of uniform weight was selected from each replicate group to collect blood from the subwing vein and separate the serum for testing. The broilers were then anesthetized by injecting sodium pentobarbital (50 mg / kg live body weight) into the subwing vein, slaughtered, and dissected to obtain ileum tissue for sectioning and observation. Ileum samples were then collected for further analysis.

[0091] 2. Data Analysis

[0092] Experimental data were analyzed using SPSS 23.0 statistical software in a two-way ANOVA. The main effects of the model included farnesol, co-infection with coccidia and Clostridium perfringens, and their interaction. When an interaction was found, Duncan's method was used for multiple comparisons between groups. P < 0.05 was considered statistically significant, and P < 0.01 was considered highly significant. Results are expressed as mean ± standard error.

[0093] 3. Test Results

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

[0095] Table 5 shows the effect of farnesol on the growth performance of broilers co-infected with coccidia and Clostridium perfringens. In the table, a, b, and c are the significance markers, and different letters on the data indicate significant differences (P < 0.05). -CCP indicates no co-infection with coccidia and Clostridium perfringens, +CCP indicates co-infection with coccidia and Clostridium perfringens, -HH indicates no farnesol added to the diet, and +HH indicates the addition of farnesol to the diet. SEM represents the standard error.

[0096] Table 5

[0097]

[0098] Table 5 shows that co-infection with coccidia and Clostridium perfringens increases the feed conversion ratio, while the addition of farnesol to the diet can alleviate the negative impact of co-infection with coccidia and Clostridium perfringens on the feed conversion ratio of broilers. The results indicate that farnesol can improve the growth performance of broilers co-infected with coccidia and Clostridium perfringens.

[0099] 3.2 Effects of farnesol on antioxidant function of blood and intestine in broiler chickens co-infected with coccidia and Clostridium perfringens

[0100] Table 6 shows the effect of farnesol on the antioxidant function of blood in broilers co-infected with coccidia and Clostridium perfringens. a and b indicate statistical significance, with different letters above the data indicating significant differences (P < 0.05). -CCP indicates no co-infection with coccidia and Clostridium perfringens, +CCP indicates co-infection with coccidia and Clostridium perfringens, -HH indicates no farnesol added to the diet, and +HH indicates farnesol added to the diet. SEM represents the standard error; T-AOC represents antioxidant capacity, CAT represents catalase, H2O2 represents hydrogen peroxide, and MDA represents malondialdehyde. All indicators in the table were measured using a kit (purchased from Nanjing Jiancheng Bioengineering Institute), and the final values ​​were determined using total protein as the denominator.

[0101] Table 6

[0102]

[0103] Table 7 shows the effect of farnesol on the antioxidant function of the ileum in broilers co-infected with coccidia and Clostridium perfringens. a, b, and c are statistically significant markers; different letters in the table's subscripts indicate statistically significant differences (P < 0.05). -CCP indicates no co-infection with coccidia and Clostridium perfringens, +CCP indicates co-infection with coccidia and Clostridium perfringens, -HH indicates no farnesol added to the diet, and +HH indicates farnesol added to the diet. SEM represents standard error, GSH-Px represents glutathione peroxidase, MPO represents myeloperoxidase, T-SOD represents superoxide dismutase, and MDA represents malondialdehyde. All indicators in the table were measured using a kit (purchased from Nanjing Jiancheng Bioengineering Institute), and the final values ​​were determined using total protein as the denominator.

[0104] Table 7

[0105]

[0106] Tables 6 and 7 show that co-infection with coccidia and Clostridium perfringens downregulates serum CAT levels and upregulates MDA levels; it also downregulates GSH-Px and T-SOD levels in the ileum. Adding farnesol to the diet can alleviate the abnormal changes in these indicators caused by co-infection with coccidia and Clostridium perfringens, indicating that farnesol can enhance the antioxidant function of broilers co-infected with coccidia and Clostridium perfringens.

[0107] 3.3 Effects of farnesol on gene expression in the ileum of broiler chickens co-infected with coccidia and Clostridium perfringens

[0108] Table 8 shows the effect of farnesol on gene expression in the ileum of broilers co-infected with coccidia and Clostridium perfringens. a, b, and c are statistical markers, with different letters above the data indicating significant differences (P < 0.05). -CCP indicates no co-infection with coccidia and Clostridium perfringens, +CCP indicates co-infection with coccidia and Clostridium perfringens, -HH indicates no farnesol added to the diet, and +HH indicates farnesol added to the diet. SEM represents standard error. TNF-α represents tumor necrosis factor-α, IFN-γ represents interferon-γ, IL-18 represents interleukin-18, IL-8 represents interleukin-8, IL-1β represents interleukin-1β, ZO-1 represents occludin-1, Claudin-1 represents tight junction protein 1, and occludin represents occludin.

[0109] Table 8

[0110]

[0111] Table 8 shows that co-infection with coccidia and Clostridium perfringens downregulated the transcription levels of ocludin and ZO-1 in the ileum of broilers, while upregulating the mRNA levels of pro-inflammatory cytokines TNF-α, IFN-γ, and IL-1β. Adding farnesol to the diet could alleviate the abnormal changes in these genes caused by co-infection with coccidia and Clostridium perfringens. The results indicate that farnesol has a protective effect against intestinal inflammation and barrier function impairment in broilers caused by co-infection with coccidia and Clostridium perfringens.

[0112] In summary, farnesol can be used to prepare drugs or feed additives for the treatment or prevention of necrotic enteritis in chickens. The prepared drugs or feed additives possess resistance to various pathogens that cause necrotic enteritis in chickens, including Clostridium perfringens. The dosage forms of these drugs and feed additives include oral liquids, granules, powders, and tablets.

[0113] The present application has been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present application. The descriptions of the embodiments above are only for the purpose of helping to understand the present application and its core ideas. It should be noted that those skilled in the art can make several improvements and modifications to the present application without departing from the principles of the present application, and these improvements and modifications also fall within the protection scope of the claims of the present application.

Claims

1. Application of farnesol in the preparation of drugs or feed additives for the treatment or prevention of necrotizing enteritis in chickens.

2. The application according to claim 1, characterized in that, The aforementioned necrotic enteritis in chickens is caused by Clostridium perfringens infection alone or in combination with coccidia.

3. The application according to claim 1, characterized in that, The drug or feed additive is farnesol.

4. The application according to claim 1, characterized in that, The drug or feed additive is a farnesol composition formed by adding excipients to farnesol as the active 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) is at least one of montmorillonite, maifanite, vermiculite, zeolite powder, fumed silica, defatted rice bran, corn cob powder, starch, and wheat bran.

5. The application 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 application according to claim 4, characterized in that, The farnesol composition is a powdered mixture of farnesol, wheat bran, and soluble starch, wherein the mass fraction of farnesol in the powdered mixture is 50%.

7. The application according to claim 4, characterized in that, The farnesol composition is a sheet-like mixture of farnesol with lactose or maltose and soluble starch, wherein the mass fraction of farnesol in the sheet-like mixture is 50%.

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

9. The application according to claim 4, characterized in that, The farnesol composition can be formulated in at least one of the following forms: oral liquid, granules, tablets, and powder, and can be prepared using conventional pharmaceutical formulation methods.

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