Preparation method and application of cottonseed meal extract

Abstract

The invention provides a preparation method and application of a cottonseed meal extract, and relates to the technical field of plant extracts and parasitic disease prevention and control. The content of active ingredients in the cottonseed meal extract is 60%-85% in terms of gossypol, and the preparation method comprises the following steps: extracting cottonseed meal with water, purifying an extracting solution through resin, concentrating an eluent, adjusting alkali, centrifuging and drying to obtain the cottonseed meal extract. Experimental exploration proves that the cottonseed meal extract has a new application of preventing and treating the chicken coccidiosis, a new solution is provided for developing a breeding input for preventing and treating the chicken coccidiosis, and the application range of the cottonseed meal extract is expanded.

Description

Technical Field

[0001] The invention belongs to the technical field of plant extracts and parasitic disease prevention and treatment, and particularly relates to a preparation method of a cottonseed meal extract and application thereof. Background Art

[0002] Cottonseed meal is primarily made from cottonseeds, and is a product (by-product) obtained after de-oiling using pre-pressing or direct extraction methods. Its crude protein content is approximately 22% to 56%. In my country, where protein-based feed is in short supply, cottonseed meal can be used as a feed to provide high-quality plant protein for animals. Cottonseed meal contains gossypol, a yellow polyphenolic hydroxybisnaphthaldehyde compound commonly known as cotton toxin. It is the primary anti-nutritional factor in cottonseed meal and naturally occurs in the glands, leaves, stems, roots, and seeds of cotton. It can stimulate inflammatory responses in animal tissues such as the liver and kidneys, affecting digestive enzyme activity and hindering the digestion and absorption of dietary nutrients by animals, thus limiting the large-scale application of cottonseed meal in animal husbandry.

[0003] However, no scholar or institution has yet conducted reverse research to apply gossypol as an effective active ingredient in cottonseed meal in animal production. Although anti-nutritional factors have a certain inhibitory effect on animal digestion, absorption, and metabolism, it mainly depends on the interaction between the ingested substance and the animal's digestion, and anti-nutritional factors also have different effects in different usage scenarios. Studies have shown that gossypol has anti-tumor and anti-malarial effects. The mechanism of its anti-tumor effect is mainly manifested in anti-proliferation, intervention in signal pathways, intervention in energy metabolism and phospholipid metabolism, and inhibition of Bcl-2 protein overexpression. It can effectively inhibit ssRNA viruses, herpes viruses, etc.

[0004] Globally, avian coccidiosis is one of the five major diseases seriously impacting the development of the livestock and poultry industry. It is a highly contagious disease caused by various coccidia of the genus Eimeria that parasitize the chicken's intestines. It is widespread and most susceptible to infection between 15 and 40 days of age. Mortality is extremely high, leading to significant losses in poultry meat and egg production. Chickens infected with this disease typically exhibit symptoms such as lethargy, slow growth, and bloody stools. Morbidity and mortality rates can reach as high as 80% in chicks aged 10 to 30 days or pullets aged 35 to 60 days. It is reported that this disease causes economic losses of up to $14 billion annually to the global poultry industry. Coccidia are classified into seven types: Eimeria tenella, Eimeria toxicophila, Eimeria maxima, Eimeria acervulina, Eimeria brucei, Eimeria malaise, and Eimeria precocious. Different types of coccidia have varying pathogenicity depending on where they infect the chicken's intestines. Among them, the most pathogenic are Eimeria tenella and Eimeria toxicophila, which can cause acute small intestinal or cecal coccidiosis.

[0005] Currently, the primary means of controlling coccidiosis in chickens are vaccines and anticoccidial drugs. However, vaccines have limitations in controlling coccidiosis, such as the potential for growth retardation and intestinal inflammation in chicks. Furthermore, vaccine stability, the risk of relapse, and cost still require further optimization. The widespread use of anticoccidial drugs has also led to concerns about drug residues and resistance, further complicating prevention and control. Therefore, the search for highly effective anticoccidial agents that are less susceptible to resistance, and have no toxic side effects or residues is urgent.

[0006] To sum up, considering the problem of chicken coccidiosis infection faced by the livestock and poultry farming industry, this project adopts a comprehensive resource utilization model. For the first time, the cottonseed meal obtained after cottonseed oil extraction is used to prepare cottonseed meal extract through an extraction process. Gossypol is used as the active ingredient indicator of the extract, and it is applied in poultry farming to prevent and control chicken coccidiosis, improve the immunity of chickens, promote the growth of chickens, and reduce losses in the poultry farming industry.

[0007] Technical problems at this stage:

[0008] (1) At present, anticoccidial drugs are mainly chemical drugs, such as diclazuril, nicarbazine, chlorpheniramine, etc., which are prone to drug resistance, toxic effects on the body, and drug residues.

[0009] (2) At present, no researchers at home or abroad have studied the process of extracting gossypol from cottonseed meal and preparing cottonseed meal extract using gossypol as an effective active ingredient indicator.

[0010] (3) Currently, there are no reports on the use of cottonseed meal extract in the prevention and treatment of chicken coccidiosis. Summary of the Invention

[0011] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0012] The purpose of the present invention is to provide a preparation method of a cottonseed meal extract and application thereof in preventing and treating chicken coccidiosis. The preventing and treating chicken coccidiosis includes: preventing or treating chicken coccidiosis.

[0013] Preferably, the pathogen of chicken coccidiosis is Eimeria.

[0014] Preferably, the pathogen of chicken coccidiosis is Eimeria tenella.

[0015] Preferably, the prevention or treatment of chicken coccidiosis includes: inhibiting the growth of Eimeria tenella, reducing the excretion of Eimeria tenella oocysts, and increasing the survival rate and relative weight gain rate of chickens infected with Eimeria tenella.

[0016] A process for extracting gossypol from cottonseed meal. The obtained cottonseed meal extract has an active ingredient gossypol content of 60% to 85%. The preparation method is as follows:

[0017] (1) Extraction: Grind the cottonseed meal into coarse powder, extract with ethanol solution, extract 2-3 times, 1h / time-1.5h / time, material-liquid ratio 1:6-10, extraction temperature 70℃-90℃, filter, and collect the extract.

[0018] (2) Concentration: The extract is concentrated under reduced pressure.

[0019] (3) Macroporous resin purification: The concentrated solution in step (2) is purified by passing it through a macroporous resin and eluting it with an ethanol solution.

[0020] (4) Concentration: The eluate from step (3) is concentrated under reduced pressure until there is no alcohol smell, allowed to stand and cool to room temperature, centrifuged, and the centrifugal residue is collected.

[0021] (5) Adding alkali solution to dissolve: Add alkali solution to the centrifugal residue in step (3), stir and dissolve, let it stand and cool to room temperature, centrifuge, and collect the centrifuge liquid.

[0022] (6) Add acid to the centrifuge solution to adjust the pH: Add acid to the centrifuge solution to adjust the pH, centrifuge again, and collect the centrifuge residue.

[0023] (7) Drying: The centrifugal residue from step (6) is dried at 65°C, crushed, and sieved to obtain the gossypol extract.

[0024] Preferably, in step 1, the solution is 60%-90% ethanol.

[0025] Preferably, in step 2, the ethanol concentration is concentrated to 30%-40%.

[0026] Preferably, the macroporous resin in step 3 is any one of the following: LS303, LS315, LS410, LY105, BS16, etc.

[0027] Preferably, the concentration of the ethanol solution in step 3 is 60%-80%, and the elution volume is 2BV-4BV.

[0028] Preferably, the alkali solution in step 5 is sodium hydroxide, sodium carbonate, sodium bicarbonate, etc. with a pH of 8-9, and the added amount is 4BV-8BV, heated to 60°C and stirred for 0.5h-1.5h.

[0029] Preferably, the acid in step 6 is hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, etc., and the pH is 3-7.

[0030] Preferably, a method for preparing an extract from cottonseed meal is:

[0031] (1) Extraction: Grind the cottonseed meal into coarse powder, add 80% ethanol for extraction, extract twice, 1 h each time, with a material-liquid ratio of 1:8, extraction temperature of 80°C, filter, and collect the extract.

[0032] (2) Concentration: The extract was concentrated under reduced pressure to an ethanol concentration of 40%.

[0033] (3) Macroporous resin purification: The concentrate from step (2) was adsorbed with LS410 resin and eluted with 2BV 80% ethanol.

[0034] (4) Concentration: The eluate from step (3) is concentrated under reduced pressure until there is no alcohol smell, allowed to stand and cool to room temperature, centrifuged, and the centrifugal residue is collected.

[0035] (5) Add alkali solution to dissolve: Add 6BV sodium hydroxide solution with pH=9 to the centrifugal residue, heat to 60°C, stir for 1 hour, then let it stand to room temperature, centrifuge, and collect the centrifuge liquid (discard the centrifugal residue).

[0036] (6) Add acid to the centrifuge to adjust the pH: Add hydrochloric acid to the centrifuge to adjust the pH to 6, centrifuge again, and collect the centrifuge residue (discard the centrifuge).

[0037] (7) Drying: The centrifugal residue from step (6) is dried at 65°C, crushed, and sieved to obtain the gossypol extract.

[0038] Beneficial effects of the present invention:

[0039] 1. Existing research mainly focuses on extracting protein, flavonoid glycosides, and phytic acid from cottonseed meal, while simultaneously removing gossypol. There is no research on cottonseed meal extraction processes (using gossypol as an activity indicator). The present invention uses gossypol as an activity indicator and prepares cottonseed meal extract by subjecting cottonseed meal to a series of extraction processes.

[0040] 2. Currently, there are few studies on the application of cottonseed meal extract, especially its application in aquaculture, and no research reports on the prevention and treatment of chicken coccidiosis. The present invention has experimentally demonstrated that cottonseed meal extract has a new use in the prevention and treatment of chicken coccidiosis, providing a new solution for the prevention and treatment of chicken coccidiosis and the development of aquaculture inputs, providing new ideas for the clinical prevention and treatment of chicken coccidiosis, and expanding the application field of cottonseed meal extract.

[0041] 3. The present invention realizes the reuse of cottonseed meal in a comprehensive resource utilization mode, turning waste into treasure, and giving full play to the efficacy of cottonseed meal extract in preventing and treating chicken coccidiosis, developing new green, safe, effective, and low-cost breeding inputs for my country's poultry farming industry, reducing the breeding costs of the farming industry, and at the same time opening up new application scenarios for cottonseed meal extract. DETAILED DESCRIPTION

[0042] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0043] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those skilled in the art in the field of the present invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar or equivalent to those disclosed herein can be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0044] The various raw materials, reagents, instruments and equipment used in the examples can be purchased from the market or prepared by existing methods. Unless otherwise specified, all reagents used in the present invention are of analytical grade.

[0045] Example 1 Preparation process of cottonseed meal extract 1

[0046] The cottonseed meal raw material was crushed into coarse powder, extracted with 80% ethanol, extracted twice, 1h / time, with a solid-liquid ratio of 1:8, and an extraction temperature of 80°C. The extract was filtered and collected. The extract was concentrated under reduced pressure to an ethanol concentration of 40%, adsorbed on LS410 resin, and eluted with 2BV80% ethanol. The eluate was concentrated under reduced pressure until there was no alcohol taste. The solution was allowed to stand at room temperature, centrifuged, and the centrifugal residue was collected (the centrifugal liquid was discarded). 6BV of sodium hydroxide solution with a pH of 9 was added to the centrifugal residue, heated to 60°C, stirred for 1h, and allowed to stand at room temperature again. The centrifugal residue was collected (the centrifugal residue was discarded), and the centrifugal liquid was adjusted to pH = 6 with hydrochloric acid, centrifuged again, and the centrifugal residue was collected (the centrifugal liquid was discarded). The centrifugal residue was dried at 65°C, crushed, and sieved to obtain a gossypol extract (gossypol content was 85.93%).

[0047] Example 2 Preparation process of cottonseed meal extract 2

[0048] The cottonseed meal raw material was crushed into coarse powder, extracted with 60% ethanol, extracted 3 times, 1h / time, with a material-liquid ratio of 1:6, and an extraction temperature of 70°C. The extract was filtered and collected. The extract was concentrated under reduced pressure to an ethanol concentration of 30%, adsorbed on LY105 resin, and eluted with 3BV70% ethanol. The eluate was concentrated under reduced pressure until there was no alcohol taste. The solution was allowed to stand at room temperature, centrifuged, and the centrifugal residue was collected (the centrifugal liquid was discarded). The centrifugal residue was added with 4BV PH = 9 sodium hydroxide solution, heated to 60°C, stirred for 1.5h, and then allowed to stand at room temperature, centrifuged, and the centrifugal liquid was collected (the centrifugal liquid was discarded). The centrifugal liquid was adjusted to pH = 3 with hydrochloric acid, centrifuged again, and the centrifugal residue was collected (the centrifugal liquid was discarded). The centrifugal residue was dried at 65°C, crushed, and sieved to obtain a gossypol extract (gossypol content was 60.46%).

[0049] Example 3 Preparation process of cottonseed meal extract 3

[0050] The cottonseed meal raw material was crushed into coarse powder, extracted with 90% ethanol, extracted twice, 1.5h / time, with a material-liquid ratio of 1:10, and an extraction temperature of 90°C. The extract was filtered and collected. The extract was concentrated under reduced pressure to an ethanol concentration of 30%, adsorbed on BS16 resin, and eluted with 4BV60% ethanol. The eluate was concentrated under reduced pressure until there was no alcohol taste. The solution was allowed to stand at room temperature, centrifuged, and the centrifugal residue was collected (the centrifugal liquid was discarded). 8BV of sodium hydroxide solution with a pH of 8 was added to the centrifugal residue, heated to 60°C, stirred for 0.5h, and allowed to stand at room temperature again. The centrifugal liquid was centrifuged and the centrifugal liquid was collected (the centrifugal liquid was discarded). The centrifugal liquid was adjusted to pH = 5 with hydrochloric acid, centrifuged again, and the centrifugal residue was collected (the centrifugal liquid was discarded). The centrifugal residue was dried at 65°C, crushed, and sieved to obtain a gossypol extract (the gossypol content was 73.16%).

[0051] Example 4 Preparation of sporulated oocysts of Eimeria tenella

[0052] Ten 14-day-old SPF chicks were selected and each chicken was orally inoculated with 100 μL of oocyst suspension (i.e., 6,000 oocysts). After the inoculation, the chicks were fed with feed dried in a constant temperature drying oven at 80°C and boiled water in a coccidia-free environment.

[0053] Collect feces 6-7 days after infestation. Place the collected feces in a 5L measuring cup and add 2000mL of water. Stir thoroughly. Filter the feces through 80, 100, and 200 mesh sieves in sequence. Repeat the filtration 2-3 times and collect the filtrate. Centrifuge the collected filtrate for 5 minutes, discard the supernatant, and retain the fecal sediment. Add 20mL of water to the centrifuge cup and mix thoroughly using a vortex instrument. Centrifuge for 5 minutes and repeat the washing 3-4 times until the supernatant color shows no significant change. Add 3 times the volume of saturated salt water to the centrifuge cup containing the fecal sediment at the bottom. Use a vortex instrument to stir thoroughly and transfer the mixture to a 500mL glass beaker. Let it stand for 5 minutes. Then, divide the entire solution into 50mL centrifuge tubes, centrifuge for 5 minutes, and collect the supernatant. Dilute the supernatant with at least 7 times the volume of water, divide the solution into 50 mL centrifuge tubes, centrifuge for 5 min, and discard the supernatant. Rinse the oocysts at the bottom of each centrifuge tube with 20 mL of 2.5% potassium dichromate solution, collect the oocysts into culture bottles, and place them in a shaker at 28°C for aeration culture. After culture, store them at 4°C for later use.

[0054] Example 5 Evaluation of the Anti-coccidia Effect of Cottonseed Meal Extract

[0055] 5.1 Test reagents and main instruments

[0056] 5.2 Experimental Animals

[0057] Experimental animals: 200 one-day-old yellow-feathered broiler chickens were purchased from Hunan Animal Husbandry and Veterinary Research Institute.

[0058] Feeding conditions: Strictly kept in a clean animal henhouse without coccidia, free access to food, sufficient light, and managed by a dedicated person.

[0059] 5.3 Reagents and materials

[0060] 5.3.1 Trial Drugs

[0061] Positive drug: 0.5% diclazuril premix, add 200 mg of diclazuril premix to every 1 kg of feed.

[0062] Test drug: cottonseed meal extract, gossypol content 85.93%.

[0063] Feed: It does not contain any anticoccidial drugs and is made by Hunan Animal Husbandry and Veterinary Research Institute.

[0064] 5.4 Test methods

[0065] 5.4.1 Trial Grouping

[0066] 300 one-day-old three-yellow chickens were fed to 13 days of age and weighed, and overweight and underweight chicks were eliminated. The remaining chicks were randomly divided into 6 groups, 24 chicks / group, and 3 replicates per group (n=8). There should be no statistical difference in body weight among the groups.

[0067] 5.4.2 Dosage and coccidia infection

[0068] The drugs were mixed into the feed according to the requirements in Table 1. At 13 days old, except for the non-infected and non-medicated group and the positive control group, all other groups were given drugs until the end of the experiment. At 14 days old, except for the non-infected and non-medicated group, each chicken in the other groups was orally infected with 1×10 5 After infection, the mental state, clinical manifestations, and mortality of chickens in each group were observed and recorded. Death was immediately followed by autopsy to determine whether the cause of death was coccidiosis. Based on this, the survival rate of each group was calculated.

[0069] Table 1 Grouping of the gossypol coccidia control test

[0070]

[0071] 5.4.3 Index detection method

[0072] 5.4.4 Determination of the Anticoccidial Index (ACI)

[0073] The anticoccidial index (ACI) is calculated based on the statistical analysis of relative weight gain, survival rate, cecal lesions, and oocyst count. The formula is: ACI = (relative weight gain + survival rate) - (oocyst count + lesion count). According to the anticoccidial efficacy criteria in the "Coccidiology of Chickens": ACI < 120 indicates ineffective anticoccidial efficacy; 120 ≤ ACI < 160 indicates low anticoccidial efficacy; 160 ≤ ACI < 180 indicates moderate anticoccidial efficacy; and ACI ≥ 180 indicates high anticoccidial efficacy.

[0074] 5.4.5 Weight gain index

[0075] The chicks were weighed on an empty stomach at 14 and 21 days of age, with the weight at 14 days of age recorded as the initial weight and the weight at 21 days of age recorded as the final weight, and the relative weight gain rate was calculated.

[0076] Average weight gain = average final weight - average initial weight,

[0077] Relative weight gain rate = average weight gain of chicks in each group / average weight gain of negative control group × 100%.

[0078] 5.4.6 Mortality

[0079] During the whole process, the death of chickens was observed, and they were dissected immediately after death. The cecal lesions were observed to determine whether they died of coccidiosis and calculate the mortality rate.

[0080] 5.4.7 Cecal lesion scoring

[0081] On the 8th day after artificial infection with sporulated eggs of Eimeria tenella, the necropsy was performed and the side with obvious cecal lesions was selected for scoring. Details are shown in Table 2. Cecal lesion score = average cecal lesion score of chicks in each group × 10

[0082] Table 2 Lesion scoring rules

[0083]

[0084]

[0085] 5.4.8 Oocyst value

[0086] Starting on day 6 after gavage with sporulated Eimeria tenella oocysts, feces were collected from each group until the end of the experiment. The feces from each group were thoroughly mixed. 2 g of feces to be tested were placed in a beaker. 10 mL of deionized water, followed by 50 mL of saturated saline, were added. After thorough mixing, 1 mL of the fecal solution was pipetted into a McMaster counting chamber and allowed to stand for 5 minutes. The number of eggs in two graduated chambers was counted under a microscope. The average of the two chambers' egg counts multiplied by 200 was the number of eggs per gram of feces. At day 21 of age, 2 g of cecal contents from each group of test chickens were collected and thoroughly mixed. Oocysts were counted under a microscope using a McMaster counting chamber. The number of oocysts per gram of feces (OPG) was calculated and averaged over three replicates.

[0087] The oocyst ratio = OPG of the test chickens in each treatment group / OPG of the test chickens in the positive control group × 100%, and the oocyst value is calculated based on the oocyst ratio.

[0088] Table 3 Calculation of oocyst value

[0089] Oocyst ratio / % 0～1％ 1～26％ 27～50％ 51～75％ 76～100％ Oocyst value 0 5 10 20 40

[0090] 5.4.9 Scoring bloody stools

[0091] On the 5th, 6th and 7th days after the oral administration of sporulated oocysts of Eimeria tenella, the bloody stools of each test group were observed and scored: 0 points means 100% of the stools were free of blood; 1 point means 25% of the stools were bloody; 2 points means 50% of the stools were bloody; 3 points means 75% of the stools were bloody; 4 points means 100% of the stools were bloody.

[0092] 5.5 Test results

[0093] 5.5.1 Relative weight gain rate

[0094] Table 4 Effect of cottonseed meal extract on body weight of experimental chickens

[0095]

[0096] The results showed that the average weight gain and relative weight gain rate of chickens in the control group were significantly lower than those in the blank group (P≤0.05); the average weight gain and relative weight gain rate of chickens in the cottonseed meal extract group were significantly higher than those in the control group (P≤0.05), indicating that cottonseed meal extract can effectively improve the symptoms of weight loss in chickens caused by coccidiosis.

[0097] 2.5.2 Mortality

[0098] Table 5 Effect of cottonseed meal extract on mortality of experimental chickens

[0099]

[0100] The results showed that compared with the 79.2% mortality rate of the control group treated with poison, the diclazuril treatment group and the high-dose cottonseed meal extract treatment group had a 100% survival rate, and the medium and low-dose cottonseed meal extract groups had a survival rate of 95.8% and 91.7%, respectively, indicating that cottonseed meal extract can effectively alleviate the death of chickens caused by coccidia infection.

[0101] 2.5.3 Cecal lesion scoring

[0102] Table 6 Effects of cottonseed meal extract on cecal lesions in experimental chickens

[0103]

[0104] The results showed that the lesion value of the challenge control group was 35.6, which was significantly higher than that of the blank group, diclazuril group and cottonseed meal extract group (P≤0.05), indicating that cottonseed meal extract can effectively reduce the cecal mucosal damage caused by chicken coccidiosis.

[0105] 5.5.4 Oocyst value

[0106] Table 7 Effect of cottonseed meal extract on oocyst value of experimental chickens

[0107]

[0108] The results showed that the diclazuril group had an oocyst reduction rate of 99.9%, while the high-, medium-, and low-dose cottonseed meal extract groups had oocyst reduction rates of 77.3%, 68.2%, and 54.8%, respectively. The results suggest that cottonseed meal extract can effectively reduce oocyst excretion in infected chickens, but the effect is not as good as diclazuril.

[0109] 5.5.5 Scoring bloody stools

[0110] Table 9 Effect of cottonseed meal extract on bloody stool scores of experimental chickens

[0111]

[0112] The results showed that the control group had obvious bloody stool discharge, which was significantly higher than that of the cottonseed meal extract treatment group (P < 0.05), indicating that cottonseed meal extract can effectively alleviate the intestinal bleeding symptoms of chickens caused by chicken coccidia.

[0113] 5.5.6 ACI Calculation

[0114] Table 8 Effect of cottonseed meal extract on ACI of experimental chickens

[0115]

[0116] The results showed that the ACI in the diclazuril-treated group was above 180, while that in the infected group was below 120, indicating that the positive drug control group and model group were established. The ACI in the high-dose cottonseed meal extract group was between 160 ≤ ACI and < 180, indicating a moderate anticoccidial effect. The ACI in the medium- and low-dose cottonseed meal extract groups was between 120 ≤ ACI and < 160, indicating a low anticoccidial effect. This suggests that cottonseed meal extract, when added to feed at a dose of 1g / kg, can achieve the effect of a moderate anticoccidial drug.

[0117] 5.6 Test Conclusion

[0118] Cottonseed meal extract can treat chicken coccidiosis by reducing chicken mortality, oocyst excretion, cecal mucosal damage, bloody stool excretion and restoring weight gain. When added to the feed at a dose of 1g / kg, the ACI is 166.8, which is a medium-effective anticoccidial drug.

[0119] The foregoing descriptions of specific exemplary embodiments of the present invention are for purposes of illustration and description. These descriptions are not intended to limit the invention to the precise forms disclosed, and it is apparent that many variations and modifications are possible in light of the foregoing teachings. The exemplary embodiments have been selected and described for the purpose of explaining the specific principles of the invention and their practical application, thereby enabling those skilled in the art to realize and utilize a variety of exemplary embodiments of the invention and various options and modifications. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A method for preparing a cottonseed meal extract and its application in preventing and treating chicken coccidiosis, characterized in that: The active ingredient in the cottonseed meal extract is gossypol.

2. According to claim 1, it is characterized in that The cottonseed meal extract has a gossypol content of 60% to 85%.

3. The cottonseed meal extract according to claim 1-2, wherein the preparation method comprises the following steps: (1) Extraction: Grind the cottonseed meal into coarse powder, extract with ethanol solution, extract 2-3 times, 1h / time-1.5h / time, material-liquid ratio 1:6-10, extraction temperature 70℃-90℃, filter, and collect the extract. (2) Concentration: The extract is concentrated under reduced pressure. (3) Macroporous resin purification: The concentrated solution in step (2) is purified by passing it through a macroporous resin and eluting it with an ethanol solution. (4) Concentration: The eluate from step (3) is concentrated under reduced pressure until there is no alcohol smell, allowed to stand and cool to room temperature, centrifuged, and the centrifugal residue is collected. (5) Adding alkali solution to dissolve: Add alkali solution to the centrifugal residue in step (3), stir and dissolve, and let it stand and cool to room temperature. Centrifuge and collect the centrifuge fluid. (6) Add acid to the centrifuge solution to adjust the pH: Add acid to the centrifuge solution to adjust the pH, centrifuge again, and collect the centrifuge residue. (7) Drying: The centrifugal residue from step (6) is dried at 65°C, crushed, and sieved to obtain the gossypol extract.

4. According to claim 3, it is characterized in that The ethanol solution in step 1 is 60%-90% ethanol solution; and the solution is concentrated under reduced pressure to an ethanol concentration of 30%-40% in step 2.

5. According to claim 3, it is characterized in that In step 3, the macroporous resin is any one of the following: LS303, LS315, LS410, LY105, BS16, etc.; the ethanol concentration is 60%-80%, and the elution volume is 2BV-4BV.

6. According to claim 3, it is characterized in that In step 5, add alkali solution to dissolve: the alkali solution can be sodium hydroxide, sodium carbonate, sodium bicarbonate, etc. with a pH of 8-9. Add 4BV-8BV, heat to 60°C and stir for 0.5h-1.5h.

7. According to claim 3, it is characterized in that In step 5, acid is added to adjust the pH: the acid is hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, etc., and the pH is 3-7.

8. According to claim 3, it is characterized in that The preparation method of the cottonseed meal extract comprises the following steps: grinding the cottonseed meal raw material into coarse powder, adding 80% ethanol for extraction, extracting twice, 1 hour each time, with a material-liquid ratio of 1:8 and an extraction temperature of 80°C, filtering, collecting the extract, concentrating the extract under reduced pressure to an ethanol concentration of 40%, adsorbing the extract with LS410 resin, eluting with 2BV 80% ethanol, concentrating the eluate under reduced pressure until there is no alcohol smell, allowing the solution to stand at room temperature, centrifuging, collecting centrifugal residue (discarding the centrifugal liquid), adding 6BV sodium hydroxide solution with a pH of 9 to the centrifugal residue, heating to 60°C, stirring for 1 hour, allowing the centrifugal residue to stand at room temperature, centrifuging, collecting the centrifugal liquid (discarding the centrifugal residue), adjusting the centrifugal liquid to a pH of 6 with hydrochloric acid, centrifuging again, collecting the centrifugal residue (discarding the centrifugal liquid), drying the centrifugal residue at 65°C, crushing, and sieving to obtain the cottony phenol extract.

9. According to claims 1-8, it is characterized in that The control of chicken coccidiosis includes: preventing or treating chicken coccidiosis.

10. According to claims 1-9, it is characterized in that The pathogen of chicken coccidiosis is Eimeria tenella.

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