WO2007003068A1

WO2007003068A1 - Feed additive, its preparation method and application

Info

Publication number: WO2007003068A1
Application number: PCT/CN2005/000960
Authority: WO
Inventors: Xin Wang; Manli Wang; Yanjun Cheng
Original assignee: Hangzhou Lixin Biotechnology Co. Ltd.
Priority date: 2005-06-30
Filing date: 2005-06-30
Publication date: 2007-01-11

Abstract

The present invention provides a feed additive, its preparation method and application. The feed additive is oligomeric proanthocyanidins, the oligomeric proanthocyanidins are bi-, ter-or tetra-polymers. The preparation method of the feed additive is: using condensed tannin or extract having condensed tannin as material and 20-60% hydrogen peroxide or potassium chlorate as oxidizer, carrying out oxidizing reaction 15-120 minutes under the condition of pH6-7 and temperature 30-50°C to yield a subtransparent liquid with a color of yellow to brown which has then separated and purified to obtain the feed additive. The feed additive can be used in feed for preventing and treating animal coccidiosis and its concentration in the feed is 5-600 ppm. The feed additive can be used in feed for chicken, duck, quail, pigeon, rabbit and pig. It also can be used together with other anticoccidiosis drugs, antioxidant, or enzyme preparation.

Description

A word additive and its preparation and application

The invention relates to a feed additive and its preparation and use. Specifically, the feed additive is an oligomeric proanthocyanidin which can effectively reduce the infection rate and mortality of avian animals against coccidiosis at an added amount of 5-600 ppm. Background technique

In the field of phytochemistry, proanthocyanidins are substances that produce anthocyanidin in the treatment of hot acid, including monomeric proanthocyanidins and polymer proanthocyanidins. The monomeric proanthocyanidin corresponds to flavan-4-ol and xanthine-3,4-alcohol, and the polymer proanthocyanidin corresponds to condensed tannin, red powder and phenolic acid.

Polymer proanthocyanidins are new health products and medicines that have become more and more popular in the public in recent years. It is highly resistant to oxidation and exhibits outstanding effects in terms of antiviral and anti-cancer effects. Proanthocyanidins are widely found in natural plants and are the main active constituents of plant defense systems. They are also the main active components in many Chinese herbal medicines. Chinese herbal medicines such as betel nut, crane grass and pomegranate peel contain a large amount of polymer proanthocyanidins. . Among them, the protective effect of grape seed tannin and tea polyphenol on the human cardiovascular system and the special physiological and biochemical activities of antibacterial, antiviral, antioxidation and anticancer have been scientifically confirmed. The extraction of proanthocyanidins from plants, especially grape seed and pine bark, has resulted in large-scale production.

Another major product of polymer proanthocyanidins is various types of tannin extracts extracted from bark, such as bayberry tannin, lozengein, black wattle, and eucalyptus. These polymeric proanthocyanidins, known as condensed tannins or tannins, are products of the condensation of dimeric anthocyanins with flavan-3,4-alcohol. The black wattle bark tannin structural unit is the original ring thorn of the A ring which is a resorcinol type, and the average degree of polymerization is 4 to 5. The structural unit of larch tannin is pyrogallol-like proanthocyanidin, and the average degree of polymerization is 9-10. The structural unit of Maoyangmei tannin is the original vermilion, with an average degree of polymerization of 12. Condensed tannins or tannins have a high average degree of polymerization, and low levels of oligomeric proanthocyanidins are the main cause of protein precipitation and palate.

Common methods for taking proanthocyanidins include water extraction, organic solvent extraction, and supercritical carbon dioxide extraction. Due to the different extraction methods, the degree of polymerization of proanthocyanidins produced is different. The physiological activity of oligomeric proanthocyanidins (2~4mers) is much larger than that of proanthocyanidins (5mers or more), and the patent application number is 01109225. 4 and 03117429. The invention patent of the invention is to use an organic solvent extraction method such as ethyl acetate to separate oligomers and polymers of proanthocyanidins from a mixture of anthocyanins, but the oligomeric proanthocyanidins obtained by the extraction method are obtained. The rate is low, and it is difficult to separate. At the same time, the use of a large amount of organic solvents is liable to cause environmental pollution. Therefore, it is urgent to find a method for rapidly converting high-polymer proanthocyanidins into oligomeric proanthocyanidins. Confirmation Coccidiosis is a very serious disease in poultry farming. Chicks usually have the highest incidence in 15-50 days of age, and the mortality rate in severe cases can reach more than 80%. The diseased chickens grow slowly and develop poorly, and the daily weight gain and egg production rate are affected. Prior to the present invention, a conventional method for preventing coccidiosis was to add anticoccidial drugs to feed or drinking water. Common anticoccidial drugs include sodium salinomycin, monensin, sedummycin, and amprolium. , sulfamethoxidine, chlorpheniramine, sulfaquinoxaline, diclazuril, nicarbazin and benzophenone. These anticoccidial drugs have obvious curative effect, moderate price and convenient use, and have been widely used in the poultry farming industry. However, the main problem with the current use of anticoccidial drugs is -

1. Coccidiosis drugs produce resistance problems: Coccidia are highly resistant to drugs, making many anticoccidial drugs short-lived. According to the investigation of resistance to coccidia in Guangdong in 1997, 10 anticoccidial drugs commonly used in production (ball net, diclazuril, clopidol, chlorpyrifos, amprolium hydrochloride, chlorine chloride) Among phenylhydrazine, maduramycin, lasamycin, salinomycin, and sindiramycin, 58% of the strains are sensitive to only one drug, and more than 80% of the strains are one or two Drugs are sensitive, and 14% of the strains are not sensitive to all drugs. It can be seen that the resistance of coccidia is very common, and the room for effective therapeutic treatment is getting smaller and smaller. This is also the coccidia in many areas. The main reason why the disease is difficult to control.

2. Coccidiosis problems in laying hens and breeders: Due to the continuous use of anticoccidial drugs during the growing season and the growing period, the flocks may not have the opportunity to develop anti-coccidial immunity, but they cannot use anti-cocci during the laying period. Insecticides, which can cause severe coccidiosis in non-immune flocks.

3. Drug residue problems: Coccidiostats are generally chemical synthetic drugs or antibiotics. The residues of these drugs in the carcass affect the export of poultry products and meat safety in China.

In recent years, with the increasing concern about food safety issues, the anti-coccidial effect of Chinese herbal extracts has been increasingly favored by the breeding industry. Compared with chemically synthesized anticoccidial drugs, the anti-coccidial effect of Chinese herbal medicine is non-toxic, no residue, no side effects and is not easy to cause coccidia resistance. It is a new feed additive that is green, environmentally friendly and safe. Commonly used Chinese herbal anticoccidial drugs include verbena, Tianmingjing, Changshan, Huanghua, Sophora, and Pulsatilla.

Multimeric anthocyanins, tannins for the prevention and treatment of avian coccidiosis have been published in several patents (US Patent No. 5,135,746, US 6,106,838, WO 00/72858), but the proanthocyanidins involved in the above patents are multimeric flowers. Natural tannins, and tannins are condensed tannins extracted directly from plants. Such condensed tannins have a higher degree of polymerization because they combine with proteins to form water-insoluble complexes. The combination of tannins and protein in the feed results in the absorption and utilization of proteins in the feed by the animal, while tannins and Intestinal digestive enzyme binding reduces the activity of enzymes in the gut of animals. Therefore, from the perspective of animal nutrition, it is not suitable for use in animal feed. Because of the negative effects of tannins in animal nutrition, tannins have always been considered as feeds. Important anti-nutritional factors. Therefore, the direct use of condensed tannins from plant extracts as anticoccidial drugs added to the feed will affect animal growth and development. Summary of the invention

In order to solve the problem that the multimeric anthocyanins having the anticoccidial function in the prior art are not suitable for being added into the feed, and the existing oligomeric anthocyanins have low preparation rate and complicated operation, the present invention provides a low The polymer proanthocyanidin is an active ingredient feed additive, and provides a method for rapidly and efficiently converting multimeric proanthocyanidins into oligomeric proanthocyanidins, and a method for using such feed additives in controlling animal coccidiosis.

Accordingly, it is an object of the present invention to provide a feed additive which is non-toxic, environmentally friendly, has no adverse effects on the activity of the digestive amylase, is inexpensive to manufacture, and is simple to use.

It is still another object of the present invention to provide a method of preparing the feed additive.

Another object of the present invention is to provide the use of the feed additive for the preparation of a feed for the prevention and treatment of coccidiosis in animals.

In order to achieve the object of the invention, the technical solution adopted by the invention is:

A feed additive, wherein the feed additive is an oligomeric proanthocyanidin, and the oligomeric proanthocyanidin is a 2 to 4 polymer.

In the present invention, the proanthocyanidins (PC) are a general term for a large class of polyphenol compounds widely found in nature. The simplest proanthocyanidins are dimers of catechins, epicatechins, or catechins and epicatechins. Usually, the two, three, and tetramers are called oligomers (Oligomeric). Proanthocyanidins (abbreviated as 0PC), which is called a polymer in the form of a polymer.

(Polymers Proanthocyanidins, referred to as PPC). The proanthocyanidins specifically referred to in the present invention are oligomeric proanthocyanidins (0PC), and are polymers of di-, tri-, tetra-mers which are specifically linked to the condensation reaction of flavan-3-ol and flavan-3,4-ol. High polymer proanthocyanidins are not included in the present invention.

The oligomeric proanthocyanidins to which the present invention relates can be obtained by conventional, well-reported extraction methods. These methods include water extraction, organic solvent extraction, and carbon dioxide supercritical extraction. Extracted objects include Grape Seeds, Grape Skin, Pine Bark, Pine Needles

(Abies siberica), Peanut Skins, Sorghum vulgare Pers., Apple (fruit of Rosaceae plant Malus pumilaMicc.), Cocoa and other beans and wild plants such as rose hips ( Rose Hips), _cerasus , raspberry, blackberry, raspberry, strawberry (Fragaria vesca L.), sea buckthorn (Hippophae rhamnoides L.), etc. But it is not limited to the plants mentioned. Oligomeric proanthocyanidin It is selected from grape seeds because it is the most abundant in grape seeds. The original flower extracted from grape seed and pine bark in the present invention«, in Chinese patent

It is mentioned in CN99111574. 0, CN03117429. 9, CN03117429. 9, CN03137933. 8, CN200410053240. 4, CN200410054008. 2, the products produced by these patents can be applied in the present invention.

The proanthocyanidins present in the grape seed extract are composed of two monomers, one is catechin and the other is epicatechin. The chemical structures of the two monomers are already in the literature of phytochemistry. Widely reported. Two-tetramer oligomeric proanthocyanidins formed by polymerization of these two monomers are used in the present invention.

The proanthocyanidins present in the pine bark extract are composed of three monomers, namely catechin, epicatechin and taxol, and the chemical structures of the three monomers are already in the literature of phytochemistry. Widely reported. The di-tetramer oligomeric proanthocyanidins formed by the polymerization of these three monomers are used in the present invention.

The oligomeric proanthocyanidins of the present invention can also be obtained from oligocyanidins and condensed tannins by oxidative degradation. The method is specifically as follows: the condensed tannin or the condensed tannin-containing extract is used as a raw material, and the condensed tannin is oxidatively degraded by using hydrogen peroxide or potassium chlorate at a concentration of 20 to 60% as an oxidizing agent, and the oxidation reaction condition is The pH is adjusted to 6 to 7, the temperature is controlled at 30 to 50 ° C, the oxidative degradation time is 15 to 120 minutes, and a yellow to brownish yellow translucent liquid is obtained, and the oligomeric proanthocyanidin is obtained by separation and purification.

The invention relates to a method for preparing the feed additive, that is, an oligomeric proanthocyanidin, which is prepared by using a condensed tannin or an extract containing condensed tannin as a raw material, and a hydrogen peroxide or potassium chlorate having a concentration of 20 to 60% as an oxidant. The oxidation reaction is carried out at a pH of 6 to 7, at a temperature of 30 to 50 ° C for 15 to 120 minutes to obtain a yellow to brownish yellow translucent liquid, which is removed by conventional physical or biochemical methods, and then dried to obtain an oligomeric proanthocyanidin. The amount of oxidizing agent added is usually from 1 to 4 times the volume of the condensed tannin or the extract containing the condensed tannin.

In the preparation method of the oligomeric proanthocyanidin, the condensed tannin is a polymer of flavanol, and the molecular skeleton is C ₆ · C ₃ · C ₆ , which is a polymer of 2 to 5 flavanol monomers. The molecular weight distribution is between 500 and 3000 angstroms.

The condensed tannin may also be a tannin-rich vegetable tannin.

The condensed tannin may also be an extract of condensed tannin. Specifically, the method is as follows -

(1) Extraction: Take the plant parts rich in tannins and add 4 to 6 times the weight of water, heat at 80 °; L00 °C for 0. 5~1 hours, remove the residue by filtration, and use the same extraction as above. The conditions are repeated 1 to 2 times; the filtrate is collected and concentrated to a relative density of 1. 205~1. 332 condensed tannin extract;

(2) oxidative degradation: to the extract of the condensed tannin obtained in the step (1), adding 2 times the volume of 27.3% to 50% hydrogen peroxide, adjusting 11 to 6.7, degrading at 40 ° C for 30 minutes, Yellow to yellow brown translucent liquid is Degradation products;

(3) Separation and purification: The degradation products are concentrated under reduced pressure to evaporate the remaining hydrogen peroxide and dry to obtain oligomeric proanthocyanidins.

The feed additive can be applied to the prevention and treatment of coccidiosis in animals.

The addition of the oligomeric proanthocyanidins to animal feed can effectively reduce the infection rate and mortality of avian animals against coccidiosis, and has no negative impact on the growth and development of avian animals, so it can be used as a feed for treating coccidiosis in animals. additive. This kind of feeding additive can treat clinical symptoms such as blood in the stool and intestinal mucosal damage caused by coccidiosis infection, and reduce the mortality after coccidiosis infection. The oligomeric proanthocyanidin is added in an amount of 5 to 600 ppm in a full-price material, and more preferably in an amount of 5 to 400 ppm. When used as a therapeutic drug, 5-600 ppm of the product of the present invention can be added to the feed.

The feed additive can also be added to 1% to 5% of the premix. The amount added in the premix can be calculated based on the active ingredient of the compound feed. At the same time, the feed additive oligomer proanthocyanidins can be mixed with anti-coccidial drugs or other anti-stress feed additives which have been supplied in large quantities on the market to achieve the best effect against coccidia. The purpose of mixing with other anticoccidial drugs is to reduce the concentration of antibiotics or chemically synthesized anticoccidial drugs in the feed to reduce the residues of antibiotics or chemically synthesized anticoccidial drugs in animal caries. . Antibiotic compounds or chemically synthesized anticoccidial drugs include sedummycin, nicarbazin, salinomycin, mandeltamycin, monensin, etc., but are not limited to only a few mentioned Within the anticoccidial drugs. The oligomeric proanthocyanidin is added in an amount of 5 to 400 ppm, and the amount of the other antibiotic compound or the chemical synthetic anticoccidial drug is half or less of the amount added alone.

The feed additive can also be used in combination with other antioxidants to enhance anti-coccidial efficacy. Antioxidants include ascorbic acid, vitamin strontium, omega-3 fatty acids having antioxidant functions, and the like. 005%〜0. 1%。 The amount of the oligo procyanidin is 0. 005%~0. 1%.

The feed additive can also be used in combination with an enzyme preparation. The enzyme preparation includes a variety of enzyme preparations conventionally used in feeds such as amylase, protease, xylanase, glucanase and the like. It also includes bromelain and the like which have anti-disease and therapeutic effects. Among them, the mixed use with bromelain is particularly effective. 001%〜1%。 The amount of the oligosaccharide proanthocyanidin is 5 to 200 ppm, and the amount of the enzyme is different depending on the enzyme species and the activity level, and is generally 0.001% to 1%.

The feed additive can be used in the following animals: 1 chicken, 2 ducks, 3 babies, 4 pigeons, 5 free, 6 pigs, or ruminants.

The feed additive is for E. tenella, which is parasitic in the cecum, E. maxima, which is parasitic in the small intestine, and E. maxima, which is parasitic in the duodenum and the small intestine. E. acervulina and E. mitis has both preventive and therapeutic effects. Especially for the E. coli which is parasitic in the cecum, it has better therapeutic and preventive effects.

The beneficial effects of the feed additive and the preparation and application thereof according to the invention are mainly as follows: (1) Proanthocyanidins are plant extracts, which are widely used as antioxidants in human health foods as a feed additive for anticoccidial effects. It has the characteristics of non-toxic, no side effects, no residue in the body, and is not easy to produce drug resistance, which meets the requirements of today's society for green, environmentally friendly and pollution-free additives. (2) Proanthocyanidins are chemically clear in plant extracts. A class of compounds that have strong antioxidant and free radical scavenging functions. Antioxidant and scavenging free radicals are one of the mechanisms of anti-coccidial action of natural extracts compared to other plant extracts that do not have clear chemical composition and function (Allen & Fetterer Cinical Microbiology Review 2002 15 : 58-65 ) ; 3) Compared with tannin, the oligomerization proanthocyanidins have a protein deposition capacity of almost 100% at a certain concentration, and the taste is degraded, which has no adverse effect on the amylase activity of the digestive tract and has no negative impact on animal growth and development. (4) The production cost is low, the method of use is simple, and the breeding cost is not greatly increased. detailed description

The present invention is further described below in conjunction with specific examples, but the scope of protection of the present invention is not limited thereto: Example 1: Decomposition of larch gum to produce oligomeric proanthocyanidins

1 kg of larch gum is added with water 1 kg, then add 2 kg of 50% hydrogen peroxide, adjust the pH to 7 with sodium hydroxide solution, and heat to 30 °C for 30 minutes. After the degradation is completed, continue to heat and stir, add 1~ 2 kg of water to replenish, volatilize clean hydrogen peroxide, and concentrate to 2 kg to obtain the oligomeric proanthocyanidins of golden or brownish yellow products. Example 2: Decomposition of black wattle bark tannin to produce oligomeric proanthocyanidins

1 kg of black wattle bark tannin plus 1 kg of water, then add 2 kg of 50% hydrogen peroxide, adjust the pH to 6 with sodium hydroxide, and heat to 40 ° C for 30 minutes. After the degradation is completed, continue to heat and stir, add 1 - 2 kg of water to replenish, volatilize clean hydrogen peroxide, and concentrate to 2 kg to obtain a golden or brownish yellow product oligomeric proanthocyanidin. Example 3: Extraction and decomposition of pine needles tannin from Pinus koraiensis. 10 kg of fresh pine needles are kneaded into small sections, adding 50 kg to 60 kg of water, and heating at 90 to 100 ° C for 30 minutes. The residue is removed by filtration, and the supernatant is heated and concentrated to a relative density of 0.25. The obtained condensed tannin extract is 20 kg, and 10 kg of Pinus massoniana condensed tannin extract is used, and the remaining 10 kg plus 27.3%. Hydrogen peroxide, adjust the pH to 7 with sodium hydroxide solution, heat to 50 ^: degrade for 30 minutes. After the degradation is completed, the mixture is heated and stirred to evaporate clean hydrogen peroxide, and concentrated to 3 kg to obtain a golden or brownish yellow product oligomeric proanthocyanidin. Example 4: Determination of average molecular weight of oligomeric proanthocyanidins

The average molecular weight of oligomeric proanthocyanidins was determined by internationally accepted thin layer chromatography (TLC). The specific method was as follows: The molecular weight was measured using a TLC60F silica gel plate manufactured by Merk. Prior to analysis, the silica gel plates were pretreated with isopropanol and then dried in a 130 ° oven for 2 hours. The oligomeric proanthocyanidins were concentrated under reduced pressure, and the solids were lyophilized, and 40 mg of the solid was dissolved in 10 ml of methanol. Standards (epicatechin, EGCE, proanthocyanidin B-1, proanthocyanidin C-1 and larch tannin) 40 mg were dissolved in 10 ml of methanol. 10 ul of degradant samples and standards were spotted on silica gel plates. The developing solvent was acetone: water: formic acid = 90: 5: 5. After separation, the silica gel plate was mixed with a 1% solution of 4-dimethylaminocinnamaldehyde in ethanol, and a 37% hydrochloric acid solution of 1:1, and uniformly sprayed to reveal a gray-blue spot of proanthocyanidins.

The molecular weight of the oligomeric proanthocyanidins produced in the above tests was determined to be between 400 MW and 1000 Å, and most of the degradation products had a molecular weight of about 800 MW. Example 5: Effect of oligomeric proanthocyanidins on protein precipitation

Experimental steps:

1. Take 1 gram of larch gum to a volume of 20 ml, which is No. 1 solution; take 1 gram of Example 10PC to a volume of 20 ml, which is No. 2 solution; commercially available oligomeric proanthocyanidins extracted from grape seeds (including 2~4 mer 60% or more) 1 gram dissolved in 20 ml of water to obtain 5% light brown solution, which is No. 3 solution;

2. Take 7 tubes, add the above 1st solution lml, 0. 5ml, 0. 25ml, 0. 125ml, 0. 063ml, 0. 031ml, 0. 016ml, add dh ₂ o water 0ml, 0. 5ml , 0. 75ml, 0. 875ml, 0. 937ml, 0. 969ml, 0. 984ml (that is, add 1ml of water to each tube), then add 1ml of lmg/ml gelatin solution, and make up to 10ml. The equivalent concentration of tannins is 5. 00g / l, 2. 50g / l, 1. 25g / l, 0. 63g / l, 0. 31g / l, 0. 16g / l, 0. 08g / l, After the shock was checked, at a constant temperature of 3 (TC was allowed to stand for 24 hr, turbidity was measured. Each group of control materials replaced 1 ml of gelatin solution with 1 ml of dh ₂ o, and the other operations were the same as before.

3. Seven tubes were also taken, and the No. 2 solution was used as described above to measure the turbidity. 4. Seven tubes were also taken, and the No. 3 solution was used as described above to measure the turbidity.

5. After measuring the turbidity of each group of tubes, centrifuge at 5000r/min, and then dry the precipitate, and weigh the weight: The experimental data are shown in Table 1:

Table 1: Effect of oligomeric proanthocyanidins (0PC) and larch tannin on protein precipitation

From the above table, we can see that the oligomeric proanthocyanidins (example 1 0PC and grape seed 0PC) have a significantly reduced ability to precipitate proteins compared to deciduous rubber, and larch gum is a commercially available product. Example 6: Effect of oligomeric proanthocyanidins (0PC) on intestinal enzyme activity in chicken

5 20-day-old chicks were removed from the small intestine immediately after slaughter to prepare the 15% phosphoric acid buffer suspension. After centrifugation, the supernatant was taken as the enzyme solution to be tested. Another 1% soluble starch is used as a substrate. Example 1 The prepared oligomeric proanthocyanidins (Example 10PC) and commercially available grape seed extract (Grape seed 0PC) containing more than 60% of 0PC were made 0.1% aqueous solution, and Example 3 was prepared to obtain Pinus massoniana condensation. 1%水溶液。 The tannin extract was also formulated into a 0.1% aqueous solution. The test tube was added with 2 ml of substrate, 1 ml of enzyme solution, and 0, 0.02%, 0.04%, 0.06%, 0.08%, 0.1% of oligomeric proanthocyanidins (each of which is an example 10PC or Grape seed 0PC) or Example 3 prepared a solution of Pinus massoniana condensed tannin extract, which was separately mixed and then incubated in a 37 ° C water bath for 60 minutes. A 1 ml sample was taken at 0 hours and 1 hour, and immediately heated in a 100 Torr water bath for 30 minutes to inactivate the enzyme, cooled, and the reducing sugar content was determined by 3,5-dinitrosalicylic acid colorimetric method. The protein content was determined by Coomassie Brilliant G250 colorimetric method. Amylase activity (IU) was calculated as: micromolar reducing sugar / microgram protein. min. The experimental data is shown in Table 2.

Table 2: Effect of Different Concentrations of Oligomer Proanthocyanidins and Masson Pine Condensed Tannin Extracts on Intestinal Amylase Activity in Chickens

It can be seen from Table 2 that the effect of oligomeric proanthocyanidins (Example 10PC and Grape Seed 0PC) and Pinus massoniana condensed tannin extract on intestinal enzyme activity in chicks was significantly reduced. Example 7: Therapeutic effect of oligomeric proanthocyanidins on chicken coccidiosis

80 animals were tested and broiler chickens (chicks) were isolated from the 11-day-old weighing group, 8 rats in each group, randomized, the body weight was similar, the error between groups was ± 10 g, the stool was checked before the test, and the infection was confirmed. Coccidia chickens were used as test chickens. The sample to be tested was evenly mixed with the feed. The drug-containing material was fed 24 hours before inoculation, and fed for 7 days. The sample to be tested was 130 ppm of the oligomeric proanthocyanidins and salinomycin prepared in Example 1 respectively.

At 12 days of age, Eimeria tenella was inoculated, except for the non-attack group, each of the other groups was infected with 10 X 10 ⁴ sporulated oocysts. The spirit, appetite and feces were examined daily after infection, and feces were collected on the 7th day, oocysts were counted, and culling was performed on the 8th day to observe the cecal lesions. The experimental data are shown in Table 3, and the production performance is shown in Table 4.

table 3

As can be seen from Table 3, the effect of weight gain was better in the salinomycin group, from bloody stool scores, lesion values, oocyst values, In terms of anticoccidial index (ACI), the oligomeric proanthocyanidin group (Example 10PC) was the best and reached an excellent level. Table 4 Production performance statistics days 9 days

It can be seen from Table 4 that the daily weight gain and the meat content of the oligomeric proanthocyanidin group are better than those of the attack group, and are worse than the salinomycin. Example 8: Therapeutic effect of oligomeric proanthocyanidins on chicken coccidiosis

Test animals AA broiler chickens (chickens) 190 pigeons, isolation and flattening to 11-day-old weighing randomized group, 10 rats in each group, the error between groups was ± 10 g, check the feces before the test, and determine that the uninfected coccidia chicken was used as the experimental chicken. . The sample to be tested (see Table 5) was evenly mixed with the feed, and the medicated feed was started 24 hours before the inoculation, and fed continuously for 7 days. At 12 days of age, Eimeria tenella was inoculated (reinforced by chicken before use). Except for the non-attack group, each group of chickens was infected with 10 X 10 ⁴ sporulated oocysts. Check mental, appetite and fecal changes daily after infection, collect feces on day 7 and perform oocyst counts,

Weighed and culled in 8 days to observe the cecal lesions. The experimental data is shown in Table 5:

Table 5: Comparison of anti-coccidial index of proanthocyanidins and antibiotics

Note: Grape seed 0PC contains more than 60% oligomeric proanthocyanidins; pine bark 0PC contains about 30% oligomerization Protoplast anthocyanins, which are commercially available products.

As can be seen from Table 5, the oligomeric proanthocyanidin group (Example 1 0PC, Grape Seed 0PC) was better than salinomycin, sea 'Nanthomycin, in bloody stool score, lesion value, oocyst value, and anticoccidial index (ACI). The effect of the maduramycin group was good. The oligomeric proanthocyanidin group and nicarbazin (62. 5ppm) were combined with the anticoccidial index close to the total addition of nicarbazin (125ppm).

Table 6: Production performance

Group Drug Name Average Starting Weight Average Ending Average Weight Gain Relative Increase 曰 Increase (g) Weight (g) Weight (g) Rate% Weight%%

(g/d)

1 0. 4%. Example 1 0PC 166. 5 248. 3 81. 8 49. 1 75. 7 10. 23

2 0. 6%. Example 1 0PC 166. 0 237. 4 71. 4 43. 0 66. 1 8. 93

3 Pine bark 0PC20 ppm 166. 0 244. 9 78. 9 47. 5 73. 1 9. 86

4 Pine bark OPC 40ppm 164. 5 243. 3 78. 8 47. 9 73. 0 9. 85

5 Grape seeds 0PC 10 ppm 165. 0 255. 9 90. 9 55. 1 84. 2 11. 36

6 Grape seed OPC 20 ppm 164. 0 259. 6 95. 6 58. 3 88. 5 11. 95

7 Example 1 OPC 0. 4% + 166. 0 246. 0 80. 0 48. 2 74. 1 10. 0

Sedummycin 15ppm

8 Pine bark 0PC 20 ppra+ 166. 5 252. 6 86. 1 51. 7 79. 7 10. 76

Sedummycin 15ppm

9 Grape seed OPC 20 ppm+ 165. 5 256. 0 90. 5 54. 7 83. 8 11. 31

Sedummycin 15ppm

10 Nicarbazine 125ppm 165. 0 267. 6 102. 6 62. 2 95. 0 12. 83

11 Example 1 0PC 0. 4%. + 165. 0 256. 6 91. 6 55. 5 84. 8 11. 45

Nicarbazine 62. 5ppm

12 Pine bark 0PC 20 ppm+ 167. 0 268. 4 101. 4 60. 7 93. 9 12. 68

Nicarbazine 62. 5ppra

13 Grape seed OPC 20 ppm+ 164. 0 263. 1 99. 1 60. 4 91. 8 12. 39

Nicarbazine 62. 5ppm

14 赛杜mycin 25ppm 166. 0 260. 0 94. 0 56. 6 87. 0 11. 75

15 Salinomycin 60 ppm 167. 0 250. 3 83. 3 49. 9 77. 1 10. 41 16 Hainanmycin 7. 5ppm 165. 5 249. 8 84. 3 50. 9 78. 1 10. 54

17 Maduramycin 5ppm 166. 0 244. 0 78. 0 47. 0 72. 2 9. 75

18 infection without medication 165. 0 247. 6 82. 6 50. 1 76. 5 10. 33

19 No infection without drugs 165. 5 273. 5 108. 0 65. 3 100. 0 13. 5

As can be seen from Table 6, the daily weight gain of the oligomeric proanthocyanidin group (Grape Seed 0PC) was better than that of salinomycin, Hainanmycin, Maduramycin, and Sedummycin. The oligomeric proanthocyanidins and nicarbazin (62. 5ppm) compounded daily weight gain rate close to the full addition of nicarbazin (125ppm). Example 9: Therapeutic effect of oligomeric proanthocyanidins and Vc or bromelain on chicken coccidiosis test Animals AA broiler (chick) 140 pigeons, isolated and raised to 11 days old weighing random group, each group of 10, group The error was ± 10 g, and the feces were checked before the test, and the chickens that were not infected with coccidia were determined as test chickens. The sample to be tested (see Table 7) was evenly mixed with the feed, and the medicated feed was started 24 hours before the inoculation, and fed continuously for 7 days. At 12 days of age, Eimeria tenella was inoculated (reinforced by chicken before use). Except for the non-attack group, each group of chickens was infected with 10 X 10 ⁴ sporulated oocysts. Mental, appetite and fecal changes were examined daily after infection, and feces were collected on day 7 for oocyst counts, and culling was performed on day 8 to observe cecal lesions. The experimental data is shown in Table 7:

Table 7: Comparison of oligomeric proanthocyanidins and Vc or bromelain complexes with antibiotics and chemical synthetic anticoccidial indices

Note: Grape seed 0PC contains more than 60% oligomeric proanthocyanidins.

As can be seen from Table 7, the oligomeric proanthocyanidin group was used in combination with vitamin C or bromelain, and its anticoccidial index was close to or better than that of salinomycin. Table 8 Production performance

It can be seen from Table 8 that the daily gain of the oligomeric proanthocyanidin group (Grape seed 0PC) and the Vc or Bromelain combination group is closer to or better than the salinomycin 60 ppm and the domestic Nika ball 125 ppm.

Claims

Rights request

A feed additive characterized in that the feed additive is an oligomeric proanthocyanidin, and the oligomeric proanthocyanidin is a 2 to 4 polymer.

2. The term additive according to claim 1, wherein the oligomeric proanthocyanidin is obtained by extracting from any of the following plants: grape seed, grape skin, pine bark, pine needle, peanut skin, sorghum, Apple, cocoa beans, rose hips, cherries, raspberries, blackberries, cranberries, strawberries, sea buckthorn.

3. A feed additive according to claim 2 wherein said oligomeric proanthocyanidins are extracted from grape seeds.

The feed additive according to claim 1, wherein the oligomer proanthocyanidin is prepared by using condensed tannin or an extract containing condensed tannin as a raw material in a concentration of 20 to 60% of hydrogen peroxide or potassium chlorate. The oxidizing agent oxidatively degrades the condensed tannin, and the oxidation reaction conditions are: pH is adjusted to 6 to 7, temperature is controlled at 30 to 50 ° C, and oxidative degradation time is 15 to 120 minutes, and yellow to brownish yellow translucent is obtained. The liquid is obtained by separation and purification.

A method for preparing a servo additive according to claim 1, wherein the method comprises using a condensed tannin or an extract containing condensed tannin as a raw material, and a hydrogen peroxide having a concentration of 20 to 60%. Or potassium chlorate as an oxidizing agent, at pH 6~7, at a temperature of 30~50 ° C for oxidation reaction 15~: L for 20 minutes, to obtain a yellow to brown yellow translucent liquid, by conventional physical or biochemical methods to remove excess oxidant, and then dried Oligomer proanthocyanidins.

The method for preparing a feed additive according to claim 5, wherein the condensed tannin is a polymer of flavanol, and the molecular skeleton is C ₆ · C ₃ · C ₆ , which is 2 to 5 yellow. A polymer of an alkanol monomer having a molecular weight distribution between 500 and 3000 MW.

The method for preparing an oligomeric proanthocyanidin according to claim 5, wherein the condensed tannin is a tannin-rich vegetable tannin.

The method of preparing a feed additive according to claim 5, wherein the method is carried out as follows:

(1) Extraction: Take the plant parts rich in tannins, add 4 to 6 times the weight of water, heat at a temperature of 80 to 100 ° C for 0. 5~1 hours, remove the residue by filtration, and use the same extraction conditions as above. Repeated 1~2 times; the filtrate was collected and concentrated to a relative density of 1. 205~1. 332 condensed tannin extract;

(2) Oxidative degradation: To the extract of the condensed tannin obtained in the step (1), adding 2 times the volume of 27.3% to 50% hydrogen peroxide, adjusting 11 to 6.7, degrading at 40 ° C for 30 minutes, Yellow to yellow brown translucent liquid Degraded product;

Use of the feed additive according to any one of claims 1 to 5 for the preparation of a feed for the prevention and treatment of coccidiosis in animals.

10. Use of a feed additive according to claim 9 for the preparation of a feed for the prevention and treatment of coccidiosis in animals, characterized in that the amount of the feed additive added to the full price is from 5 to 600 ppm.

The use of the word additive according to claim 9 for the preparation of a feed for the prevention and treatment of coccidiosis in an animal, characterized in that the feed additive is added in an amount of 5 to 200 ppm in the full price.

12. Use of a feed additive according to claim 6 for the preparation of a feed for the prevention and treatment of coccidiosis in an animal, characterized in that said application is said feed additive and anticoccidial other than said oligomeric proanthocyanidins Use together with drugs, or antioxidants, or enzyme preparations.

13. Use of a feed additive according to claim 6 for the preparation of a medicament for the prevention and treatment of coccidiosis in animals, characterized in that said feed additive is applied to the following animals:

1 chicken; 2 ducks; 3 鹌鹑; 4 pigeons; 5 rabbits; 6 pigs.