WO1995001104A1 - Feed additive of tea origin and animal feed containing the same - Google Patents

Feed additive of tea origin and animal feed containing the same Download PDF

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Publication number
WO1995001104A1
WO1995001104A1 PCT/JP1994/001037 JP9401037W WO9501104A1 WO 1995001104 A1 WO1995001104 A1 WO 1995001104A1 JP 9401037 W JP9401037 W JP 9401037W WO 9501104 A1 WO9501104 A1 WO 9501104A1
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WO
WIPO (PCT)
Prior art keywords
animal feed
tea
feed additive
animal
milk
Prior art date
Application number
PCT/JP1994/001037
Other languages
French (fr)
Japanese (ja)
Inventor
Noriyuki Ishihara
Souichi Mamiya
Shigemitsu Akachi
Noriko Tsuji
Senji Sakanaka
Izumi Ozeki
Ken Tsuda
Original Assignee
Taiyo Kagaku Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP18913993A external-priority patent/JP3361148B2/en
Priority claimed from JP5243865A external-priority patent/JPH0767544A/en
Priority claimed from JP5277490A external-priority patent/JPH07107919A/en
Application filed by Taiyo Kagaku Co., Ltd. filed Critical Taiyo Kagaku Co., Ltd.
Priority to AU69839/94A priority Critical patent/AU6983994A/en
Publication of WO1995001104A1 publication Critical patent/WO1995001104A1/en

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Classifications

    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/70Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75Feeding-stuffs specially adapted for particular animals for birds for poultry
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/30Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/116Heterocyclic compounds
    • A23K20/121Heterocyclic compounds containing oxygen or sulfur as hetero atom
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/10Feeding-stuffs specially adapted for particular animals for ruminants

Definitions

  • the present invention relates to a feed additive derived from tea and an animal feed comprising the same.
  • feed additives and animal feed derived from tea extract which are useful for improving milk quality and increasing milk yield of raw milk produced by livestock;
  • a tea leaf-derived feed additive that promotes the intestinal action of animals such as livestock and poultry, reduces the occurrence of diarrhea, increases the amount of milking, improves feed efficiency, and eliminates the odor of feces and urine from animals.
  • Animal feed
  • Livestock and poultry diarrhea is a serious problem in livestock management.
  • digestive diseases such as diarrhea that occur in cattle during the lactation and rearing periods are not only fatal due to this, but also have a major effect on physical exhaustion and subsequent growth of healthy internal organs. Is an important issue to give (Kazunori Hashimoto, Beef Cattle Journal, Vol. 5 , No. 11, 38 (1988).
  • the incidence of gastrointestinal illness mainly due to diarrhea during lactation, in beef cattle breeding is 13.3% (Akio Nakane, Livestock Research, Vol. 33, No. 1, 37 (19779)).
  • the mortality rate of dairy cattle during lactation and breeding has reached 11.9%, mainly due to diarrhea and pneumonia (Hiroyuki Tojo et al., Livestock Research, Vol. 4). 0, No. 1, 51, (1966)).
  • diarrhea occurs, the loss of milk yield, milk quality, and meat quality is observed due to reduced feed efficiency without death, resulting in a large loss of profit.
  • Diarrheal chicks generally have poor growth, lose weight, increase the range of weight fluctuation, adversely affect subsequent productivity, and decrease the commercialization rate in broilers and the egg production rate in laying hens. Is a factor. It is also known that the antibody viability after vaccination decreases.
  • the prevention or treatment of infectious diarrhea has been an effective means of administering antibiotics to livestock, poultry and pets, but the types and amounts of antibiotics that can be used safely are limited.
  • the emergence and resistance of antibiotic-resistant bacteria have led to questions about their effectiveness.
  • various natural antibacterial substances have come to be used, and it has been disclosed that the tea extract has an anti-infective microbial activity or a virus activity (Japanese Patent Application Laid-open No. Hei 12-265203). No., JP-A-2-277652).
  • the tea extract alone is used for infectious diarrhea or when a feed containing the same is used, the effect is weak, and practical use is questioned.
  • a specific antibody is used alone, there are actually various types of pathogenic microorganisms, and due to these specificities, a diarrhea-preventing effect cannot be expected, but only a therapeutic effect. Not practically effective.
  • Japanese Patent Publication No. 3-1 7469 discloses a composition for adapting livestock to reduce stress, but does not expect to suppress non-infectious diarrhea. The same applies to the method disclosed in Japanese Patent Publication No. 3-7045-8.
  • the method of parentheses is limited to the breeding time and environment. Under these circumstances, no effective feed additive or animal feed containing the same has been found at present, which suppresses the occurrence of non-infectious diarrhea in livestock, poultry and pets.
  • raw milk produced by livestock is ranked and traded in terms of milk fat percentage, non-fat solids percentage, cell number, and milk protein percentage.
  • A In the case of rank raw milk, it is required that the milk fat ratio is 3.5% or more, the non-fat solid content ratio is 8.5 or more, the number of cells is 300,000 or less, and the milk protein ratio is 3.0% or more. If one of these figures is not reached, normal transactions will not be made, and raw milk that does not achieve the figures will be penalized.
  • the percentage of milk fat, the percentage of non-fat solids, the number of cells, and the percentage of milk protein show certain values. For this reason, reduced milk quality has resulted in significant economic losses for dairy farmers. The same can be said for a decrease in milk yield, which leads to an economic loss due to a decrease in productivity.
  • reproductive rate for example, in the case of dairy cows, when the individual's milk production is increased, mismanagement of breeding management can quickly cause breeding problems for dairy cows. This can cause reproductive harm to large numbers of cattle in a short period of time.
  • reproductive disorders Livestock Research Vol. 44, No. 10, 1143-1149 (1990) Toshihiko Nakao.
  • livestock such as dairy cows, beef cattle, pigs, and chickens have been increasingly used in recent years and tend to increase the economic effect by providing concentrated feed as described above in order to improve productivity.
  • the burden on the gastrointestinal tract is greater than that of raw feed, causing problems such as reduced livestock growth, reduced milk production, and deterioration of meat quality.
  • feed additives and animal feeds that are effective against non-infectious and infectious diarrhea in livestock, poultry, and pet animals; 2) Feed additives and animal feeds that show the effects of improving milk quality and increasing milk yield of raw milk produced by livestock. (3) Feed additives and animals that show the effect of improving the reproduction rate of animals such as livestock and poultry. And (4) feed additives and animal feeds that promote the intestinal action of animals, reduce the occurrence of diarrhea, increase the amount of milking, improve feed efficiency, and eliminate the odor of feces and urine. Diligently studied.
  • tea leaves obtained by extracting and removing soluble components from tea leaves using water, alcohol, acetate, or a mixture thereof are added to the feed and used, the intestinal action of animals can be reduced. Acceleration, reduction of diarrhea episodes, increase of milking volume, improvement of feed efficiency, and elimination of odor in animal feces and urine.
  • the gist of the present invention lies in the following points.
  • an animal feed additive comprising an extract of tea
  • the animal feed additive according to the above (4) comprising one or more selected compounds.
  • each polyfunol compound in the polyfunol compound is (+)-catechin 0.2 to 6.5%, (+)-gallocatechin 2.0
  • An animal feed comprising the animal feed additive according to any one of (1) to (7) above, which is used for prevention or treatment of diarrhea;
  • the specific antibody is an infectious microorganism or virus, or
  • An animal feed comprising the animal feed additive according to any one of the above (1) to (7), which is used for increasing milk yield.
  • An animal feed comprising the animal feed additive according to any one of (1) to (7), which is used for improving milk quality.
  • An animal feed comprising the animal feed additive as described in any one of (1) to (7) above, which is used for improving the reproduction rate of animals.
  • An animal feed additive comprising, as an active ingredient, a tea leaf fiber obtained by removing water, alcohol, acetone, or a soluble component extracted from a mixture thereof from tea leaves,
  • the present invention relates to a feed additive derived from tea and an animal feed comprising the same. This includes a method using a tea extract and a method using a tea extraction residue.
  • the tea extract used in the present invention refers to a tea extract containing the following polyphenol compound, which is extracted from tea leaves using a solvent such as water, alcohol, or ethyl acetate.
  • the polyphenol compound in the present invention is (+)-force techin, (+)-gallocatechin, (1) -gallocatechin gallate, (1) -g Picatechin, (1) epigallocatechin, (1) epigallocatechin, (1) epigallocatechin gallate, free theaflavin
  • Tannins such as theaflavin monogallate A, theaflavin monogallet B, and theaflavin digallate.
  • the above-mentioned polyphenol compound is preferably a component of a hot water extract of tea such as green tea, black tea, and oolong tea.Ethyl acetate-soluble fraction of tea water or alcohol extract or an ultrafiltration membrane and reverse osmosis It can also be obtained by membrane treatment.
  • the method for extracting such a polyphenol compound from tea is not particularly limited, and for example, the method described below can be used.
  • the tea may be used in a crushed state or in a non-ground state.
  • the solvent to be used is not particularly limited, and water, ethanol, acetone, and the like can be used. These may be used alone, or may be used in a mixture at an arbitrary ratio.
  • the solvent may be the previously warmed (3 0 ⁇ 9 5 ° C) , may be heated in a stirred (3 0 ⁇ 9 5 e C) .
  • the stirring time may be in the range of 30 minutes to 7 hours. If the length is shorter than this range, the extraction efficiency is poor. If the length is longer than this range, the corresponding effect cannot be obtained.
  • the separated solution may be dried as it is, or may be dried after being concentrated.
  • a drying method a commonly used method such as spray drying, freeze drying and hot air drying may be used. Concentration can be performed using an ultrafiltration membrane or a reverse osmosis membrane. These may be used alone or in combination. Further, by redistributing the obtained extract, the purity of the polyphenol compound can be increased. At this time, in the system of water ethyl acetate Partitioning is preferred, and the polyphenols are concentrated in the ethyl acetate fraction.
  • tea leaves are extracted with water, alcohol and / or ethyl acetate for 30 to 95, 0.5 to 7 hours, and the obtained extract is spray-dried
  • Tea leaves are extracted with water, alcohol and / or ethyl acetate for 30 to 95 hours and 0.5 to 7 hours, and the resulting extract is subjected to ultrafiltration with a cut-off molecular weight of 300 to 600. Concentrate with a membrane and then with a reverse osmosis membrane.
  • Examples of the method include removing the obtained ethyl acetate layer.
  • the tea extract thus obtained usually contains two or more kinds of polyphenol compounds.
  • these polyphenol compounds may be isolated and used alone. Alternatively, they may be used as a mixture of two or more. Further, the extract thus obtained may be used as it is as a crude extract containing a polyphenol compound.
  • the compound is not particularly limited, and any of the polyphenol compounds in the present invention may be used. It is preferable to use gallocatechin gallate.
  • the compound is not limited to a tea-derived compound. That is, the compound may be derived from other organisms, for example, persimmon shibu or a chemically synthesized product.
  • the method for isolating the compound is not particularly limited, and a general method may be used.
  • it can be isolated by dissolving a tea extract in ethyl acetate or the above-mentioned ethyl acetate fraction by silica gel chromatography and eluting with a suitable solvent. Then, even higher purity compounds can be obtained by using HPLC.
  • the animal feed additive of the present invention refers to one containing a polyphenol compound derived from the tea extract thus obtained, and the content of the polyphenol compound contained in the animal feed additive of the present invention. Is usually 5 to 80%, preferably 10 to 50%, and more preferably 20 to 35%.
  • each polyphenol compound in the animal feed additive is as follows: (+)-0.2-6.5% of force techin, (+)-2.0-18.0% of garoka techin, (1) -Gallocatechin gallate 1.0 to 15.0%, (1) -Epicatechin 0.5 to 10.0%, (1) -Epicatechin gallate 0.3 to 8.0%, (1) -epigallocatechin 2.0 to 18.0%, (1) -epigallocatechin gallate 3.0 to 21.0%, free theaflavin 0 to 20.0%, theaflavin monogallate AO 5.5.0%, theaflavin monogallate B 0 5.5.0% and theaflavin digallate 0 ⁇ 5.0%.
  • This polyfunol compound is contained in a large amount as a component of tea, and is effective in preventing caries, hyperlipidemia, cancer, and the like. Its safety is very high because it is already used in food.
  • the animal feed additive of the present invention is administered to livestock, poultry, and companion animals as a feed containing the polyphenol compound by being added to the animal feed.
  • Feed is provided to livestock for the purpose of supplying nutrients, and is generally classified into roughage, concentrated feed, and specialty feed.
  • the feed to which the feed additive of the present invention is to be added is not particularly limited, but generally high-protein, high-energy concentrated feed is commonly used in livestock. When given in an amount, diarrhea is particularly likely to occur. Therefore, it is desirable to add the feed additive of the present invention to such a concentrated feed.
  • the feed additive of the present invention contains a polyphenol compound, and it is also known that these compounds have an antioxidant effect, and from the viewpoint of the antioxidant effect of fat components. Should also be added to concentrated feed.
  • the feed additive of the present invention When the feed additive of the present invention is added to the feed, it may be added together with the ingredients of the feed at the time of mixing the feed in advance, or may be added together at the time of feeding the feed to livestock without any problem.
  • the method and timing of addition are not particularly limited. There is no particular problem when used in combination with other feed additives such as probiotic agents and antibiotics.
  • the amount of the animal feed additive added to the animal feed of the present invention is not particularly limited, and is appropriately adjusted.
  • livestock and poultry are, for example, red sea lions, bushes, poma, goats, hidge, foxes, mink, nits, turkeys, ducks, zebras, guinea fowls, ducklings, and wild birds.
  • Geese, pigeons and other animals bred for industrial purposes, and beloved animals are animals bred for personal tastes such as dogs and cats.
  • the animal feed additive of the present invention and the animal feed containing the same can be used for non-infection caused by sudden changes in the environment of livestock, poultry, companion animals, etc., changes in feed composition, and inaccuracies in breeding management. It is useful for preventing and treating sexually transmitted diarrhea or infectious diarrhea induced by pathogens such as viruses or bacteria.
  • non-infectious diarrhea refers to stress-induced diarrhea caused by stress given to livestock, poultry, and companion animals, or unexplained diarrhea not involving infectious microorganisms.
  • Infectious diarrhea refers to diarrhea caused by pathogenic bacteria or viruses. You.
  • the dose for livestock, poultry, and pets for obtaining the effect of suppressing non-infectious diarrhea is usually 0.5 to 50 mg / kg of body weight per day in terms of polyphenol compound. Preferably it is between 2 and 20 m body weight kg.
  • the dose is lower than 0.5 mgZ kg body weight, the effect of suppressing non-infectious diarrhea cannot be expected, and when the dose exceeds 50 mgZ kg body weight, the antibacterial action of the polyphenol compound is not expected. Therefore, it also has an effect on the gut microbes that exist in livestock, poultry and pets.
  • iso-epigallocatechin gallate is used after isolation or the like, it is usually 0.03 to 5 mg / kg of body weight per day, more preferably 0.1 to 3 mg / kg of body weight. .
  • the dose when used in combination with a specific antibody is usually 0.3 to 25 per day as a polyphenol compound. mg / kg body weight, more preferably 2 to 15 mg gZ body weight kg. If the dose is less than 0.3 m body weight kg, the effect of suppressing infectious diarrhea cannot be expected.
  • the polyphenol compound can be represented by the amount of tannin determined by the official tannin analysis method (Chakenho V 01.71, 43-74 (1990)).
  • the specific antibody in the present invention refers to an antibody that specifically binds to infectious microorganisms or viruses of livestock, poultry and pets, or toxins produced by these.
  • the type of antibody include egg antibodies obtained from eggs of laying hens hyperimmunized with infectious microorganisms or viruses of livestock, poultry and companion animals, or toxins produced by them, and milk antibodies obtained from milk of mammals Say etc.
  • the antibody purity is not limited. That is, the antibody may be pure or chicken
  • the egg antibody is not particularly limited, but may be whole egg, egg yolk, whole egg solution, egg yolk solution, whole egg powder, egg yolk powder or a powder of a water-soluble protein fraction of egg yolk containing the egg antibody.
  • the case of milk antibody is not particularly limited, but may be whole fat milk powder, skim milk powder or whey protein powder containing it.
  • Infectious microorganisms in livestock, poultry, and pets are not particularly limited, but include, for example, Clostridium perfringens A, Clostridium perfringens C, Salmonella typhimurium, and Salmonella. Evening Blind, Campylopactor Jeju 21, Campylopactor Collie, E. coli 0-88, E. coli 0-99, E. coli 9887P, Staphylococcus oleus, Rotavirus KK-3, Oral virus NCDV Pathogenic bacteria and diarrhea virus.
  • the animal that is hyperimmunized with the microorganism or the toxin produced by the microorganism may be any animal that can produce a specific antibody against the microorganism or the toxin.
  • Livestock, poultry, and pets which are the objects of the present invention, may be used. From the practical point of view of antibodies against infectious microorganisms in animals and the application of this antibody composition, laying hens or mammals such as cows, goats and sheep, which can produce large amounts of specific antibodies, are particularly desirable. Among them, a method of immunizing a laying hen with the microorganism or a toxin produced by the microorganism and obtaining an antibody from the hen egg from the viewpoints of hyperimmunity workability, antibody production ability, and animal breeding cost. Most preferred.
  • One method of hyperimmunizing laying hens is to raise specific antibodies in hen eggs by repeatedly immunizing laying hens with the above-mentioned microorganisms or viruses from livestock, poultry and pets, or toxins produced by them. It is good to add.
  • Methods for hyperimmunizing mammals include cattle and cattle. It is advisable to increase the amount of specific antibodies in milk by repeatedly immunizing animals such as giants and sheep with the antigen.
  • the antigen used in this case may be prepared by a known method. For example, after infectious microorganisms of livestock, poultry, and companion animals are cultured in large amounts, attenuated or inactivated by a known method, and used as an antigen.
  • any method such as intramuscular injection, subcutaneous injection, intravenous injection, intraperitoneal injection or oral immunization by drinking water may be used.
  • the immunization of the antigen is repeated until the antibody titer reaches the maximum value, while examining the specific antibody titer appearing in chicken eggs or milk by a method such as enzyme immunoassay.
  • the antibody titer can be maintained at a certain level or more throughout the laying period or the period of colostrum secretion by repeatedly immunizing a laying hen or mammal with an antigen at appropriate intervals. .
  • the amount of antigen to be immunized depends on the type of animal to be immunized, the type of antigen, etc., so it is necessary to select it in a timely preliminary test, etc.
  • As the virus amount 10 g to 1 mg Z feathers
  • the antigen amount of Z times is selected.
  • a specific antibody is prepared by immunizing a laying hen or mammal with an infectious disease microorganism of livestock, poultry, and companion animals or a toxin produced thereby as an antigen, and then hen eggs or chicken eggs containing a specific antibody against the antigen. This can be done by collecting milk.
  • milk When milk is used, it contains a specific antibody to the antigen used by sterilizing the milk or skim milk obtained by separating lipid components in the milk with a cream separator and then drying with hot air or freeze-drying. A whole milk powder or skim milk powder is obtained. From the milk or the skim milk, whey protein powder having a higher specific antibody purity to the used antigen or a pure product of the specific antibody can be prepared by a known method.
  • the amount of the specific antibody to be used in combination with the polyfuninol compound in the present invention has an antibody titer of at least 1.5 times that of the blank in an enzyme immunoassay. It is preferable to adjust the dose to 1 mg or more, and more preferably 5 mgZ kg or more. If the dose is less than 11113 / body weight 1 ⁇ g, the expected effect cannot be obtained, which is not preferable.
  • the present invention can be applied to all livestock, poultry and pets, it can be easily consumed by herbivores irrespective of the bitterness of the polyphenol compound from the viewpoint of effect or palatability.
  • herbivorous animals are preferable, and cows such as dairy cows and beef cattle, goats, sheep, deer and the like, which are particularly useful in industry, are preferable.
  • animal feed additive of the present invention and the animal feed containing the same are useful for improving the milk quality of raw milk produced by livestock and increasing the amount of milk.
  • Dosage for livestock is usually 0.5 to 50 mg / kg body weight per day, more preferably 2 to 20 mg / kg body weight, in terms of polyphenol compound.
  • the dose is lower than 0. S mgZ body weight kg, the effect of the present invention cannot be expected.
  • the dose exceeds 5 O mgZ body weight kg, it is resident in livestock and poultry due to the antibacterial action of the polyphenol compound. Adversely affect the digestive tract microbes.
  • the polyphenol compound can be represented by the amount of tannin determined by the official tannin analysis method described above.
  • iso-epigallocatechin gallate is used by isolation or the like, it is usually 0.03 to 5 mg / kg of body weight per day, more preferably 0.1 to 3 mg / kg of body weight.
  • the livestock to which the polyphenol compound is administered in the present invention may be any animal used for raw milk production, such as dairy cows, goats, horses, and sheep, and the type thereof is not particularly limited. Furthermore, it is particularly preferable to use dairy cows in consideration of palatability and the like due to the bitterness of the polyphenol compound. .
  • animal feed additive of the present invention and the animal feed prepared by blending the same are effective in improving the reproduction rate of livestock and poultry.
  • the dose for obtaining the effect of improving the reproductive rate is usually 0.5 to 50 mg / kg body weight per day, more preferably 2 to 2 mg / kg body weight in terms of polyphenol compound. . If the dose is lower than this range, the effects of the present invention cannot be expected. If the dose exceeds this range, the antimicrobial action of the polyphenol compound will adversely affect microorganisms resident in the gastrointestinal tract.
  • For the quantification of the polyphenol compound use the official Yunnin analysis method described above. (1) When iso-epigallocatechin galleate is used after isolation, it is usually 0.03 to 5 mgZ body weight per day. g, more preferably 0.1 to 3 mg / kg of body weight.
  • the type of livestock and poultry to which the polyphenol compound is administered is not particularly limited, but is preferably used for dairy cows from an industrial viewpoint.
  • the reproductive rate can be evaluated by measuring the period required for postpartum conception and the number of artificial inseminations of dairy cows that have been confirmed to be pregnant.
  • the present invention provides a feed additive and an animal feed containing tea leaf fiber as an active ingredient.
  • animals such as livestock and poultry are fed animal feeds to which feed additives containing tea leaf fiber are added
  • the intestinal microflora of animals is improved, thereby promoting intestinal regulation and causing diarrhea. It reduces milk production, increases milk yield, improves feeding efficiency, and reduces and eliminates stool and urine odors.
  • This increases the number of useful microorganisms such as bifidobacteria and lactic acid bacteria among the indigenous microorganisms in the gastrointestinal tract due to the combined effect of cellulose, which is tea leaf fiber, and the borofuynol compound remaining in tea leaf fiber, which is extraction residue.
  • the number of harmful bacteria such as pathogenic Escherichia coli and Welsh bacteria is reduced, and the above effects are achieved.
  • the polyphenol compound remaining in the tea leaf fiber usually contains about 2 to 5%.
  • the tea leaf fiber used in the present invention can be obtained as an extraction residue obtained by removing a soluble component obtained by extracting ground tea leaves with 5 to 20 times the amount of water, alcohol, acetate or a mixture thereof.
  • the alcohol is not particularly limited, but methanol, ethanol, propanol, isopropanol, butanol, and the like are preferable.
  • the tea leaves used as raw materials can be from any stage of the normal manufacturing process, from fresh leaves to finished tea (dried tea), and can be used as unfermented tea, semi-fermented tea, or fermented tea regardless of the degree of fermentation. Heat during extraction There is no particular problem.
  • the extraction residue is 20 to 40 kg and the extract of Brix 2 to 10 'depending on the amount of water is 40 to 1 60 kg is obtained.
  • the obtained extraction residue can be dried by a commonly used method, for example, hot-air drying, freeze-drying, or the like, and then pulverized and mixed with feed.
  • 6 to 8 kg of tea leaf fiber is obtained from 1 O kg of tea leaf.
  • the resulting tea leaf fiber has about 70% or more of the power-fein and tea polyphenol compounds removed from the raw tea leaves, so that bitterness and astringency are reduced and taste is improved. It becomes more suitable as feed for use.
  • the production of tea leaf fiber in the present invention can be combined with the above-mentioned method for producing a polyfunor compound, and thereby, the productivity of each production method can be enhanced. There are two.
  • the amount of the tea leaf fiber (including the polyfuninol compound remaining in the tea leaf fiber) to be added to livestock feed is 0.01% or more by weight.
  • the method of adding the tea leaf fiber to the feed can be appropriately added during each production process, and any method may be used as long as it is an orally administered method.
  • the component composition of the obtained polyphenol compound is (+)-1.2% of kinetechin, (+)-5.0% of gallocatechin, (1) -gallocatechin gallate 3.9%, (1) -eppicatechin 2.3%, (1) -epigallocatechin gallate 1.5%, (1) -epigallocatechin gallate 5.0% and (1) -epigallocatechin gallate 6.1%.
  • the content of the polyphenol compound in the extract was determined by the aforementioned official Yunnin analysis method, and the composition of each component was determined by HPLC analysis.
  • High performance liquid chroma DOO graph device uses 6 5 5 A- 1 1 LC manufactured by Hitachi, column: J 'sphere ODS- M 8 0 (( Ltd.) Waiemushi Ltd. I), solvent: 2 0 mM KH 2 P 0 4 - H 3 P 0 4 (p H 2. 4) drink ethanol (7 5: 2 5, vZv :), flow rate: 0. 8 ml / min, detection: 2 8 0 nm of UV, of various HPLC analysis was performed under the conditions.
  • the content of the polyphenol compound and the composition of each component were determined by the methods described above.
  • the component composition of the obtained polyphenol compound is as follows: (+) — 1.4% strength, (+) — 5.8% gallocatechin, (1) -gallocatechin gallate 4 .5 (1) Epicatechin gallate 2.7%, (I) Epicatechin gallate 1.8%, (I) Epigallocatechin 5.8% and (I) Epigallocatechin gallate 7.0% Met.
  • Example 2 To 350 g of the hot water extract obtained in Example 1, 8 liters of water was added and dissolved, and the mixture was sequentially partitioned with hexane and chloroform. Add 10 liters of ethyl acetate to the aqueous layer after partitioning, vigorously stir, and allow to stand. Separate the ethyl acetate layer, evaporate the ethyl acetate, dry, and dry 70 g of the ethyl acetate-soluble fraction. Obtained (purity 74.5% as a mixture of polyphenol compounds) o
  • the content of all polyphenol compounds in the ethyl acetate-soluble fraction was 74.5%, and the ratio of each polyphenol compound was (+)-3.5%, (+)-gallocatechin 14. 8%, (1) -gallocatechin gallate 11.6%, (1) -epicatechin 7%, (1) 1-epicatechin gallate 4.6%, (1) -epigallocatechin 15.0% and (1) 1) Yepigallocatechin gallate 18.0%.
  • Salmonella tabulin, Escherichia coli 0-88, Escherichia coli 0-99, and Escherichia coli 988P which are known as causative bacteria of infectious diarrhea in cattle, were cultured in brain heart infusion medium.
  • MA10 cells derived from rhesus monkeys cultured on bovine rotavirus KK-13 and bovine rotavirus NCDV, respectively, in an Eagle MEM medium were cultured mainly.
  • Laying hens were hyperimmunized with the respective bacteria or viruses as the antigen. From 10 kg of hen egg yolk produced by the laying hen, 45 g of a specific hen egg antibody against the bacterium was obtained.
  • Example 5 10 g of the ethyl acetate-soluble fraction obtained in Example 3 was applied to a silica gel column (5 ⁇ 80 cm), and eluted with a mixed solution of chloroform and methanol in a mouth. Elution was performed in two steps. That is, elution was first carried out with the mixture at a mixture ratio of 20: 1 (vZv), and then with the mixture at a mixture ratio of 10: 1 (v / v). Each of the obtained polyphenol compounds is further separated and purified by recycling HPLC (LC-908, GS-320 column, manufactured by Nippon Bunka Kogyo Co., Ltd., using methanol as the solvent) to obtain higher purity polyphenol compounds.
  • HPLC LC-908, GS-320 column, manufactured by Nippon Bunka Kogyo Co., Ltd., using methanol as the solvent
  • the obtained polyphenol compound contained (+)-0.3 g of force techin, 1.2 g of (+)-gallocatechin, 0.9 g of (-1) -gallocatechin gallate, 0.9 g of (-1) -epeicatechin 0 5 g, (1) -epigallocatechin gallate 0.38 g, (1) -epigallocatechin gallate 1.2 g and (1) -epigallocatechin gallate 1.5 g.
  • the 30 calves were divided into 3 groups of 10 calves, and the group that received only the artificial milk was Group A, the group that received 0.7 g of the tea extract prepared in Example 1 per day was Group B, The group to which 1.5 g of the tea extract prepared in Example 1 was administered per day was bred as Group C for 5 weeks.
  • the tea extract was administered after being mixed with the artificial milk. Water was provided ad libitum.
  • the artificial milk used was a calf top made by All Dairy. In the following test examples, calf tops were used as the artificial milk.
  • Fecal scores and intestinal flora were measured from 0 to 5 weeks from the start of the test as indices indicating non-infectious diarrhea.
  • the stool score is a quantification of the stool properties, with normal stool being 0, loose stool being 1, muddy stool being 2, and watery stool being 3. The results are shown in Tables 1 and 2, respectively. table 1
  • the numerical values indicate the average of 10 animals, with 0 for normal stool, 1 for loose stool, 2 for muddy stool, and 3 for watery stool.
  • the numerical values are the average of 5 animals, with 0 for normal stool, 1 for loose stool, 2 for muddy stool, and 3 for watery stool. Table 5
  • the figures indicate the average of 10 animals, 0 for normal stools and 1 for loose stools.
  • Mud-like stool was set to 2 and watery stool was set to 3.
  • the values are the average of 10 animals and are expressed as logarithmic values per 1 g of stool.
  • Salmonella 15 adult cattle infected with evening primrose were divided into 3 groups of 5 cows, the group fed the basic diet shown in Table 3 was group J, and the tea extract prepared in Example 1 was used per day.
  • Group K received 7.5 g and group L received 7.5 g / day of the tea extract prepared in Example 1 and 6 g / day of the specific chicken egg antibody prepared in Example 4.
  • the group to which 6 g of the specific chicken egg antibody prepared in Example 4 was administered per day was bred for 5 weeks as the K2 group.
  • the tea extract or specific chicken egg antibody was administered as a mixture with the basic feed shown in Table 3. Water was provided ad libitum.
  • the figures indicate the average of 5 animals, 0 for normal stool and 1 for loose stool
  • Tables 8 and 9 show that adult cats in group L had lower fecal scores and fewer bacteria of Salmonella evening primin in feces than adult cows in groups J, K and K2. Was. Therefore, it was found that the tea extract of the present invention effectively suppressed infectious diarrhea. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, similar tests were performed using tea extracts derived from black tea and oolong tea, and similar results were obtained. Similar tests were performed using Escherichia coli 088, Escherichia coli 0-99, Escherichia coli 987P, Ushiguchi Yu virus KK-13 and Bovine rotavirus NCDV, and similar results were obtained.
  • Healthy 30 dairy cows were divided into 3 groups of 10 cows each, and the group that received 15 g of the tea extract prepared in Example 1 per day was Group A, and the tea extract prepared in Example 1 was The group administered 7.5 g per day was bred for 5 weeks as group B, and the group without administration as group C.
  • the tea extract was administered by mixing with a normal feed. Milk fat ratio, non-fat solids ratio, cell number, milk protein ratio for milked milk are used as indicators for improving milk quality and increasing milk yield.
  • the amount of milk per day was measured from 0 to 5 weeks after the start of the test according to a conventional method. The results are shown in Tables 10 to 14, respectively.
  • three and five weeks after the start of the test 15 panelists conducted a sensory test of milk. The results are shown in Table 15.
  • Table 10 Table 10
  • the numerical value indicates the average of 10 animals, and the unit is 1 ⁇ 10 Zm 1.
  • the figures are the average of 15 people, 5 points for very good, 4 points for good, 3 points for normal, 2 points for bad, and 1 point for very bad.
  • milk expressed from dairy cows in Group A was higher than milk expressed from dairy cows in Groups B and C, in terms of milk fat ratio, nonfat solids ratio, milk protein ratio and Increased milk yield per day and decreased cell number were observed.
  • the milk fat ratio was 3.5%
  • the non-fat solids ratio was 8.5%
  • the milk protein ratio exceeded 3.0% within 2 weeks after administration, and the conditions for qualification as Rank A were given. Was found to meet. Therefore, it was found that the tea extract of the present invention was effective for improving milk quality and increasing milk yield.
  • Tables 16 and 17 show that dairy cows in group A have a shorter time to conception after parturition than cows in group B, and that the number of artificial fertilizations of dairy cows that have been confirmed to be pregnant decreases, and that the reproductive rate is lower. Improvement was observed. Therefore, it was found that the tea extract of the present invention had an effect of improving the reproductive rate of dairy cows. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, similar tests were performed using tea extracts derived from black tea and Wu Kuang tea, and similar results were obtained.
  • the 30 calves were divided into three groups of 10 calves each.
  • Group A prepared in Example 5 (+)-90 mg / day of dytechin was administered to group B, prepared in Example 5 (1) 90 mg perepigallocatechin gallate per day
  • the administered group was reared as Group C for 5 weeks.
  • the above two polyphenol compounds were administered by mixing with artificial milk. Water was provided ad libitum.
  • the fecal score and intestinal microflora were measured from 0 to 5 weeks from the start of the test as indices indicating non-infectious diarrhea. The results are shown in Tables 18 and 19, respectively.
  • the figures indicate the average of 10 animals, 0 for normal stools and 1 for loose stools.
  • Mud-like stool was set to 2 and watery stool was set to 3.
  • the figures indicate the average of 5 animals, 0 for normal stool and 1 for loose stool
  • the numerical values represent the average of 10 animals and were expressed as logarithmic values per 1 g of feces. From Tables 22 and 23, it was confirmed that the fecal score and the number of Salmonella tabulin in feces were reduced in Group I compared to calves in Group G and H. Therefore, it was found that (1) epigallocatechin gallate of the present invention effectively suppressed infectious diarrhea. Similar infectious disease diarrhea tests were performed with E. coli 0-88, E. coli 0-99, E. coli 987P, bovine rotavirus KK-13 and bovine rotavirus NCDV. Was done.
  • Salmonella 15 adult cattle infected with evening brine were divided into 3 groups of 5 cows each, and the group fed the basic diet shown in Table 3 was prepared in Group J and prepared in Example 5.
  • (1) -epigallocatechin The group receiving 50 mg of gallate per day was group K, prepared in Example 5.
  • (1) 500 mg of epigallocatechin gallate per day and the specific chicken antibody prepared in Example 4 was administered for 5 weeks as L group.
  • (1) Epigallocatechin gallate or specific chicken egg antibody was administered by mixing with the basic feed shown in Table 3. Water was provided ad libitum.
  • the fecal score and the number of Salmonella evening brilli in feces were determined from 0 to 5 weeks from the start of the test as indicators of diarrhea. The results are shown in Table 24 and Table 25, respectively. Indicated
  • the figures indicate the average of 5 animals, 0 for normal stool and 1 for loose stool
  • Mud-like stool was set to 2 and watery stool was set to 3.
  • the figures represent the average of 5 animals and are expressed in logarithms per g of feces.
  • Healthy 30 dairy cows were divided into 3 groups of 10 cows each and prepared in Example 5.
  • the (+)-forced techin prepared in 5 was administered 9 0 0 111 per day as a 1 ⁇ group, and a group fed with the above-mentioned two types of polyphenol-free normal feed as a 0 group, and bred for 5 weeks. did.
  • the above-mentioned polyfunool compound was administered by mixing with a normal feed. Milk fat ratio, nonfat solids ratio, cell count, milk protein ratio, and milk yield per day for milked milk were used as indicators of milk quality improvement and milk yield. Measured until week. The results are shown in Tables 26 to 30 respectively. In addition, three and five weeks after the start of the test, sensory tests of milk were performed by 15 panelists. The results are shown in Table 31. Table 26
  • Numbers shows the average of 1 0 head, the unit is shown in 1 0 4 1.
  • the figures are the average of 15 people, with 5 points for very good, 3 points for good, 2 points for bad, and 1 point for very bad.
  • milk expressed from cows in Group M was higher than milk expressed from cows in Groups N and 0 compared with milk expressed in milk, fat-free solids, milk protein and milk per day. An increase in the amount and a decrease in the number of cells were observed. Specifically, in the M group, the milk fat ratio exceeded 3.5%, the nonfat solid content ratio was 8.5%, and the milk protein ratio exceeded 3.0% within 2 weeks after administration, satisfying the criteria for A rank It has been found. From the above results, it was found that (1) -epigallocatechin gallate of the present invention was effective for improving milk quality and increasing milk yield of dairy cows. In addition, from a functional test conducted with 15 panelists, milking from cows in the M group was evident from Table 31. Milk was found to have good aroma, tongue taste, and mouthfeel.
  • Sencha and oolong tea mixture (3: 1) 200 g, crushed to 2-3 mm in diameter, 50.
  • Four liters of a mixture of water, ethanol, and acetate (5: 3: 2 (v / v)) heated in C was added and extracted for 2 hours.
  • the mixture was subjected to solid-liquid separation, and 320 g of the obtained wet residue was dried with hot air at 80, followed by pulverization to obtain 150 g of tea leaf fiber.
  • the content of caffeine and polyphenol in this tea leaf fiber was as low as 0.7 g and 3.5 g, respectively, in 100 g, with little bitterness and bitterness, and which could be sufficiently added to feed. 8
  • poultry feed was prepared by the usual method
  • the feed contains 0.03 as a polyphenol compound.
  • a feed for dairy cows was prepared by a conventional method according to the above mixing ratio.
  • the value before the start of the control was expressed as 100.
  • the group administered with the additive of the present invention was effective in suppressing the decrease in milking amount during the test period, and an increase in milking amount was observed.
  • Example 8 Twenty pigs (adults) bred in a pig house were fed with the solid feed for swine raising of the present invention of Example 8 and the same feed without the additive of the present invention as a control for 6 weeks, respectively.
  • the ammonia concentrations were compared. Ammonia concentration was measured using a gas detector tube at a height of about 1 m above the floor at four locations in each pig house, and the average value was shown.
  • Example 8 The effect on piglet rearing performance of 30 piglets each raised for 20 days using the pig feed of the present invention shown in Example 8 and the same feed containing no additives of the present invention as a control.
  • Food and water were available ad libitum.
  • the body weight at the start, the body weight at the end, and the feed intake were measured, and the breeding performance was evaluated by calculating the daily weight gain and the feed demand rate from the average values.
  • Feed demand Feed intake Period gained weight
  • Table 36 piglets reared on the diet containing the additive of the present invention showed an increase in daily weight gain and a lower feed demand rate than piglets in the control group. It has been reduced and feed efficiency has been improved.
  • Example 10 Using the dairy cattle feed shown in Example 10, five dairy cows were preliminarily reared with the basic feed shown in Table 3 for 10 days, and then fed the additive-containing feed of the present invention and reared for 20 days. They were kept on the basic diet for another 10 days. Food and water were available ad libitum. Before the start of the test, on the 10th and 20th days of the test, and at 10 days after the end of the test, the rate of increase in the number of bacteria in the feces before the start of the test for Bifidobacterium, before the start of the test for Clostridium genus The percentage of decrease in the number of bacteria, ⁇ , was measured, and the intestinal control effect was evaluated. Table 3 7
  • Bifidobacterium and Clostridium bacteria were expressed as 100 before the start.
  • the product of the present invention has a great effect on non-infectious diarrhea and infectious diarrhea of livestock, poultry and pets, as well as improving milk quality, increasing milk yield and reproductive rate of livestock. Contribute to
  • the tea fiber of the present invention is derived from tea that is consumed daily and has extremely high safety.When administered to livestock, it promotes the intestinal action of the livestock, reduces the occurrence of diarrhea, increases the amount of milking, It is extremely convenient for the livestock industry because it improves feed efficiency and has the action of eliminating odors from feces and urine. Further, the production of feed additives according to the present invention is carried out in combination with a method for producing tea bolifenol, which has recently been established as an industrially important material. It is one of the features of the present invention that the productivity of each manufacturing method can be enhanced.

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Abstract

An animal feed additive comprising tea extracts, an animal feed additive containing tea leaf fibers as the active ingredient, and an animal feed containing these additives. The tea extracts of the invention are remarkably efficacious in preventing and treating both of noninfectious and infectious diarrheas of domestic animal, poultry and pet, improving raw milk quality, increasing milk quantity, and enhancing breeding efficiency. The tea leaf fibers of the invention have the effects of promoting the control of intestinal functions of domestic animals, relieving diarrhea, increasing milk yield, improving feed efficiency, and deodorizing excreta.

Description

明 細 書  Specification
茶由来の飼料添加物及びそれを含んでなる動物用飼料  Tea-derived feed additive and animal feed comprising the same
技術分野 Technical field
本発明は茶由来の飼料添加物及びそれを含んでなる動物用飼料に 関する。 さらに詳しく は、  The present invention relates to a feed additive derived from tea and an animal feed comprising the same. For more information,
1 ) 家畜、 家禽及び愛玩動物等の環境の急変及び飼料組成の変化、 飼養管理の失宜などの影響により生じる非感染性下痢症、 あるいは ウィルス若しく は細菌等の病原菌により誘発される感染性下痢症を 予防、 治療するのに有用な茶抽出物由来の飼料添加物及び動物用飼 料、  1) Non-infectious diarrhea caused by sudden changes in the environment of livestock, poultry, companion animals, etc., changes in feed composition, inadequate feeding management, or infectious diarrhea induced by pathogens such as viruses or bacteria. Feed additives and animal feeds derived from tea extracts useful for preventing and treating
2 ) 家畜の生産する生乳の乳質を改善し、 乳量を増加させるのに有 用な茶抽出物由来の飼料添加物及び動物用飼料、  2) feed additives and animal feed derived from tea extract, which are useful for improving milk quality and increasing milk yield of raw milk produced by livestock;
3 ) 家畜、 家禽等の動物の繁殖率の向上に有用な茶抽出物由来の飼 料添加物及び動物用飼料、 並びに  3) Feed additives and animal feed derived from tea extracts useful for improving the reproduction rate of animals such as livestock and poultry, and
4 ) 家畜、 家禽等の動物の整腸作用の促進、 下痢発症の低減、 搾乳 量の増加、 飼料効率の改善、 及び動物の糞便や尿の悪臭消去作用を 有する、 茶葉由来の飼料添加物及び動物用飼料、  4) A tea leaf-derived feed additive that promotes the intestinal action of animals such as livestock and poultry, reduces the occurrence of diarrhea, increases the amount of milking, improves feed efficiency, and eliminates the odor of feces and urine from animals. Animal feed,
に関する。 背景技術 About. Background art
畜産経営において、 家畜、 家禽の下痢の発生は深刻な問題になつ ている。 例えば、 牛に関して述べると哺育、 育成期に牛に発生する 下痢等の消化器疾病は、 これが原因の死亡は言うに及ばず、 体力の 消耗やこの後の健全な内臓器官の成長に大きな影響を与えるために 重要な問題となっている (橋本和典、 肉牛ジャーナル、 V o l . 5 、 N o . 1 1、 3 8 ( 1 9 8 8 ) ) 。 実際に肉用牛の育成における 哺育期の下痢を主症状とする消化器官の疾病の発生率は 1 3. 3 % である (中根叔夫、 畜産の研究、 V o l . 3 3、 N o . 1、 3 7 ( 1 9 7 9 ) ) 。 また、 哺育、 育成期での乳用牛の死廃率は 1 1 . 9 %に達しているが、 この主な原因は下痢と肺炎である (東條博之ら 、 畜産の研究、 V o l . 4 0、 N o . 1、 5 1、 ( 1 9 8 6 ) ) 。 さらに、 成牛に関しても、 下痢が発生すると、 死亡にいたらなく と も飼料効率が低下することにより乳量、 乳質、 肉質の低下が認めら れ、 利益の損失は多大なものとなる。 Livestock and poultry diarrhea is a serious problem in livestock management. For example, in the case of cattle, digestive diseases such as diarrhea that occur in cattle during the lactation and rearing periods are not only fatal due to this, but also have a major effect on physical exhaustion and subsequent growth of healthy internal organs. Is an important issue to give (Kazunori Hashimoto, Beef Cattle Journal, Vol. 5 , No. 11, 38 (1988). In fact, the incidence of gastrointestinal illness, mainly due to diarrhea during lactation, in beef cattle breeding is 13.3% (Akio Nakane, Livestock Research, Vol. 33, No. 1, 37 (19779)). The mortality rate of dairy cattle during lactation and breeding has reached 11.9%, mainly due to diarrhea and pneumonia (Hiroyuki Tojo et al., Livestock Research, Vol. 4). 0, No. 1, 51, (1966)). In addition, in the case of adult cattle, when diarrhea occurs, the loss of milk yield, milk quality, and meat quality is observed due to reduced feed efficiency without death, resulting in a large loss of profit.
豚若しく は愛玩動物 (ペッ ト) についても同様なことが言え、 一 般に出生直後より母子を分離して育生することが一般的飼育形態に なり、 これら初生獣は抗原性が極めて低く、 また腸内細菌叢が未形 成であるため病原菌の定着が容易となり、 さらに種々のス ト レスに 対する抵抗性も弱く、 これらが相互に作用し下痢症に陥り、 へい死 率が極めて高く、 へい死を免れてもその後の発育に多大な影響を与 え、 発育不良をきたすこととなる。  The same can be said for pigs or pets. In general, it is a common breeding method that mothers and children are separated and raised immediately after birth, and these primary animals have extremely low antigenicity. In addition, since the intestinal flora is not formed, colonization of pathogenic bacteria is easy, and resistance to various stresses is weak.These interact and cause diarrhea, resulting in extremely high mortality and death. Even if they escape, they can have a significant effect on subsequent growth, leading to poor growth.
また、 鶏も同様で、 下痢雛は一般に発育が悪く、 体重低下、 体重 変動幅が増大し、 その後の生産性に悪影響を与え、 ブロイラーにお いては商品化率、 産卵鶏では産卵率の低下の要因となる。 さらにヮ クチン接種後の抗体產生能も減少することが知られている。  The same is true for chickens.Diarrheal chicks generally have poor growth, lose weight, increase the range of weight fluctuation, adversely affect subsequent productivity, and decrease the commercialization rate in broilers and the egg production rate in laying hens. Is a factor. It is also known that the antibody viability after vaccination decreases.
ところで、 これらの下痢の発生要因は、 非常に複雑でクロス ト リ ジゥム ノ、。一フ リ ンジエ ンス A型、 クロス ト リ ジゥム パ一フ リ ンジエンス C型、 サルモネラ ティ フィ ムリ ウム、 サルモネラ タブリ ン、 カムピロパクター ジェジュ二一、 カムピロバクタ一 コ リー、 大腸菌 0 - 8 8、 大腸菌 0 - 9 9、 大腸菌 9 8 7 P 、 スタフイ ロコ ッカス ォゥ レウス、 口夕ウィルス KK一 3、 口 夕ウィルス N C D V等の病原性細菌、 下痢症ウィルスの産生する 毒素又は感染による腸内細菌叢の変化によって生じる感染性下痢だ けでなく、 家畜、 家禽及び愛玩動物に与えられたス ト レスが原因で 発生するス トレス性下痢等の感染性微生物の関与していない原因不 明の下痢といった非感染性下痢も数多く発生している。 By the way, the cause of these diarrhea is very complicated and cross-trimming. 1F-A, Cross-Stream P.F.-C, Salmonella typhimurium, Salmonella tabulin, Campylobacter Jeju 21, Campylobacter coli, Escherichia coli 0-88, Escherichia coli 0-9 9, Escherichia coli 9887P, Staphylococcus oleus, oral virus KK-1 3, oral Evening virus Pathogenic bacteria such as NCDV, toxins produced by diarrhea virus, or infectious diarrhea caused by changes in intestinal flora caused by infection, as well as stress given to livestock, poultry and pets Many non-infectious diarrhea such as diarrhea of unknown cause that do not involve infectious microorganisms, such as stress diarrhea occurring in Japan, have occurred.
これら家畜、 家禽及び愛玩動物のス ト レス原因は、 環境の変化、 栄養状態の変化、 心理的な変化の 3要因であることが知られている 。 例えば、 過密飼育、 運動不足、 労力不足による畜舎の衛生管理の 不備、 分娩による飼料組成の変化、 離乳期における長距離の輸送な どの様々なス ト レスの原因が存在している。  It is known that the causes of stress in livestock, poultry, and companion animals are three factors: environmental changes, nutritional status changes, and psychological changes. For example, there are various causes of stress, such as overcrowding, lack of exercise, inadequate barn hygiene due to lack of labor, changes in feed composition due to parturition, and long distance transport during weaning.
さらに、 出生後の初生獣は一定期間母親の保護下で成育させるの が自然である。 しかし、 畜産経営上、 生産性又は経済性を向上させ る目的のため、 一般に肉用牛を除いて、 出生直後から母子を分離し て育成することが一般的飼養形態となった。 母親から分離された初 生獣の大部分は生後一週間前後に哺育施設などに集められ、 集団的 に育成される。 この時、 生後まもない初生獣は抗原性が極めて低い 、 また腸内細菌叢が未形成であるため、 病原菌の定着が容易となり 、 さらに種々のス ト レスに対する抵抗性も弱く、 これらが相互に作 用し合い容易に下痢性に陥ると考えられる。  Furthermore, it is natural for postnatal primary animals to grow for a period of time under the care of their mother. However, for the purpose of improving productivity or economy in livestock management, it has become common practice to separate mothers and babies immediately after birth, except for beef cattle in general, for rearing. Most of the primary animals separated from their mothers are collected at nursing facilities around one week after birth, and are bred collectively. At this time, the early-born animals have very low antigenicity, and since the intestinal flora is not formed, colonization of pathogenic bacteria becomes easy, and resistance to various stresses is weak. It is thought that diarrhea easily occurs when working together.
従来、 感染性下痢に対する予防あるいは治療は、 家畜、 家禽及び 愛玩動物に抗生物質を投与することが有効な手段であつたが、 安全 に使用できる抗生物質の種類、 量には制限があり、 また、 抗生物質 に対する耐性菌の出現、 安全性の面から、 その有効性は疑問視され るようになった。 これに代わる方法として各種天然の抗菌性物質が 用いられるようになり、 茶抽出物に抗感染性微生物活性あるいはゥ ィルス活性があることが開示されている (特開平 1 一 2 6 5 0 2 3 号、 特開平 2 — 2 7 6 5 6 2号) 。 しかしながら、 茶抽出物を単独 で感染性下痢症に用いた場合あるいはこれを配合した飼料を用いた 場合、 効果が弱く、 実用面で疑問視されている。 また、 特異的抗体 を単独で用いる場合、 実際には、 病原性微生物は様々な種類が存在 するため、 これらの有する特異性のために、 下痢の予防効果は期待 できず、 治療効果のみとなり、 実用上効果的でない。 Conventionally, the prevention or treatment of infectious diarrhea has been an effective means of administering antibiotics to livestock, poultry and pets, but the types and amounts of antibiotics that can be used safely are limited. However, the emergence and resistance of antibiotic-resistant bacteria have led to questions about their effectiveness. As an alternative, various natural antibacterial substances have come to be used, and it has been disclosed that the tea extract has an anti-infective microbial activity or a virus activity (Japanese Patent Application Laid-open No. Hei 12-265203). No., JP-A-2-277652). However, when the tea extract alone is used for infectious diarrhea or when a feed containing the same is used, the effect is weak, and practical use is questioned. In addition, when a specific antibody is used alone, there are actually various types of pathogenic microorganisms, and due to these specificities, a diarrhea-preventing effect cannot be expected, but only a therapeutic effect. Not practically effective.
—方、 非感染性下痢症に対する予防、 治療法については、 初生獣 に対し、 出生後より初乳を十分量給与する。 また、 動物にス ト レス が生じると、 結果として感染性微生物が感染しやすい環境になる等 、 腸内細菌叢に変化が生じることから、 感染性下痢症と同様の方法 あるいは各種生菌剤、 オリゴ糖が用いられている。 しかしながら、 これらの方法は対処療法にすぎない。 また、 生菌剤、 オリゴ糖につ いては、 効果の持鐃性に問題がある。 また、 ス ト レスの予防方法と しては、 豚の退屈感を除くために、 豚房内に鉄製の鎖や古タイヤを つり下げて豚に自由に咬ませる方法や、 敷きわらを毎日豚房に入れ て豚に咬ませることによって退屈感をまぎらわせる方法があるが、. これは消極的予防法であり、 かつ家畜、 家禽及び愛玩動物の種類も 限定される。 また、 特公平 3 — 1 7 4 6 9号ではス ト レス低減に家 畜の馴致用組成物が開示されているが非感染性下痢症を抑制するこ とを期待したものではない。 さらに、 特公平 3 — 7 0 4 5 8号に開 示されている方法でも同様であり、 かっこの方法は飼育時期、 飼育 環境にも限定される。 このようなことから家畜、 家禽及び愛玩動物 の非感染性下痢症の発生を抑制する有効な飼料添加物又はこれを配 合した動物用飼料が未だ見出されていないのが現状である。  —On the other hand, for prevention and treatment of non-infectious diarrhea, a sufficient amount of colostrum should be fed to newborn animals after birth. In addition, when stress occurs in animals, the intestinal flora changes as a result, for example, resulting in an environment that is more susceptible to infection by infectious microorganisms. Oligosaccharides have been used. However, these methods are only coping. In addition, the probiotic agent and the oligosaccharide have a problem in the cyclability of the effect. To prevent stress, there are several methods to prevent the pig from feeling bored, such as hanging an iron chain or old tires inside the pig's tress and allowing the pig to bite it freely. There is a method of disturbing boring by placing the pigs in their bunches and biting them. However, this is a passive prevention method and the types of livestock, poultry and pets are limited. In addition, Japanese Patent Publication No. 3-1 7469 discloses a composition for adapting livestock to reduce stress, but does not expect to suppress non-infectious diarrhea. The same applies to the method disclosed in Japanese Patent Publication No. 3-7045-8. The method of parentheses is limited to the breeding time and environment. Under these circumstances, no effective feed additive or animal feed containing the same has been found at present, which suppresses the occurrence of non-infectious diarrhea in livestock, poultry and pets.
また、 家畜の生産する生乳は、 乳脂率、 無脂固形分率、 細胞数、 及び乳蛋白率でランク付けされ、 取り引きされている。 例えば、 A ランクの生乳の場合では、 乳脂率 3 . 5 %以上、 無脂固形分率 8 . 5 以上、 細胞数 3 0万個以下、 乳蛋白率 3 . 0 %以上であること が要求されており、 これらの数値を一つでも下回ると正常な取り引 きが成されなくなり、 数値を達成していない生乳にはペナルティが かけられている。 すなわち、 Aランクの生乳として認定されるため には、 乳脂率、 無脂固形分率、 細胞数、 乳蛋白率が、 一定以上の値 を示すことが必要である。 このようなことから、 乳質の低下は酪農 家に多大の経済的損失を与えている。 また、 乳量の低下も同様のこ とが言え、 生産性の低下による経済的損失につながる。 In addition, raw milk produced by livestock is ranked and traded in terms of milk fat percentage, non-fat solids percentage, cell number, and milk protein percentage. For example, A In the case of rank raw milk, it is required that the milk fat ratio is 3.5% or more, the non-fat solid content ratio is 8.5 or more, the number of cells is 300,000 or less, and the milk protein ratio is 3.0% or more. If one of these figures is not reached, normal transactions will not be made, and raw milk that does not achieve the figures will be penalized. In other words, in order to be certified as A-rank raw milk, it is necessary that the percentage of milk fat, the percentage of non-fat solids, the number of cells, and the percentage of milk protein show certain values. For this reason, reduced milk quality has resulted in significant economic losses for dairy farmers. The same can be said for a decrease in milk yield, which leads to an economic loss due to a decrease in productivity.
このようなことから、 乳質の改善、 乳量の増加のために濃厚飼料 を中心とした改善の試みがなされている (特開平 2 — 1 4 5 1 5 4 号、 特開平 2 - 2 8 6 0 4 7号) 。  For these reasons, attempts have been made to improve milk quality and increase milk yield, mainly concentrated feed (Japanese Patent Application Laid-Open Nos. 2-145,154 and 2-2886). 0 4 7).
しかしながら、 上記方法による対策では期待する効果が十分では なく、 より一層の改善策が望まれているのが現状である。  However, the expected effect is not sufficient with the measures by the above method, and further improvement measures are currently desired.
また、 繁殖率に関しては、 例えば乳牛の場合、 個体の乳量生産を 増加させた時に、 飼養管理の失宜がすぐに乳牛の繁殖障害を引き起 こすことがある。 このことにより、 短期間のうちに多数の牛に繁殖 障害が発生することがある。 しかしながら現在のところ、 繁殖障害 に対する有効な治療法及び有効な予防法は皆無である (畜産の研究 Vo l . 44, No. 1 0, 1 143- 1 149 ( 1 990)中尾敏彦) 。  Regarding the reproductive rate, for example, in the case of dairy cows, when the individual's milk production is increased, mismanagement of breeding management can quickly cause breeding problems for dairy cows. This can cause reproductive harm to large numbers of cattle in a short period of time. However, at present, there are no effective treatments or effective preventive measures for reproductive disorders (Livestock Research Vol. 44, No. 10, 1143-1149 (1990) Toshihiko Nakao).
また、 乳牛、 肉牛、 豚、 鶏等の家畜は、 近年その利用度が高まり 、 生産性向上のため、 前記のように濃厚飼料を与え経済効果を高め る傾向にあるが、 従来行われていた素材型の飼料と比べ胃腸にかか る負担が大き く、 家畜の成長率の低下、 搾乳量の減少、 肉質の劣化 などの問題を生じている。  In addition, livestock such as dairy cows, beef cattle, pigs, and chickens have been increasingly used in recent years and tend to increase the economic effect by providing concentrated feed as described above in order to improve productivity. The burden on the gastrointestinal tract is greater than that of raw feed, causing problems such as reduced livestock growth, reduced milk production, and deterioration of meat quality.
また、 前記のように飼育環境の集約化のため、 ス ト レス等の諸要 因により細菌性、 あるいはウイルス性の下痢を発症する機会も増し 、 畜舎の維持に障害をもたらす等、 業界にとっても大きな問題を生 じている。 さらに疾病に対する抗生物質や強力な抗菌剤の投与は、 家畜の消化器官、 腸内の細菌叢を攪乱させ新たな疾病を誘発するな ど問題が多く、 また乳中、 食肉中へのこれら薬品の残留が指摘され るなど商品価値の低下や安全面の不安がある。 In addition, as described above, stresses such as stress are required to consolidate the breeding environment. For this reason, the chances of developing bacterial or viral diarrhea also increase, causing problems in the maintenance of stables, causing serious problems for the industry. In addition, the administration of antibiotics and powerful antibacterial agents against diseases has many problems, such as disrupting the digestive tract and intestinal microflora of livestock and inducing new diseases.In addition, the use of these drugs in milk and meat is problematic. There are concerns about residual products, such as a decrease in product value and safety concerns.
—方、 畜舎から発生する糞尿臭等が周囲の環境に及ぼす影響も大 き く、 特に下痢便では悪臭が強くなり、 悪臭公害を引き起こすなど 大きな問題であるが、 現時点ではこのような飼育環境を改善する有 効な解決法が見出せない状況である。 発明の開示  —On the other hand, the smell of manure generated from the barn has a large effect on the surrounding environment, especially with diarrheal stool, which is a serious problem, such as causing odor pollution. There is no effective solution to improve. Disclosure of the invention
上述の状况に鑑み、 本発明者らは、 ( 1 ) 家畜、 家禽及び愛玩動 物にみられる非感染性及び感染性下痢症に対して効果のある飼料添 加物及び動物用飼料について、 ( 2 ) 家畜の生産する生乳の乳質改 善効果及び乳量増加効果を示す飼料添加物及び動物用飼料について 、 ( 3 ) 家畜、 家禽等の動物の繁殖率の向上効果を示す飼料添加物 及び動物用飼料について、 並びに ( 4 ) 動物の整腸作用の促進、 下 痢発症の低減、 搾乳量の増加、 飼料効率の改善、 並びに糞便及び尿 の悪臭消去作用を有する飼料添加物及び動物用飼料について鋭意研 究した。  In view of the situation described above, the present inventors have determined that (1) feed additives and animal feeds that are effective against non-infectious and infectious diarrhea in livestock, poultry, and pet animals; 2) Feed additives and animal feeds that show the effects of improving milk quality and increasing milk yield of raw milk produced by livestock. (3) Feed additives and animals that show the effect of improving the reproduction rate of animals such as livestock and poultry. And (4) feed additives and animal feeds that promote the intestinal action of animals, reduce the occurrence of diarrhea, increase the amount of milking, improve feed efficiency, and eliminate the odor of feces and urine. Diligently studied.
その結果、 以下の点を見いだし、 本発明を完成するに至った。  As a result, they have found the following points, and have completed the present invention.
( 1 ) 茶抽出物中のポリ フエノール化合物である (+ ) —力テキン 、 (+ ) —ガロカテキン、 (一) ーガロカテキンガレー ト、 (一) ーェピカテキン、 (一) 一ェピカテキンガレー ト、 (一) ーェピガ ロカテキン、 (一) ーェピガロカテキンガレー ト、 遊離型テアフラ ビン、 テアフラ ビンモノガレー ト八、 テアフラ ビンモノガレ一 ト B 、 及びテアフラビンジガレー ト等の夕ンニン類を飼料に添加して用 いると非感染性下痢症に対し効果があること、 また、 病原性細菌若 しく はウィルスに対する特異的抗体又はこれらの産生する毒素に対 する特異的抗体を該ポリ フエノール化合物と共に飼料に添加して用 いると、 該ポリ フエノール化合物単独で用いる場合と比較して感染 性下痢症に対し優れた効果を発揮すること、 (1) Polyphenolic compounds in tea extract (+)-force techin, (+)-gallocatechin, (1) -gallocatechin gallate, (1) -epicatechin, (1) -epicatechin gallate, (1) -epigallocatechin gallate, (1) -epigallocatechin gallate, free theafla The use of ninnins such as bottles, theaflavin monogallate 8, theaflavin monogallet B, and theaflavin digallate in the feed is effective against non-infectious diarrhea. Alternatively, when a specific antibody against a virus or a specific antibody against a toxin produced by the virus is added to the feed together with the polyphenol compound, the infectious diarrhea is higher than when the polyphenol compound is used alone. To exert an excellent effect on
( 2 ) 該ボリ フエノール化合物を飼料に添加して用いると家畜の生 産する生乳の乳質改善効果及び乳量増加効果を示すこと、  (2) that when the polyphenol compound is added to a feed and used, it shows an effect of improving milk quality and an effect of increasing milk yield of raw milk produced by livestock;
( 3 ) 該ポリ フユノール化合物を飼料に添加して用いると家畜、 家 禽等の動物の繁殖率の向上効果を示すこと、 並びに  (3) When the polyfunol compound is added to feed and used, it shows an effect of improving the reproduction rate of animals such as livestock and poultry; and
( 4 ) 茶葉を水、 アルコール、 アセ ト ン又はこれらの混合液を用い て可溶性成分を茶葉から抽出除去して得られる茶葉織維を飼料に添 加して用いると、 動物の整腸作用の促進、 下痢発症の低減、 搾乳量 の増加、 飼料効率の改善、 並びに動物の糞便及び尿の悪臭消去作用 がみられるこ と。  (4) If tea leaves obtained by extracting and removing soluble components from tea leaves using water, alcohol, acetate, or a mixture thereof are added to the feed and used, the intestinal action of animals can be reduced. Acceleration, reduction of diarrhea episodes, increase of milking volume, improvement of feed efficiency, and elimination of odor in animal feces and urine.
即ち、 本発明の要旨は、 以下の点にある。  That is, the gist of the present invention lies in the following points.
( 1 ) 茶の抽出物よりなる動物用飼料添加物、  (1) an animal feed additive comprising an extract of tea,
( 2 ) 茶が緑茶、 ウーロン茶若しく は紅茶である前記 ( 1 ) 記載の 動物用飼料添加物、  (2) The animal feed additive according to (1), wherein the tea is green tea, oolong tea or black tea,
( 3 ) 茶の抽出物が茶葉を水、 アルコール及び 又は酢酸ェチルで 抽出したものである前記 ( 1 ) 記載の動物用飼料添加物、  (3) The animal feed additive according to (1), wherein the tea extract is obtained by extracting tea leaves with water, alcohol, and / or ethyl acetate.
( 4 ) 茶の抽出物がボリ フエノ一ル化合物を含有する前記 ( 1 ) 記 載の動物用飼料添加物、  (4) The animal feed additive according to (1), wherein the tea extract contains a polyphenol compound,
( 5 ) ポリ フヱノール化合物の含量が 5〜 8 0 %である前記 ( 4 ) 記載の動物用飼料添加物、 ( 6 ) ボリ フヱノール化合物が (+ ) —カテキン、 (+ ) —ガロカ テキン、 (一) ーガロカテキンガレー ト、 (一) ーェピカテキン、(5) The animal feed additive according to (4), wherein the content of the polyphenol compound is 5 to 80%, (6) Polyphenol compounds are (+)-catechin, (+)-gallocatechin, (1) -gallocatechin gallate, (1) -eppicatechin,
(一) 一ェピカテキンガレー ト、 (一) 一ェピガロカテキン、 (一 ) ーェピガロカテキンガレー ト、 遊離型テアフラビン、 テアフラビ ンモノ ガレー ト八、 テアフラ ビンモノガレー ト B及びテアフラ ビン ジガレー トよりなる群から選ばれる 1種又は 2種以上の化合物を含 有する前記 ( 4 ) 記載の動物用飼料添加物、 (1) one epicatechin gallate, (1) one epigallocatechin gallate, (1) one epigallocatechin gallate, free theaflavin, theaflavin monogallate 8, eight theaflavin monogallate B, and theaflavin digallate The animal feed additive according to the above (4), comprising one or more selected compounds.
( 7 ) ポリ フユノール化合物中の各ポリ フユノール化合物の含量が ( + ) -カテキン 0. 2〜 6. 5 %、 ( + ) —ガロカテキン 2. 0 (7) The content of each polyfunol compound in the polyfunol compound is (+)-catechin 0.2 to 6.5%, (+)-gallocatechin 2.0
〜 1 8. 0 %, (一) ーガロカテキンガレー ト 1. 0〜 1 5. 0 % 、 (一) ーェピカテキン 0. 5〜 1 0. 0 %、 (一) 一ェピカテキ ンガレー ト 0. 3〜 8. 0 %、 (一) ーェピガロカテキン 2. 0〜 1 8. 0 %、 (一) ーェピガロカテキンガレー ト 3. 0〜 2 1. 0 %、 遊離型テアフラ ビン 0〜 2 0. 0 %、 テアフラ ビンモノガレー ト A 0〜 5. 0 %、 テアフラ ビンモノガレ一 ト B 0〜 5. 0 %及び テアフラビンジガレ一 ト 0〜 5. 0 %である前記 ( 6 ) 記載の動物 用飼料添加物、 -18.0%, (1) -gallocatechin gallate 1.0-15.0%, (1) eppicatechin 0.5-10.0%, (1) 1-epicatechin gallate 0.3- 8.0%, (1) -epigallocatechin 2.0-18.0%, (1) -epigallocatechin gallate 3.0-21.0%, free theaflavin 0-2 The animal feed according to the above (6), which is 0.0%, theaflavin monogallate A 0 to 5.0%, theaflavin monogallet B 0 to 5.0%, and theaflavin digallate 0 to 5.0%. Additive,
( 8 ) 下痢の予防又は治療に用いる前記 ( 1 ) 〜 ( 7 ) いずれか記 載の動物用飼料添加物を含有してなる動物用飼料、  (8) An animal feed comprising the animal feed additive according to any one of (1) to (7) above, which is used for prevention or treatment of diarrhea;
( 9 ) 下痢が非感染性下痢症である前記 ( 8 ) 記載の動物用飼料、 (9) The animal feed according to (8), wherein the diarrhea is non-infectious diarrhea.
( 1 0 ) 非感染性下痢症がス ト レス性下痢又は感染性微生物の関与 していない原因不明の下痢である前記 ( 9 ) 記載の動物用飼料、(10) The animal feed according to (9), wherein the non-infectious diarrhea is stress diarrhea or unexplained diarrhea not involving infectious microorganisms.
( 1 1 ) 感染性微生物又はウィルス、 あるいはそれらの産生する毒 素に対する特異的抗体をさらに含有する前記 ( 8 ) 記載の動物用飼 料、 (11) The animal feed according to the above (8), further comprising a specific antibody against an infectious microorganism or virus, or a toxin produced by the infectious microorganism or virus.
( 1 2 ) 特異的抗体が感染性微生物又はウィルスで、 あるいはそれ らが産生する毒素で過免疫された産卵鶏の卵より得られる鶏卵抗体 である前記 ( 1 1 ) 記載の動物用飼料、 (12) The specific antibody is an infectious microorganism or virus, or The animal feed according to the above (11), which is a chicken egg antibody obtained from eggs of laying hens hyperimmunized with a toxin produced by them.
( 1 3 ) 特異的抗体が感染性微生物又はウィルスで、 あるいはそれ らが産生する毒素で過免疫された哺乳類の乳汁より得られる乳汁抗 体である前記 ( 1 1 ) 記載の動物用飼料、  (13) The animal feed according to the above (11), wherein the specific antibody is a milk antibody obtained from milk of a mammal hyperimmunized with an infectious microorganism or virus or a toxin produced thereby.
( 1 4 ) 特異的抗体の配合量が酵素免疫測定法でブランクに対し 1 (14) The amount of the specific antibody was 1
. 5倍以上の抗体価を有したものを 11118< 体重1^ g以上の投与量 となるように調製された前記 ( 1 1 ) 記載の動物用飼料、 The animal feed according to the above (11), wherein the animal feed having an antibody titer of 5 times or more is prepared so as to have a dose of 11118 < 1 ^ g or more of body weight.
( 1 5 ) 乳量増加に用いる前記 ( 1 ) 〜 ( 7 ) いずれか記載の動物 用飼料添加物を含有してなる動物用飼料、  (15) An animal feed comprising the animal feed additive according to any one of the above (1) to (7), which is used for increasing milk yield.
( 1 6 ) 乳質改善に用いる前記 ( 1 ) 〜 ( 7 ) いずれか記載の動物 用飼料添加物を含有してなる動物用飼料、  (16) An animal feed comprising the animal feed additive according to any one of (1) to (7), which is used for improving milk quality.
( 1 7) 動物の繁殖率向上に用いる前記 ( 1 ) 〜 ( 7 ) いずれか記 載の動物用飼料添加物を含有してなる動物用飼料、  (17) An animal feed comprising the animal feed additive as described in any one of (1) to (7) above, which is used for improving the reproduction rate of animals.
( 1 8 ) 茶葉から水、 アルコール、 アセ トン又はこれらの混合液に より抽出される可溶性成分を除去して得られる茶葉織維を有効成分 とする動物用飼料添加物、  (18) An animal feed additive comprising, as an active ingredient, a tea leaf fiber obtained by removing water, alcohol, acetone, or a soluble component extracted from a mixture thereof from tea leaves,
( 1 9 ) 腸内細菌叢の改善に用いる前記 ( 1 8 ) 記載の動物用飼料 添加物を含有してなる動物用飼料、  (19) An animal feed comprising the animal feed additive according to (18), which is used for improving intestinal flora,
( 2 0 ) 悪臭の低減 ' 消去に用いる前記 ( 1 8 ) 記載の動物用飼料 添加物を含有してなる動物用飼料、  (20) An animal feed comprising the additive for animal feed according to the above (18), which is used for elimination of malodor.
( 2 1 ) 飼料効率の改善に用いる前記 ( 1 8 ) 記載の動物用飼料添 加物を含有してなる動物用飼料、  (21) An animal feed comprising the animal feed additive according to (18), which is used for improving feed efficiency,
( 2 2 ) 動物が家畜、 家禽又は愛玩動物である前記 ( 8:) 〜 ( 1 7 ) . ( 1 9 ) 〜 ( 2 1 ) いずれか記載の動物用飼料、  (22) The animal feed according to any one of (8 :) to (17). (19) to (21), wherein the animal is a domestic animal, poultry, or a companion animal.
( 2 3 ) 茶葉を 3 0〜 9 5 、 0. 5〜7時間、 水、 アルコール及 び Z又は酢酸ェチルで抽出し、 得られた抽出液を噴霧乾燥するこ と よりなる前記 ( 1 ) 〜 ( 7 ) 記載の動物用飼料添加物の製造方法、(23) Tea leaves 30-95, 0.5-7 hours, water, alcohol and The method for producing an animal feed additive according to any one of the above (1) to (7), comprising extracting the extract with Z or Z or ethyl acetate, and spray-drying the obtained extract.
( 2 4 ) 茶葉を 3 0〜 9 5。C、 0. 5〜 7時間、 水、 アルコール及 び 又は酢酸ェチルで抽出し、 得られた抽出液を分画分子量 3 0 0 0〜 6 0 0 0の限外濾過膜にて濃縮し、 次いで逆浸透膜で濃縮する ことよりなる前記 ( 1 ) 〜 ( 7 ) 記載の動物用飼料添加物の製造方 法、 (24) 30-95 tea leaves. C, for 0.5 to 7 hours, extracted with water, alcohol and / or ethyl acetate, and the obtained extract was concentrated with an ultrafiltration membrane having a cut-off molecular weight of 300 to 600, then The method for producing an animal feed additive according to any one of the above (1) to (7), comprising concentrating with a reverse osmosis membrane.
( 2 5 ) 茶葉を 3 0〜 9 5て、 0. 5〜 7時間、 水及び 又はアル コールで抽出し、 さらに酢酸ェチルで分配して得られた酢酸ェチル 層を除去することよりなる前記 ( 1 ) 〜 ( 7 ) 記載の動物用飼料添 加物の製造方法、  (25) The method comprising extracting tea leaves for 30 to 95, extracting them with water and / or alcohol for 0.5 to 7 hours, and further removing the ethyl acetate layer obtained by partitioning with ethyl acetate. 1) The method for producing an animal feed additive according to any one of (7) to (7),
( 2 6 ) (—) ーェピガロカテキンガレー トを有効成分とする動物 用飼料添加物、  (26) (—) Animal feed additive containing epigallocatechin gallate as an active ingredient,
( 2 7 ) 前記 ( 2 6 ) 記載の動物用飼料添加物を含有してなる動物 用飼料。 発明を実施するための最良の形態  (27) An animal feed comprising the animal feed additive according to the above (26). BEST MODE FOR CARRYING OUT THE INVENTION
本発明は茶由来の飼料添加物及びそれを含んでなる動物用飼料に 関する。 これには茶の抽出物を使用するものと茶の抽出残渣を使用 する態様とがある。  The present invention relates to a feed additive derived from tea and an animal feed comprising the same. This includes a method using a tea extract and a method using a tea extraction residue.
まず、 茶の抽出物を使用する態様について説明する。 本発明に用 いられる茶抽出物とは、 水、 アルコール、 酢酸ェチル等の溶媒を用 いて茶葉より抽出される、 次に述べるポリ フエノール化合物を含有 するものを指す。  First, an embodiment using a tea extract will be described. The tea extract used in the present invention refers to a tea extract containing the following polyphenol compound, which is extracted from tea leaves using a solvent such as water, alcohol, or ethyl acetate.
本発明におけるボリ フエノール化合物とは (+ ) —力テキン、 ( + ) —ガロカテキン、 (一) ーガロカテキンガレー ト、 (一) ーェ ピカテキン、 (一) ーェピカテキンガレ一 ト、 (一) ーェピガロカ テキン、 (一) ーェピガロカテキンガレー ト、 遊離型テアフラ ビンThe polyphenol compound in the present invention is (+)-force techin, (+)-gallocatechin, (1) -gallocatechin gallate, (1) -g Picatechin, (1) epigallocatechin, (1) epigallocatechin, (1) epigallocatechin gallate, free theaflavin
、 テアフラ ビンモノガレー ト A、 テアフラ ビンモノ ガレー B、 テ アフラビンジガレー ト等のタンニン類を指す。 Tannins such as theaflavin monogallate A, theaflavin monogallet B, and theaflavin digallate.
上記のポリ フヱノール化合物は、 緑茶、 紅茶、 ウーロン茶等の茶 の熱水抽出物の成分が好ましいが、 茶の水若しく はアルコール抽出 物の酢酸ェチル可溶画分又は限外濾過膜及び逆浸透膜処理により得 ることもできる。 このようなボリ フヱノール化合物の茶からの抽出 方法としては、 特に限定されるものではなく、 例えば以下に記述す る方法を用いることができる。  The above-mentioned polyphenol compound is preferably a component of a hot water extract of tea such as green tea, black tea, and oolong tea.Ethyl acetate-soluble fraction of tea water or alcohol extract or an ultrafiltration membrane and reverse osmosis It can also be obtained by membrane treatment. The method for extracting such a polyphenol compound from tea is not particularly limited, and for example, the method described below can be used.
例えば、 茶に溶媒を加え、 攪拌を行う。 次いで溶液と残渣とを分 離し、 得られた溶液を乾燥することにより、 求めるポリ フエノール 化合物を得ることができる。 ここで、 茶は粉砕したものを用いても よく、 粉砕しないものを用いてもよい。 用いる溶媒は特に限定され るものではなく、 水、 エタノール、 アセ トン等が使用でき、 これら は単独で用いてもよく、 任意の割合で混合したものを用いてもよい 。 また、 溶媒はあらかじめ加温 ( 3 0〜 9 5 °C ) していてもよく、 攪拌中に加温 ( 3 0〜 9 5 eC ) してもよい。 撹拌時間は 3 0分〜 7 時間の範囲で行えばよい。 この範囲より短いと抽出の効率が悪く、 この範囲より長くても、 応じた効果は得られない。 分離した溶液は 、 そのまま乾燥してもよく、 濃縮した後に乾燥してもよい。 乾燥方 法としては、 噴霧乾燥、 凍結乾燥、 熱風乾燥といった通常用いる方 法でよい。 濃縮は、 限外濾過膜や、 逆浸透膜を用いて行う ことがで きる。 これらは単独で使用してもよく、 組み合わせて使用してもよ い。 また、 得られた抽出物を再分配することでボリ フエノール化合 物の純度を高めることができる。 このとき、 水一酢酸ェチルの系で 分配するのが好ま しく、 酢酸ェチル画分中にボリ フエノ一ル化合物 が濃縮される。 For example, add a solvent to tea and stir. Next, the solution and the residue are separated, and the obtained solution is dried to obtain the desired polyphenol compound. Here, the tea may be used in a crushed state or in a non-ground state. The solvent to be used is not particularly limited, and water, ethanol, acetone, and the like can be used. These may be used alone, or may be used in a mixture at an arbitrary ratio. The solvent may be the previously warmed (3 0~ 9 5 ° C) , may be heated in a stirred (3 0~ 9 5 e C) . The stirring time may be in the range of 30 minutes to 7 hours. If the length is shorter than this range, the extraction efficiency is poor. If the length is longer than this range, the corresponding effect cannot be obtained. The separated solution may be dried as it is, or may be dried after being concentrated. As a drying method, a commonly used method such as spray drying, freeze drying and hot air drying may be used. Concentration can be performed using an ultrafiltration membrane or a reverse osmosis membrane. These may be used alone or in combination. Further, by redistributing the obtained extract, the purity of the polyphenol compound can be increased. At this time, in the system of water ethyl acetate Partitioning is preferred, and the polyphenols are concentrated in the ethyl acetate fraction.
さらに具体的には、 例えば ( 1 ) 茶葉を 3 0〜 9 5 、 0 . 5〜 7時間、 水、 アルコール及び/又は酢酸ェチルで抽出し、 得られた 抽出液を噴霧乾燥する、 ( 2 ) 茶葉を 3 0〜 9 5で、 0 . 5〜 7時 間、 水、 アルコール及び 又は酢酸ェチルで抽出し、 得られた抽出 液を分画分子量 3 0 0 0〜 6 0 0 0の限外濾過膜にて濃縮し、 次い で逆浸透膜で濃縮する、 ( 3 ) 茶葉を 3 0〜 9 5で、 0 . 5〜 7時 間、 水及びノ又はアルコールで抽出し、 さらに酢酸ェチルで分配し て得られた酢酸ェチル層を除去する等の方法が例示される。  More specifically, for example, (1) tea leaves are extracted with water, alcohol and / or ethyl acetate for 30 to 95, 0.5 to 7 hours, and the obtained extract is spray-dried, (2) Tea leaves are extracted with water, alcohol and / or ethyl acetate for 30 to 95 hours and 0.5 to 7 hours, and the resulting extract is subjected to ultrafiltration with a cut-off molecular weight of 300 to 600. Concentrate with a membrane and then with a reverse osmosis membrane. (3) Extract tea leaves for 30 to 95, 0.5 to 7 hours with water and ethanol or alcohol, and partition with ethyl acetate Examples of the method include removing the obtained ethyl acetate layer.
このようにして得られる茶の抽出物中には通常 2種以上のポリ フ エノ一ル化合物を含有するものであるが、 本発明ではこれらのボリ フエノール化合物を単離して単独で用いてもよく、 あるいは 2種以 上の混合物として使用してもよい。 さらに、 このようにして得られ る抽出物をポリ フエノール化合物を含む粗抽出物としてそのまま使 用してもよい。 特定のポリ フエノール化合物を単独で用いる場合、 その化合物は特に限定されるものではなく、 本発明におけるポリ フ ェノール化合物のうちのどの化合物を用いてもかまわないが、 なか でも (一) ーェピガロカテキンガレー トを用いるのが好適である。 またこの場合、 当該化合物は茶由来の化合物に限定されるものでは ない。 すなわち、 当該化合物が他の生物由来、 例えば柿しぶ由来の もの又は化学合成品であつても差し支えない。  The tea extract thus obtained usually contains two or more kinds of polyphenol compounds.In the present invention, these polyphenol compounds may be isolated and used alone. Alternatively, they may be used as a mixture of two or more. Further, the extract thus obtained may be used as it is as a crude extract containing a polyphenol compound. When a specific polyphenol compound is used alone, the compound is not particularly limited, and any of the polyphenol compounds in the present invention may be used. It is preferable to use gallocatechin gallate. In this case, the compound is not limited to a tea-derived compound. That is, the compound may be derived from other organisms, for example, persimmon shibu or a chemically synthesized product.
化合物の単離方法としては特に限定されるものではなく、 一般的 な方法を用いればよい。 例えば、 茶抽出物を酢酸ェチルに溶解した もの又は前述の酢酸ェチル画分をシリカゲルクロマ トグラフィーに 付し、 適切な溶媒を用いて溶出することで単離することができる。 次いで H P L Cを用いることによりさらに高純度の化合物を得るこ ともできる。 The method for isolating the compound is not particularly limited, and a general method may be used. For example, it can be isolated by dissolving a tea extract in ethyl acetate or the above-mentioned ethyl acetate fraction by silica gel chromatography and eluting with a suitable solvent. Then, even higher purity compounds can be obtained by using HPLC.
本発明の動物用飼料添加物は、 このようにして得られる茶の抽出 物由来のポリ フエノール化合物を含有するものをいい、 本発明の動 物用飼料添加物中に含まれるボリ フエノール化合物の含量は、 通常 5〜 8 0 %、 好ましく は 1 0〜 5 0 さらに好ましく は 2 0〜 3 5 %である。 ここで、 動物用飼料添加物中の各ボリ フヱノール化合 物の含量は、 (+ ) —力テキン 0. 2〜 6. 5 %、 ( + ) —ガロカ テキン 2. 0〜 1 8. 0 %、 (一) ーガロカテキンガレー ト 1 . 0 〜 1 5. 0 %、 (一) ーェピカテキン 0. 5〜 1 0. 0 %、 (一) ーェピカテキンガレー ト 0. 3〜 8. 0 %、 (一) ーェピガロカテ キン 2. 0〜 1 8. 0 %、 (一) ーェピガロカテキンガレー ト 3. 0〜 2 1 . 0 %、 遊離型テアフラビン 0〜 2 0. 0 %、 テアフラビ ンモノガレー ト A O〜 5. 0 %、 テアフラビンモノガレー ト B 0〜 5. 0 %及びテアフラビンジガレ一 ト 0〜 5. 0 %である。  The animal feed additive of the present invention refers to one containing a polyphenol compound derived from the tea extract thus obtained, and the content of the polyphenol compound contained in the animal feed additive of the present invention. Is usually 5 to 80%, preferably 10 to 50%, and more preferably 20 to 35%. Here, the content of each polyphenol compound in the animal feed additive is as follows: (+)-0.2-6.5% of force techin, (+)-2.0-18.0% of garoka techin, (1) -Gallocatechin gallate 1.0 to 15.0%, (1) -Epicatechin 0.5 to 10.0%, (1) -Epicatechin gallate 0.3 to 8.0%, (1) -epigallocatechin 2.0 to 18.0%, (1) -epigallocatechin gallate 3.0 to 21.0%, free theaflavin 0 to 20.0%, theaflavin monogallate AO 5.5.0%, theaflavin monogallate B 0 5.5.0% and theaflavin digallate 0〜5.0%.
このポリ フユノール化合物は、 茶の成分として多量に含まれてい ることや、 う蝕、 高脂血症、 ガン予防等に効果があることから、 こ れらに関与する疾病予防のために数多くの食品に既に使用されてい るため、 その安全性は非常に高い。  This polyfunol compound is contained in a large amount as a component of tea, and is effective in preventing caries, hyperlipidemia, cancer, and the like. Its safety is very high because it is already used in food.
本発明の動物用飼料添加物は、 動物用飼料に添加することにより 、 ポリ フエノール化合物を含有する飼料として家畜、 家禽及び愛玩 動物に投与される。  The animal feed additive of the present invention is administered to livestock, poultry, and companion animals as a feed containing the polyphenol compound by being added to the animal feed.
飼料とは、 栄養素の供給を目的として家畜に給与するものであり 、 一般に粗飼料、 濃厚飼料、 特殊飼料に分類される。 本願発明の飼 料添加物の添加されるべき飼料としては、 特に限定されるものでは ないが、 一般に高タンパク質、 高エネルギーの濃厚飼料を家畜に多 量に与えた場合、 特に下痢が発生しやすいため、 このような濃厚飼 料に本願発明の飼料添加物を添加することが望ましい。 このことは 本願発明の飼料添加物はポリ フ ノ一ル化合物を含有するものであ り、 これらの化合物は抗酸化作用を有することも知られており、 脂 質成分の抗酸化作用の観点からも濃厚飼料に添加されることが望ま しい。 本願発明の飼料添加物を飼料に添加する場合、 予め飼料配合 時に飼料の成分と共に一緒に配合してもよく、 家畜への飼料給与時 に一緒に与えても特に問題はなく、 飼料への添加方法、 添加時期は 特に限定されない。 又、 生菌剤、 抗生物質等の他の飼料添加物と併 用しても特に問題はない。 Feed is provided to livestock for the purpose of supplying nutrients, and is generally classified into roughage, concentrated feed, and specialty feed. The feed to which the feed additive of the present invention is to be added is not particularly limited, but generally high-protein, high-energy concentrated feed is commonly used in livestock. When given in an amount, diarrhea is particularly likely to occur. Therefore, it is desirable to add the feed additive of the present invention to such a concentrated feed. This means that the feed additive of the present invention contains a polyphenol compound, and it is also known that these compounds have an antioxidant effect, and from the viewpoint of the antioxidant effect of fat components. Should also be added to concentrated feed. When the feed additive of the present invention is added to the feed, it may be added together with the ingredients of the feed at the time of mixing the feed in advance, or may be added together at the time of feeding the feed to livestock without any problem. The method and timing of addition are not particularly limited. There is no particular problem when used in combination with other feed additives such as probiotic agents and antibiotics.
本発明の動物用飼料に添加される動物用飼料添加物の添加量は、 特に限定されるものではなく、 適宜調整される。  The amount of the animal feed additive added to the animal feed of the present invention is not particularly limited, and is appropriately adjusted.
本明細書において、 家畜、 家禽とは、 例えばゥシ、 ブ夕、 ゥマ、 ャギ、 ヒッジ、 キツネ、 ミ ンク、 ニヮ ト リ、 七面鳥、 カモ、 ゥズラ 、 ホロホロ鳥、 ァヒル、 ノくリゲン、 ガチョウ、 ハ ト等の産業上飼育 する動物のことであり、 愛 ¾動物とは例えば犬、 猫等の個人の趣味 で飼育する動物のことである。  In the present specification, livestock and poultry are, for example, red sea lions, bushes, poma, goats, hidge, foxes, mink, nits, turkeys, ducks, zebras, guinea fowls, ducklings, and wild birds. , Geese, pigeons and other animals bred for industrial purposes, and beloved animals are animals bred for personal tastes such as dogs and cats.
本発明の動物用飼料添加物、 およびそれを含有してなる動物用飼 料は、 家畜、 家禽及び愛玩動物等の環境の急変及び飼料組成の変化 、 飼養管理の失宜などの影響により生じる非感染性下痢症、 あるい はウィルス若しく は細菌等の病原菌により誘発される感染性下痢症 を予防、 治療するのに有用である。  The animal feed additive of the present invention and the animal feed containing the same can be used for non-infection caused by sudden changes in the environment of livestock, poultry, companion animals, etc., changes in feed composition, and inaccuracies in breeding management. It is useful for preventing and treating sexually transmitted diarrhea or infectious diarrhea induced by pathogens such as viruses or bacteria.
本発明において非感染性下痢症とは、 家畜、 家禽及び愛玩動物に 与えられたス トレスが原因で発生するス ト レス性下痢、 又は感染性 微生物の関与していない原因不明の下痢のことを指し、 感染性下痢 症とは病原性細菌若しく はウィルスが原因で生じた下痢のことを指 す。 In the present invention, non-infectious diarrhea refers to stress-induced diarrhea caused by stress given to livestock, poultry, and companion animals, or unexplained diarrhea not involving infectious microorganisms. Infectious diarrhea refers to diarrhea caused by pathogenic bacteria or viruses. You.
本発明において非感染性下痢症を抑制する効果を得るための、 家 畜、 家禽並びに愛玩動物に対する投与量は、 ポリ フ ノール化合物 換算で通常 1 日当たり 0. 5〜 5 0 m g/体重 k g、 さらに好まし く は、 2〜 2 0 m 体重 k gである。 投与量が 0. 5 m gZ体重 k gより低い場合、 非感染性下痢症を抑制する効果が期待できず、 また、 5 0 m gZ体重 k gを越える場合、 ポリ フ ヱ ノール化合物の 有する抗菌作用のため家畜、 家禽及び愛玩動物に常在する消化管内 微生物に対しても影響を及ぼす。 尚、 (一) ーェピガロカテキンガ レー トを単離等して用いる場合は通常 1 日当たり 0. 0 3〜 5 m g ノ体重 k g、 さらに好ましくは 0. 1 〜 3 m g/体重 k gである。 In the present invention, the dose for livestock, poultry, and pets for obtaining the effect of suppressing non-infectious diarrhea is usually 0.5 to 50 mg / kg of body weight per day in terms of polyphenol compound. Preferably it is between 2 and 20 m body weight kg. When the dose is lower than 0.5 mgZ kg body weight, the effect of suppressing non-infectious diarrhea cannot be expected, and when the dose exceeds 50 mgZ kg body weight, the antibacterial action of the polyphenol compound is not expected. Therefore, it also has an effect on the gut microbes that exist in livestock, poultry and pets. In addition, (1) When iso-epigallocatechin gallate is used after isolation or the like, it is usually 0.03 to 5 mg / kg of body weight per day, more preferably 0.1 to 3 mg / kg of body weight. .
—方、 感染性下痢症を抑制する効果を期待するために、 特異的抗 体と併用して使用する場合の投与量としては、 ポリ フエノール化合 物として通常、 1 日当たり、 0. 3〜 2 5 m g/体重 k g、 さらに 好ましく は 2〜 1 5 m gZ体重 k gである。 投与量が 0. 3 m 体重 k gより低い場合、 感染性下痢症を抑制する効果が期待できな い。 なお、 ポリ フエノール化合物は公定タンニン分析法 (茶研報 V 01.71, 43-74(1990)) により定量されるタンニンの量をもって表す ことができる。 On the other hand, in order to expect the effect of suppressing infectious diarrhea, the dose when used in combination with a specific antibody is usually 0.3 to 25 per day as a polyphenol compound. mg / kg body weight, more preferably 2 to 15 mg gZ body weight kg. If the dose is less than 0.3 m body weight kg, the effect of suppressing infectious diarrhea cannot be expected. The polyphenol compound can be represented by the amount of tannin determined by the official tannin analysis method (Chakenho V 01.71, 43-74 (1990)).
本発明における特異的抗体とは、 家畜、 家禽及び愛玩動物の感染 性微生物又はウィルス、 あるいはこれらの産生する毒素に対して特 異的に結合する抗体を言う。 抗体の種類としては、 家畜、 家禽及び 愛玩動物の感染性微生物又はウイルス、 あるいはこれらの産生する 毒素で過免疫された産卵鶏の卵より得られる鶏卵抗体、 並びに哺乳 類の乳汁より得られる乳汁抗体等を言う。 当該抗体純度については 限定されるものではない。 即ち、 抗体の純品であってもよいし、 鶏 卵抗体の場合は特に限定されないが、 これを含む全卵、 卵黄、 全卵 液、 卵黄液、 全卵粉末、 卵黄粉末若しく は卵黄の水溶性蛋白質画分 粉末であってもよい。 また、 乳汁抗体の場合も特に限定されないが これを含む全脂粉乳、 脱脂粉乳若しく は乳清蛋白質粉末であっても よい。 The specific antibody in the present invention refers to an antibody that specifically binds to infectious microorganisms or viruses of livestock, poultry and pets, or toxins produced by these. Examples of the type of antibody include egg antibodies obtained from eggs of laying hens hyperimmunized with infectious microorganisms or viruses of livestock, poultry and companion animals, or toxins produced by them, and milk antibodies obtained from milk of mammals Say etc. The antibody purity is not limited. That is, the antibody may be pure or chicken The egg antibody is not particularly limited, but may be whole egg, egg yolk, whole egg solution, egg yolk solution, whole egg powder, egg yolk powder or a powder of a water-soluble protein fraction of egg yolk containing the egg antibody. Also, the case of milk antibody is not particularly limited, but may be whole fat milk powder, skim milk powder or whey protein powder containing it.
家畜、 家禽及び愛玩動物の感染性微生物は特に限定されるもので はないが、 たとえばクロス ト リ ジゥム パーフ リ ンジエンス A型 、 クロス ト リ ジゥム パーフ リ ンジエンス C型、 サルモネラ テ ィ フィ ムリ ウム、 サルモネラ 夕ブリ ン、 カムピロノくクタ一 ジェ ジュ二一、 カムピロパクター コ リー、 大腸菌 0— 8 8、 大腸菌 0— 9 9、 大腸菌 9 8 7 P、 スタフイ ロコ ッカス ォゥレウス 、 ロタウィルス K K— 3、 口夕ウィルス N C D V等の病原性細 菌、 下痢症ウィルスがあげられる。 当該微生物又は当該微生物が産 生する毒素で過免疫される動物としては、 当該微生物又は毒素に対 する特異的抗体を産生できる動物であればよいが、 本発明の目的で ある家畜、 家禽及び愛玩動物の感染性微生物に対する抗体及びこの 抗体組成物の応用という実用的観点から考えると、 大量の特異的抗 体を産生できる産卵鶏又は牛、 ャギ、 羊等の哺乳類が特に望ましい 。 これらの中でも、 過免疫の作業性、 抗体産生能力さらに動物飼育 コス ト等の観点から、 産卵鶏を当該微生物若しく は当該微生物の産 生する毒素により免疫し、 この鶏卵より抗体を得る方法が最も好ま しい。  Infectious microorganisms in livestock, poultry, and pets are not particularly limited, but include, for example, Clostridium perfringens A, Clostridium perfringens C, Salmonella typhimurium, and Salmonella. Evening Blind, Campylopactor Jeju 21, Campylopactor Collie, E. coli 0-88, E. coli 0-99, E. coli 9887P, Staphylococcus oleus, Rotavirus KK-3, Oral virus NCDV Pathogenic bacteria and diarrhea virus. The animal that is hyperimmunized with the microorganism or the toxin produced by the microorganism may be any animal that can produce a specific antibody against the microorganism or the toxin. Livestock, poultry, and pets, which are the objects of the present invention, may be used. From the practical point of view of antibodies against infectious microorganisms in animals and the application of this antibody composition, laying hens or mammals such as cows, goats and sheep, which can produce large amounts of specific antibodies, are particularly desirable. Among them, a method of immunizing a laying hen with the microorganism or a toxin produced by the microorganism and obtaining an antibody from the hen egg from the viewpoints of hyperimmunity workability, antibody production ability, and animal breeding cost. Most preferred.
産卵鶏を過免疫する方法としては、 家畜、 家禽及び愛玩動物の上 記微生物又はウィルス、 あるいはこれらが産生する毒素を抗原とし て産卵鶏に繰り返し免疫することにより、 鶏卵中に特異的抗体を増 加させるとよい。 また、 哺乳類を過免疫する方法としては、 牛、 ャ ギ、 羊等の動物に当該抗原を繰り返し免疫することにより、 乳汁中 に特異的抗体を増加させるとよい。 One method of hyperimmunizing laying hens is to raise specific antibodies in hen eggs by repeatedly immunizing laying hens with the above-mentioned microorganisms or viruses from livestock, poultry and pets, or toxins produced by them. It is good to add. Methods for hyperimmunizing mammals include cattle and cattle. It is advisable to increase the amount of specific antibodies in milk by repeatedly immunizing animals such as giants and sheep with the antigen.
この場合用いられる抗原の調製については、 公知の方法で行えば よい。 例えば、 家畜、 家禽及び愛玩動物の感染性微生物を大量培養 した後、 公知の方法により弱毒化又は不活化を行い、 抗原として使 用すれば良い。  The antigen used in this case may be prepared by a known method. For example, after infectious microorganisms of livestock, poultry, and companion animals are cultured in large amounts, attenuated or inactivated by a known method, and used as an antigen.
抗原を産卵鶏又は哺乳類に免疫する方法としては、 筋肉注射、 皮 下注射、 静脈注射、 腹腔内注射又は飲水による経口免疫等、 いずれ の方法でも良い。  As a method for immunizing a laying hen or mammal with an antigen, any method such as intramuscular injection, subcutaneous injection, intravenous injection, intraperitoneal injection or oral immunization by drinking water may be used.
抗原の免疫は、 鶏卵中又は乳汁中に現れる特異的抗体価を酵素免 疫測定法などの方法で調べながら、 該抗体価が最大値になるまで繰 り返し実施される。 なお、 該抗体価は、 適当な間隔で産卵鶏若しく は哺乳類に抗原を繰り返し免疫することにより、 産卵期間又は初乳 分泌期間を通してある一定レベル以上の特異的抗体量を維持するこ とができる。  The immunization of the antigen is repeated until the antibody titer reaches the maximum value, while examining the specific antibody titer appearing in chicken eggs or milk by a method such as enzyme immunoassay. The antibody titer can be maintained at a certain level or more throughout the laying period or the period of colostrum secretion by repeatedly immunizing a laying hen or mammal with an antigen at appropriate intervals. .
免疫される抗原の量は、 被免疫動物の種類、 抗原の種類等により 異なるため、 適時予備試験等により選択する必要があるが、 産卵鶏 への免疫では、 一般的には、 例えばウィルスの場合、 ウィルス量と して 1 0 g〜 1 m g Z羽 Z回の抗原量が選択される。  The amount of antigen to be immunized depends on the type of animal to be immunized, the type of antigen, etc., so it is necessary to select it in a timely preliminary test, etc. As the virus amount, 10 g to 1 mg Z feathers The antigen amount of Z times is selected.
特異的抗体を調製は、 家畜、 家禽及び愛玩動物の感染症微生物若 しく はこれらの産生する毒素を抗原として産卵鶏若しく は哺乳類に 免疫した後、 該抗原に対する特異的抗体を含有する鶏卵又は乳汁を 集めることにより行なうことができる。  A specific antibody is prepared by immunizing a laying hen or mammal with an infectious disease microorganism of livestock, poultry, and companion animals or a toxin produced thereby as an antigen, and then hen eggs or chicken eggs containing a specific antibody against the antigen. This can be done by collecting milk.
鶏卵を用いる場合、 割卵後、 全卵液若しく は卵黄液を分離し、 ホ モジナイザー等で均質化した後、 殺菌し熱風乾燥又は凍結乾燥によ り用いた抗原に対する特異的抗体含有全卵粉末又は卵黄粉末を得る ことができる。 また、 該卵黄液又は卵黄粉末より公知の鶏卵抗体精 製法 (特開昭 6 4 - 3 8 0 9 8号、 Ag r i c. B i o l . C h e m. 、 V o l . 5 4、 N o. 1 0, 2 5 3 1 - 2 5 3 5 ( 1 9 9 0 ) 等) により、 用いた抗原に対する特異的抗体純度を高めた卵黄水 溶性蛋白質粉末又は特異的抗体の純品粉末等が調製できる。 When using chicken eggs, whole eggs or egg yolks are separated after egg breaking, homogenized with a homogenizer, etc., sterilized, and whole eggs containing a specific antibody against the antigen used by hot air drying or freeze drying are used. Obtain powder or egg yolk powder be able to. Further, a known method for purifying a chicken egg antibody from the egg yolk solution or yolk powder (Japanese Patent Application Laid-Open No. 1988-1988, Agric. Biol. Chem., Vol. 54, No. 1 0, 25 3 1-25 35 (199 9 0) etc.) can be used to prepare a yolk water-soluble protein powder or a pure antibody specific antibody powder with enhanced specific antibody purity for the antigen used. .
乳汁を用いる場合、 該乳汁若しく は該乳汁中の脂質成分をク リ一 ムセパレーター等で分離した脱脂乳を殺菌後、 熱風乾燥又は凍結乾 燥することにより用いた抗原に対する特異的抗体を含有する全脂粉 乳又は脱脂粉乳が得られる。 また該乳汁若しく は該脱脂乳より、 公 知の方法で、 用いた抗原に対する特異的抗体純度を高めたホェ一夕 ンパク質粉末又は特異的抗体の純品等も調製できる。  When milk is used, it contains a specific antibody to the antigen used by sterilizing the milk or skim milk obtained by separating lipid components in the milk with a cream separator and then drying with hot air or freeze-drying. A whole milk powder or skim milk powder is obtained. From the milk or the skim milk, whey protein powder having a higher specific antibody purity to the used antigen or a pure product of the specific antibody can be prepared by a known method.
本発明においてポリ フニノール化合物と併用する特異的抗体の配 合量は、 感染性下痢症の抑制効果を期待するためには、 酵素免疫測 定法でブランクに対し 1. 5倍以上の抗体価を有したものを 1 mg ノ体重 k g以上、 さらに好ましく は 5 m gZ体重 k g以上の投与量 になるよう調製するのが好ましい。 11113 /体重1^ gより少ない投 与量の場合、 期待する効果が得られず好ましくない。  In the present invention, in order to expect an effect of suppressing infectious diarrhea, the amount of the specific antibody to be used in combination with the polyfuninol compound in the present invention has an antibody titer of at least 1.5 times that of the blank in an enzyme immunoassay. It is preferable to adjust the dose to 1 mg or more, and more preferably 5 mgZ kg or more. If the dose is less than 11113 / body weight 1 ^ g, the expected effect cannot be obtained, which is not preferable.
本発明はすべての家畜、 家禽及び愛玩動物に適用することができ るが、 効果若しく は嗜好性の面から草食動物はポリ フエノール化合 物の苦みに関係なく容易に摂取することができるため、 特に草食動 物への適用が好ま しく、 特に産業上有用である乳牛、 肉牛等の牛、 山羊、 めん羊、 鹿等が好ましい。  Although the present invention can be applied to all livestock, poultry and pets, it can be easily consumed by herbivores irrespective of the bitterness of the polyphenol compound from the viewpoint of effect or palatability. Particularly, application to herbivorous animals is preferable, and cows such as dairy cows and beef cattle, goats, sheep, deer and the like, which are particularly useful in industry, are preferable.
また、 本発明の動物用飼料添加物、 およびそれを含有してなる動 物用飼料は、 家畜の生産する生乳の乳質を改善し、 乳量を増加させ るのに有用である。  In addition, the animal feed additive of the present invention and the animal feed containing the same are useful for improving the milk quality of raw milk produced by livestock and increasing the amount of milk.
本発明において家畜の生乳の乳質改善作用、 乳量増加作用の効果 を得るための、 家畜に対する投与量は、 ボリ フ ノール化合物換算 で通常 1 日当たり 0. 5〜 5 0 mgZ体重 k g、 さらに好ましく は 、 2〜 2 0 m g/体重 k gである。 投与量が 0. S m gZ体重 k g より低い場合、 本発明の効果が期待できず、 また、 5 O m gZ体重 k gを越える場合、 ポリ フエノール化合物の有する抗菌作用のため 家畜、 家禽に常在する消化管内微生物に対して悪影響を及ぼす。 ボ リ フエノール化合物は前記の公定タンニン分析法により定量される タンニンの量をもって表すことができる。 尚、 (一) ーェピガロカ テキンガレー トを単離等して用いる場合は通常 1 日当たり 0. 0 3 〜 5 mg/体重 k g、 さらに好ましく は 0. l〜 3 mg/体重 k g 、め 。 Effect of improving milk quality and increasing milk yield of raw milk of livestock in the present invention Dosage for livestock is usually 0.5 to 50 mg / kg body weight per day, more preferably 2 to 20 mg / kg body weight, in terms of polyphenol compound. When the dose is lower than 0. S mgZ body weight kg, the effect of the present invention cannot be expected. When the dose exceeds 5 O mgZ body weight kg, it is resident in livestock and poultry due to the antibacterial action of the polyphenol compound. Adversely affect the digestive tract microbes. The polyphenol compound can be represented by the amount of tannin determined by the official tannin analysis method described above. In addition, (1) When iso-epigallocatechin gallate is used by isolation or the like, it is usually 0.03 to 5 mg / kg of body weight per day, more preferably 0.1 to 3 mg / kg of body weight.
本発明においてボリ フ ノール化合物を投与する家畜は、 乳牛、 山羊、 馬、 めん羊等の生乳生産のために使用されるいずれの動物で も良く、 その種類は特に限定されないが、 乳業産業上、 さらにボリ フエノ一ル化合物の有する苦みのため嗜好性等から考慮すると乳牛 に用いるのが特に好ましい。.  The livestock to which the polyphenol compound is administered in the present invention may be any animal used for raw milk production, such as dairy cows, goats, horses, and sheep, and the type thereof is not particularly limited. Furthermore, it is particularly preferable to use dairy cows in consideration of palatability and the like due to the bitterness of the polyphenol compound. .
さらに、 本発明の動物用飼料添加物及びこれを配合してなる動物 用飼料は、 家畜、 家禽の繁殖率の向上にも効果がある。  Furthermore, the animal feed additive of the present invention and the animal feed prepared by blending the same are effective in improving the reproduction rate of livestock and poultry.
当該繁殖率の向上効果を得るための投与量は、 ボリ フエノール化 合物換算で通常 1 日あたり 0. 5〜5 0 mgZ体重 k g、 さらに好 ま しく は 2〜2 O m gノ体重 k gである。 投与量がこの範囲より低 い場合、 本発明の効果が期待できず、 この範囲を越えると、 ポリ フ ェノール化合物のもつ抗菌作用によって、 消化管に常在する微生物 に悪影響をおよぼす。 ポリ フエノール化合物の定量は前述の公定夕 ンニン分析法を用いる。 尚、 (一) ーェピガロカテキンガレ一 トを 単離等して用いる場合は通常 1 日当たり 0. 0 3〜5 mgZ体重 k g、 さらに好ましく は 0 . 1 〜 3 m g /体重 k gである。 The dose for obtaining the effect of improving the reproductive rate is usually 0.5 to 50 mg / kg body weight per day, more preferably 2 to 2 mg / kg body weight in terms of polyphenol compound. . If the dose is lower than this range, the effects of the present invention cannot be expected. If the dose exceeds this range, the antimicrobial action of the polyphenol compound will adversely affect microorganisms resident in the gastrointestinal tract. For the quantification of the polyphenol compound, use the official Yunnin analysis method described above. (1) When iso-epigallocatechin galleate is used after isolation, it is usually 0.03 to 5 mgZ body weight per day. g, more preferably 0.1 to 3 mg / kg of body weight.
本発明においてポリ フエノール化合物を投与する家畜、 家禽の種 類は特に限定されないが、 産業上の観点から乳牛に用いるのが好ま しい。 また乳牛を対象とした場合、 繁殖率の評価は分娩後受胎に要 する期間と妊娠の確認された乳牛の人工受精回数を測定することに より行う ことができる。  In the present invention, the type of livestock and poultry to which the polyphenol compound is administered is not particularly limited, but is preferably used for dairy cows from an industrial viewpoint. For dairy cows, the reproductive rate can be evaluated by measuring the period required for postpartum conception and the number of artificial inseminations of dairy cows that have been confirmed to be pregnant.
次に、 本発明の他の態様として、 本発明は茶葉繊維を有効成分と して含有する飼料添加物及び動物用飼料を提供するものである。 茶葉織維を含有する飼料添加物の添加された動物用飼料を家畜、 家禽等の動物に与えた場合、 動物の腸内細菌叢が改善されることに より整腸作用を促進し、 下痢発症を低減し、 搾乳量を増加させ、 飼 料効率を改善し、 さらに糞便、 尿の悪臭が低減 · 消去される。 これ は茶葉繊維であるセルロースと抽出残渣である茶葉繊維中に残存す るボリ フユノール化合物の併用効果により、 消化管内における常在 微生物のうちビフィ ズス菌、 乳酸菌といった有用菌の菌数を上昇さ せ、 病原性大腸菌、 ウエルシュ菌といった有害菌の菌数が低下する こととなり、 上記効果が達成されるものである。 茶葉繊維中に残存 するポリ フ ノール化合物は、 通常 2〜 5 %程度含まれる。  Next, as another embodiment of the present invention, the present invention provides a feed additive and an animal feed containing tea leaf fiber as an active ingredient. When animals such as livestock and poultry are fed animal feeds to which feed additives containing tea leaf fiber are added, the intestinal microflora of animals is improved, thereby promoting intestinal regulation and causing diarrhea. It reduces milk production, increases milk yield, improves feeding efficiency, and reduces and eliminates stool and urine odors. This increases the number of useful microorganisms such as bifidobacteria and lactic acid bacteria among the indigenous microorganisms in the gastrointestinal tract due to the combined effect of cellulose, which is tea leaf fiber, and the borofuynol compound remaining in tea leaf fiber, which is extraction residue. However, the number of harmful bacteria such as pathogenic Escherichia coli and Welsh bacteria is reduced, and the above effects are achieved. The polyphenol compound remaining in the tea leaf fiber usually contains about 2 to 5%.
本発明に用いられる茶葉繊維は、 粉砕した茶葉を 5〜 2 0倍量の 水、 アルコール、 アセ ト ン又はこれらの混合液を用いて抽出した可 溶性成分を除去した抽出残渣として得ることができる。 ここでアル コールとは特に限定されるものではないが、 メタノール、 エタノー ル、 プロパノール、 イソプロパノール、 ブ夕ノール等が好ましい。 原料に用いる茶葉は生葉から仕上げ茶 (乾燥茶) まで、 通常の製造 工程のいずれの段階のものでも良く、 かつ発酵の程度に関係なく不 発酵茶、 半発酵茶、 発酵茶いずれでも使用できる。 抽出時、 加熱す ることは特に問題はなく、 水抽出の場合は、 むしろ 6 0 °C以上、 好 ましく は 7 5〜 9 0でで 0. 5〜7 h r加熱するのが望ましい。 通 常、 茶葉 1 O k gを 5 0〜2 0 0 k gの水で抽出した場合、 抽出残 渣 2 0〜4 0 k gと加水量に応じて B r i x 2〜 l 0 'のエキス 4 0 〜 1 6 0 k gが得られる。 得られた抽出残渣は、 通常用いられる方 法、 例えば熱風乾燥、 凍結乾燥などにより乾燥後、 粉砕し飼料に混 合することができる。 このようにして茶葉 1 O k gから茶葉繊維 6 〜 8 k gが得られる。 得られる茶葉織維は原料の茶葉と比較して力 フェイン、 茶ボリ フヱノール化合物の約 7 0 %以上が除去されるた め、 苦み、 渋みが低減し味覚的に改善され、 飼料添加物及び動物用 飼料としてより好適となる。 The tea leaf fiber used in the present invention can be obtained as an extraction residue obtained by removing a soluble component obtained by extracting ground tea leaves with 5 to 20 times the amount of water, alcohol, acetate or a mixture thereof. . Here, the alcohol is not particularly limited, but methanol, ethanol, propanol, isopropanol, butanol, and the like are preferable. The tea leaves used as raw materials can be from any stage of the normal manufacturing process, from fresh leaves to finished tea (dried tea), and can be used as unfermented tea, semi-fermented tea, or fermented tea regardless of the degree of fermentation. Heat during extraction There is no particular problem. In the case of water extraction, heating at 60 ° C or more, preferably at 75 to 90, for 0.5 to 7 hours is desirable. Normally, when 1 O kg of tea leaves is extracted with 50 to 200 kg of water, the extraction residue is 20 to 40 kg and the extract of Brix 2 to 10 'depending on the amount of water is 40 to 1 60 kg is obtained. The obtained extraction residue can be dried by a commonly used method, for example, hot-air drying, freeze-drying, or the like, and then pulverized and mixed with feed. Thus, 6 to 8 kg of tea leaf fiber is obtained from 1 O kg of tea leaf. The resulting tea leaf fiber has about 70% or more of the power-fein and tea polyphenol compounds removed from the raw tea leaves, so that bitterness and astringency are reduced and taste is improved. It becomes more suitable as feed for use.
ここで本発明における茶葉繊維の製造は、 前記のボリ フユノール 化合物の製造方法と組み合わせることが可能であり、 これによりそ れぞれの製造方法の生産性が高められることも本発明の特徴の一^つ である。  Here, the production of tea leaf fiber in the present invention can be combined with the above-mentioned method for producing a polyfunor compound, and thereby, the productivity of each production method can be enhanced. There are two.
家畜飼料に添加する茶葉織維 (茶葉織維中に残存するポリ フニノ —ル化合物を含む。 ) の量は通常重量で 0. 0 1 %以上とすること が望ましい。 茶葉織維の飼料への添加方法に関しては特に制限はな く、 それぞれの製造工程中適宜添加することができ、 経口的に投与 する方法であればいずれの方法でもよい。  Normally, it is desirable that the amount of the tea leaf fiber (including the polyfuninol compound remaining in the tea leaf fiber) to be added to livestock feed is 0.01% or more by weight. There is no particular limitation on the method of adding the tea leaf fiber to the feed, and it can be appropriately added during each production process, and any method may be used as long as it is an orally administered method.
以下、 実施例及び試験例により本発明をさらに詳しく説明するが 、 本発明はこれらの実施例等によりなんら限定されるものではない o  Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples, but the present invention is not limited to these Examples and the like.
実施例 1 Example 1
綠茶 1 k gに約 1 5 リ ッ トルの水を加え攪拌し、 8 0 で 3時間 抽出した。 濾過により得られる抽出液を噴霧乾燥し、 純度 2 5 %の ポリ フエノール化合物を含む緑茶の熱水抽出物 3 5 0 gを得た。 得 られたポリ フヱノール化合物の成分組成は、 (+ ) —力テキン 1 . 2 %、 ( + ) —ガロカテキン 5. 0 %、 (一) ーガロカテキンガレ ー ト 3. 9 %、 (一) ーェピカテキン 2. 3 %、 (一) ーェピカテ キンガレー ト 1 . 5 %、 (一) ーェピガロカテキン 5. 0 %及び ( 一) ーェピガロカテキンガレー ト 6. 1 %であった。 抽出物中のポ リ フ ノール化合物の含量は前述の公定夕ンニン分析法によって定 量し、 各成分組成は H P L C分析により求めた。 高速液体クロマ ト グラフ装置は日立製作所製の 6 5 5 A— 1 1 L Cを使用し、 カラ ム : J ' s p h e r e O D S— M 8 0 ( (株) ヮイエムシ一製) 、 溶媒 : 2 0 mM KH2 P 04 — H3 P 04 ( p H 2. 4 ) ノメ タノール ( 7 5 : 2 5、 vZv:) 、 流速 : 0. 8 m l /m i n、 検 出 : 2 8 0 n mの UV、 の諸条件で H P L C分析を行った。 以下の 実施例において、 ポリ フエノール化合物の含量及び各成分組成は上 記の方法で求めたものである。 綠 Approximately 15 liters of water was added to 1 kg of tea, stirred, and extracted with 80 for 3 hours. The extract obtained by filtration is spray-dried to a purity of 25%. 350 g of a hot water extract of green tea containing a polyphenol compound was obtained. The component composition of the obtained polyphenol compound is (+)-1.2% of kinetechin, (+)-5.0% of gallocatechin, (1) -gallocatechin gallate 3.9%, (1) -eppicatechin 2.3%, (1) -epigallocatechin gallate 1.5%, (1) -epigallocatechin gallate 5.0% and (1) -epigallocatechin gallate 6.1%. The content of the polyphenol compound in the extract was determined by the aforementioned official Yunnin analysis method, and the composition of each component was determined by HPLC analysis. High performance liquid chroma DOO graph device uses 6 5 5 A- 1 1 LC manufactured by Hitachi, column: J 'sphere ODS- M 8 0 (( Ltd.) Waiemushi Ltd. I), solvent: 2 0 mM KH 2 P 0 4 - H 3 P 0 4 (p H 2. 4) drink ethanol (7 5: 2 5, vZv :), flow rate: 0. 8 ml / min, detection: 2 8 0 nm of UV, of various HPLC analysis was performed under the conditions. In the following examples, the content of the polyphenol compound and the composition of each component were determined by the methods described above.
実施例 2 Example 2
綠茶 2 0 0 gを 8 5 の熱水 4 リ ッ トルで 3 0分攪拌しながら抽 出し、 茶葉を濾過により除き 2. 5 リ ッ トルの抽出液を得た。 この 液を限外濾過装置 (D D S社製、 膜タイプ G R - 8 1 P P、 分画分 子量 6 0 0 0 ) を用いて通過液 2 リ ッ トルを得た。 濃縮残液に水 1 リ ッ トルを加え同様に操作し、 通過液 1 . 2 リ ッ トルを得た。 両液 を合わせ逆浸透膜 (D D S社製、 膜タイプ H C— 5 0 ) により濃縮 し 2 0 0 ミ リ リ ッ トルとした後、 凍結乾燥し、 純度 2 9 %のポリ フ エノ一ル化合物を含む茶の熱水抽出物 4 8. 6 gを得た。 得られた ボリ フヱノール化合物の成分組成は、 (+ ) —力テキン 1. 4 %、 (+ ) —ガロカテキン 5. 8 %、 (一) ーガロカテキンガレー ト 4 . 5 (一) ーェピカテキン 2. 7 %、 (一) 一ェピカテキンガ レー ト 1. 8 %、 (一) ーェピガロカテキン 5. 8 %及び (一) 一 ェピガロカテキンガレー ト 7. 0 %であった。 2200 g of tea was extracted with 4 liters of 85 hot water while stirring for 30 minutes, and the tea leaves were removed by filtration to obtain a 2.5 liter extract. This solution was subjected to ultrafiltration (DDS, membrane type GR-81 PP, fractional molecular weight: 600) to obtain 2 liters of the passing solution. One liter of water was added to the concentrated residue, and the same operation was performed to obtain 1.2 liters of the passing solution. The two solutions were combined, concentrated using a reverse osmosis membrane (DDS, membrane type HC-50) to give 200 milliliters, and then lyophilized to obtain a 29% pure polyphenol compound. 48.6 g of a hot water extract of tea was obtained. The component composition of the obtained polyphenol compound is as follows: (+) — 1.4% strength, (+) — 5.8% gallocatechin, (1) -gallocatechin gallate 4 .5 (1) Epicatechin gallate 2.7%, (I) Epicatechin gallate 1.8%, (I) Epigallocatechin 5.8% and (I) Epigallocatechin gallate 7.0% Met.
実施例 3 Example 3
実施例 1で得られた熱水抽出物 3 5 0 gに水 8 リ ッ トルを加え溶 解後、 へキサン及びクロ口ホルムで順次分配した。 分配後の水層に 酢酸ェチル 1 0 リ ツ トルを加えて激しく攪拌、 静置後、 酢酸ェチル 層を分離し、 酢酸ェチルを留去後、 乾燥し、 酢酸ェチル可溶画分 7 0 gを得た (ボリ フヱノール化合物の混合物として純度 7 4. 5 % ) o  To 350 g of the hot water extract obtained in Example 1, 8 liters of water was added and dissolved, and the mixture was sequentially partitioned with hexane and chloroform. Add 10 liters of ethyl acetate to the aqueous layer after partitioning, vigorously stir, and allow to stand. Separate the ethyl acetate layer, evaporate the ethyl acetate, dry, and dry 70 g of the ethyl acetate-soluble fraction. Obtained (purity 74.5% as a mixture of polyphenol compounds) o
本酢酸ェチル可溶画分の全ボリ フ ノール化合物の含量は 7 4. 5 %であり、 各ポリ フヱノール化合物の割合は (+ ) —力テキン 3 . 5 %、 ( + ) —ガロカテキン 1 4. 8 %、 (一) ーガロカテキン ガレー ト 1 1. 6 %、 (一) ーェピカテキン 7 %、 (一) 一ェピカ テキンガレー ト 4. 6 %、 (一) ーェピガロカテキン 1 5. 0 %及 ぴ (一) ーェピガロカテキンガレー ト 1 8. 0 %であった。  The content of all polyphenol compounds in the ethyl acetate-soluble fraction was 74.5%, and the ratio of each polyphenol compound was (+)-3.5%, (+)-gallocatechin 14. 8%, (1) -gallocatechin gallate 11.6%, (1) -epicatechin 7%, (1) 1-epicatechin gallate 4.6%, (1) -epigallocatechin 15.0% and (1) 1) Yepigallocatechin gallate 18.0%.
実施例 4 Example 4
牛の感染性下痢症の原因菌として知られているサルモネラ タブ リ ン、 大腸菌 0 - 8 8、 大腸菌 0— 9 9及び大腸菌 9 8 7 P をそれぞれブレインハー トインフュージョ ン培地で培養した。 また 、 牛ロタウィルス KK一 3、 牛ロタウィルス N C D Vをそれぞれィ ーグル MEM培地で培養したァカゲザル由来の M A 1 0 細胞を宿 主として培養した。 それぞれの細菌又はウィルスを該抗原として、 産卵鶏に過免疫した。 この産卵鶏が産する鶏卵卵黄 1 0 k gより該 菌に対し、 特異的な鶏卵抗体 4 5 gをそれぞれ得た。  Salmonella tabulin, Escherichia coli 0-88, Escherichia coli 0-99, and Escherichia coli 988P, which are known as causative bacteria of infectious diarrhea in cattle, were cultured in brain heart infusion medium. In addition, MA10 cells derived from rhesus monkeys cultured on bovine rotavirus KK-13 and bovine rotavirus NCDV, respectively, in an Eagle MEM medium were cultured mainly. Laying hens were hyperimmunized with the respective bacteria or viruses as the antigen. From 10 kg of hen egg yolk produced by the laying hen, 45 g of a specific hen egg antibody against the bacterium was obtained.
実施例 5 実施例 3で得られた酢酸ェチル可溶画分 1 0 gをシリカゲルカラ ム ( 5 X 8 0 c m) に付し、 クロ口ホルム一メタノール混液で溶出 した。 溶出は 2段階に分けて行った。 すなわち、 最初に 2 0 : 1 の 混合比 ( vZv ) の当該混液で溶出し、 次いで 1 0 : 1 の混合比 ( v/v) の当該混液で溶出した。 得られた各ボリ フエノール化合物 をリサイクル H P L C (日本分折工業製 L C一 9 0 8、 G S— 3 2 0カラム、 溶媒はメタノールを使用) によってさらに分離精製し、 より高純度のボリ フエノール化合物を得た。 得られたボリ フエノー ル化合物は、 (+ ) —力テキン 0. 3 g、 ( + ) —ガロカテキン 1 . 2 2 g、 (一) ーガロカテキンガレー ト 0. 9 g、 (一) ーェピ カテキン 0. 5 g、 (一) ーェピカテキンガレー ト 0. 3 8 g、 ( 一) ーェピガロカテキン 1. 2 g及び (一) ーェピガロカテキンガ レー ト 1 . 5 gである。 Example 5 10 g of the ethyl acetate-soluble fraction obtained in Example 3 was applied to a silica gel column (5 × 80 cm), and eluted with a mixed solution of chloroform and methanol in a mouth. Elution was performed in two steps. That is, elution was first carried out with the mixture at a mixture ratio of 20: 1 (vZv), and then with the mixture at a mixture ratio of 10: 1 (v / v). Each of the obtained polyphenol compounds is further separated and purified by recycling HPLC (LC-908, GS-320 column, manufactured by Nippon Bunka Kogyo Co., Ltd., using methanol as the solvent) to obtain higher purity polyphenol compounds. Was. The obtained polyphenol compound contained (+)-0.3 g of force techin, 1.2 g of (+)-gallocatechin, 0.9 g of (-1) -gallocatechin gallate, 0.9 g of (-1) -epeicatechin 0 5 g, (1) -epigallocatechin gallate 0.38 g, (1) -epigallocatechin gallate 1.2 g and (1) -epigallocatechin gallate 1.5 g.
試験例 1 Test example 1
3 0頭の子牛を 1 0頭ずつ 3群に分け、 人工乳のみを与えた群を A群、 実施例 1 で調製した茶抽出物を 1 日当たり 0. 7 g投与した 群を B群、 実施例 1 で調製した茶抽出物を 1 日当たり 1 . 5 g投与 した群を C群として 5週間飼育した。 茶抽出物は人工乳と混合して 投与した。 なお、 水は自由に摂取させた。 人工乳は全酪連製のカー フ トップを用いた。 以下の試験例においても、 人工乳はカーフ トツ プを用いた。 非感染性下痢症を示す指標として、 糞便スコアと腸内 細菌叢を試験開始 0週〜 5週まで測定した。 なお、 糞便スコアとは 糞便の性状を数値化したものであり、 正常便を 0、 軟便を 1、 泥様 便を 2、 水様便を 3 とする。 この結果をそれぞれ表 1 と表 2に示し た。 表 1 The 30 calves were divided into 3 groups of 10 calves, and the group that received only the artificial milk was Group A, the group that received 0.7 g of the tea extract prepared in Example 1 per day was Group B, The group to which 1.5 g of the tea extract prepared in Example 1 was administered per day was bred as Group C for 5 weeks. The tea extract was administered after being mixed with the artificial milk. Water was provided ad libitum. The artificial milk used was a calf top made by All Dairy. In the following test examples, calf tops were used as the artificial milk. Fecal scores and intestinal flora were measured from 0 to 5 weeks from the start of the test as indices indicating non-infectious diarrhea. The stool score is a quantification of the stool properties, with normal stool being 0, loose stool being 1, muddy stool being 2, and watery stool being 3. The results are shown in Tables 1 and 2, respectively. table 1
糞便スコア  Fecal score
Figure imgf000027_0001
Figure imgf000027_0001
数値は、 1 0頭の平均を示し、 正常便を 0、 軟便を 1 泥様便を 2、 水様便を 3 とした。 The numerical values indicate the average of 10 animals, with 0 for normal stool, 1 for loose stool, 2 for muddy stool, and 3 for watery stool.
表 2 Table 2
腸内細菌叢 Intestinal flora
Figure imgf000028_0001
Figure imgf000028_0001
数値は、 1 0頭の平均を求め、 糞便 1 g当たりの対数値で示した 表 1 と表 2より、 C群の子牛は A群及び B群の子牛と比較して、 糞便スコアの低下、 ウエルシュ菌、 大腸菌群の減少、 ビフィ ズス菌 、 乳酸菌の菌数の減少割合の抑制が認められた。 従って、 本発明の 茶抽出物は子牛の非感染性下痢を効果的に抑制することがわかった 。 また、 実施例 2及び実施例 3で得られた茶抽出物を用いて同様の 試験を行ったところ、 同様の結果が得られた。 さらに、 紅茶、 ウー ロン茶由来の茶抽出物を用いて同種の試験を行ったところ、 同様の 結果が得られた。 Values were averaged for 10 animals and expressed as logarithmic value per 1 g of stool From Tables 1 and 2, calves in group C had lower fecal scores, reduced numbers of Welsh and Escherichia coli groups, and decreased percentages of Bifidobacterium and lactic acid bacteria compared to groups A and B calves. Suppression was observed. Therefore, it was found that the tea extract of the present invention effectively suppressed non-infectious diarrhea in calves. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, similar tests were performed using tea extracts derived from black tea and oolong tea, and similar results were obtained.
試験例 2 Test example 2
1 5頭の分娩直後の成牛を 5頭ずつ 3群に分け、 表 3の基本飼料 を与えた群を D群、 実施例 1 で調製した茶抽出物を 1 日当たり 1 0 g投与した群を E群、 実施例 1 で調製した茶抽出物を 1 日当たり 1 5 g投与した群を F群とし 5週間飼育した。 茶抽出物は表 3の基本 飼料に混合したものを投与した。 なお、 水は自由に摂取させた。 そ して、 非感染性下痢症を示す指標として試験例 1 と同様に糞便スコ ァと腸内細菌叢を試験開始 0.週〜 5週まで測定した。 この結果をそ れぞれ表 4 と表 5に示した。 (1) Five adult calves immediately after calving were divided into three groups of five, and the group that received the basic diet shown in Table 3 was group D, and the group that received the tea extract prepared in Example 1 was 10 g per day. Group E, a group to which 15 g of the tea extract prepared in Example 1 was administered per day was designated as Group F and bred for 5 weeks. The tea extract was administered as a mixture with the basic feed shown in Table 3. Water was provided ad libitum. Then, fecal score and intestinal flora were measured from 0 to 5 weeks from the start of the test as indices indicating the infectious diarrhea in the same manner as in Test Example 1. The results are shown in Tables 4 and 5, respectively.
表 3 Table 3
基本飼料の組成  Composition of basic feed
Figure imgf000030_0001
Figure imgf000030_0001
表 4 Table 4
糞便スコア Fecal score
Figure imgf000030_0002
Figure imgf000030_0002
数値は、 5頭の平均を示し、 正常便を 0、 軟便を 1 泥様便を 2、 水様便を 3 とした。 表 5 The numerical values are the average of 5 animals, with 0 for normal stool, 1 for loose stool, 2 for muddy stool, and 3 for watery stool. Table 5
腸内細菌叢 Intestinal flora
Figure imgf000031_0001
Figure imgf000031_0001
数値は、 5頭の平均を求め、 糞便 1 g当たりの対数値で示した, 表 4 と表 5 より、 F群の成牛は D群及び E群で飼育した成牛と比 較して、 糞便スコアの低下、 ウエルシュ菌、 大腸菌群の減少、 及び ビフィズス菌、 乳酸菌の増加が認められた。 従って、 本発明の茶抽 出物は、 分娩によって生じたス ト レス性 (非感染性) の下痢を効果 的に抑制することがわかった。 また、 実施例 2及び実施例 3で得ら れた茶抽出物を用いて同様の試験を行ったところ、 同様の結果が得 られた。 さらに、 紅茶、 ウーロン茶由来の茶抽出物を用いた同種の 試験を行ったところ、 同様の結果が得られた。 The values were averaged for 5 animals and expressed as a logarithmic value per g of feces. Tables 4 and 5 show that adult cats in group F had lower fecal scores, reduced numbers of Welsh and Escherichia coli, and increased numbers of bifidobacteria and lactic acid bacteria compared to adult cats reared in groups D and E. Admitted. Therefore, it was found that the tea extract of the present invention effectively suppressed stress (non-infectious) diarrhea caused by parturition. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, similar tests were performed using tea extracts derived from black tea and oolong tea, and similar results were obtained.
試験例 3 Test example 3
サルモネラ 夕ブリ ンで感染させた 3 0頭の子牛を 1 0頭ずつ 3 群に分け、 人工乳のみを与えた群を G群、 実施例 1 で調製した茶抽 出物を 1 日当たり 1 . 5 g投与した群を H群、 実施例 1で調製した 茶抽出物を 1 日当たり 1 . 5 g及び実施例 4で調製した特異的鶏卵 抗体を 1 日当たり 4 0 0 m g投与した群を I群、 実施例 4で調製し た特異的鶏卵抗体を 1 日当たり 4 0 O m g投与した群を H 2群とし て 5週間飼育した。 茶抽出物又は特異的鶏卵抗体は人工乳に混合し たものを投与した。 下痢症を示す指標として糞便スコアと糞便中の サルモネラ 夕ブリ ンの菌数を試験開始 0週〜 5週まで求めた。 こ の結果をそれぞれ表 6 と表 7に示した。 30 calves infected with Salmonella Evenin were divided into 3 groups of 10 calves, the group that received only the artificial milk was Group G, and the tea extract prepared in Example 1 was used at 1.0 mg / day. Group H received 5 g, and group I received 1.5 g of the tea extract prepared in Example 1 per day and 400 mg of the specific chicken egg antibody prepared in Example 4 per day. A group to which the specific egg antibody prepared in Example 4 was administered at 40 mg / day was bred for 5 weeks as an H2 group. The tea extract or the specific chicken egg antibody was administered as a mixture with artificial milk. As an indicator of diarrhea, the fecal score and the number of Salmonella evening spores in the feces were determined from 0 to 5 weeks from the start of the test. The results are shown in Tables 6 and 7, respectively.
表 6 Table 6
糞便スコア  Fecal score
Figure imgf000033_0001
Figure imgf000033_0001
数値は、 1 0頭の平均を示し、 正常便を 0、 軟便を 1  The figures indicate the average of 10 animals, 0 for normal stools and 1 for loose stools.
泥様便を 2、 水様便を 3 とした。  Mud-like stool was set to 2 and watery stool was set to 3.
糞便中のサルモネラ 夕ブリ ンの菌数 Salmonella count in evening feces in feces
Figure imgf000033_0002
Figure imgf000033_0002
数値は、 1 0頭の平均を示し、 糞便 1 g当たりの対数値で 示した。  The values are the average of 10 animals and are expressed as logarithmic values per 1 g of stool.
表 6 と表 7から、 I群は G群、 H群及び H 2群の子牛と比較して 糞便スコアの低下及び糞便中のサルモネラ 夕プリ ンの菌数の減少 が認められた。 従って、 本発明の茶抽出物は感染性の下痢を効果的 に抑制することがわかった。 また、 実施例 2及び実施例 3で得られ た茶抽出物を用いて同様の試験を行ったところ、 同様の結果が得ら れた。 さらに、 紅茶、 ウーロン茶由来の茶抽出物を用いて同様の試 験を行ったところ、 同様の効果が得られた。 また、 大腸菌 0— 8 8、 大腸菌 0— 9 9、 大腸菌 9 8 7 P、 牛ロタウィルス K K 一 3及び牛ロタウィルス N C D Vで同様の感染症下痢の試験を行つ たところ同様の結果が得られた。 From Table 6 and Table 7, it was found that the fecal score and the number of Salmonella duopurin in feces were reduced in Group I compared to the calves in Groups G, H and H2. Therefore, it was found that the tea extract of the present invention effectively suppressed infectious diarrhea. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, a similar test was performed using a tea extract derived from black tea and oolong tea, and a similar effect was obtained. E. coli 0-8 8. Escherichia coli 0-99, Escherichia coli 987P, bovine rotavirus KK-13 and bovine rotavirus NCDV were tested for similar infectious diseases and diarrhea, and similar results were obtained.
試験例 4 Test example 4
サルモネラ 夕プリ ンで感染させた 1 5頭の成牛を 5頭ずつ 3群 に分け、 表 3に示した基本飼料を与えた群を J群、 実施例 1 で調製 した茶抽出物を 1 日当たり 7 . 5 g投与した群を K群、 実施例 1 で 調製した茶抽出物を 1 日当たり 7 . 5 g及び実施例 4で調製した特 異的鶏卵抗体を 1 日当たり 6 g投与した群を L群、 実施例 4で調製 した特異的鶏卵抗体を 1 日当たり 6 g投与した群を K 2群として 5 週間飼育した。 茶抽出物又は特異的鶏卵抗体は表 3の基本飼料に混 合したものを投与した。 なお、 水は自由に摂取させた。 そして、 試 験例 3 と同様に下痢症を示す指標として糞便スコアと糞便中のサル モネラ タブリ ンの菌数を試験開始 0週〜 5週まで求めた。 この結 果をそれぞれ表 8 と表 9に示した。  Salmonella 15 adult cattle infected with evening primrose were divided into 3 groups of 5 cows, the group fed the basic diet shown in Table 3 was group J, and the tea extract prepared in Example 1 was used per day. Group K received 7.5 g and group L received 7.5 g / day of the tea extract prepared in Example 1 and 6 g / day of the specific chicken egg antibody prepared in Example 4. The group to which 6 g of the specific chicken egg antibody prepared in Example 4 was administered per day was bred for 5 weeks as the K2 group. The tea extract or specific chicken egg antibody was administered as a mixture with the basic feed shown in Table 3. Water was provided ad libitum. In the same manner as in Test Example 3, the fecal score and the number of Salmonella tabulin in the feces were determined from 0 to 5 weeks from the start of the test as indices indicating diarrhea. The results are shown in Tables 8 and 9, respectively.
表 8 Table 8
糞便スコア  Fecal score
Figure imgf000034_0001
Figure imgf000034_0001
数値は、 5頭の平均を示し、 正常便を 0、 軟便を 1  The figures indicate the average of 5 animals, 0 for normal stool and 1 for loose stool
泥様便を 2、 水様便を 3 とした。 表 9 Mud-like stool was set to 2 and watery stool was set to 3. Table 9
糞便中のサルモネラ 夕ブリ ンの菌数  Salmonella count in evening feces in feces
Figure imgf000035_0001
Figure imgf000035_0001
—数値は 5頭の平均を示し、 糞便 1 g当たりの対数値で  —The figures are the average of 5 animals and are expressed in logarithms per g of feces
不した。 表 8 と表 9 より、 L群の成牛は J群、 K群及び K 2群の成牛と比 較して糞便スコアの低下及び糞便中のサルモネラ 夕プリ ンの菌数 の減少が認められた。 従って、 本発明の茶抽出物は感染性の下痢を 効果的に抑制することがわかった。 また、 実施例 2及び実施例 3で 得られた茶抽出物を用いて同様の試験を行ったところ、 同様の結果 が得られた。 さらに、 紅茶、 ウーロン茶由来の茶抽出物を用いて同 種の試験を行ったところ、 同様の結果が得られた。 また大腸菌 0 一 8 8、 大腸菌 0— 9 9、 大腸菌 9 8 7 P、 牛口夕ウィルス K K一 3及び牛ロタウィルス N C D Vを用いて同様の試験を行った ところ、 同様の結果が得られた。  I didn't. Tables 8 and 9 show that adult cats in group L had lower fecal scores and fewer bacteria of Salmonella evening primin in feces than adult cows in groups J, K and K2. Was. Therefore, it was found that the tea extract of the present invention effectively suppressed infectious diarrhea. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, similar tests were performed using tea extracts derived from black tea and oolong tea, and similar results were obtained. Similar tests were performed using Escherichia coli 088, Escherichia coli 0-99, Escherichia coli 987P, Ushiguchi Yu virus KK-13 and Bovine rotavirus NCDV, and similar results were obtained.
試験例 5 Test example 5
健康な 3 0頭の乳牛を 1 0頭ずつ 3群に分け、 実施例 1 で調製し た茶抽出物を 1 日当たり 1 5 g投与した群を A群、 実施例 1 で調製 した茶抽出物を 1 日当たり 7 . 5 g投与した群を B群、 無投与の群 を C群、 として 5週間飼育した。 なお、 茶抽出物は通常の飼料と混 合して投与した。 そして、 乳質改善、 乳量増加を示す指標として、 搾乳した牛乳について、 乳脂率、 無脂固形分率、 細胞数、 乳蛋白率 及び 1 日当たりの乳量を常法に従い試験開始 0週〜 5週まで測定し た。 その結果を表 1 0から表 1 4にそれぞれ示した。 さらに、 試験 開始 3週間後と 5週間後に牛乳の官能検査をパネラー 1 5人により 行った。 その結果を表 1 5に示した。 表 1 0 Healthy 30 dairy cows were divided into 3 groups of 10 cows each, and the group that received 15 g of the tea extract prepared in Example 1 per day was Group A, and the tea extract prepared in Example 1 was The group administered 7.5 g per day was bred for 5 weeks as group B, and the group without administration as group C. The tea extract was administered by mixing with a normal feed. Milk fat ratio, non-fat solids ratio, cell number, milk protein ratio for milked milk are used as indicators for improving milk quality and increasing milk yield. In addition, the amount of milk per day was measured from 0 to 5 weeks after the start of the test according to a conventional method. The results are shown in Tables 10 to 14, respectively. In addition, three and five weeks after the start of the test, 15 panelists conducted a sensory test of milk. The results are shown in Table 15. Table 10
乳脂率  Milk fat percentage
Figure imgf000036_0001
Figure imgf000036_0001
数値は、 1 0頭の平均を示し、 単位は、 %で示した, 表 1 1  Numerical values indicate the average of 10 animals, and the unit is%.
無脂固形分率  Non-fat solid content
Figure imgf000036_0002
Figure imgf000036_0002
数値は、 1 0頭の平均を示し、 単位は、 %で示した, 表 1 2 Numerical values indicate the average of 10 animals, and the unit is%. Table 1 2
細胞数  Cell count
Figure imgf000037_0001
Figure imgf000037_0001
数値は、 1 0頭の平均を示し、 単位は 1 X 1 0 個 Z m 1 で示した。  The numerical value indicates the average of 10 animals, and the unit is 1 × 10 Zm 1.
表 1 3 Table 13
乳蛋白率  Milk protein rate
Figure imgf000037_0002
Figure imgf000037_0002
数値は、 1 0頭の平均を示し、 単位は、 %で示した,  Numerical values indicate the average of 10 animals, and the unit is%.
表 1 4 Table 14
1 日当たりの乳量  Milk yield per day
Figure imgf000037_0003
Figure imgf000037_0003
数値は、 1 0頭の平均を示し、 単位は、 k gで示した, 表 1 5 Numerical values indicate the average of 10 animals, and the unit is kg. Table 15
官能検査試験  Sensory test
Figure imgf000038_0001
Figure imgf000038_0001
数値は、 15人の平均を示し、 非常に良いを 5点、 良いを 4点 普通を 3点、 悪いを 2点、 非常に悪いを 1 点とした。 表 1 0から表 1 4 より明らかなように、 A群の乳牛より搾乳した 牛乳は B群及び C群の乳牛より搾乳した牛乳と比較して乳脂率、 無 脂固形分率、 乳蛋白率及び 1 日当たりの乳量の増加、 細胞数の減少 が認められた。 具体的には、 A群では投与後 2週間で乳脂率は 3 . 5 %、 無脂固形分率は 8 . 5 %、 乳蛋白率は 3 . 0 %を越え、 Aラ ンク としての認定条件を満たすことが判明した。 したがって、 本発 明の茶抽出物は乳質改善、 乳量増加に有効であることがわかった。 さらにパネラー 1 5人を用いて行った官能検査からも表 1 5 より明 らかなように A群の乳牛より搾乳した牛乳は香気、 舌味、 口当たり が良好であることが認められた。 また、 実施例 2及び実施例 3で得 られた茶抽出物を用いて同様の試験を行ったところ、 同様の結果が 得られた。 さらに、 紅茶、 ウーロン茶由来の茶抽出物を用いて同様 の試験を行ったところ、 同様の結果が得られた。  The figures are the average of 15 people, 5 points for very good, 4 points for good, 3 points for normal, 2 points for bad, and 1 point for very bad. As evident from Tables 10 to 14, milk expressed from dairy cows in Group A was higher than milk expressed from dairy cows in Groups B and C, in terms of milk fat ratio, nonfat solids ratio, milk protein ratio and Increased milk yield per day and decreased cell number were observed. Specifically, in Group A, the milk fat ratio was 3.5%, the non-fat solids ratio was 8.5%, and the milk protein ratio exceeded 3.0% within 2 weeks after administration, and the conditions for qualification as Rank A were given. Was found to meet. Therefore, it was found that the tea extract of the present invention was effective for improving milk quality and increasing milk yield. Furthermore, a sensory test conducted with 15 panelists revealed that milk expressed by milk from Group A cows had good aroma, tongue taste, and mouthfeel, as is clear from Table 15. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. Furthermore, similar tests were performed using tea extracts derived from black tea and oolong tea, and similar results were obtained.
試験例 6 Test example 6
分娩直後の健康な 2 0頭の乳牛を 1 0頭ずつ 2群に分け、 実施例 1 で調製した茶抽出物を 1 日当たり 1 5 g投与した群を A群、 無投 与の群を B群として 1年間飼育した。 なお、 茶抽出物は通常の飼料 と混合して投与した。 そして、 繁殖率を示す指標として、 分娩後受 胎に要する期間と妊娠確認された乳牛の人工受精回数を調べた。 そ の結果を表 1 6 と表 1 7にそれぞれ示した。 表 1 6 Healthy 20 dairy cows immediately after parturition were divided into 2 groups of 10 cows, and the group that received 15 g of the tea extract prepared in Example 1 per day was Group A, and the group that received no milk was Group B. Bred for one year. In addition, tea extract is a normal feed And administered in a mixture. Then, as an indicator of the reproductive rate, the time required for conception after delivery and the number of artificial inseminations of dairy cows confirmed to be pregnant were examined. The results are shown in Table 16 and Table 17, respectively. Table 16
分娩後受胎に要する期間  Time required for conception after delivery
Figure imgf000039_0001
Figure imgf000039_0001
表 1 7  Table 17
妊娠確認された乳牛の人工受精回数  Number of artificial inseminations of dairy cows confirmed to be pregnant
Figure imgf000039_0002
表 1 6 と表 1 7より、 A群の乳牛は B群の乳牛と比較して、 分娩 後受胎に要する期間が短縮され、 妊娠確認された乳牛の人工受精回 数が減少し、 繁殖率の向上が認められた。 したがって、 本発明の茶 抽出物は乳牛の繁殖率を向上させる効果があることがわかった。 ま た、 実施例 2及び実施例 3で得られた茶抽出物を用いて同様の試験 を行なったところ、 同様の結果が得られた。 さらに、 紅茶、 ウー口 ン茶由来の茶抽出物を用いて同様の試験を行なったところ、 同様の 結果が得られた。
Figure imgf000039_0002
Tables 16 and 17 show that dairy cows in group A have a shorter time to conception after parturition than cows in group B, and that the number of artificial fertilizations of dairy cows that have been confirmed to be pregnant decreases, and that the reproductive rate is lower. Improvement was observed. Therefore, it was found that the tea extract of the present invention had an effect of improving the reproductive rate of dairy cows. Similar tests were performed using the tea extracts obtained in Example 2 and Example 3, and similar results were obtained. In addition, similar tests were performed using tea extracts derived from black tea and Wu Kuang tea, and similar results were obtained.
試験例 7 Test example 7
3 0頭の子牛を 1 0頭ずつ 3群に分け、 人工乳のみを与えた群を A群、 実施例 5で調製した (+ ) —力テキンを 1 日当たり 9 0 m g 投与した群を B群、 実施例 5で調製した (一) ーェピガロカテキン ガレー トを 1 日当たり 9 0 m g投与した群を C群として 5週間飼育 した。 上記 2種類のボリ フエノール化合物は人工乳に混合して投与 した。 なお、 水は自由に摂取させた。 そして、 非感染性下痢症を示 す指標として糞便スコアと腸内細菌叢を試験開始 0週〜 5週まで測 定した。 この結果をそれぞれ表 1 8 と表 1 9に示した。 The 30 calves were divided into three groups of 10 calves each. Group A, prepared in Example 5 (+)-90 mg / day of dytechin was administered to group B, prepared in Example 5 (1) 90 mg perepigallocatechin gallate per day The administered group was reared as Group C for 5 weeks. The above two polyphenol compounds were administered by mixing with artificial milk. Water was provided ad libitum. The fecal score and intestinal microflora were measured from 0 to 5 weeks from the start of the test as indices indicating non-infectious diarrhea. The results are shown in Tables 18 and 19, respectively.
表 1 8  Table 18
糞便スコァ  Fecal score
Figure imgf000040_0001
Figure imgf000040_0001
数値は、 1 0頭の平均を示し、 正常便を 0、 軟便を 1  The figures indicate the average of 10 animals, 0 for normal stools and 1 for loose stools.
泥様便を 2、 水様便を 3 とした。 Mud-like stool was set to 2 and watery stool was set to 3.
表 1 9 Table 19
腸内細菌叢 Intestinal flora
Figure imgf000041_0001
Figure imgf000041_0001
数値は、 1 0頭の平均を求め、 糞便 1 g当たりの対数値で示した 表 1 8 と表 1 9 より、 C群の子牛は糞便スコアの低下、 ゥエルシ ュ菌、 大腸菌群の減少及びビフィズス菌、 乳酸菌の菌数の減少割合 の抑制が認められ、 A群及び B群の子牛と比較して非感染性下痢が 効果良く抑制されたことがわかった。 したがって、 本発明の (一) ーェピガロカテキンガレー トは非感染性下痢を効果的に抑制するこ とがわかった。 Values were averaged for 10 animals and expressed as logarithmic value per 1 g of stool According to Tables 18 and 19, calves in group C showed a decrease in fecal score, a decrease in the number of Bacillus cerevisiae and Escherichia coli, and a decrease in the number of Bifidobacteria and lactic acid bacteria, and a decrease in the numbers of groups A and B. It was found that non-infectious diarrhea was more effectively suppressed as compared to calves in Japan. Therefore, it was found that (1) epigallocatechin gallate of the present invention effectively suppressed non-infectious diarrhea.
試験例 8 Test example 8
1 5頭の分娩直後の成牛を 5頭ずつ 3群に分け、 表 3の基本飼料 を与えた群を D群、 実施例 5で調製した (+ ) —力テキンを 1 日当 たり 9 0 0 111 8投与した群を£群、 実施例 5で調製した (一) ーェ ピガロカテキンガレー トを 1 日当たり 9 0 0 111 2投与した群を?群 とし 5週間飼育した。 上記 2種類のボリ フエノール化合物は表 3の 基本飼料に混合して投与した。 なお、 水は自由に摂取させた。 そし て、 非感染性下痢症を示す指標として試験例 1 と同様に糞便スコア と腸內細菌叢を試験開始 0週〜 5週まで測定した。 この結果をそれ ぞれ表 2 0 と表 2 1 に示した。  1 Five calves immediately after calving were divided into 3 groups of 5 cows. The group fed the basic diet shown in Table 3 was prepared in group D, and prepared in Example 5 (+) — 90% per day. The group to which 0 111 8 was administered was prepared in the £ group, and the group prepared in Example 5 (1) The group to which 9 0 0 11 12 was administered per day with pigallocatechin gallate? Groups were reared for 5 weeks. The above two polyphenol compounds were mixed with the basic feed shown in Table 3 and administered. Water was provided ad libitum. The fecal score and intestinal microflora were measured from 0 to 5 weeks from the start of the test in the same manner as in Test Example 1 as indexes indicating non-infectious diarrhea. The results are shown in Table 20 and Table 21 respectively.
表 2 0 Table 20
糞便スコァ  Fecal score
Figure imgf000042_0001
Figure imgf000042_0001
数値は、 5頭の平均を示し、 正常便を 0、 軟便を 1  The figures indicate the average of 5 animals, 0 for normal stool and 1 for loose stool
泥様便を 2、 水様便を 3 とした。 表 2 1 Mud-like stool was set to 2 and watery stool was set to 3. Table 2 1
腸内細菌叢 Intestinal flora
Figure imgf000043_0001
Figure imgf000043_0001
数値は、 5頭の平均を求め、 糞便 1 g当たりの対数値で示した 表 2 0 と表 2 1 より、 F群の成牛は D群及び E群で飼育した成牛 と比較して、 糞便スコアの低下、 ウエルシュ菌、 大腸菌群の減少、 及びビフィズス菌、 乳酸菌の増加が認められた。 したがって、 本発 明の (一) ーェピガロカテキンガレー トは分娩によって生じたス ト レス性 (非感染性) の下痢を効果的に抑制することがわかった。 試験例 9 The values were averaged for 5 animals and expressed as a logarithmic value per g of feces Tables 20 and 21 show that adult cats in group F had lower fecal scores, reduced numbers of Welsh and Escherichia coli groups, and increased numbers of bifidobacteria and lactic acid bacteria compared to adult cats reared in groups D and E. Was observed. Therefore, it was found that the (1) -epigallocatechin gallate of the present invention effectively suppressed stress (non-infectious) diarrhea caused by parturition. Test example 9
サルモネラ タブリ ンで感染させた 3 0頭の子牛を 1 0頭ずつ 3 群に分け、 人工乳のみを与えた群を G群、 実施例 5で調製した (- ) ーェピガロカテキンガレー トを 1 日当たり 9 O m g投与した群を H群、 実施例 5で調製した (一) ーェピガロカテキンガレー トを 1 日当たり 9 0 m g及び実施例 4で調製した特異的鶏卵抗体を 1 日当 たり 4 0 0 m g投与した群を I群とし 5週間飼育した。 上記 2種類 のポリ フヱノール化合物は人工乳に混合して投与した。 そして、 下 痢症を示す指標として糞便スコアと糞便中のサルモネラ 夕ブリ ン の菌数を試験開始 0週〜 5週まで求めた。 この結果をそれぞれ表 2 2 と表 2 3に示した。  30 calves infected with Salmonella tabulin were divided into 3 groups of 10 cows each, and the group fed with only artificial milk was prepared in group G, prepared in Example 5 (-)-epigallocatecate gallate. Group was administered in 9 Hmg / day, group H, prepared in Example 5 (1) 90 mg of epigallocatechin gallate per day and the specific hen egg antibody prepared in Example 4 were applied per day. The group administered 400 mg / day was group I and bred for 5 weeks. The above two types of polyphenol compounds were mixed with artificial milk and administered. Then, as an index indicating diarrhea, the fecal score and the number of Salmonella duo in the feces were determined from 0 to 5 weeks from the start of the test. The results are shown in Tables 22 and 23, respectively.
表 2 2 Table 2 2
糞便スコァ  Fecal score
Figure imgf000044_0001
Figure imgf000044_0001
数値は、 1 0頭の平均を示し、 正常便を 0、 軟便を 1 泥様便を 2、 水様便を 3 とした。 表 2 3 The numerical values indicate the average of 10 animals, with 0 for normal stool, 1 for loose stool, 2 for muddy stool, and 3 for watery stool. Table 23
糞便中のサルモネ 夕ブリ ンの菌数  Number of bacteria in feces of salmonone evening
Figure imgf000045_0001
Figure imgf000045_0001
数値は、 1 0頭の平均を示し、 糞便 1 g当たりの対数値 で示した。 表 2 2 と表 2 3から、 I群は G群及び H群の子牛と比較して、 糞 便スコアの低下、 糞便中のサルモネラ タブリ ンの菌数の減少が認 められた。 したがって、 本発明の (一) ーェピガロカテキンガレー トは感染性の下痢を効果的に抑制することがわかった。 また、 大腸 菌 0 - 8 8、 大腸菌 0 - 9 9、 大腸菌 9 8 7 P、 牛ロタウイ ルス K K一 3及び牛ロタウィルス N C D Vで同様の感染症下痢の 試験を行ったところ、 同様の結果が得られた。  The numerical values represent the average of 10 animals and were expressed as logarithmic values per 1 g of feces. From Tables 22 and 23, it was confirmed that the fecal score and the number of Salmonella tabulin in feces were reduced in Group I compared to calves in Group G and H. Therefore, it was found that (1) epigallocatechin gallate of the present invention effectively suppressed infectious diarrhea. Similar infectious disease diarrhea tests were performed with E. coli 0-88, E. coli 0-99, E. coli 987P, bovine rotavirus KK-13 and bovine rotavirus NCDV. Was done.
試験例 1 0 Test example 10
サルモネラ 夕ブリ ンで感染させた 1 5頭の成牛を 5頭ずつ 3群 に分け、 表 3の基本飼料を与えた群を J群、 実施例 5で調製した ( 一) ーェピガロカテキンガレー トを 1 日当たり 5 0 O m g投与した 群を K群、 実施例 5で調製した (一) ーェピガロカテキンガレー ト を 1 日当たり 5 0 0 m g及び実施例 4で調製した特異的鶏卵抗体を 1 日当たり 6 g投与した群を L群として 5週間飼育した。 (一) 一 ェピガロカテキンガレー ト又は特異的鶏卵抗体は表 3の基本飼料に 混合して投与した。 なお、 水は自由に摂取させた。 下痢症を示す指 標として糞便スコアと糞便中のサルモネラ 夕ブリ ンの菌数を試験 開始 0週〜 5週まで求めた。 この結果をそれぞれ表 2 4 と表 2 5に 示した Salmonella 15 adult cattle infected with evening brine were divided into 3 groups of 5 cows each, and the group fed the basic diet shown in Table 3 was prepared in Group J and prepared in Example 5. (1) -epigallocatechin The group receiving 50 mg of gallate per day was group K, prepared in Example 5. (1) 500 mg of epigallocatechin gallate per day and the specific chicken antibody prepared in Example 4 Was administered for 5 weeks as L group. (1) Epigallocatechin gallate or specific chicken egg antibody was administered by mixing with the basic feed shown in Table 3. Water was provided ad libitum. The fecal score and the number of Salmonella evening brilli in feces were determined from 0 to 5 weeks from the start of the test as indicators of diarrhea. The results are shown in Table 24 and Table 25, respectively. Indicated
表 2 4 糞便スコア Table 2-4 Fecal score
Figure imgf000046_0001
Figure imgf000046_0001
数値は、 5頭の平均を示し、 正常便を 0、 軟便を 1  The figures indicate the average of 5 animals, 0 for normal stool and 1 for loose stool
泥様便を 2、 水様便を 3 とした。  Mud-like stool was set to 2 and watery stool was set to 3.
表 2 5 Table 25
糞便中のサルモネラ ダブリ ンの菌数  Salmonella Dublin counts in feces
Figure imgf000046_0002
Figure imgf000046_0002
数値は 5頭の平均を示し、 糞便 1 g当たりの対数値で  The figures represent the average of 5 animals and are expressed in logarithms per g of feces.
示した。  Indicated.
表 2 4 と表 2 5 より、 L群の成牛は J群、 K群及び K 2群の成牛 と比較して、 糞便スコアの低下、 糞便中のサルモネラ 夕プリ ンの 菌数の減少が認められた。 したがって、 本発明の (一) ーェピガ口 力テキンガレー トは感染性の下痢を効果的に抑制することがわかつ た。 また、 大腸菌 0— 8 8、 大腸菌 0— 9 9、 大腸菌 9 8 7 P、 牛口夕ウィルス K K— 3及び牛ロタウィルス N C D Vを用い て同様の試験を行ったところ、 同様の結果が得られた。 試験例 1 1 According to Tables 24 and 25, adult cats in group L showed lower fecal scores and fewer bacteria in Salmonella duo in feces compared to adult cows in groups J, K and K2. Admitted. Therefore, it was found that the (1) epiga oral techin gallate of the present invention effectively suppressed infectious diarrhea. Similar tests were performed using Escherichia coli 0-88, Escherichia coli 0-99, Escherichia coli 987P, Ushiguchi Yu virus KK-3 and Bovine rotavirus NCDV, and similar results were obtained. . Test example 1 1
健康な 3 0頭の乳牛を 1 0頭ずつ 3群に分け、 実施例 5で調製し た (一) ーェピガロカテキンガレー トを 1 日当たり 9 0 O m g投与 した群を M群、 実施例 5で調製した (+ ) —力テキンを 1 日当たり 9 0 0 111 投与した群を1^群、 上記 2種類のポリ フヱノール化合物 を含まない通常の飼料を与えた群を 0群とし、 5週間飼育した。 上 記ボリ フユノール化合物は通常の飼料と混合して投与した。 乳質改 善、 乳量増加を示す指標として、 搾乳した牛乳について、 乳脂率、 無脂固形分率、 細胞数、 乳蛋白率及び 1 日当たりの乳量を常法に従 い試験開始 0週〜 5週まで測定した。 その結果を表 2 6から表 3 0 にそれぞれ示した。 さらに、 試験開始 3週間後と 5週間後に牛乳の 官能検査をパネラー 1 5人により行った。 その結果を表 3 1 に示し た。 表 2 6  Healthy 30 dairy cows were divided into 3 groups of 10 cows each and prepared in Example 5. (1) Group administered M-epigallocatechin gallate at 90 mg / day to the M group. The (+)-forced techin prepared in 5 was administered 9 0 0 111 per day as a 1 ^ group, and a group fed with the above-mentioned two types of polyphenol-free normal feed as a 0 group, and bred for 5 weeks. did. The above-mentioned polyfunool compound was administered by mixing with a normal feed. Milk fat ratio, nonfat solids ratio, cell count, milk protein ratio, and milk yield per day for milked milk were used as indicators of milk quality improvement and milk yield. Measured until week. The results are shown in Tables 26 to 30 respectively. In addition, three and five weeks after the start of the test, sensory tests of milk were performed by 15 panelists. The results are shown in Table 31. Table 26
乳脂率  Milk fat percentage
Figure imgf000047_0001
Figure imgf000047_0001
数値は、 1 0頭の平均を示し、 単位は、 %で示した。 表 2 7 The numerical values indicate the average of 10 animals, and the unit is expressed in%. Table 27
無脂固形分率  Non-fat solid content
Figure imgf000048_0001
Figure imgf000048_0001
数値は、 1 0頭の平均を示し、 単位は、 %で示した,  Numerical values indicate the average of 10 animals, and the unit is%.
表 2 8 Table 28
細胞数  Cell count
Figure imgf000048_0002
Figure imgf000048_0002
数値は、 1 0頭の平均を示し、 単位は 1 0 4 個 1 で示した。 Numbers, shows the average of 1 0 head, the unit is shown in 1 0 4 1.
表 2 9 Table 2 9
乳蛋白率  Milk protein rate
Figure imgf000048_0003
Figure imgf000048_0003
数値は、 1 0頭の平均を示し、 単位は、 %で示した, 表 3 0 Numerical values indicate the average of 10 animals, and the unit is%. Table 30
1 日当たりの乳量  Milk yield per day
Figure imgf000049_0001
Figure imgf000049_0001
数値は、 1 0頭の平均を示し、 単位は、 k gで示した,  Numerical values indicate the average of 10 animals, and the unit is kg.
表 3 1 Table 3 1
官能検査試験  Sensory test
Figure imgf000049_0002
Figure imgf000049_0002
数値は、 1 5人の平均を示し、 非常に良いを 5点、 良いを 点、 普通を 3点、 悪いを 2点、 非常に悪いを 1 点とした。  The figures are the average of 15 people, with 5 points for very good, 3 points for good, 2 points for bad, and 1 point for very bad.
表 2 6から表 3 0 より、 M群の乳牛より搾乳した牛乳は N群及び 0群の乳牛より搾乳した牛乳と比較して乳脂率、 無脂固形分率、 乳 蛋白率及び 1 日当たりの乳量の増加、 細胞数の減少が認められた。 具体的に述べると、 M群では投与後 2週間で乳脂率は 3 . 5 %、 無 脂固形分率 8 . 5 %、 乳蛋白率 3 . 0 %を越え、 Aランクとしての 認定条件を満たすことが判明した。 以上の結果から、 本発明の (一 ) ーェピガロカテキンガレー トは、 乳牛の乳質改善、 乳量増加に有 効であることがわかった。 さらにパネラー 1 5人を用いて行った官 能検査からも、 表 3 1 より明らかなように M群の乳牛より搾乳した 牛乳は香気、 舌味、 口当たりが良好であることが認められた。 From Table 26 to Table 30, milk expressed from cows in Group M was higher than milk expressed from cows in Groups N and 0 compared with milk expressed in milk, fat-free solids, milk protein and milk per day. An increase in the amount and a decrease in the number of cells were observed. Specifically, in the M group, the milk fat ratio exceeded 3.5%, the nonfat solid content ratio was 8.5%, and the milk protein ratio exceeded 3.0% within 2 weeks after administration, satisfying the criteria for A rank It has been found. From the above results, it was found that (1) -epigallocatechin gallate of the present invention was effective for improving milk quality and increasing milk yield of dairy cows. In addition, from a functional test conducted with 15 panelists, milking from cows in the M group was evident from Table 31. Milk was found to have good aroma, tongue taste, and mouthfeel.
試験例 1 2 Test example 1 2
分娩直後の健康な 3 0頭の乳牛を 1 0頭ずつ 3群に分け、 実施例 5で調製した (一) ーェピガロカテキンガレー トを 1 日当たり 9 0 O m g投与した群を P群、 実施例 5で調製した (+ ) —力テキンを 1 日当たり 9 0 O m g投与した群を Q群、 通常の飼料を与えた群を R群として 1年間飼育した。 なお、 上記ボリ フエノール化合物は通 常の飼料と混合して投与した。 繁殖率を示す指標として、 分娩後受 胎に要する期間と妊娠確認された乳牛の人工受精回数を調べた。 そ の結果を表 3 2 と表 3 3にそれぞれ示した。 表 3 2  Healthy 30 dairy cows immediately after parturition were divided into 3 groups of 10 cows each, and the group that received 90 mg Oepigallocatechin gallate prepared in Example 5 (1) -epigallocatechin gallate was treated as group P, A group to which 90 mg of (+)-forced techin prepared in Example 5 was administered per day was bred for 1 year as a Q group, and a group fed with a normal diet as a R group. The above polyphenol compound was administered after being mixed with a normal feed. As an indicator of the reproductive rate, the time required for conception after delivery and the number of artificial inseminations of dairy cows confirmed to be pregnant were examined. The results are shown in Tables 32 and 33, respectively. Table 3 2
分娩後受胎に要する期間  Time required for conception after delivery
Figure imgf000050_0001
Figure imgf000050_0001
表 3 3  Table 3 3
妊娠確認された乳牛の人工受精回数  Number of artificial inseminations of dairy cows confirmed to be pregnant
Figure imgf000050_0002
Figure imgf000050_0002
表 3 2 と表 3 3 より明らかなように、 P群の乳牛は、 Q群及び R 群の乳牛と比較して、 分娩後受胎に要する期間の短縮及び妊娠確認 された乳牛の人工受精回数の減少が認められた。 したがって、 本発 明の (一) ーェピガロカテキンガレー トは乳牛の繁殖率の向上に効 果のあることがわかった。 As is clear from Tables 3 and 3, the dairy cows in group P Compared with the cows in the herd, the time required for postpartum conception and the number of artificial inseminations in cows confirmed to be pregnant were reduced. Therefore, it was found that (1) -epigallocatechin gallate of the present invention was effective in improving the reproductive rate of dairy cows.
実施例 6 Example 6
パワーミルで粉砕し、 直径 1 . 5 mmのフィルターを通した茶葉 1 k gに水 1 0 リ ッ トルを加え、 混合物を 9 0〜 9 5 で 3 0分間 加熱した。 混合物を固液分離し湿潤残渣 2. 4 k gを得た。 湿潤残 渣は 7 0てで一晚乾燥した後、 粉砕し茶葉織維 7 0 0 gを得た。 こ の茶葉織維中の力フヱイン及びボリ フエノールの含量は、 1 0 0 g 中それぞれ 0. 6 g、 2. 7 gと低く、 渋み、 苦み共にほとんどな く飼料へ充分添加し得るものであった。  10 liters of water was added to 1 kg of tea leaves which had been ground with a power mill and passed through a filter having a diameter of 1.5 mm, and the mixture was heated at 90 to 95 for 30 minutes. The mixture was subjected to solid-liquid separation to obtain 2.4 kg of a wet residue. The wet residue was dried at 70 ° C and then pulverized to obtain 700 g of tea leaf fabric. The content of power phenol and polyphenol in this tea leaf fabric was low at 0.6 g and 2.7 g, respectively, in 100 g, and both astringency and bitterness could be added to feed with little bitterness and bitterness. Was.
実施例 7 Example 7
煎茶、 ウーロン茶の混合物 ( 3 : 1 ) 2 0 0 gを径 2〜 3 mmに 粉砕し、 5 0。Cに加温した水、 エタノール、 アセ ト ン混液 ( 5 : 3 : 2 ( v/v) ) 4 リ ッ トルを加え 2時間抽出した。 混合物を固液 分離し、 得られた湿潤残渣 3 2 0 gを 8 0でで熱風乾燥した後、 粉 砕し茶葉織維 1 5 0 gを得た。 この茶葉繊維中のカフ イン及びボ リ フエノール含量は、 1 0 0 g中それぞれ 0. 7 g、 3. 5 gと低 く、 渋み、 苦み共にほとんどなく飼料へ充分添加し得るものであつ 実施例 8  Sencha and oolong tea mixture (3: 1) 200 g, crushed to 2-3 mm in diameter, 50. Four liters of a mixture of water, ethanol, and acetate (5: 3: 2 (v / v)) heated in C was added and extracted for 2 hours. The mixture was subjected to solid-liquid separation, and 320 g of the obtained wet residue was dried with hot air at 80, followed by pulverization to obtain 150 g of tea leaf fiber. The content of caffeine and polyphenol in this tea leaf fiber was as low as 0.7 g and 3.5 g, respectively, in 100 g, with little bitterness and bitterness, and which could be sufficiently added to feed. 8
養豚用固形飼料  Solid feed for pig farming
成 分 配合割合 (重量%)  Component Mixing ratio (% by weight)
脱脂粉乳 3 2. 6 Skim milk powder 32.6
小麦粉 2 9. 9 パン粉 7 . 0 Flour 2 9. 9 Breadcrumbs 7.0
脱脂大豆粕 5 . 0 Defatted soybean meal 5.0
魚粉 7 . 0 Fishmeal 7.0
砂糖 4 0 Sugar 4 0
ぶとう糖 8 0 Glucose 8 0
動物性油脂 2 0 Animal fats and oils 2 0
ォリゴ糖 1 0 Oligo sugar 1 0
ビタ ミ ン、 ミ ネラル類 3 0 Vitamin, minerals 30
実施例 6で得られた茶葉繊維 0 5 Tea leaf fiber obtained in Example 6 0 5
(飼料中にポリ フエノール化合物として 0 . 0 1 %含有) 上記配合割合に従い、 常法により養豚用飼料を調製した。 実施例 9  (Polyphenol compound is contained in the feed at 0.01%) A feed for swine raising was prepared according to the above mixing ratio by an ordinary method. Example 9
養鶏用飼料  Poultry feed
成 分 配合割合 (重量%)  Component Mixing ratio (% by weight)
とう もろこ し 5 8 . 0 Corn 58.0
大豆粕 1 5 9 Soybean meal 1 5 9
魚粉 6 0 Fish meal 6 0
ふすま 5 0 Bran 5 0
アルフ ァルファー 3 0 Alpha 3 0
炭酸カルシウム 0 Calcium carbonate 0
リ ン酸カルシウム 1 6  Calcium phosphate 1 6
食塩 0 4 Salt 0 4
ビタ ミ ン、 ミ ネラル類 0 1  Vitamin, minerals 0 1
大豆油 2 0 Soybean oil 2 0
実施例 7で得られた茶葉織維 1 0 The tea leaves obtained in Example 7 10
(飼料中にポリ フエノール化合物として 0 . 0 4 %含有) 上記配合割合に従い、 常法により養鶏用飼料を調製した (0.04% as polyphenol compound in feed) According to the above mixing ratio, poultry feed was prepared by the usual method
実施例 1 0 Example 10
乳牛用飼料  Feed for dairy cows
成 分 配合割合 (重量  Component Mixing ratio (weight
トゥモロコシ 2 4 . 4  Tomorrow 24.4
ライ麦 1 3 0 Rye 1 3 0
大豆粕 2 1 5 Soybean meal 2 1 5
ナタネ柏 5 2 Rape Kashiwa 5 2
コーングルテンフィー ド 8 0 Corn gluten feed 8 0
麦ヌカ 8 0 Mugi Nuka 8 0
アルファルファ 1 0 Alfalfa 1 0
綿実柏 1 0 Cotton seed Kashiwa 1 0
糖蜜 3 5 Molasses 3 5
炭酸カルシウム 4 4 Calcium carbonate 4 4
第 2 リ ン酸カルシウム 3 8 Calcium phosphate 2 3 8
食塩 2 4 Salt 2 4
酵母 0 4 Yeast 0 4
ビタ ミ ン、 ミ ネラル類 2 4 Vitamin, minerals 2 4
実施例 6で得られた茶葉繊維 1 0 Tea leaf fiber 10 obtained in Example 6
(飼料中にボリ フエノール化合物として 0 . 0 3 含有) 上記配合割合に従い、 常法により乳牛用飼料を調製した。  (The feed contains 0.03 as a polyphenol compound.) A feed for dairy cows was prepared by a conventional method according to the above mixing ratio.
試験例 1 3 Test example 1 3
開放畜舎で飼育されている乳牛 (成体) に 2 0頭を 2分し、 実施 例 1 0の乳牛用飼料と、 対照として本発明の添加物を含まない同飼 料を搾乳量の減少し始めた時期から 6 0 日間自由摂取で与えた。 試 験終了後は再び通常の飼料を与えた。 表 3 4 Twenty dairy cows (adults) bred in open stalls were divided into 2 cows, and the milk for cows of Example 10 and the feed without additives of the present invention as a control began to decrease in milking amount. The animals were given free access for 60 days. After the test, normal feed was given again. Table 3 4
搾乳量に与える影響  Effect on milking volume
Figure imgf000054_0001
Figure imgf000054_0001
対照区の開始前を 100として表した。 表 3 4 に示すように、 対照区と比較し本発明の添加物投与区では 試験期間中の搾乳量減少の抑制に効果があり、 搾乳量の増加が認め られた。  The value before the start of the control was expressed as 100. As shown in Table 34, compared with the control group, the group administered with the additive of the present invention was effective in suppressing the decrease in milking amount during the test period, and an increase in milking amount was observed.
試験例 1 4 Test example 1 4
豚舍で飼育されている豚 (成体) 各 2 0頭に各々実施例 8の本発 明の養豚用固形飼料と対照として本発明の添加物を含まない同飼料 を 6週間与え、 豚舎内のアンモニア濃度を比較した。 アンモニア濃 度の測定はガス検知管を用い、 床上約 1 mの高さで各豚舍 4力所ず つ測定し、 その平均値を示した。 Twenty pigs (adults) bred in a pig house were fed with the solid feed for swine raising of the present invention of Example 8 and the same feed without the additive of the present invention as a control for 6 weeks, respectively. The ammonia concentrations were compared. Ammonia concentration was measured using a gas detector tube at a height of about 1 m above the floor at four locations in each pig house, and the average value was shown.
表 3 5 Table 35
豚舎内のァンモニァ濃度  Ammonia concentration in piggery
Figure imgf000055_0001
Figure imgf000055_0001
表 3 5に示すように、 本発明の飼料添加物を与えた豚舎のアンモ ニァ濃度は対照区と比べ明らかに減少し、 飼育環境の改善が認めら れた。 As shown in Table 35, the concentration of ammonia in the piggery fed with the feed additive of the present invention was clearly reduced as compared with the control group, and improvement in the breeding environment was observed.
試験例 1 5 Test example 1 5
実施例 8に示した本発明の養豚用飼料と対照として本発明の添加 物を含まない同飼料を用いて各々 3 0頭の子豚を 2 0 日間飼育し、 子豚の飼育成績に与える影響を試験した。 飼料及び水は自由に摂取 させた。 開始時体重、 終了時体重、 飼料摂取量を測定し、 各々の平 均値より日増体重及び飼料要求率を算出することで飼育成績を評価 した。 The effect on piglet rearing performance of 30 piglets each raised for 20 days using the pig feed of the present invention shown in Example 8 and the same feed containing no additives of the present invention as a control. Was tested. Food and water were available ad libitum. The body weight at the start, the body weight at the end, and the feed intake were measured, and the breeding performance was evaluated by calculating the daily weight gain and the feed demand rate from the average values.
表 3 6 Table 3 6
子豚の飼育成績に与える影響  Effect on rearing performance of piglets
Figure imgf000056_0001
Figure imgf000056_0001
飼料要求率 =飼料摂取量 期間増体重 表 3 6に示すように本発明の添加物含有飼料で飼育した子豚は、 対照区の子豚に比べ、 日増体重が増加し、 飼料要求率が減少し飼料 効率の改善が認められた。  Feed demand = Feed intake Period gained weight As shown in Table 36, piglets reared on the diet containing the additive of the present invention showed an increase in daily weight gain and a lower feed demand rate than piglets in the control group. It has been reduced and feed efficiency has been improved.
試験例 1 6 Test Example 1 6
実施例 1 0で示した乳牛用飼料を用いて、 5頭の乳牛を表 3の基 本飼料で 1 0 日間予備飼育した後、 本発明の添加物含有飼料を与え 2 0 日間飼育した。 更に 1 0 日間基本飼料で飼育した。 飼料及び水 は自由に摂取させた。 試験開始前、 試験 1 0 日目、 2 0 日目、 試験 終了 1 0 日後に糞便中のビフィ ドバクテリ ゥム属細菌の試験開始前 に対する菌数の増加割合、 クロスト リ ジケム属細菌の試験開始前に 対する菌数の減少割合、 ρ Ηを測定し、 整腸効果を評価した。 表 3 7 Using the dairy cattle feed shown in Example 10, five dairy cows were preliminarily reared with the basic feed shown in Table 3 for 10 days, and then fed the additive-containing feed of the present invention and reared for 20 days. They were kept on the basic diet for another 10 days. Food and water were available ad libitum. Before the start of the test, on the 10th and 20th days of the test, and at 10 days after the end of the test, the rate of increase in the number of bacteria in the feces before the start of the test for Bifidobacterium, before the start of the test for Clostridium genus The percentage of decrease in the number of bacteria, ρΗ, was measured, and the intestinal control effect was evaluated. Table 3 7
整腸効果  Intestinal effect
Figure imgf000057_0001
Figure imgf000057_0001
ビフィ ドバクテリゥム属細菌とクロスト リ ジゥム属細菌については 開始前を 100として表した。  Bifidobacterium and Clostridium bacteria were expressed as 100 before the start.
表 3 7に示すように本発明の添加物含有飼料を与えることにより 、 糞便 p Hが低下し、 有用菌であるビフィ ドバクテリゥム属細菌が 増加し、 有害菌であるクロス ト リ ジゥム属細菌が減少し、 整腸効果 が認められた。 As shown in Table 37, by providing the feed containing the additive of the present invention, fecal pH is reduced, useful bacteria Bifidobacterium are increased, and harmful bacteria Clostridium are reduced. A bowel control effect was observed.
産業上の利用可能性 Industrial applicability
本発明品は、 家畜、 家禽及び愛玩動物の非感染性下痢症及び感染 性下痢症並びに生乳の乳質改善、 乳量増加、 繁殖率の向上に多大な 効果があり、 畜産業界及びぺッ ト業界に貢献するところは多大であ る 0  INDUSTRIAL APPLICABILITY The product of the present invention has a great effect on non-infectious diarrhea and infectious diarrhea of livestock, poultry and pets, as well as improving milk quality, increasing milk yield and reproductive rate of livestock. Contribute to
加えて、 本発明の茶葉繊維は日常飲用する茶に由来し安全性が極 めて高く、 家畜に投与すれば、 家畜の整腸作用を促進し、 下痢発症 を低減し、 搾乳量の増加、 飼料効率の改善、 及び糞便、 尿の悪臭消 去作用を有することから畜産業界にとって極めて都合がよい。 また 、 本発明の実施による飼料添加物の製造は、 最近産業上の重要性が 確立されつつある茶ボリフエノールの製造方法と組み合わせて実行 することが可能であり、 これによりそれぞれの製造方法の生産性が 高められることも本発明の特徴の—つである。 In addition, the tea fiber of the present invention is derived from tea that is consumed daily and has extremely high safety.When administered to livestock, it promotes the intestinal action of the livestock, reduces the occurrence of diarrhea, increases the amount of milking, It is extremely convenient for the livestock industry because it improves feed efficiency and has the action of eliminating odors from feces and urine. Further, the production of feed additives according to the present invention is carried out in combination with a method for producing tea bolifenol, which has recently been established as an industrially important material. It is one of the features of the present invention that the productivity of each manufacturing method can be enhanced.

Claims

請 求 の 範 囲 The scope of the claims
1 . 茶の抽出物よりなる動物用飼料添加物。 1. Animal feed additive consisting of tea extract.
2. 茶が緑茶、 ウーロン茶若しく は紅茶である請求項 1 記載の動物 用飼料添加物。  2. The animal feed additive according to claim 1, wherein the tea is green tea, oolong tea or black tea.
3. 茶の抽出物が茶葉を水、 アルコール及び 又は酢酸ェチルで抽 出したものである請求項 1記載の動物用飼料添加物。  3. The animal feed additive according to claim 1, wherein the tea extract is obtained by extracting tea leaves with water, alcohol and / or ethyl acetate.
4. 茶の抽出物がボリ フエノール化合物を含有する請求項 1記載の 動物用飼料添加物。  4. The animal feed additive according to claim 1, wherein the tea extract contains a polyphenol compound.
5. ポリ フヱノール化合物の含量が 5〜 8 0 %である請求項 4記載 の動物用飼料添加物。  5. The animal feed additive according to claim 4, wherein the content of the polyphenol compound is 5 to 80%.
6. ボリ フエノール化合物が (+ ) —力テキン、 (+ ) —ガロカテ キン、 (一) ーガロカテキンガレー ト、 (一) ーェピカテキン、 ( 一) 一ェピカテキンガレー ト、 (一) ーェピガロカテキン、 (一) ーェピガロカテキンガレー ト、 遊離型テアフラ ビン、 テアフラ ビン モノガレ一 ト八、 テアフラ ビンモノガレー ト B及びテアフラ ビンジ ガレー トよりなる群から選ばれる 1種又は 2種以上の化合物を含有 する請求項 4記載の動物用飼料添加物。  6. The polyphenol compounds are (+)-forced, (+)-gallocatechin, (1) -gallocatechin gallate, (1) -epicatechin, (1) -epicatechin gallate, (1) -epi One or more compounds selected from the group consisting of gallocatechin, (1) epigallocatechin gallate, free theaflavin, theaflavin monogallate 8, theaflavin monogallate B and theaflavin digallate The animal feed additive according to claim 4, which comprises:
7. ボリ フヱノール化合物中の各ボリフヱノール化合物の含量が ( + ) —力テキン 0. 2〜 6. 5 %、 ( + ) —ガロカテキン 2. 0〜 1 8. 0 %、 (一) ーガロカテキンガレー ト 1 . 0〜 1 5. 0 %、 (一) ーェピカテキン 0. 5〜 1 0. 0 %、 (一) ーェピカテキン ガレ一 ト 0 , 3〜 8. 0 %、 (一) ーェピガロカテキン 2. 0〜 1 8. 0 %、 (一) ーェピガロカテキンガレー ト 3. 0〜 2 1 . 0 % 、 遊離型テアフラ ビン 0〜 2 0. 0 %、 テアフラ ビンモノ ガレー ト A 0〜 5. 0 %、 テアフラ ビンモノガレー ト B 0〜 5. 0 %及びテ アフラビンジガレー ト 0〜 5 . 0 %である請求項 6記載の動物用飼 料添加物。 7. The content of each borophenol compound in the borophenol compound is (+)-0.2-6.5%, (+)-gallocatechin 2.0-18.0%, (-1)-gallocatechin galley G 1.0 to 15.0%, (1) eppicatechin 0.5 to 10.0%, (1) eppicatechin galleries 0, 3 to 8.0%, (1) epigallocatechin 2 0-18.0%, (1) epigallocatechin gallate 3.0-21.0%, free theaflavin 0-20.0%, theaflavin monogallate A 0-5. 0%, theaflavin monogallate B 0-5.0% and 7. The animal feed additive according to claim 6, wherein the aflavin digallate is 0 to 5.0%.
8 . 下痢の予防又は治療に用いる請求項 1 〜 7いずれか記載の動物 用飼料添加物を含有してなる動物用飼料。  8. An animal feed containing the animal feed additive according to any one of claims 1 to 7, which is used for prevention or treatment of diarrhea.
9 . 下痢が非感染性下痢症である請求項 8記載の動物用飼料。  9. The animal feed according to claim 8, wherein the diarrhea is non-infectious diarrhea.
1 0 . 非感染性下痢症がス ト レス性下痢又は感染性微生物の関与し ていない原因不明の下痢である請求項 9記載の動物用飼料。  10. The animal feed according to claim 9, wherein the non-infectious diarrhea is stress diarrhea or unexplained diarrhea not involving infectious microorganisms.
1 1 . 感染性微生物又はウィルス、 あるいはそれらの産生する毒素 に対する特異的抗体をさらに含有する請求項 8記載の動物用飼料。  11. The animal feed according to claim 8, further comprising a specific antibody against an infectious microorganism or virus, or a toxin produced thereby.
1 2 . 特異的抗体が感染性微生物又はウィルスで、 あるいはそれら が産生する毒素で過免疫された産卵鶏の卵より得られる鶏卵抗体で ある請求項 1 1記載の動物用飼料。 12. The animal feed according to claim 11, wherein the specific antibody is an egg antibody obtained from eggs of a laying hen that is hyperimmunized with an infectious microorganism or virus or a toxin produced thereby.
1 3 . 特異的抗体が感染性微生物又はウィルスで、 あるいはそれら が産生する毒素で過免疫された哺乳類の乳汁より得られる乳汁抗体 である請求項 1 1記載の動物用飼料。  13. The animal feed according to claim 11, wherein the specific antibody is a milk antibody obtained from the milk of a mammal hyperimmunized with an infectious microorganism or virus or a toxin produced thereby.
1 4 . 特異的抗体の配合量が酵素免疫測定法でブランクに対し 1 . 5倍以上の抗体価を有したものを 1 111 £ 体重1^ g以上の投与量と なるように調製された請求項 1 1 記載の動物用飼料。  14. The amount of the specific antibody was adjusted to be more than 1.5 times that of the blank by an enzyme-linked immunosorbent assay. Item 11. An animal feed according to item 11.
1 5 . 乳量増加に用いる請求項 1 〜 7いずれか記載の動物用飼料添 加物を含有してなる動物用飼料。  15. An animal feed comprising the animal feed additive according to any one of claims 1 to 7 for use in increasing milk yield.
1 6 . 乳質改善に用いる請求項 1 〜 7いずれか記載の動物用飼料添 加物を含有してなる動物用飼料。  16. An animal feed containing the animal feed additive according to any one of claims 1 to 7, which is used for improving milk quality.
1 7 . 動物の繁殖率向上に用いる請求項 1〜 7いずれか記載の動物 用飼料添加物を含有してなる動物用飼料。  17. An animal feed containing the animal feed additive according to any one of claims 1 to 7, which is used for improving the reproduction rate of the animal.
1 8 . 茶葉から水、 アルコール、 アセ トン又はこれらの混合液によ り抽出される可溶性成分を除去して得られる茶葉繊維を有効成分と する動物用飼料添加物。 18 8. Use tea, water, alcohol, acetone, or a mixture of these An animal feed additive comprising, as an active ingredient, tea leaf fiber obtained by removing a soluble component extracted from the animal.
1 9. 腸内細菌叢の改善に用いる請求項 1 8記載の動物用飼料添加 物を含有してなる動物用飼料。  19. An animal feed comprising the animal feed additive according to claim 18, which is used for improving intestinal flora.
2 0. 悪臭の低減 · 消去に用いる請求項 1 8記載の動物用飼料添加 物を含有してなる動物用飼料。  20. An animal feed containing the additive for animal feed according to claim 18, which is used for reduction and elimination of offensive odor.
2 1 . 飼料効率の改善に用いる請求項 1 8記載の動物用飼料添加物 を含有してなる動物用飼料。  21. An animal feed comprising the animal feed additive according to claim 18, which is used for improving feed efficiency.
2 2. 動物が家畜、 家禽又は愛玩動物である請求項 8〜 1 7、 1 9 〜 2 1 いずれか記載の動物用飼料。  2 2. The animal feed according to any one of claims 8 to 17, 19 to 21 wherein the animal is a domestic animal, poultry, or pet animal.
2 3. 茶葉を 3 0〜 9 5で、 0. 5〜 7時間、 水、 アルコール及び 又は酢酸ェチルで抽出し、 得られた抽出液を噴霧乾燥することよ りなる請求項 1〜 7記載の動物用飼料添加物の製造方法。  23. The method according to claim 1, wherein the tea leaves are extracted with water, alcohol and / or ethyl acetate for 30 to 95 for 0.5 to 7 hours, and the obtained extract is spray-dried. A method for producing an animal feed additive.
2 4. 茶葉を 3 0〜 9 5 eC、 0. 5〜 7時間、 水、 アルコール及び /又は酢酸ェチルで抽出し、 得られた抽出液を分画分子量 3 0 0 0 〜 6 0 0 0の限外濾過膜にて濃縮し、 次いで逆浸透膜で濃縮するこ とよりなる請求項 1 〜 7記載の動物用飼料添加物の製造方法。 2 4. tea leaves 3 0~ 9 5 e C, 0. 5~ 7 hours, water and extracted with alcohol and / or acetic acid Echiru, fractionate the extract obtained molecular weight 3 0 0 0-6 0 0 0 The method for producing an animal feed additive according to any one of claims 1 to 7, comprising concentrating with an ultrafiltration membrane, and then concentrating with a reverse osmosis membrane.
2 5. 茶葉を 3 0〜 9 5 eC、 0. 5〜 7時間、 水及び 又はアルコ ールで抽出し、 さらに酢酸ェチルで分配して得られた酢酸ェチル層 を除去することよりなる請求項 1〜 7記載の動物用飼料添加物の製 造方法。 2 5. tea leaves 3 0~ 9 5 e C, 0. 5~ 7 hours, extracted with water and or alcohol consists in further remove acetic Echiru layer obtained was partitioned with acetic acid Echiru claims Item 10. The method for producing an animal feed additive according to any one of Items 1 to 7.
2 6. (—) ーェビガロカテキンガレー トを有効成分とする動物用 飼料添加物。  2 6. (—)-Animal feed additive containing ebigalocatechin gallate as an active ingredient.
2 7. 請求項 2 6記載の動物用飼料添加物を含有してなる動物用飼 料。  2 7. An animal feed comprising the animal feed additive according to claim 26.
PCT/JP1994/001037 1993-06-30 1994-06-27 Feed additive of tea origin and animal feed containing the same WO1995001104A1 (en)

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EP0724839A1 (en) * 1995-01-31 1996-08-07 Nippon Formula Feed Mfg. Co., Ltd. Method of improving quality of eggs
EP0746985A1 (en) * 1995-06-07 1996-12-11 Nagata Agriculture Laboratory Inc. A method of producing a green tea egg
WO1997044407A1 (en) * 1996-05-23 1997-11-27 Ian Alexander Gilmour Process for extraction of proanthocyanidins from botanical material
EP0951838A1 (en) * 1997-11-11 1999-10-27 Inaba Shokuhin Co. Ltd. Cat food
EP1129628A1 (en) * 1998-11-09 2001-09-05 Taiyo Kagaku Co., Ltd. Poultry producibility improver and poultry producibility improvement method
WO2003094878A1 (en) * 2002-05-10 2003-11-20 Suntory Limited Gallocatechin gallate-containing composition
KR100556633B1 (en) * 1996-04-08 2006-04-21 소니 가부시끼 가이샤 Information processing systme and apparatus

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JPH0319656A (en) * 1989-06-15 1991-01-28 Nippon Mining Co Ltd Pet food
JPH04103539A (en) * 1990-08-24 1992-04-06 Taiyo Kagaku Co Ltd Whitened egg yolk containing specific antibody and specific antibody-containing composition
JPH0551A (en) * 1991-06-21 1993-01-08 Taiyo Kagaku Co Ltd Feed additive

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JPS61119144A (en) * 1984-11-16 1986-06-06 Osaka Chem Lab Feed for improving meat quality
JPH0297353A (en) * 1988-09-30 1990-04-09 Kotaro Shiozaki Extraction of essence in tea leaf and essence of tea
JPH0319656A (en) * 1989-06-15 1991-01-28 Nippon Mining Co Ltd Pet food
JPH04103539A (en) * 1990-08-24 1992-04-06 Taiyo Kagaku Co Ltd Whitened egg yolk containing specific antibody and specific antibody-containing composition
JPH0551A (en) * 1991-06-21 1993-01-08 Taiyo Kagaku Co Ltd Feed additive

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0724839A1 (en) * 1995-01-31 1996-08-07 Nippon Formula Feed Mfg. Co., Ltd. Method of improving quality of eggs
US5766595A (en) * 1995-01-31 1998-06-16 Nippon Formula Feed Mfg. Co., Ltd. Method of improving quality of eggs by feeding tea polyphenol
EP0746985A1 (en) * 1995-06-07 1996-12-11 Nagata Agriculture Laboratory Inc. A method of producing a green tea egg
KR100556633B1 (en) * 1996-04-08 2006-04-21 소니 가부시끼 가이샤 Information processing systme and apparatus
WO1997044407A1 (en) * 1996-05-23 1997-11-27 Ian Alexander Gilmour Process for extraction of proanthocyanidins from botanical material
AU727283B2 (en) * 1996-05-23 2000-12-07 Enzo Nutraceuticals Limited Process for extraction of proanthocyanidins from botanical material
EP0951838A1 (en) * 1997-11-11 1999-10-27 Inaba Shokuhin Co. Ltd. Cat food
EP0951838A4 (en) * 1997-11-11 2000-08-23 Inaba Shokuhin Co Ltd Cat food
EP1129628A1 (en) * 1998-11-09 2001-09-05 Taiyo Kagaku Co., Ltd. Poultry producibility improver and poultry producibility improvement method
EP1129628A4 (en) * 1998-11-09 2005-04-13 Taiyo Kagaku Kk Poultry producibility improver and poultry producibility improvement method
WO2003094878A1 (en) * 2002-05-10 2003-11-20 Suntory Limited Gallocatechin gallate-containing composition

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