JP2009173558A - Anti-canine periodontal disease composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an effective and highly safe means for preventing and/or treating canine periodontal diseases. <P>SOLUTION: Provided are a composition for preventing and/or treating the canine periodontal diseases, containing egg laid by a bird immunized with a protease originated from Porphyromonas gingivalis, and/or its treated product, and a method for preventing and/or treating the canine periodontal diseases, comprising administering an effective dose of the egg or its treated product to a dog. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a composition for preventing and / or treating periodontal disease in dogs and a method for preventing and / or treating periodontal disease in dogs.

  Periodontal disease is an infectious disease caused by bacteria in the subperipheral plaque, and is one of the most prevalent infectious diseases in modern people. One of the reasons why periodontal disease has recently attracted attention is not only the greatest cause of tooth loss, leading to deterioration of oral function and various neurological functions, but also bad breath, premature birth, and birth of low-weight infants. It is also a cause, and it becomes a high risk factor for systemic diseases such as cardiovascular disease, diabetes, endocarditis and osteoporosis. Therefore, overcoming periodontal disease is an important issue that is urgently required not only in the dental field but also in other clinical fields. However, despite many previous studies, it has not been sufficient to elucidate the etiology of periodontal disease and establish measures for prevention and treatment.

  Thus, the inventors have been researching the molecular target of this disease by paying attention to two proteases produced by Porphyromonas gingivalis, a major periodontopathic bacterium. One is Arg-gingipain (Rgp), which specifically recognizes arginine residues and cleaves at the C-terminal side, and the other is Lys-, which specifically recognizes lysine residues and cleaves at the C-terminal side. Gindipine (Kgp). Together, these two protease activities account for over 85% of the total protease activity produced by this bacterium. They are both membrane-bound and secreted.

  While these gingipains degrade proteins in the oral environment and supply amino acids essential for the growth and growth of gingivalis as nutrients, they play important roles such as adhesion of gingivalis, hemagglutination, and heme iron acquisition. In addition, various biological proteins are strongly degraded against the host, and various pathogenic functions are exhibited.

  Based on these technical informations, the inventors have found that chicken egg antibodies purified from chicken eggs obtained by immunizing chickens with gingipain Rgp and Kgp derived from Gingivalis are effective against periodontal disease in adults. And it was clarified that the safety is high (Patent Document 1).

  Recently, it has been reported that Porphyromonas species are often isolated from dog and cat plaques affected by periodontal disease, and the frequency of separation increases with age (Non-patent Document 1). The periodontal symptoms of these pets are almost the same, and it is not uncommon for tooth loss to occur between the ages of 8 and 10. Treatment is generally carried out by scaling planning and then administering antibiotics such as minocycline or clindamycin, but the emergence of resistant bacteria is a concern (Non-patent Documents 2 and 3).

  J. et al. Hardham et al. Have reported a study of a pet periodontal disease vaccine (Non-patent Document 4). They made an inactivated vaccine using Porphyromonas gulae strain B43, immunized with two subcutaneous injections into Balb / cCyJ mice, and then challenged twice with a homo or hetero strain (P. denticanis, B106). The effectiveness of this vaccine was evaluated by scoring the degree of mandibular alveolar bone resorption in mice. As a result, P. a homozygous strain. P. gulae B43 strain or hetero strain P. The effectiveness could be verified only when challenged with salivosa B104. In contrast, P. a homozygous strain. Gulae B69 or P. a heterozygous strain When challenged with denticanis B106, the effectiveness could not be verified. P. Since the monovalent inactivated vaccine produced using the galae B43 strain did not show a cross-protective effect, gulae, P.M. salivosa and P.I. The necessity of a triple vaccine consisting of denticanis was suggested.

JP 2006-36661 A H. Isogai et al. Vet. Med B 46, 467-473 (1999) M.M. Hirasawa et al., Am. J. et al. Vet. Res, 61, 1349-1352 (2000) Tetsuya Nakade et al., Animal Antibacterial Bulletin, 26, 45-51 (2004) J. et al. Hardham et al., Vet. Microbiol, 106, 119-128 (2005)

  An object of the present invention is to provide an effective and highly safe means for preventing and / or treating periodontal disease in dogs.

  The present inventors have found that a chicken egg antibody obtained by immunizing birds with a protease derived from Porphyromonas gingivalis as an antigen is effective against periodontal disease in adults, but not effective against periodontal disease in cats. That is, it was found that the chicken egg antibody is effective for periodontal disease in specific mammals. The inventors have also found that the chicken egg antibody is effective against periodontal disease in dogs, although the periodontal disease symptoms in dogs and cats are almost the same. Further, the chicken egg antibody is a P. aeruginosa derived from a dog. gingivalis, P.M. gulae, P.M. denticanis and P.I. It has been found that it has a cross-protective effect against salivosa and can effectively prevent and / or treat periodontal disease in dogs, and has completed the present invention.

That is, the present invention includes the following inventions.
(1) A composition for preventing and / or treating canine periodontal disease, comprising an egg produced by birds immunized with a protease derived from Porphyromonas gingivalis and / or a processed product thereof.
(2) The composition according to (1), wherein Arg-gingipain, Lys-gingipain or a combination thereof is used as a protease.
(3) A method for preventing and / or treating canine periodontal disease, comprising administering an effective amount of an egg produced by a bird immunized with a protease derived from Porphyromonas gingivalis or a processed product thereof to the dog.
(4) The method according to (3), wherein Arg-gingipain, Lys-gingipain or a combination thereof is used as a protease.

  According to the present invention, a causal factor that induces periodontal disease in dogs can be controlled, and treatment and prevention of periodontal disease in dogs can be carried out effectively and safely.

  In the present invention, birds immunized with a protease derived from Porphyromonas gingivalis are not particularly limited, and usually include poultry such as chickens, pupae and ducks. From the viewpoint of mass production of antibodies, it is preferable to use chickens, particularly egg-laying species.

  P. used in the present invention. The gingivalis may be collected from mammals, collected from humans, collected from dogs, and can be purchased from, for example, the American Type Culture Collection (ATCC). Specifically, P.I. gingivalis (ATCC 33277) can be used.

  P. Examples of the protease derived from gingivalis include Arg-gingipain (Rgp) and Lys-gingipain (Kgp). Rgp and Kgp may be used alone to immunize birds, or may be used in combination to immunize birds. These proteases are described in, for example, P.I. The cultured cells of gingivalis can be purified by sonication and salting out with ammonium sulfate or the like. Desirably, the salted out precipitate is purified with a Mono-Q affinity column or the like.

  As a method for immunizing birds, it is possible to immunize antigens alone or in combination of two or more thereof, in which case immunization subcutaneously, intramuscularly, nasally or with an adjuvant, orally immunized with feed or water, etc. Ordinary methods can be employed. When immunizing birds with the above-mentioned antigens, adjuvants such as Freund's complete adjuvant (FCA), Freund's incomplete adjuvant (FIA), oil adjuvant or aluminum adjuvant can be used as necessary. The interval between immunizations is not particularly limited, and 3 to 10 immunizations are performed at intervals of several days to several weeks. Usually, antibodies reacting specifically with the administered antigen within a few weeks after the first immunization are obtained in eggs, especially yolk. The amount of antigen to be inoculated is preferably a protein amount of 0.01 to 10 mg.

  Antibody titer in eggs can be measured by enzyme-linked immunosorbent assay (ELISA), bacterial cell agglutination, toxin or virus neutralization test, etc. Chicken eggs are collected at intervals of about 2 weeks after immunization. The transition of antibody titer can be traced by measuring. Usually, a high antibody titer can be obtained over about 4 months. When a decrease in antibody titer is observed after immunization, the antibody titer can be kept high by appropriately boosting at appropriate intervals.

  In the present invention, the processed egg product is not particularly limited as long as it contains an antibody against the immunogen used for immunization of birds, for example, whole eggs, yolks and egg whites of immunized eggs, these egg liquids and solutions, Moreover, the extract etc. which extracted egg liquid using propanol and chloroform are contained. From the viewpoint of the amount of antibody contained, it is preferable to contain an egg yolk component. Also included are those powdered by spray drying or freeze drying. In addition, after removing the lipid component by adding water to the egg yolk or removing the egg yolk lipid component from the egg yolk by a method using hydroxypropylmethylcellulose phthalate, polyethylene glycol, dextran sulfate, or an organic solvent such as propanol, ethanol, hexane, etc. Powdered products are also included. Such a powder in a paste or liquid form is also included. Further, in the present invention, the processed egg product includes an antibody itself purified from eggs by a known method such as ammonium sulfate salting out, sodium sulfate salting out, low temperature ethanol precipitation, ion exchange chromatography, gel filtration, affinity chromatography, etc. Are also included. In order to improve the preservability of the processed product, it is preferable to sterilize the sterilized whole egg yolk or egg yolk liquor by spray drying or freeze drying. Hereinafter, P.I. Eggs produced by birds immunized with a protease derived from gingivalis and processed products thereof are referred to as eggs and processed eggs of the present invention.

The egg and egg processed product of the present invention are characterized in that the antibody titer is 1.2 × 10 ⁵ times or more, preferably 2.5 to 5.0 × 10 ⁵ times. The antibody titer can be measured by ELISA.

  The egg and egg processed product obtained as described above are effective for the prevention and / or treatment of periodontal disease in dogs. Therefore, a composition for treating or preventing periodontal disease in dogs can be produced by blending the egg of the present invention and / or a processed product thereof into a food such as a food for specified health use or a pharmaceutical composition. it can. In the present invention, periodontal disease refers to an infection in tissues around teeth, such as gums and bones, and includes gingivitis, periodontitis and alveolar pus leakage.

  The form in particular of the composition of this invention is not restrict | limited, For example, it can be set as the form of a feed, a supplement, and a pharmaceutical composition. Examples of the dosage form of the composition include tablets, capsules, granules, powders, syrups, dry syrups, oral preparations such as liquids, suspensions and inhalants, enteral preparations such as suppositories, ointments and creams. Preparations, gels, external preparations such as patches, drops, injections and the like. Of these, oral preparations and external preparations are preferable. A composition of such a dosage form can be produced according to a conventional method by blending conventional additives in the egg and / or processed egg of the present invention according to the dosage form.

  Examples of the additive include an excipient, a binder, a disintegrant, a lubricant, an antioxidant, a coloring agent, and a corrigent, which are used as necessary. In order to achieve sustained release so that it can act in the oral cavity for a long time, it can be coated with a known retarder or the like. Excipients include, for example, sodium carboxymethylcellulose, agar, light anhydrous silicic acid, gelatin, crystalline cellulose, sorbitol, talc, dextrin, starch, lactose, sucrose, glucose, mannitol, magnesium aluminate metasilicate, calcium hydrogen phosphate Etc. can be used. Examples of the binder include gum arabic, sodium alginate, ethanol, ethyl cellulose, sodium caseinate, sodium carboxymethyl cellulose, agar, purified water, gelatin, starch, tragacanth, lactose, hydroxycellulose, hydroxymethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, etc. Is mentioned. Examples of the disintegrant include carboxymethyl cellulose, carboxymethyl cellulose sodium, carboxymethyl cellulose calcium, crystalline cellulose, starch, hydroxypropyl starch and the like. Examples of the lubricant include stearic acid, calcium stearate, magnesium stearate, talc, hydrogenated oil, sucrose fatty acid ester, waxes and the like. Examples of the antioxidant include tocopherol, gallic acid ester, dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA), ascorbic acid and the like.

  The composition of the present invention further includes various fats and oils, crude drugs, amino acids, polyhydric alcohols, natural polymers, vitamins, minerals, dietary fibers, surfactants, stabilizers, pH adjusters, antioxidants, sweeteners, A taste ingredient, a sour agent, a fragrance | flavor, etc. may be mix | blended. Other active ingredients such as antibiotics and anti-TNF-α antibodies may be added.

  In the composition of the present invention, the active ingredient egg and / or egg processed product is usually 0.01 to 1% by mass for feed and 0.1% for supplement based on the dry mass, based on the mass of the antibody. Although it is preferable to mix | blend 0.5-10 weight% normally in a pharmaceutical composition -1.0 mass%, it can also mix | blend less or more than the said range according to the objective.

  By administering an effective amount of the egg of the present invention and / or its processed product to a dog, periodontal disease in the dog can be prevented and / or treated. The egg and / or processed product thereof of the present invention can be administered as it is or in the form of the composition as described above. Effective amount means an amount sufficient to elicit the physiological or medical response of an animal, for example as sought by a physician, and more specifically corresponds to not receiving such an amount By an amount that results in an improved or improved treatment, cure, prevention, or improvement in a disease, disorder, or side effect or a decrease in the rate of progression of the disease or disorder when compared to a subject.

  The dose of the egg of the present invention and / or its processed product is not particularly limited, but is usually 25 to 45 mg, preferably 90 to 180 mg based on the dry mass based on the mass of the antibody per 1 kg of body weight per day. It is.

  The dog to be administered with the egg of the present invention and / or its processed product is not particularly limited, but preferably a dog suffering from periodontal disease, a dog possibly suffering from periodontal disease, for example, 3 years old It is the above dog.

  EXAMPLES Hereinafter, the present invention will be specifically described with reference to examples. However, the present invention is not limited to the following examples.

Example 1: Anti-P. cross-reactivity of gingivalis gingipain chicken egg antibody
(1) Preparation of protease (gingipain) gingivalis (ATCC 33277) was anaerobically cultured using 10% sterile equine defibrinated blood-treated Brucella HK agar (Kyokuto Pharmaceutical). The grown colonies were picked and transplanted to trypticase soy broth liquid medium (BD) supplemented with yeast extract, hemin, L-cysteine and menadione. The culture medium was shaken at 37 ° C. for 48 hours while blowing carbon dioxide (50 rpm).

The cultured cells were collected by centrifugation at 10,000 rpm for 30 minutes, then washed three times with PBS, and the pellet was suspended in Tris buffer (0.05M Tris-1M CaCl ₂ , pH 7.5). Sonication was performed to extract gingipain (1 minute at 150 watts, this was repeated 6 times), and purification was carried out roughly by salting out with ammonium sulfate, followed by purification with a Mono-Q affinity column (Pharmacia). did. As the elution buffer, the above buffer solution with 1M NaCl added was used. The major peak collected accounted for over 80% of the total protease activity. By this preparation method, the gingivalis, P.M. gulae, P.M. denticanis and P.I. Gindipain could also be efficiently recovered from salivosa cells.

(2) Anti-P. Preparation of gingivalis gingipain egg antibody The immunized chicken was a white leghorn species (Highline W36, Gen Corporation) about 5 months old. As the immunogen, purified gingipain added with an oil adjuvant and emulsified was used. The first immunization was performed by injecting 0.5 ml each (protein amount: 1 mg) into the left and right pectoral muscles of the chicken. Booster immunization was carried out in the same manner 8 weeks after the first immunization, and immunized eggs were collected 2 weeks later. The immune egg separates egg white and egg yolk and removes the yolk lipid component. After adding PBS (pH 7.5) equal to the egg yolk weight and stirring, further adding an equal volume of chloroform and stirring well, Centrifugation gave a water soluble fraction. The antibody was purified by salting out using ammonium sulfate. After desalting, about 10 kg of purified antibody powder was obtained by lyophilization (hereinafter referred to as anti-gingipaine chicken egg antibody). The following examples were carried out using this sample. As a control, P.P. Anti-H. pylori urease purified antibody (hereinafter referred to as negative chicken egg antibody) that was confirmed not to show specificity for gingivalis-derived gingipain was used.

(3) ELISA
The specificity of the anti-gingipain chicken egg antibody prepared in (2) for various gingipain derived from dogs was verified by ELISA.

  Each gingipain was added with 100 μl of an antigen (5 μg / ml) dissolved in 0.05 M carbonate buffer (pH 9.6) to each well of a 96-well microplate, and then allowed to stand at 4 ° C. for 18 hours to solidify. . The antigen solution in each well was removed, and in order to block non-specific reaction, 150 μl of 3% bovine serum albumin-added PBS (pH 7.5) was added to each well and allowed to stand at 37 ° C. for 60 minutes. After removing the blocking solution in each well, the wells were washed 3 times with 0.02% Tween 20-PBS. Next, an anti-gingipaine chicken egg antibody and a negative chicken egg antibody were dissolved in the washing solution, 100 μl was added to each well, and the plate was reacted at 37 ° C. for 60 minutes. After washing each well three times, horseradish peroxidase-labeled anti-chicken IgY antibody as a secondary antibody was diluted 1: 8,000 with the above washing solution, added to each well, and then at 25 ° C. for 30 minutes. Reacted. Thereafter, o-phenylenediamine and dihydrochloride were added as color formers, and 3N sulfuric acid was added to stop the reaction after 20 minutes. The data was obtained by measuring the absorbance at 490 nm.

  As a result, the anti-gingipaine chicken egg antibody gingivalis, P.M. gulae, P.M. salivosa and P.I. It showed 100%, 94%, 90% and 86% cross-reactivity for gingipain purified from denticanis, respectively. On the other hand, the negative chicken egg antibody showed no cross-reactivity.

(4) Enzyme inhibition test The anti-gingipain chicken egg antibody prepared in (2) was used to verify the protease activity inhibitory effect of various canine-derived gindipines.

  P. gingivalis, P.M. gulae, P.M. salivosa and P.I. Gin dipine purified from denticanis, respectively, and anti-gingipain egg antibody or negative egg egg antibody (750, 375, 188, 99 and 47 μg / ml, respectively) dissolved in 100 mM Tris-HCl buffer (pH 7.5) were mixed. Sensitized for 60 minutes at 4 ° C. 50 μl of the enzyme reaction was added to 150 μl of a reaction mixture consisting of Tris-HCl buffer containing 5 mM dithiothreitol, 5 mM L-cysteine and 1 mM N-α-benzoyl-L-arginine-p-nitroanilide (BapNA). This mixture was sensitized at 37 ° C. for 25 minutes, and 20% acetic acid was added to stop the enzyme reaction. The release of P-nitroaniline was measured by absorbance at 405 nm. One unit of protease activity of gingipain was the amount of enzyme that released 1 μmol of p-nitroaniline per minute of the reaction mixture, and was measured in U / ml.

  The anti-gingipaine chicken egg antibody gingivalis, P.M. gulae, P.M. salivosa and P.I. Enzyme inhibitory effects of 100%, 90%, 84% and 80% were observed for gingipain purified from denticanis, respectively. In contrast, the negative chicken egg antibody did not show an enzyme inhibitory effect.

(5) Cytotoxicity inhibition test Human pharyngeal epithelial cells (FaDu) were cultured for 18 hours in Eagle's minimum essential medium (EMEM) supplemented with 10% FBS and antibiotics. Antibody or negative chicken egg antibody and various gingipain antigens were added and sensitized for 60 minutes at 37 ° C. Cells in each well were peeled off, suspended in EMEM, and then stained with trypan blue. The number of adherent cells was calculated by microscopic examination.

  The anti-gingipaine hen egg antibody of the present invention comprises gingivalis, P.M. gulae, P.M. salivosa and P.I. Cell damage by gingipain purified from denticanis was suppressed by 88%, 78%, 75% and 70%, respectively. On the other hand, the negative chicken egg antibody did not show the cytotoxic effect.

  From the above test results, the anti-gingipaine hen egg antibody of the present invention was found to be derived from dog-derived p. gingivalis, P.M. gulae, P.M. salivosa and P.I. It was confirmed to show a remarkable cross-reactivity with Denticanis gingipain.

Example 2: Efficacy and safety of anti-gingipaine hen egg antibody against periodontal disease in dogs The effectiveness and safety of the anti-gingipain hen egg antibody prepared in (2) of Example 1 were verified. Anti-gingipaine chicken egg antibody powder is added to the feed (dry pet food) and administered to dogs continuously for 8 weeks (feed addition test) and once a day at a weekly interval, filling the periodontal groove pocket four times in total In the test (direct application test) in which direct application and administration were carried out until the end, the effect on periodontal disease and the safety for dogs were examined.

Test animals:
Breed: Beagle, Miniature Dachshund, Shiba, Cavalier, Pug Age: 5 years 6 months-7 years 5 months and breeding history 5 years or older dogs of unknown age Number of test heads: 18 Selection criteria: Left and right oral gingival sulcus Dogs with pockets in the mouth and a halitosis of medium (obvious smell) or higher

Rearing conditions:
Breeding environment: Breeding by individual in cage of open system barn [565 x 795 x 635 (H) mm] Type of feed: Solid feed for commercial dogs Feeding amount and feeding method: Place per day calculated from body weight Dosing once in the morning Drinking water: Free intake of private water supply with automatic water dispenser

Administration method:
A diet supplemented with 0.1% of anti-gingipaine chicken egg antibody powder was administered for 8 weeks. The test group consists of two groups: a group administered with an anti-gingipaine chicken egg antibody-added diet of 0.1% body weight once a day (hereinafter referred to as an administration group) and a control group administered with an antibody-free diet. The internal observation score was assigned to be approximately the same, and the administration was continued for 8 weeks for observation. In addition, after the feed addition test was completed, a periofiel gel containing 20% anti-gingipaine chicken egg antibody (Showa Showa) was used with a root canal syringe for two of the control group with two severe symptoms and other dogs with severe periodontal disease. Yakuhin Kako Co., Ltd.) was applied directly. Administration was performed once a day at weekly intervals until the gingival crevice pocket was filled, and observation was performed for 4 weeks after the first administration. The observation items are as follows.

Observation Items 1) Observation of general condition: Daily observations were made on vitality, appetite, fecal properties and the like.
2) Observation in the oral cavity: Observation in the oral cavity was performed 3 times every 4 weeks in the feed addition test, and 5 times before each administration and 1 week after the final administration in the direct administration test. In addition, according to the score of following Table 1, the state in an oral cavity evaluated separately right and left. In addition, the depth of the periodontal pocket was measured and recorded. In addition, if there is a marked change in the administration site, the details will be recorded.
3) Bacterial examination in the oral cavity: A sample was collected from a specific periodontal groove pocket using a paper point and stored at −30 ° C. until measurement. Bacteria were detected using the PCR invader method, and 16S rRNA of bacteria was used as a measurement target, and the number of total anaerobic bacteria and Porphyromonas was measured. The inspection was commissioned to BML Co., Ltd.
4) Body weight measurement: Measurement was performed immediately before the start of administration in the feed addition test, 4 and 8 weeks after the start of administration, and in the direct administration test, immediately before the first administration and after 4 weeks.
5) Feed intake: The remaining feed was confirmed for a fixed amount of feed.
6) Photographing: Photographs of the gingival sulcus (left and right) were taken immediately before administration in the feed addition test and 4 weeks and 8 weeks after the start of administration, and in the direct administration test before each administration and 1 week after the end of the final administration.

  The test schedule is summarized in Tables 2 and 3 below, and the test categories are summarized in Table 4 below.

Results In the feed addition test, the oral score was significantly decreased in the administration group compared to the pre-administration group and the control group, and oral inflammation and periodontal pocket symptoms were improved by continuous administration of anti-gingipaine chicken egg antibody for 8 weeks. (FIGS. 1 and 2). Although no significant change was observed in the total anaerobic bacteria count in the periodontal groove pockets of the administration group and the control group at the 8th week, the number of Porphyromonas genus bacteria, which are considered to be periodontal disease-causing bacteria, There was a significant decrease compared to the control group (FIG. 3). In the direct application test, the administration group showed a significant decrease in the oral score and periodontal pocket depth compared to the control group, confirming the effect of direct application of anti-gingipaine chicken egg antibody (Fig. 4 and FIG. 5). On the other hand, the number of Porpyromonas bacteria in the antibody-administered group at 4 weeks was significantly reduced as compared with that in the control group. On the other hand, there was no significant difference in the total anaerobic bacteria count in the periodontal groove pockets of both groups (FIG. 6). During the test period, no abnormalities were observed in the general condition of the test animals, confirming the safety of the anti-gingipaine chicken egg antibody.

  Based on the above, anti-P. It was confirmed that an improvement effect on periodontal disease can be obtained by administering gingivalis gingipaine hen's egg anti-feed and / or direct coating administration. In addition, since no abnormalities were observed in the general condition of the test animals during the test period, safety for dogs when used as pet oral care was confirmed.

Comparative Example: Efficacy and safety of anti-gingipaine chicken egg antibody against periodontal disease in cats Anti-gingipaine chicken egg antibody powder prepared in (1) of Example 1 was added to the feed (dry pet food) and continuously administered to cats for 8 weeks. In this study, the effect on feline periodontal disease and the safety for cats were examined.

Test animals:
Animal species: Cats Breeds: Hybrid Age: Unknown cats older than 5 years Number of test animals: 10 Selection criteria: Cats with periodontal disease have pockets in the left and right oral gingival sulcus and moderate halitosis A cat that is over)

Rearing conditions:
Rearing environment: reared individually in cages in open barns.
Feed: A commercially available cat chow was used, and the daily dose calculated from the body weight was fed once in the morning.
Drinking water: Self-administered tap water was freely consumed by an automatic water dispenser.

Administration method:
A feed supplemented with 0.1% anti-gingipaine chicken egg antibody powder was fed for 8 weeks. The test group consists of an administration group in which 0.1% of the body weight of anti-gingipaine chicken egg antibody-added feed is administered once a day and a control group in which antibody-free feed is administered. Five animals were assigned so that they would be similar.

Observations:
1) Observation of general condition: Vigor, appetite, fecal properties, etc. were observed.
2) Intraoral examination was performed in each group before administration and at 4 weeks and 8 weeks after administration. The oral clinical score was based on the criteria used in dogs.

result:
In the administration group, no significant difference was observed when the data before administration and at 4th and 8th weeks of administration were compared. Furthermore, there was no significant difference between the groups. During the test period, no clinical abnormality was observed in the general condition of the test animals. Based on the above, although the species of Porphyromonas spp. Were isolated from both dog and cat plaques affected by periodontal disease and the periodontal disease symptoms were almost the same, anti-P. It was confirmed that the gingivalis gingipain chicken egg antibody is not effective for cats but effective for dogs. That is, it was confirmed that the antibody is effective against a specific mammal.

The fluctuation | variation of the intraoral inflammation of an anti- gingipaine hen egg antibody administration group and a control group in a feed addition test is shown. The fluctuation | variation of the depth of the periodontal pocket of an anti- gingipaine hen egg antibody administration group and a control group in a feed addition test is shown. In the feed addition test, the total anaerobic bacteria count and the number of Porphyromonas spp. In the periodontal groove pockets of the anti-gingipaine hen egg antibody administration group and the control group are shown. The fluctuation | variation of the intraoral inflammation of an anti- gingipaine hen egg antibody administration group and a control group in a direct application | coating test is shown. The reduction | decrease of the periodontal pocket depth of an anti- gingipaine hen egg antibody administration group and a control group in a direct application | coating test is shown. The total anaerobic bacteria number and the number of Porphyromonas bacteria in the periodontal groove pocket of the anti-gingipaine hen egg antibody administration group and the control group in the direct application test are shown.

Claims (4)

  A composition for preventing and / or treating canine periodontal disease, comprising an egg produced by birds immunized with a protease derived from Porphyromonas gingivalis and / or a processed product thereof.   The composition according to claim 1, wherein Arg-gingipain, Lys-gingipain or a combination thereof is used as a protease.   A method for preventing and / or treating canine periodontal disease, comprising administering to a dog an effective amount of an egg produced by a bird immunized with a protease derived from Porphyromonas gingivalis or a processed product thereof.   The method according to claim 3, wherein Arg-gingipain, Lys-gingipain or a combination thereof is used as the protease.

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