阅读说明：本技术 动物用的口腔用组合物及使用了它的动物用的牙周病预防剂、传染病预防剂及口臭预防剂 (Oral composition for animal, and agent for preventing periodontal disease, infectious disease, and halitosis for animal using the same ) 是由 渡边到 于 2019-07-04 设计创作，主要内容包括：本发明的技术问题在于提供一种动物用的口腔用组合物,其通过使用乳香树脂和/或乳香精油而抑制制造成本,且特别是对狗或猫等玩赏动物所特有的牙周病相关细菌显示作用。所述动物用的口腔用组合物用于灭杀牙周病细菌,其含有乳香树脂作为有效成分,所述牙周病细菌为齿周卟啉单胞菌(Porphyromonas circumdentaria)。(The present invention addresses the problem of providing an animal oral composition that is capable of suppressing the production cost by using a mastic resin and/or mastic essential oil, and that exhibits an effect on periodontal disease-related bacteria specific to pet animals such as dogs and cats in particular. The composition for oral cavity use for animals is used for killing periodontal disease bacteria, which contains mastic resin as an active ingredient, wherein the periodontal disease bacteria is Porphyromonas circumdentia (Porphyromonas circumdentia).)

1. An oral composition for animals, which is used for killing periodontal bacteria and infectious bacteria, characterized by comprising mastic resin as an active ingredient, wherein the periodontal bacteria is Porphyromonas gingivalis (Porphyromonas circumdentia).

2. The oral composition for animals according to claim 1, wherein said infectious pathogen is Pasteurella multocida (Pasteurella multocida).

3. The composition for oral cavity use in animals according to claim 1 or 2, wherein said mastic resin is used as a mastic resin solution having a concentration of 10 to 60%.

4. The animal oral composition according to claim 3, wherein the mastic resin solution is prepared by dissolving mastic resin in a solvent, and the solvent for dissolving the mastic resin is any one selected from ethanol, glycerol, dipropylene glycol, 1, 3-butylene glycol, triglycerol fatty acid ester triglyceride (caprylic/capric) glyceride having 8-18 carbon atoms derived from fatty acid, fatty acid monoglyceride having 8-18 carbon atoms derived from fatty acid, glyceryl monocaprate, fatty acid ester having 8-18 carbon atoms derived from fatty acid, isopropyl myristate, ethyl isooctanoate, octyldodecanol myristate, higher alcohol having 8-22 carbon atoms, oleyl alcohol, sorbitan fatty acid ester having 8-18 carbon atoms derived from fatty acid, and sucrose fatty acid ester having 8-18 carbon atoms derived from fatty acid.

5. The composition for oral cavity of animal as claimed in claim 3 or 4, wherein the content of said mastic resin liquid is 0.1-50%.

6. The oral composition for animals according to any one of claims 1 to 5, further comprising mastic essential oil as an active ingredient.

7. The composition for oral cavity of animal as claimed in claim 6, wherein the mastic essential oil is contained in an amount of 0.01-1.0%.

8. The oral composition for an animal according to any one of claims 1 to 7, wherein the subject animal is a dog or a cat.

9. The oral composition for animals according to any one of claims 1 to 8, wherein the retention agent is one selected from the group consisting of liquid paraffin, gelled hydrocarbon which is a mixture of liquid paraffin and polyethylene, vegetable oil, and beeswax.

10. The oral composition for animals according to any one of claims 1 to 8, wherein the retention agent is two or more selected from the group consisting of liquid paraffin, gelled hydrocarbon which is a mixture of liquid paraffin and polyethylene, vegetable oil, and beeswax.

11. A prophylactic agent for periodontal disease in an animal, which comprises the oral composition for an animal according to any one of claims 1 to 10.

12. An agent for preventing an infectious disease in an animal, which comprises the oral composition for an animal according to any one of claims 1 to 10.

13. An agent for preventing halitosis in an animal, which comprises the oral composition for an animal according to any one of claims 1 to 10.

Technical Field

The present invention relates to an oral composition for animals, which contains mastic resin and/or mastic essential oil as an active ingredient, and which is particularly effective for pet animals (pets) such as dogs and cats. The present invention also relates to a prophylactic agent for periodontal disease, a prophylactic agent for infectious disease, and a prophylactic agent for halitosis for animals, each using the oral composition for animals.

Background

In general, in humans, the adhesion of dental plaque (tartar) is one of the causes of dental caries (caries) or periodontal disease, and it has been pointed out that in oral hygiene, the removal or prevention of dental plaque, i.e., plaque control, is important. With regard to the mechanism of plaque formation, microorganisms in the oral cavity, particularly glucosyltransferases, which are extracellular enzymes of Streptococcus mutans (Streptococcus mutans), synthesize adhesive and insoluble glucans using sucrose as a substrate, and the glucans adhere to the tooth surface to form plaque as an agglomerated mass of bacteria.

As a method for controlling dental plaque, mechanical removal of dental plaque by a toothbrush or the like and oral sterilization using an oral bactericide are mainly used. However, when the dental plaque is mechanically removed by a toothbrush or the like, the dental plaque cannot be sufficiently removed unless a trained and skilled tooth brushing method is performed for a long time. In addition, in the method using an oral disinfectant, there is a problem that the disinfectant component cannot penetrate into the interior of the bacterial aggregated mass such as dental plaque, and thus the effect thereof cannot be sufficiently exhibited. Therefore, measures such as increasing the concentration of the bactericide component or prolonging the treatment time are required. In addition, since the antibacterial agent acts on all bacteria in the oral cavity, normal oral flora and bacteria useful for the human body are also killed, and thus it is not satisfactory from the viewpoint of safety, economy and effectiveness.

In addition, periodontal diseases are diseases with symptoms of inflammation occurring in periodontal tissues of teeth, such as gingivitis, and alveolar pyorrhea. Bacteria such as Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Candida are known as pathogenic bacteria of periodontal disease. Periodontal disease is mainly a disease caused by dental plaque, and like the prevention of dental caries, plaque control is useful for the prevention of periodontal disease, but causes the same concerns as those described above. In addition, in periodontal disease, if the disease or condition is not further worsened by dental plaque, it is necessary to receive professional treatment (mainly chemotherapy using antibiotics, tooth extraction, etc.) in dental or oral surgery hospitals, and various digestive system diseases may be caused as side effects depending on the administered drugs, for example.

In order to eliminate the concern of preventing dental caries and periodontal disease in humans as described above, various oral compositions have been developed using materials (e.g., natural products, etc.) which exhibit effects with a small amount of active ingredients and whose ingredients can be easily handled. As an example thereof, japanese patent No. 3389556 (patent document 1) and japanese patent application laid-open No. 2012 and 97018 (patent document 2) disclose oral compositions using mastic gum (mainly its sap). Olibanum refers to Pistacia Lentiscus (Pistacia Lentiscus) of Pistacia of Anacardiaceae planted on Greek Grosk island, and its sap is mainly used for preventing dental caries or periodontal disease. In addition, it is also known that Boswellia serrata juice not only prevents human dental caries and periodontal disease but also has antibacterial effect against helicobacter pylori or Campylobacter. In addition, as main components of Olibanum tree juice, Olibanum essential oil, Olibanum dienoic acid, IsoOlibanum dienoic acid, triterpenes, aldehydes, alcohols, poly beta-myrcene, etc. can be mentioned.

Further, as an oral composition containing a material which exhibits an effect by a small amount of an active ingredient and whose treatment with the ingredient is simple, japanese patent application laid-open No. 2014-166992 (patent document 3) discloses an oral composition mainly containing Lactobacillus brevis, which is one of plant lactic acid bacteria. The plant lactic acid bacteria used in patent document 3 are used as dead bacteria, with the cell length of about 0.1 to 5 μm. Thus, an oral composition with a low content and a suppressed production cost is provided.

Furthermore, it is essential to prevent dental caries and periodontal disease in animals other than humans, particularly pet animals (pets) such as dogs and cats. As in the case of humans, pet animals are also subjected to mechanical plaque control by a breeder using a toothbrush, or chemical treatment by mouth rinsing using a prophylactic or therapeutic agent for periodontal disease for animals. In addition, treatment using pet food or chewing gum containing these therapeutic agents, administration of drugs, and preventive treatment by vaccination are also performed. Further, although depending on living environment, pathogens causing periodontal diseases and infectious diseases of dogs and cats include, for example, Porphyromonas gingivalis (Porphyromonas gingivalis), Prevotella intermedia (Prevotella intermedia), Campylobacter rectus (Campylobacter rectus), and Tenbetween (tannorella forsythensis) in common with humans. However, there are infectious diseases of dogs and cats such as Pasteurella, Porphyromonas salivarius, Porphyromonas odonta (Porphyromonas Circumdentaria), edobacter denticans, or Pasteurella multocida (Pasteurella multocida), Pasteurella canis (Pasteurella canis), Pasteurella damascena (Pasteurella dagmatis), Pasteurella oralis (Pasteurella stomatis), etc. which are not found in humans.

For example, japanese patent No. 4099213 (patent document 4) discloses a prophylactic vaccine against Porphyromonas gulae and the like, which are pathogenic bacteria of periodontal disease parasitic on dogs or cats. Further, as an oral composition against Porphyromonas gulae and the like, Japanese patent laid-open publication No. 2015-67539 (patent document 5) discloses an oral composition derived from Angelica keiskei Koidz extract. Further, as an oral composition for porphyryromonas gulae and the like, japanese patent No. 6468559 (patent document 6) discloses an oral composition using mastic gum.

As a pathogenic bacterium of infectious diseases other than humans such as dogs and cats, Pasteurella multocida (Pasteurella multocida) and other Pasteurella bacteria are described, but infection by Pasteurella has been the most common cause of periodontal disease and pneumonia in dogs and cats. However, it is known that when a person is infected with pasteurella-carrying dogs or cats, there is a high possibility that pasteurella disease is induced, and eventually cellulitis or sepsis progresses. For example, japanese patent application laid-open No. 2018-58875 (patent document 7) discloses a drug for preventing such bacterial infection by pasteurella multocida or the like and a treatment method using the same. The invention of patent document 7 discloses that the composition shows an effect on infectious diseases such as humans, dogs, cats, cows, sheep, goats, pigs, birds, fish and horses regardless of the types of gram-negative or gram-positive bacteria.

Documents of the prior art

Patent document

Patent document 1: japanese patent No. 3389556

Patent document 2: japanese laid-open patent publication No. 2012-97018

Patent document 3: japanese patent laid-open No. 2014-166992

Patent document 4: japanese patent No. 4099213

Patent document 5: japanese laid-open patent publication No. 2015-67539

Patent document 6: japanese patent No. 6468559

Patent document 7: japanese patent laid-open publication No. 2018-58875

Disclosure of Invention

Technical problem to be solved by the invention

However, patent documents 1,2 and 3 are only for human subjects, and although they show an effective effect on, for example, Porphyromonas gingivalis (Porphyromonas gingivalis), there is no description or suggestion that such an effect is shown on pathogenic bacteria of periodontal disease parasitizing dogs or cats, such as Porphyromonas gulae, Porphyromonas salivarius, odonobacterium denticola, and the like.

The invention of patent document 4 is mainly vaccination, and although not directly described, needs veterinary treatment. In other words, it is not described or suggested that a breeder who does not qualify as a veterinarian can care pets such as dogs and cats. Of course, patent document 4 does not describe or suggest the use of mastic as in patent documents 1 and 2, and does not describe or suggest the use of plant lactic acid bacteria as in patent document 3.

In patent document 5, a preparation such as a paste or a gel is used for the convenience of handling by a breeder. In addition, the use of mastic is described. Based on the above, an oral composition for animals based on the findings of patent documents 1 to 5 can be considered.

However, in patent document 5, only the angelica keiskei koidzumi extract is used as an active ingredient for Porphyromonas gulae. Although patent document 5 describes the use of mastic, it does not describe or suggest that mastic is used only as a flavoring agent and shows an effect on pathogenic bacteria of periodontal disease which parasitize dogs or cats.

Further, as described above, since pasteurella causes infectious diseases of other mammals, birds, and the like, it is considered that prevention and sterilization are required at the stage of parasitizing dogs or cats, that is, prevention and sterilization by oral care are required. However, patent document 6 describes that the antibacterial effect against Porphyromonas gulae is observed, but it is not clear whether or not the antibacterial effect against pasteurella is exerted. In addition, patent document 7 acts only when infectious diseases (pasteurellosis) are infected, and since the active ingredient is robenidine (1, 2-bis [ (E) - (4-chlorophenyl) methyleneamino ] guanidine) and robenidine or the like is produced by synthesis, the cost for molecular design and synthesis is high, and the risk of side effects is higher than that of natural products. Patent document 7 does not disclose or suggest whether robenidine can be used as an oral composition. Further, even when the inventions of patent documents 1 to 6 and the invention of patent document 7 are combined, there is no description or suggestion as to whether or not frankincense is effective against pasteurella.

The present invention has been made in view of the above circumstances, and provides an oral composition for animals which can suppress the production cost by using a mastic resin and/or a mastic essential oil and which exhibits an effect on periodontal disease-related bacteria specific to pet animals such as dogs and cats in particular.

Means for solving the problems

The above object of the oral composition for animals of the present invention is achieved by: an oral composition for animals for killing periodontal bacteria and infectious bacteria, which contains mastic resin as effective component, wherein the periodontal bacteria is Porphyromonas dentata (Porphyromonas circumdentaria).

Further, the above object of the animal oral composition of the present invention can be more effectively achieved by: rendering said infectious pathogen Pasteurella multocida (Pasteurella multocida); or preparing the mastic resin into mastic resin liquid with the concentration of 10-60% for use; or the mastic resin liquid is prepared by dissolving mastic resin in a solvent, wherein the solvent for dissolving mastic resin is selected from ethanol, glycerol, dipropylene glycol, 1, 3-butylene glycol, triglyceride fatty acid ester tri (caprylic/capric) glyceride having 8 to 18 carbon atoms of a part derived from fatty acid, fatty acid monoglyceride having 8 to 18 carbon atoms of a part derived from fatty acid, monodecanoic glyceride, fatty acid ester having 8 to 18 carbon atoms of a part derived from fatty acid, any one of isopropyl myristate, ethyl isooctanoate, octyldodecanol myristate, higher alcohol having 8 to 22 carbon atoms, oleyl alcohol, sorbitan fatty acid ester having 8 to 18 carbon atoms derived from fatty acid, or sucrose fatty acid ester having 8 to 18 carbon atoms derived from fatty acid; or the content of the frankincense resin liquid is 0.1-50%; or further comprises Olibanum essential oil as effective component; or the content of the frankincense essential oil is 0.01-1.0%; or making the subject animal a dog or cat; or the retention agent is one selected from the group consisting of liquid paraffin, gelled hydrocarbon which is a mixture of liquid paraffin and polyethylene, vegetable oil, and beeswax; or the retention agent is two or more selected from the group consisting of liquid paraffin, gelled hydrocarbon which is a mixture of the liquid paraffin and polyethylene, vegetable oil, and beeswax.

The present invention can also be effectively achieved by an animal periodontal disease preventive agent, an animal halitosis preventive agent, or an animal infectious disease preventive agent, each using the animal oral composition.

Effects of the invention

It is found that the use of the mastic resin and/or the mastic essential oil in the composition for oral cavity for animals according to the present invention can suppress the production cost, and particularly, the composition exhibits an effect on bacteria (e.g., pasteurella multocida) related to periodontal disease and infectious disease, which are specific to pet animals such as dogs and cats.

Further, the use of the oral composition for animals of the present invention can be applied to a prophylactic agent for periodontal disease for animals or a prophylactic agent for halitosis for animals depending on the formulation.

Drawings

Fig. 1 is a graph showing the change in the score of the gingival index of example (test example 3).

FIG. 2 is a graph showing the change in the number of bacteria in the oral cavity.

FIG. 3 is a graph showing the results of measurement of halitosis.

Detailed Description

The oral composition for animals of the present invention will be described in detail below. In the present application, the "boswellia tree sap" refers to a sap collected from Pistacia lentiscus (Pistacia lentinus) of Pistacia of Anacardiaceae, and as described in the background art, the main components thereof are boswellia dienoic acid, iso-boswellia dienoic acid, triterpenes, aldehydes, alcohols, poly-beta-myrcene, and the like. "mastic resin" refers to a substance obtained by naturally drying and solidifying mastic tree juice. The "Olibanum essential oil" refers to a substance obtained by refining volatile components (mainly terpenes) in Olibanum tree juice or Olibanum resin by steam distillation or dry distillation. In addition, "%" is a weight percentage unless otherwise specified.

First, the mastic resin will be described. In the oral composition for animals of the present invention, mastic resin is an important component that exhibits an antibacterial effect against periodontal disease-related bacteria (in particular, Porphyromonas gulae). As described above, mastic resin is obtained by naturally drying mastic tree juice. The time for natural drying is not particularly limited as long as it is 1 day or more.

In addition, in this composition, the mastic resin is dissolved in a solvent and used. The mastic resin itself is insoluble in water, and compatibility with various additives when the composition is in various dosage forms such as gel or liquid is studied, so that the mastic resin is dissolved in a solvent.

As the solvent for dissolving the mastic resin, there can be used ethanol, glycerin, dipropylene glycol, 1, 3-butylene glycol, triglycerol fatty acid ester (the number of carbon atoms of the fatty acid-derived portion is about 8 to 18, preferably about 8 to 12), tricaprylin/capric acid glyceride, fatty acid monoglyceride (the number of carbon atoms of the fatty acid-derived portion is about 8 to 18, preferably about 8 to 12), monocaprylic acid glyceride, fatty acid ester (the number of carbon atoms of the fatty acid-derived portion is about 8 to 18, preferably about 8 to 12), isopropyl myristate, ethyl isooctanoate, octyldodecanol myristate, higher alcohol (the number of carbon atoms is about 8 to 22), oleyl alcohol, sorbitan fatty acid ester (the number of carbon atoms of the fatty acid-derived portion is about 8 to 18, preferably about 8 to 12), A solvent such as sucrose fatty acid ester (the fatty acid-derived portion has about 8 to 18 carbon atoms, preferably 8 to 12 carbon atoms).

When the mastic resin is used in the oral composition for animals of the present invention, it is sufficient to use a homogeneous system solution with respect to the solvent. Hereinafter, the homogeneous system solution is referred to as "mastic resin solution". The concentration of the mastic resin solution is preferably 5 to 60%. If the concentration is 5% or less, the antibacterial effect against Porphyromonas gulae cannot be obtained, and if the concentration is 60% or more, the antibacterial effect against Porphyromonas gulae is reduced, although the antibacterial effect is not reduced more than that when the concentration is 5% or less.

The preparation method of the mastic resin solution may be performed by a conventional method, as long as the concentration is maintained. The temperature at which the mastic resin is dissolved may be appropriately increased in consideration of the boiling point of the solvent, and may be room temperature. Further, it is preferable to use a mastic resin solution obtained by dissolving mastic resin in a solvent and then filtering the solution.

In addition, the content of the mastic resin liquid is preferably 0.1 to 50% based on the total amount of the animal oral composition of the present invention. In addition, when the content of the mastic resin liquid is 0.1% or less in terms of mastic resin itself, the bactericidal effect against Porphyromonas gulae is reduced or not exhibited. If the content of the mastic resin solution is 50% or more, even if the bactericidal effect against Porphyromonas gulae is sufficient, there is a concern that tissues around the affected part of the pet may cause some inflammation or allergic reaction, and in some cases, the bactericidal effect against Porphyromonas gulae may be reduced from that in the above concentration range.

Next, the essential oil of Olibanum will be explained. As described above, the mastic essential oil may be obtained by converting volatile components (mainly terpenes) in mastic tree juice or resin into essential oils by steam distillation or dry distillation. In addition, the essential oil is prepared by a conventional method.

The content of the mastic essential oil is preferably 0.01 to 1.0% based on the total amount of the animal oral composition of the present invention. If the content of the mastic essential oil is 1.0% or more, there is a possibility that tissues around the affected part of the pet, etc. may cause some inflammation or allergic reaction, and the bactericidal effect against Porphyromonas gulae may be weakened, as in the case of mastic resin solution. Further, even when the composition for oral cavity for animal of the present invention does not contain mastic oil, i.e., mastic resin alone, it shows bactericidal effect on Porphyromonas gulae, but when mastic oil is contained, more excellent bactericidal effect can be obtained. In addition, the essential oil of Olibanum is added to prevent halitosis. In addition, it is because the effect of preventing halitosis is not exhibited when the content of the mastic essential oil is 0.01% or less.

The composition for the oral cavity for animals of the present invention is established by further containing lactic acid bacteria having a mode of particle size distribution of 1.0 μm or less as an active ingredient. The "mode of particle size distribution" as used herein is a value indicating an index of the size of bacteria (cell length), and means a particle size at which the relative frequency in the particle size distribution is the maximum when the particle size of the cells (cell length) is measured. In other words, the term "mode of 1.0 μm or less" means a bacterium having a cell length in the range of 0.1 to 5 μm. The length of the cells can be measured by a known technique such as electron microscopy. When the mode is 1.0 μm or more, the inactivation effect is exhibited on Porphyromonas gulae, but the number of bacteria incorporated is drastically reduced, and therefore, it is preferably 1.0 μm or less. The lactic acid bacterium used in the present invention may be prepared by a known technique (see, for example, international patent publication No. 2009/157073).

Examples of lactic acid bacteria used in the oral composition for animals of the present invention include Lactobacillus species such as Lactobacillus brevis (Lactobacillus brevis), Lactobacillus brevis subspecies coagulans (l.breves coemulans), Lactobacillus acidophilus (l.acidophilus), Lactobacillus gasseri (l.gasseri), Lactobacillus mareli (l.mali), Lactobacillus plantarum (l.plantarum), Lactobacillus buchneri (l.buchneri), Lactobacillus casei (l.casei), Lactobacillus johnsonii (l.johnsonii), Lactobacillus gallinarum (l.gallinarum), Lactobacillus amylovorans (l.amylovorus), Lactobacillus rhamnosus (l.rhamnosus), Lactobacillus kefir (l.kefir), Lactobacillus paracasei (l.paracasei), and Lactobacillus crispus (l.ispassius); bacteria belonging to the genus Lactococcus such as Lactococcus lactis; enterococcus bacteria such as Enterococcus faecalis (Enterococcus faecalis) and Enterococcus faecium (e.faecalis); bifidobacterium bacteria such as Bifidobacterium bifidum (Bifidobacterium bifidum), Bifidobacterium longum (b.longum), Bifidobacterium adolescentis (b.adolescentis), Bifidobacterium infantis (b.infantis), Bifidobacterium breve (b.breve), and Bifidobacterium catenulatum (b.catenulatum). Among them, bacteria belonging to the genus Lactobacillus are preferable, and among them, a strain of Lactobacillus brevis (L.brevis) FERM BP-4693 is preferable. In addition, dead bacteria are preferably used as the lactic acid bacteria. This is because the lactic acid bacteria used in the present invention can be easily produced and can exhibit a sufficient bactericidal effect even when the bacteria are dead.

In the oral composition for animals of the present invention, the lactic acid bacteria are preferably contained in an amount of 0.01 to 1.0% based on the total amount of the oral composition for animals. If the content is less than 0.01%, the lactic acid bacteria cannot exhibit the effect of inactivating Porphyromonas gulae. In addition, if the content is 1.0% or more, the number of bacteria incorporated is affected.

The formulation of the oral composition for animals of the present invention may be selected from toothpaste, gel, liquid toothpaste, powder toothpaste, mouthwash, film, chewing gum, additive for pet food, and pasta (pasta).

Furthermore, a prophylactic agent for periodontal disease for animals and a prophylactic agent for halitosis for animals can be further prepared under the same blending conditions as those of the oral composition for animals of the present invention.

The oral composition for animals of the present invention, and the agent for preventing periodontal disease for animals and the agent for preventing halitosis for animals using the same can be implemented as described above, but may contain various additives. Hereinafter, the additive will be described.

As an example of the additive, the oral composition for animals of the present invention is prepared by further blending papaya extract and/or chitosan. These substances show a synergistic effect with the lactic acid bacteria mentioned above.

The papaya extract is an extract from natural papaya fruits, which is extracted by grinding and soaking the natural papaya fruits in a solvent such as ethanol, and the papaya fruits may be ripe papaya or incompletely ripe papaya in a cyan state. The fructus Chaenomelis extract can be used as humectant for keeping oral cavity moist, and especially immature fructus Chaenomelis is rich in papain. The papain can easily remove tartar on the surface of teeth or between teeth and gingiva, and can more effectively inactivate Porphyromonas gulae by using lactic acid bacteria. The amount of the papaya extract to be blended is not particularly limited, but is preferably 0.005% to 10% based on the total amount of the oral composition of the present invention. If the ratio is less than 0.005%, the above-mentioned effect cannot be exerted, and if the ratio exceeds 10%, the effect of lactic acid bacteria on Porphyromonas gulae may be reduced.

In contrast, chitosan is obtained by boiling chitin obtained from the exoskeletons of crustaceans such as crabs and shrimps with strong alkali or the like. These polysaccharides are sometimes used as an adhesive, but also have an antibacterial effect and an effect of exhibiting a coating effect on the surface of teeth. In addition, the above lactic acid bacteria or papain can be retained on the surface of the teeth for a longer time. This can further inactivate Porphyromonas gulae. The amount of the papaya extract to be blended is not particularly limited, but is preferably 0.005% to 10% based on the total amount of the oral composition of the present invention. If the ratio is less than 0.005%, the above-mentioned effect cannot be exerted, and if the ratio exceeds 10%, the effect of lactic acid bacteria on Porphyromonas gulae may be reduced.

Further, chitosan and papaya extract may be blended at the same time. Therefore, the chitosan can prolong the time of the papain and the lactic acid bacteria staying on the surface of the teeth or between the teeth and the gingiva, and can further exert the effect of inactivating Porphyromonas gulae by combining the sterilization effect brought by the chitosan. The amount of chitosan and papaya extract added is not particularly limited, but is preferably 0.005% to 10%, respectively. If the ratio is less than 0.005%, the above-mentioned effect cannot be exerted, and if the ratio exceeds 10%, the effect of lactic acid bacteria on Porphyromonas gulae may be reduced.

Examples of the polishing agent include silica-based polishing agents such as silica gel, precipitated silica, additive silica, hydrated silicic acid, silica, zeolite, aluminosilicate, and zirconium silicate, crystalline cellulose, dibasic calcium phosphate dihydrate, anhydrous dibasic calcium phosphate, calcium pyrophosphate, magnesium phosphate, calcium phosphate, aluminum hydroxide, alumina, light calcium carbonate, heavy calcium carbonate, magnesium carbonate, zirconium silicate, and synthetic resin polishing agents. One or two or more of them can be used. The amount of these abrasives blended is usually 0 to 60% based on the total amount of the oral composition of the present invention.

Examples of the humectant include polyhydric alcohols such as glycerin, concentrated glycerin, diglycerin, sorbitol, maltitol, dipropylene glycol, propylene glycol, 1, 3-butylene glycol, xylitol, and polyethylene glycol, and one or two or more of these polyhydric alcohols can be used. In addition, these wetting agents may also be used as a solvent for dissolving the mastic resin.

Examples of the binder (thickener) include alginic acids and derivatives thereof such as carrageenan, alginic acid, sodium alginate, propylene glycol alginate, calcium-containing sodium alginate, potassium alginate, calcium alginate, and ammonium alginate, xanthan gum, guar gum, gelatin, agar, sodium carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, sodium polyacrylate, and pullulan, and one or two or more of them can be used. In addition, the tackifier also acts as a gelling (gel) agent.

Examples of the foaming agent include sodium lauryl sulfate, sodium lauroyl sarcosinate, sodium sulfosuccinate, sodium coconut fatty acid monoglyceride sulfonate, sodium α -olefin sulfonate, N-acyl amino acid salts such as N-acyl glutamate, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazoline betaine, maltitol fatty acid esters, sucrose fatty acid esters, polyglycerol fatty acid esters, fatty acid diethanolamides, polyoxyethylene sorbitan monostearate, polyoxyethylene hydrogenated castor oil, polyoxyethylene fatty acid esters, and glycerin fatty acid esters, and one or two or more of these may be used.

Examples of the pH (hydrogen ion concentration) adjuster include citric acid, sodium (mono-or di) -citrate, malic acid, sodium (mono-or di) -malate, gluconic acid, sodium (mono-or di) -gluconate, succinic acid, sodium succinate, lactic acid, sodium (mono-or di) -lactate, potassium carbonate, and sodium hydrogen carbonate, and one or two or more of them may be used.

As a retention agent for retaining (retaining) the active ingredient of the animal oral composition of the present invention, liquid paraffin, gelled hydrocarbon which is a mixture of liquid paraffin and polyethylene, vegetable oil, beeswax, or the like can be used, and one or two or more of them can be used at the same time. In addition, the gelled hydrocarbon also serves as a gelling agent.

Examples of the sweetener include saccharin sodium, aspartame, trehalose, stevioside, stevia extract, p-methoxycinnamaldehyde, neohesperidin dihydrochalcone (neohesperidin dihydrochalcone), perillartine, and xylitol.

As the preservative, there are parabens such as methyl paraben, ethyl paraben, propyl paraben and butyl paraben, sodium benzoate, phenoxyethanol, alkyldiaminoethylglycine hydrochloride and the like.

As the flavor component, one or two or more of L-menthol, anethole, menthone, eucalyptol, limonene, carvone, methyl salicylate, ethyl butyrate, eugenol, thymol, cinnamaldehyde, trans-2-hexenal, and the like can be used. These components may be blended singly, or essential oils containing these components and the like may be used.

The amount of each component such as the humectant, the binder, the foaming agent, the retention agent, the sweetener, the preservative, and the flavor component is not particularly limited, and is usually in the range of 0.001 to 20% relative to the total amount of the oral composition for animals.

In addition to the above-mentioned perfume components, perfume components such as aliphatic alcohols and esters thereof, terpenoids or terpenoid alcohols, phenol ethers, aldehydes, ketones, and lactones, and essential oils may be blended as long as the effects of the present invention are not hindered. The amount of the flavorant is usually in the range of 0.001 to 20% based on the total amount of the oral composition for animals of the present invention.

In addition to the above, the animal oral composition of the present invention may further contain an active ingredient. Examples of such active ingredients include lysozyme chloride, sodium fluoride, potassium fluoride, sodium monofluorophosphate, potassium nitrate, sodium polyphosphate, polyethylene glycol, polyvinylpyrrolidone, hinokitiol (hinokitiol), ascorbic acid (vitamin C), ascorbate, chlorhexidine salts, cetylpyridinium chloride, benzalkonium chloride, benzethonium chloride, bisabolol, triclosan, 3-methyl-4-isopropylphenol, tocopherol acetate, epsilon-aminocaproic acid, tranexamic acid, aluminum hydroxyallantoate, aluminum lactate, dihydrocholesterol, glycyrrhetinic acid salts, copper chlorophyllin salt, sodium chloride, guaiazulene sulfonate, dextranase, pyridoxine hydrochloride, and medicinal hydroxyapatite, and one or two or more of these may be blended. The content of the active ingredient is usually in the range of 0.001 to 20% based on the total amount of the oral composition of the present invention.

In addition, when the mastic resin (which is a mastic resin solution), the mastic essential oil, the lactic acid bacteria, the additive, and the like are mixed in the above numerical range with respect to the total amount of the animal oral composition of the present invention, the remaining part thereof may be a solvent (for example, a solvent for dissolving the mastic resin) or a gelling agent. The additive can be used in the same manner as the animal oral composition of the present invention, and can be used as a prophylactic agent for periodontal disease or a prophylactic agent for halitosis for animals.

The oral composition for animals of the present invention can be prepared according to a conventional method, and the preparation method thereof is not particularly limited.

Further, although various modes have been described for the animal oral composition of the present invention in the case of Porphyromonas gulae, it is considered that the animal oral composition of the present invention exhibits an effect on periodontal disease bacteria existing in mammals other than humans, for example, Porphyromonas salivarius, odonobacteria nodericanus, pasteurella, and the like.

Further, the oral composition for animals of the present invention has been described with respect to an embodiment in which the composition is directed to a dog or cat, which is an animal other than human, but it may be applied to other rabbits, hamsters, guinea pigs, and the like which are enjoyed as pets.

While the embodiments of the oral composition for animals of the present invention have been described above, the present invention is not limited to the above embodiments, and various embodiments may be adopted without departing from the scope of the claims and the description of the present specification.

Examples

In order to support the above-described embodiments of the oral composition of the present invention, examples are further described. In the examples described herein, the effects of mastic resin, mastic essential oil, and lactic acid bacteria having a particle size distribution mode of 1.0 μm or less (hereinafter, referred to as "nano-type lactic acid bacteria" in the present examples) on periodontal disease bacteria (Porphyromonas gulae in the present examples; hereinafter, referred to as "p.gulae" in the present examples) as components of the oral composition for animals of the present invention were examined.

[ PREPARATION EXAMPLE 1] preparation of Olibanum resin solution and Olibanum essential oil

First, a sap (hereinafter referred to as "boswellia tree sap") was collected from Pistacia lentiscus (Pistacia lentiscus) of the genus Pistacia of the family Anacardiaceae, produced in Greek, Grosvenor island, and naturally dried for 1 day. After drying, the volatile component and the resinous material are separated by steam distillation, and the resinous material is referred to as "mastic resin" used in the examples of the present application. On the other hand, the volatile components were separated by steam distillation and then refined into "mastic essential oil" used in this example. In addition, the essential oil refining was carried out according to a conventional method.

Next, a 30% solution of mastic resin was prepared using a tri (caprylic/capric) glyceride as a solvent. The resulting solution was used as "boswellia resin solution" used in this example.

[ preparation example 2] preparation of Nano-type lactic acid bacterium

According to international patent publication No. 2009/157073, the nano-type lactic acid bacteria used in the present example were prepared.

First, as the lactic acid bacteria, Lactobacillus brevis (strain FERM BP-4693; hereinafter, sometimes referred to simply as "Brevibacterium") which is a plant lactic acid bacteria was used, and the culture was terminated at a point when glucose (glucose) was completely consumed by culturing the Brevibacterium at 36.5 ℃ while adjusting the pH (hydrogen ion concentration) at the time of culture to 6.5 using a known nutrient medium to which 5% glucose was added and using a 20% sodium hydroxide aqueous solution.

After the termination of the culture, the culture solution was heated at 80 ℃ for 10 minutes, the cells were washed with PBS (phosphate buffer solution), dextrin was added as an excipient in an amount of 4 times the weight of the cells, the mixture was dispersed in a mixer and freeze-dried to prepare a sample, and the sample was resuspended in PBS so that the cell concentration was 10 mg/mL. In addition, Brevibacterium which maintained pH at 6.5 in the processing step was used as the nano-type lactic acid bacterium (Brevibacterium) used in the present example. The nano-lactic acid bacteria are further subjected to a heating treatment to produce dead bacteria.

Further, the particle size of the produced nano-type lactic acid bacteria was measured, and as a result, the particle size of all the cells was 0.7 to 1.0 μm or less, and the mode of the particle size distribution was 1.0 μm or less. In addition, the particle size measurement was carried out according to a conventional method.

[ PREPARATION EXAMPLE 3 preparation of dilutions of Olibanum resin solution, Olibanum essential oil and Lactobacillus plantarum

Furthermore, the frankincense resin solution, the frankincense oil and the nano-type lactic acid bacteria used in the present example were diluted with glycerol tris (caprylic/capric) ester as a solvent. Specifically, the frankincense resin liquid is diluted to 10 times, and the frankincense essential oil and the nano lactic acid bacteria are respectively diluted to 1000 times. Hereinafter, in this example, the term "milk flavor resin diluent", "milk flavor essential oil diluent" and "nano type lactic acid bacteria diluent" are used, respectively. In addition, a sterilized test tube or the like is used as a test tube or the like for dilution. In addition, each dilution was prepared just prior to use.

[ preparation example 4] preparation of test solution for testing Effect against periodontal disease bacteria

As a typical example of the test solution (which may be simply referred to as "test solution") used in the test of the effect against periodontal bacteria in the present example, the test solution in example 8 described later is explained. In example 8, "boswellia resin solution 3.00%, boswellia essential oil 0.03%, and lactobacillus nanoscopic 0.01%" (see table 2), at this time, 2.7mL of the boswellia resin diluent, 2.7mL of the boswellia essential oil diluent, 1.0mL of the lactobacillus nanoscopic diluent, and 2.6mL of tri (caprylic/capric) glyceride, which were prepared in preparation example 3, were added to 1 tube which had been sterilized, and the total amount was set to 9 mL. Also, the order of mixing is not particularly required.

In each of the examples (see tables 1 to 4) described later, the amounts of each diluent and the tri (caprylic/capric) glyceride may be appropriately changed depending on the concentrations of the frankincense resin liquid, the frankincense essential oil and the nano-type lactic acid bacteria, as long as the total amount is 9 mL.

In addition, the test solution in the control test (comparative example; refer to tables 1 to 4) was only tri (caprylic/capric) glyceride, and not mixed with the frankincense resin solution, the frankincense essential oil and the nano-type lactic acid bacteria. The details will be described later.

In addition, test solutions of examples and comparative examples were also prepared immediately before use.

Preparation example 5 preparation of bacterial suspension for periodontal disease (P.gulae)

P.gulae, which is a pathogenic bacterium of periodontal disease of dogs, was selected as the periodontal disease bacterium used in test example 1 described later, and JCM13865 was used as the strain thereof^T。

Also, the strain was anaerobically cultured at 37 ℃ for 5 days using a blood agar medium. After the culture, a proper amount of the grown colonies were inoculated with platinum and suspended in sterilized physiological saline and a standard solution of Mycoplasma turbidity (No. 1). This suspension was used as "periodontal disease (p. gulae) bacterial suspension" used in this example. The bacterial strain of periodontal disease (P.gulae) has a bacterial count of about 1 to 3X 10⁸CFU/mL. And, periodontal disease (p. gulae) bacterial solution was prepared just before use.

Test example 1 one of the tests for the Effect against periodontal disease bacteria

The test was performed using the test solutions for the anti-periodontal disease bacterial effect test of each example and each comparative example prepared in preparation example 3 and the periodontal disease (p.gulae) bacterial solution prepared in preparation example 5.

First, the test solutions of the respective examples will be described. In examples 1 to 4, the boswellia resin solution was fixed at 10.0%, and the varieties (variation) of boswellia essential oil (fixed at 0.03%) and lactobacillus nano-particles (fixed at 0.01%) were changed, respectively. In examples 5 to 8, the boswellia resin solution was fixed at 3.0%, and the varieties of boswellia essential oil (fixed at 0.03%) and lactobacillus nano-particles (fixed at 0.01%) were changed, respectively. In examples 9 to 12, the boswellia resin solution was fixed at 1.0%, and the varieties of boswellia essential oil (fixed at 0.03%) and lactobacillus nano-particles (fixed at 0.01%) were changed, respectively. In examples 13 to 16, the boswellia resin solution was fixed at 0.5%, and the varieties of boswellia essential oil (fixed at 0.03%) and lactobacillus nano-particles (fixed at 0.01%) were changed, respectively. The test solutions of the examples were prepared just before use by the method shown in preparation example 3.

Next, each comparative example will be explained. "comparative example 1" as the control experiment of examples 1 and 2, "comparative example 2" as the control experiment of examples 3 and 4, "comparative example 3" as the control experiment of examples 5 and 6, "comparative example 4" as the control experiment of example 7, "comparative example 5" as the control experiment of example 8, "comparative example 6" as the control experiment of example 9, "comparative example 7" as the control experiment of example 10, "comparative example 8" as the control experiment of examples 11 and 12, "comparative example 9" as the control experiment of examples 13 and 14, and "comparative example 10" as the control experiment of examples 15 and 16 were respectively tested.

Next, the test for the effect against periodontal bacteria will be described. In short, the time point when 1mL of periodontal disease (p.gulae) bacterial suspension was added to 9mL of the test solution in each example was regarded as 0 minute, and the number of p.gulae bacteria was counted with time. The following is a detailed description.

First, 1mL of periodontal disease (p.gulae) bacterial solution was added to each of 9mL of test solutions of examples 1 to 16, and at this time point (the action time is 0 minute), 1mL of the mixed solution was aspirated by a pipette gun. To the aspirated mixed solution, 9mL of sterilized physiological saline was added and stirred, and this operation was repeated, thereby performing 10-fold gradient dilution. 0.1mL of each solution considered to be at the appropriate dilution gradient was pipetted and smeared onto 2 pieces of blood agar medium. After the smearing, each medium was subjected to anaerobic culture at 37 ℃ for 5 days, and the number of colonies in each medium was counted, and the number of bacteria in p.gulae was counted when the action time of each example was 0 minute.

The numbers of bacteria in p.gulae were counted after the test solutions of examples 1 to 16 were allowed to act on the periodontal disease (p.gulae) bacterial suspension for 5 minutes, 10 minutes, 15 minutes, and 30 minutes under the same conditions. However, for some reasons, the action time of example 7 was not measured after 5 minutes, the action time of example 9 was not measured after 5 minutes, and the action time of example 11 was not measured after 30 minutes (see tables 2 and 3 described later).

After the above counting of the number of bacteria, in examples 5 to 8, the number of bacteria of p.gulae was counted again (second time) after the test solution and the periodontal disease (p.gulae) bacterial solution were allowed to act for 0 minute, 5 minutes, 10 minutes, 15 minutes, and 30 minutes, respectively, under the same conditions. After the above counting of the number of bacteria, in examples 1 to 4, the number of bacteria of p.gulae was counted for the second time after the test solution was allowed to act on the periodontal disease (p.gulae) bacterial solution for 0 minute, 1 minute, 3 minutes, and 5 minutes under the same conditions. In examples 9 to 16, the number of p.gulae bacteria was not counted again (see tables 3 and 4 described later).

Next, in comparative examples 1 to 10, the frankincense resin solution, the frankincense oil and the nano-type lactic acid bacteria were not mixed, 1mL of periodontal disease (p.gulae) bacterial solution was added to 9mL of tricaprylin (caprylic/capric) glyceride, the acting time was taken as 0 minute, and the number of p.gulae bacteria was counted under the same dilution conditions, culture medium conditions and acting time as in each example.

Next, the changes in the number of p.gulae bacteria in examples 1 to 16 and comparative examples 1 to 10 are shown in tables 1 to 4.

[ Table 1]

TABLE 1 Olibanum resin solution (10%) and Olibanum essential oil and/or nano-type lactobacillus test solution

Bactericidal effect on p

% by weight

First, table 1 will be explained. Table 1 shows the results of counting the number of p.gulae bacteria in examples 1 to 4, in which the boswellia resin solution was fixed to 10% and the varieties of boswellia essential oil and lactobacillus nano-particles were changed, respectively. In addition, for the number of bacteria, in order to clarify the number of digits of the number of bacteria, conversion was made to a common logarithm.

In comparative example 1, the number of bacteria was reduced by one digit or almost constant with the lapse of time, that is, almost no change in the number was observed, compared to example 1 in which only the 10% frankincense resin solution was used and example 2 in which the 10% frankincense resin solution and 0.03% frankincense essential oil were used, the number was reduced by about 4 digits at the moment of action (about log [ CFU/mL ] before the test solution and the periodontal disease (p.gulae) bacterial solution acted), and reached the limit of the count range (log [ CFU/mL ] < 3.5) after 5 minutes. Further, in examples 1 and 2, the number of bacteria was measured again, and as a result, it was found that the action time became the limit of the counting range at 3 minutes ("second time" in table 1).

Next, as in comparative example 1, the number of bacteria was also reduced by one digit or almost constant in comparative example 2, that is, almost no change in the number was observed, and in comparison with example 3 using 10% of the frankincense resin solution and 0.01% of the nano-type lactic acid bacteria and example 4 using 10% of the frankincense resin solution, 0.03% of the frankincense essential oil and 0.01% of the nano-type lactic acid bacteria, the number of bacteria was not more than the limit of the count range (log [ CFU/mL ] < 3.5) in the initial measurement at an action time of 5 to 10 minutes. Further, in examples 3 and 4, the number of bacteria was measured again, and as a result, it was found that the number of bacteria was gradually decreased at an action time of 5 minutes (see "second time" in table 1), unlike the cases of examples 1 and 2.

From this, it is seen that in examples 1 to 4, when the boswellia resin solution is 10%, the number of bacteria in p.gulae tends to decrease either by itself or when it is mixed with the boswellia essential oil and/or the nano-type lactic acid bacteria, and the count range limit (log [ CFU/mL ] < 3.5) is surely reached after 30 minutes.

[ Table 2]

TABLE 2 Olibanum resin solution (3.0%) and Olibanum essential oil and/or nano-type lactobacillus test solution

Bactericidal effect on p

% by weight

N.m. ═ not determined

The following description will be made with reference to table 2. Table 2 shows the results of counting the number of p.gulae bacteria in examples 5 to 8, i.e., when the boswellia resin solution was fixed to 3.0% and the varieties of boswellia essential oil and lactobacillus nanoscales were changed, respectively. In addition, for the number of bacteria, the number of digits was converted to a common logarithm in order to clarify the number of bacteria, as in table 1.

In comparative example 3, the number of bacteria was reduced by one digit or almost constantly as time passed, that is, almost no change in the number was observed, compared to example 5 in which only the 3.0% frankincense resin solution and example 6 in which the 3.0% frankincense resin solution and 0.03% frankincense essential oil were used, the number was reduced by about 2.5 digits at the moment of action (about log [ CFU/mL ] before the test solution and periodontal disease (p.gulae) bacterial solution acted), and the count range limit (log [ CFU/mL ] < 3.5) was reached after 10 minutes. Further, as a result of measuring the number of bacteria again (for the second time) in examples 3 and 4, it was found that the action time was 15 minutes and almost became the count range limit or less, and that both examples 3 and 4 became the count range limit or less after 30 minutes (see "for the second time" in table 2).

Next, the number of bacteria in comparative examples 3 and 4 was also reduced by one digit or almost constant, that is, almost no change in the number was observed, as compared to example 7 using 3.0% of the frankincense resin solution and 0.01% of the nano-type lactic acid bacteria and example 8 using 3.0% of the frankincense resin solution, 0.03% of the frankincense essential oil and 0.01% of the nano-type lactic acid bacteria, which were almost equal to or less than the limit of the count range (log [ CFU/mL ] < 3.5) at an action time of 30 minutes in the initial measurement. However, unlike the cases of examples 5 and 6, the results of measuring the number of bacteria again in examples 7 and 8 revealed that the reaction time was 10 minutes and then gradually became not more than the limit of the counting range (log [ CFU/mL ] < 3.5) ("second time" in Table 2).

It is thus understood that in examples 5 to 8, the number of bacteria in p.gulae tends to decrease when the boswellia resin solution is 3.0%, either by itself or when it is mixed with the boswellia essential oil and/or the nano-lactic acid bacteria, but as shown in example 8, the number of bacteria in the boswellia resin solution, the boswellia essential oil and the nano-lactic acid bacteria is surely below the limit of the count range (log [ CFU/mL ] < 3.5) after 10 minutes, as compared with examples 5 to 7.

[ Table 3]

TABLE 3 Olibanum resin solution (1.0%) and Olibanum essential oil and/or nano-type lactobacillus test solution

Bactericidal effect on p

% by weight

N.m. ═ not determined

The following description will be made with reference to table 3. Table 3 shows the results of counting the number of p.gulae bacteria in examples 9 to 12, i.e., when the boswellia resin solution was fixed to 1.0% and the varieties of boswellia essential oil and lactobacillus nanoscales were changed, respectively. In addition, the number of bacteria was converted to a common logarithm in order to clarify the number of digits of the number of bacteria, as in tables 1 and 2.

In comparative examples 6 and 7, the number of bacteria was reduced by one digit or almost constant with the passage of time, i.e., no change in the number was observed, compared to example 9 using only 1.0% boswellia resin solution and example 10 using 1.0% boswellia resin solution and 0.03% boswellia essential oil, where the number of bacteria was almost constant or even reduced by log [ CFU/mL ] > 5.5.

Next, the number of bacteria was also reduced by one digit or almost constant in comparative example 8, that is, almost no change in the number was observed, as in other comparative examples, compared to example 11 using 1.0% of the frankincense resin solution and 0.01% of the nano-type lactic acid bacteria and example 12 using 1.0% of the frankincense resin solution, 0.03% of the frankincense essential oil and 0.01% of the nano-type lactic acid bacteria, which slightly reduced the number of bacteria as compared to examples 9 and 10.

From this, it is seen that in examples 9 to 12, when the boswellia resin solution is 1.0%, the number of bacteria in p.gulae tends to be slightly reduced when the nano-lactic acid bacteria are added, as compared with the cases of the boswellia resin solution and the boswellia essential oil alone. However, it is found that the reduction of the number of bacteria is slow or almost constant, unlike the examples 1 to 8 in which the concentration of the mastic resin solution is relatively high.

[ Table 4]

TABLE 4 Olibanum resin solution (0.5%) and Olibanum essential oil and/or nano-type lactobacillus test solution

Bactericidal effect on p

% by weight

The following description will discuss table 4. Table 4 shows the results of counting the number of p.gulae bacteria in examples 13-16, in which the boswellia resin solution was fixed to 0.5% and the varieties of boswellia essential oil and lactobacillus nano-devices were changed, respectively. In addition, the number of bacteria was converted to a common logarithm in order to clarify the number of digits of the number of bacteria, as in tables 1,2 and 3.

In comparative examples 9 and 10, the number of bacteria was reduced by one digit or almost constant with the lapse of time, i.e., no change in the number was observed, compared to example 13 in which only 0.5% of the frankincense resin solution was used and example 14 in which 0.5% of the frankincense resin solution and 0.03% of the frankincense essential oil were used, which were also the same as comparative examples 9 and 10, and only a slight reduction in the number of bacteria of p. In addition, the results of example 15 using 0.5% of the frankincense resin solution and 0.01% of the nano-type lactic acid bacteria and example 16 using 0.5% of the frankincense resin solution, 0.03% of the frankincense essential oil and 0.01% of the nano-type lactic acid bacteria were also almost the same as those of example 13 and example 14.

From this, it is seen that in examples 13 to 16, i.e., when the boswellia resin solution is 0.5%, the number of bacteria in p.gulae tends to be slightly reduced when the nano-lactic acid bacteria are added, as compared with the cases of the boswellia resin solution and the boswellia essential oil alone. However, it is found that the reduction of the number of bacteria is slow or almost constant, unlike the examples 1 to 8 in which the concentration of the mastic resin solution is relatively high.

As described above, in test example 1, regarding only p.gulae, there is room for study as to whether or not the antibacterial (bactericidal) properties are exhibited against periodontal disease bacteria (for example, Porphyromonas salivarius, Porphyromonas macaca, etc.) specific to other dogs or cats, and there is room for study as to detailed conditions (for example, in vivo scale), but the applicant believes that at least in test tube (in vitro) scale, the mastic resin, the mastic essential oil, and/or the nano-type lactic acid bacteria, which are components of the oral composition for animals of the present invention, sufficiently exhibit the antibacterial effects and antibacterial effects against p.gulae.

Preparation example 6 preparation of bacterial solutions for periodontal diseases (P.gulae (cat) and P.circumdentaria) and infectious diseases (pas.mutocida) in cats

Three periodontal disease bacteria, i.e., Porphyromonas Gulae (hereinafter referred to as "P.Gulae (cat)"; strain C26Db5), Porphyromonas denticola (hereinafter referred to as "P.Cicumdataria"; strain YC34b), and Pasteurella multocida (hereinafter referred to as "pas.multocida"; strain 12K), which are pathogenic bacteria of feline periodontal disease, were selected as the periodontal disease bacteria used in test example 2 described later.

Subsequently, P.gulae (cat) was anaerobically cultured at 37 ℃ for 5 days using a sheep blood agar medium (Oxioid; manufactured by BD Co., Japan) for anaerobic bacteria, CDC (Center for Disease Control and prevention, USA Center for Disease Control and prevention). P.Circumdataria was anaerobically cultured at 37 ℃ for 5 days using blood agar medium (mhTS) supplemented with 10.0ml/L hemin and 10.0ml/L menadione. We performed aerobic culture of pas. ultrascidia using soybean casein agar medium (TSA, Difco) at 37 ℃ for 1 day.

And the number of the first and second electrodes,for each of the bacteria p.gulae (cat), p.circumdentaria and pas.multocida, a proper amount of a grown colony of each bacterium was extracted by platinum loop inoculation and suspended in sterilized normal saline (concentration of "mazel turbidity standard solution No. 1") to prepare a bacterial solution. Hereinafter, a bacterial solution of each bacterium was prepared as a periodontal disease (p.gulae) bacterial solution, a periodontal disease (p.circumdentaria) bacterial solution, and an infectious disease (pas.mutocida) bacterial solution, depending on the species. In addition, the bacterial count of the bacterial liquid of each bacterium is about 1-3 multiplied by 10⁸CFU/mL。

Test example 2 second test for Effect against periodontal disease bacteria

An anti-periodontal disease bacterial effect test was performed using the periodontal disease (p.gulae (cat)) bacterial solution, periodontal disease (p.circumdentaria) bacterial solution, infectious disease (pas.mutocida) bacterial solution, and test solution (test solution) for an anti-periodontal disease bacterial effect test prepared in preparation example 6. The test solution (test solution) used in test example 2 for the effect test on periodontal disease bacteria was prepared in the same manner as in preparation example 4, except that the content of the mastic resin solution was 5.0% and the content of mastic essential oil was 0.03%. In addition, as in the above preparation example 4, the test solution (referred to as "control test solution") in the control test (control zone) was only tri (caprylic/capric) glyceride, and the boswellia resin solution and boswellia essential oil were not mixed.

First, 1mL of each bacterial solution (periodontal disease (p. gulae) bacterial solution, periodontal disease (p. circumdentaria) bacterial solution, and infectious disease (pas. mucocida) bacterial solution) was added to 9mL of the test solution, and the number of bacteria was counted with time.

For counting the number of bacteria, after each bacterial solution was added to the test solution and stirred (this was assumed to be 0 minute in action time), 1mL of a mixed solution of the stirred test solution and each bacterial solution was immediately sucked by a pipette gun, 9mL of sterilized physiological saline was further added and stirred, and the operation of further adding 9mL of sterilized physiological saline to 1mL of a diluted solution and stirring was repeated to dilute each bacterial solution by 10-fold gradient. After dilution, 0.1mL of each of the bacterial solutions was applied to 2 blood agar medium. Furthermore, using this medium, periodontal disease (p.gulae (cat)) bacterial suspension and periodontal disease (p.circumdentaria) bacterial suspension were anaerobically cultured at 37 ℃ for 5 days, and infectious disease (pas. mucotida) bacterial suspension was aerobically cultured at 37 ℃ for 1 day. After the culture of each bacterial suspension, the number of bacteria in the test solution (mixed with each bacterial suspension) was determined from the number of colonies in each medium. The limit of the number of bacteria was 3.5(log [ cfu/mL ]), and the number of bacteria below the limit was < 3.5(log [ cfu/mL ]).

Then, for test solutions 1 minute, 3 minutes, 5 minutes, 10 minutes, or 5 minutes, 10 minutes, 15 minutes, and 30 minutes after the respective bacterial solutions were added and stirred, sterilized saline was added to each test solution and diluted by 10-fold gradient dilution in the same manner as in the case where the action time was 0 minute, and the number of bacteria after culture was counted. As a control, bacterial liquid was added to the control test solution, and the number of bacteria was determined in the same manner as the time-dependent change.

The results of the anti-periodontal disease bacteria effect test based on the mixing and stirring of each bacterial solution (periodontal disease (p. gulae (cat)) bacterial solution, periodontal disease (p. circumdentaria) bacterial solution, and infectious disease (pas. mucocida) bacterial solution) with the test solution are shown in tables 5 to 7, respectively. The "test area" in tables 5 to 7 is a solution obtained by adding each bacterial solution to a test solution.

[ Table 5]

TABLE 5 Bactericidal effect on periodontal disease (P. gulae (cat)) bacterial liquid

[ Table 6]

TABLE 6 bacterial liquid for periodontal disease (P. Circumdataria)

[ Table 7]

TABLE 7 bactericidal effect on infectious disease (pas. mucocida) bacterial solutions

Table 5 shows the results of the anti-periodontal disease bacteria effect test of the periodontal disease (p. gulae (cat)) bacterial solution and the test solution. As a result of adding 7.7(log [ cfu/mL ]) of the bacterial suspension to the test solution, the bacterial count was 4.6(log [ cfu/mL ]) 5 minutes after the addition (stirring), and the measurement limit was not more than 10 minutes after the addition. In contrast, in the control (control zone), the number of bacteria was almost constant at about 7.0. + -. 0.5(log [ cfu/mL ]) within 0 to 30 minutes after the stirring. When 5.2(log [ cfu/mL ]) of the bacterial suspension was added to the test solution, the measurement limit was not more than 1 minute after the addition.

Table 6 shows the results of the anti-periodontal disease bacteria effect test of the periodontal disease (p. circumdentaria) bacterial solution and the test solution. The bacterial solution was added to the test solution, and as a result, 1 minute or 3 minutes after the addition (stirring) was below the measurement limit. In contrast, in the control (control zone), the number of bacteria was 5.9 to 7.0(log [ cfu/mL ]) within 0 to 30 minutes after the stirring.

Table 7 shows the results of the anti-periodontal bacteria effect test of infectious disease (pas. mucocida) bacterial liquid and test liquid. When this bacterial suspension was added to the test solution, the concentration was 1% or less (about 4.0(log [ cfu/mL ])) at 1 minute after the addition, and the measurement limit was 3 minutes or less after the addition. In contrast, in the control (control zone), the number of bacteria was 6.3 to 8.1(log [ cfu/mL ]) within 0 to 30 minutes after the stirring.

In the above test examples, although the number of periodontal disease bacteria or infectious bacteria varies under reaction conditions such as aerobic or anaerobic conditions, when the animal oral composition of the present invention is studied using the boswellia resin solution and/or the boswellia essential oil as the active ingredient, it was found that the animal oral composition of the present invention exhibits bactericidal activity not only against Porphyromonas gulae derived from dogs and cats but also against other Porphyromonas denticola or pasteurella multocida of a different genus, which is Porphyromonas.

[ preparation example ] preparation of oral composition for animal of the present invention

Based on the findings of the test examples, the oral composition for animals (gel paste) of the present invention was prepared. In addition, as for the preparation method of the composition, it is carried out by a conventional method. The composition ratios (wt%) of the respective components are shown in table 8 below. The total amount of the oral composition was set to 25 kg.

[ Table 8]

TABLE 8 formulation of oral compositions of the present invention

In addition: 30% solution of Tris (caprylic/capric) glyceride in mastic resin

In addition: carboxymethyl cellulose

In addition: mixture of liquid paraffin and polyethylene

The gel paste-like oral composition for animals prepared in the preparation example was immersed in a toothbrush or a cotton swab and applied to the teeth or gums of animals (dogs and cats).

The above preparation examples (addition amounts) are only examples, and other components may be appropriately changed as long as the composition of the mastic resin (resin solution) and the mastic essential oil and/or the nano-type lactic acid bacteria is observed.

[ test example 3] clinical test of oral composition for animals

Clinical tests were conducted on ordinary domestic dogs (dog species are not limited; hereinafter, may be abbreviated as "dog") for the gel-paste animal oral composition (hereinafter, referred to as "mastic gel") prepared in the above preparation examples.

First, 10 dogs, which were generally kept at home, scored 0.5 or more for all gum indices, received scaling (removal of dental calculus) treatment, and had a body weight of less than 25kg and 3 years or more, and were not limited to a certain dog. And, 10 dogs were grouped into 5 control groups (not coated with mastic gel; general dental care except for coating mastic gel) and 5 test drug groups (coated with mastic gel). In addition, the dogs in the control group and the test drug group were observed for canine teeth and 4 th premolar teeth in the left, right, upper and lower jaws of the teeth of the test subjects. Here, a method of calculating the gingival index score is shown in table 9.

[ Table 9]

TABLE 9 calculation method of gingival index score

Next, the method of administering (painting and smearing) the mastic gel to the dogs of the test drug group was: about 0.5g of mastic gel was applied to the gums of the left upper jaw, left lower jaw, right upper jaw, and right lower jaw of the dog with the fingers of each breeder of the dog. Also, the application and rubbing of the masticatory gel was performed 1 time 1 day before sleep, and every day for 4 weeks. In addition, care was taken to avoid the concomitant use of supplements for dental care purposes when administering the mastic gel, and to avoid diet and drinking within 30 minutes after administration of the gel.

Then, clinical efficacy tests were performed. The effect test was performed in four ways: 1) calculation of a score for the gingival index, 2) analysis of the type of periodontal pathogenic bacteria and pili, 3) counting of the number of bacteria in the oral cavity and 4) determination of halitosis. Hereinafter, each test will be described in turn.

[ test example 3-1] calculation of a Gum index score

First, the gingival index scores of all dogs in the control group and the test drug group were visually observed and judged before (hereinafter referred to as "preoperative") and after 1 month (hereinafter referred to as "1 month") of the scaling treatment on the basis shown in table 9. In addition, in this case, it is considered that the gingival index score is not deteriorated or improved as compared with the preoperative score. The change in the gum index score of this test example 3-1 is shown in FIG. 1.

In fig. 1, differences in the value of the score of the gingival index were observed before (before) the scaling treatment and after 1 month (after 1 month) the scaling treatment for the control group and the test drug group, respectively. Further, when comparing the control group with the test drug group after 1 month of the scaling treatment (after 1 month), a decrease in the score of the test drug group (dogs using mastic gel), i.e. an improvement in gingivitis, was observed. From this, it is clear that, although there is room for statistical or detailed studies, at least the results suggest that the oral composition of the present invention exhibits an effect as a prophylactic agent for gingivitis or periodontal disease.

[ test examples 3-2] analysis of periodontopathic bacteria and pili types

Next, the detection of periodontal pathogenic bacteria and the analysis of the pilus type will be described.

First, detection of periodontal pathogenic bacteria is performed using a PCR (polymerase chain reaction) analysis method. For the collection of periodontal pathogenic bacteria, 3 cotton swabs (seed swab (registered trademark) γ 2; manufactured by Eiken Chemical co., ltd.) were used to collect the gingival margin on the side of the 4 th premolar tooth of the upper jaw and the left and right of each dog of the control group and the test drug group. Then, with respect to 3 cotton swabs, bacterial (periodontal disease) DNA was extracted from the collected cotton swab samples, PCR analysis was performed, and bacteria were detected by agarose gel electrophoresis. In addition, the fimA type of the detected bacteria and Porphyromonas gulae are as follows: a to C types of bacteria selected from the group consisting of Trastonella furathensis (tannorella forsythis), Campylobacter rectus (Campylobacter rectus), Porphyromonas gingivalis, Porphyromonas gulae, and Porphyromonas gulae fimA. Also, fimA detection was performed simultaneously with PCR analysis (refer to International patent publication No. 2013/089166). In addition, in the PCR analysis, t-test, Mann-Whitney U-test, Wilcoxon signed rank test and Fisher's exact test were performed, and a case where the P value was <0.05 was regarded as statistically significant.

The results of detection of periodontal pathogenic bacteria by PCR analysis and Porphyromonas gulae fimA are shown in table 10.

[ Table 10]

TABLE 10 analysis results of periodontal disease bacteria based on PCR analysis and fimA detection

*: has significant difference with the prior art (P <0.05)

Pgu: porphyromonas gulae, Pgi: porphyromonas gingivalis

type A: type A, type B of Pgu fimA: type B, type C of Pgu fimA: type C of Pgu fimA

Cr Campylobacter rectus and Tf Fostainer

In addition: for example, "4/5" in "preoperative" of "Pgu" means 4 out of 5.

For the PCR analysis, both in the control and test drug groups: the number of individuals analyzed for periodontal disease pathogenic bacteria was decreased after scraping teeth, and the number of individuals detected after 1 month tended to increase again (refer to table 10).

Regarding the fimA test, 2 cases of the type a carrying Porphyromonas gulae fimA were present in both groups before the scaling, but the type a of the fimA disappeared in all dogs after the surgery and was not detected after 1 month. With regard to type C, there was no significant difference between the control group and the test drug group, and with regard to type B, the effect of the test drug was unclear since no dog carried type B in the preoperative examination of the test drug group.

Test example 3-3 count of the number of bacteria in oral cavity

Regarding the number of bacteria in the oral cavity, the buccal side of the 4 th molar tooth of the left upper jaw of the dogs of the control group and the test drug group was rubbed three times with a white sterilized cotton swab, and the bacteria attached to the cotton swab were counted using an automatic bacteria counter (DU-AA01 NP-H; manufactured by Suo-Loose health medical Co., Ltd.). In addition, the t test, the mann-whitney U test, the wilcoxon signed rank test, and the fisher's exact test were performed in the same manner as the PCR count of test example 3-2, and a case where the P value was <0.05 was regarded as statistically significant.

Fig. 2 is a graph showing changes in the number of bacteria in the oral cavity. In fig. 2, "before operation" means before the scaling treatment, "after operation" means immediately after the scaling treatment, and "after 1 month" means after 1 month of the scaling treatment. In FIG. 2, the number of bacteria in the oral cavity was significantly reduced in both the control group and the test drug group by scraping teeth (P < 0.05). With respect to the number of bacteria in the oral cavity after 1 month, the number of bacteria in the oral cavity in the test drug group was significantly reduced as compared with the control group, and a significantly low value was shown as compared with the preoperative value.

[ test examples 3-4] measurement of halitosis

Next, oral malodor measurement was performed on dogs in the control group and the test drug group using orasctip (DS PHARMA ANIMAL HEALTH co., LTD). The results are shown in FIG. 3. Similarly to fig. 2, "before" in fig. 3 means before the scaling treatment, "after" means immediately after the scaling treatment, and "after 1 month" means after 1 month of the scaling treatment. The orasetrp scores before and after scaling showed lower values in the test drug group and the control group. Further, after 1 month, both groups showed lower values compared to before the operation. In addition, the orasetrp score of the test drug group was always a lower value than the control group in the control group and the test drug group.

In the above test examples 3(3-1 to 3-4), clinical trials using the oral composition of the present invention revealed that at least the mastic gum component has applications as a prophylactic agent for periodontal disease (gingivitis) or a prophylactic agent for halitosis in dogs and cats, although there is still room for various studies.

As described above, various examples of the oral composition for animals of the present invention have been described, but the present invention is not limited thereto, and various examples can be carried out without departing from the scope of the claims and the scope described in the above embodiments.

Industrial applicability

The oral composition for animals of the present invention according to the present invention is described in the above embodiments and examples by taking toothpaste as an example, but since the composition for oral cavity for animals of the present invention uses mastic resin and/or essential oil, it can be used as an antibacterial agent for animals or an agent for preventing infectious diseases (for example, against parasitic bacteria in the digestive system).