Method for delivering polyhydroxybutyric acid to large intestine of animal

Abstract

The present invention is a method for delivering polyhydroxybutyric acid to the large intestine of an animal by using algae, which are photosynthetic organisms including polyhydroxybutyric acid, as a carrier for polyhydroxybutyric acid. The algae are, for example, the green algae Chlorella or the blue-green algae Spirulina.

Description

Method for delivering polyhydroxybutyrate to the large intestine of an animal

[0001] The present invention relates to a method for delivering polyhydroxybutyrate to the large intestine of an animal.

[0002] Polyhydroxybutyrate (hereinafter referred to as "PHB") is known to suppress inflammation or infectious diseases occurring in animals. Patent Document 1 describes that PHB induces a butyric acid bacteria-dominated intestinal flora and can therefore be used as a new prebiotic. Patent Document 2 describes that PHB suppresses ulcerative colitis in mammals. Patent Document 3 describes that PHB has a growth-promoting effect in fish and suppresses infectious diseases. Patent Document 4 describes that PHB suppresses infectious diseases in crustaceans.

[0003] Japanese Patent No. 7138391 Japanese Patent Application Laid-Open No. 2019-176839 Japanese Patent Application Laid-Open No. 2022-177817 Japanese Patent Application Laid-Open No. 2008-231070

[0004] Mudtham NA, Promariya A, Duangsri C, Maneeruttanarungroj C, Ngamkala S, Akrimajirachoote N, Powtongsook S, Salminen TA, Raksajit W. Exogenous Trehalose Improves Growth, Glycogen and Poly-3-Hydroxybutyrate (PHB) Contents in Photoautotrophically Grown Arthrospira platensis under Nitrogen Deprivation. Biology (Basel). 2024 Feb 18;13(2):127.Cassuriaga APA, Freitas BCB, Morais MG, Costa JAV. Innovative polyhydroxybutyrate production by Chlorella fusca grown with pentoses. Bioresour Technol. 2018 Oct;265:456-463.Chaouachi M, Vincent S, Groussard C. A Review of the Health-Promoting Properties of Spirulina with a Focus on athletes' Performance and Recovery. J Diet Suppl. 2024;21(2):210-241.Bito T, Okumura E, Fujishima M, Watanabe F. Potential of Chlorella as a Dietary Supplement to Promote Human Health. Nutrients. 2020 Aug 20;12(9):2524.Duan Y, Zhang Y, Dong H, Zheng X, Wang Y, Li H, Liu Q, Zhang J.Effect of dietary poly-β-hydroxybutyrate (PHB) on growth performance, intestinal health status and body composition of Pacific white shrimp Litopenaeus vannamei (Boone, 1931). Fish Shellfish Immunol. 2017 Jan;60:520-528.

[0005] The problem with PHB is that its extraction is too costly. While its application in regenerative medicine, which can tolerate high costs, is progressing, its application in food and feed has not yet progressed. Therefore, in order to utilize PHB in food and feed compositions, it is important to be able to use PHB without the complicated procedures for PHB extraction.

[0006] The present invention has been made in view of these points, and has as its object to make it possible to use PHBs without performing complicated operations for extracting them.

[0007] One aspect of the present invention is a method for delivering polyhydroxybutyrate to the large intestine of an animal using algae, which are photosynthetic organisms that encapsulate polyhydroxybutyrate, as a carrier of polyhydroxybutyrate.

[0008] The algae is, for example, Chlorella, a green algae, or Spirulina, a blue-green algae.

[0009] The algae may be at least one type of chlorella selected from the group consisting of Chlorella vulgaris, Chlorella ellipsoidea, Chlorella saccarophila, Chlorella regularis, Chlorella pyrenoidosa, Chlorella fusca, and Chlorella sorokiniana.

[0010] The algae may be at least one type of spirulina selected from the group consisting of Arthrospira ardissonei, Arthrospira erdosensis, Arthrospira fusiformis, Arthrospira indica, Arthrospira innermongoliensis, Arthrospira jenneri, Arthrospira massartii, Arthrospira maxima, and Arthrospira platensis.

[0011] One aspect of the present invention is a carrier for delivering polyhydroxybutyrate to the large intestine of an animal, comprising algae encapsulating polyhydroxybutyrate.

[0012] One aspect of the present invention is a food product containing a carrier comprising algae encapsulating polyhydroxybutyrate, which delivers polyhydroxybutyrate to the large intestine of an animal.

[0013] One aspect of the present invention is a feed composition containing a carrier comprising algae encapsulating polyhydroxybutyrate, which delivers polyhydroxybutyrate to the large intestine of an animal.

[0014] One aspect of the present invention is an intracolonic propionic acid releaser containing a carrier consisting of algae encapsulating polyhydroxybutyric acid, which delivers polyhydroxybutyric acid to the large intestine of an animal.

[0015] One aspect of the present invention is an intracolonic butyrate-releasing agent containing a carrier made of algae encapsulating polyhydroxybutyrate, which delivers polyhydroxybutyrate to the large intestine of an animal.

[0016] One aspect of the present invention is an intracolonic acetic acid-releasing agent containing a carrier consisting of algae encapsulating polyhydroxybutyric acid, which delivers polyhydroxybutyric acid to the large intestine of an animal.

[0017] One aspect of the present invention is an agent for lowering pH in the large intestine, which comprises a carrier made of algae encapsulating polyhydroxybutyric acid and which delivers polyhydroxybutyric acid to the large intestine of an animal.

[0018] FIG. 1 is a diagram showing the process by which spirulina containing PHB acts in the large intestine. FIG. 2 is a diagram for explaining that PHB liberates ketone bodies. FIG. 3 is a diagram for explaining the action of PHB in the large intestine. FIG. 4 is a diagram showing the chemical formulas of PHA, PHB, PHBV, and PHBH. FIG. 5 is a diagram for explaining this experiment. FIG. 6 is a diagram showing experimental results. FIG. 7 is a diagram showing experimental results showing the survival rate of Artemia.

[0019] [Summary of the Present Embodiment] The inventors have conducted research into food and feed compositions containing Chlorella genus encapsulating a biodegradable polyester, and have found that polyhydroxyalkanoate alkanoate (hereinafter referred to as "PHA"), a type of biodegradable polyester, can be easily delivered to the large intestine by encapsulating PHA in Chlorella or Spirulina.

[0020] Examples of PHA include PHB, PHBH, and PHBV. Chlorella or spirulina containing PHA act as ketone donors in the human body, providing ketone bodies to intestinal bacteria and converting the intestinal environment to one dominated by butyric acid bacteria. Furthermore, it is possible to sustainably increase ketone body concentrations in the human body.

[0021] In one embodiment of the present invention, a new "synbiotic" is created using chlorella containing PHA, which acts as a "prebiotic," and spores of Clostridium butyricum, which acts as a "probiotic." This embodiment can effectively increase the abundance of butyric acid bacteria and Akkermansia, which are included in the bacterial group belonging to groups IV, XIVa, and XVIII of the Clostridium cluster, such as Eubacterium, Roseburia, Coprococcus, Faecalibacterium, Ruminococcus, Lachnospira, Clostridium, and Butrycoccus.

[0022] Furthermore, this embodiment facilitates the delivery of biodegradable polyesters to the large intestine, and can promote the development of feed or food compositions containing biodegradable polyesters with various physiological effects. In particular, this embodiment can promote the development of ketone donors for humans.

[0023] That is, this embodiment is characterized in that a PHA (e.g., PHB, a type of PHA) is delivered to the large intestine lumen using algae (e.g., chlorella (a green algae) or spirulina (a cyanobacterium)), a type of photosynthetic organism, as a carrier, which encapsulates the PHA. Because algae are digested before reaching the large intestine lumen, even if the PHA is ingested encapsulated in chlorella or spirulina, it is believed that the PHA can have the effects of suppressing inflammation that occurs in animals, suppressing infectious diseases, and promoting animal growth. Therefore, by delivering PHA to the large intestine lumen using algae, a type of photosynthetic organism, which encapsulates PHA, as a carrier, it is possible to effectively suppress inflammation that occurs in animals, suppress infectious diseases, and promote animal growth.

[0024] A carrier containing a photosynthetic organism (e.g., algae) containing a PHA and delivering the PHA to the large intestine of an animal can be contained in, for example, a food, a feed composition, a colonic propionic acid releaser, a colonic butyric acid releaser, a colonic acetic acid releaser, or a colonic pH lowering agent. That is, by having an animal ingest a food (including a supplement), a feed composition, a colonic propionic acid releaser, a colonic butyric acid releaser, a colonic acetic acid releaser, or a colonic pH lowering agent, it becomes possible to suppress inflammation and infections occurring in the animal and promote the growth of the animal.

[0025] In this way, when PHB is encapsulated in chlorella or spirulina and delivered to the large intestine using chlorella or spirulina as a carrier, there is no need to purify the PHB, making it possible to effectively utilize PHB while keeping costs down.

[0026] Chlorella that can be used as a carrier is, for example, at least one species selected from the group consisting of Chlorella vulgaris, Chlorella ellipsoidea, Chlorella saccarophila, Chlorella regularis, Chlorella pyrenoidosa, Chlorella fusca, and Chlorella sorokiniana.

[0027] Examples of spirulina that can be used as a carrier include at least one species selected from the group consisting of Arthrospira ardissonei, Arthrospira erdosensis, Arthrospira fusiformis, Arthrospira indica, Arthrospira innermongoliensis, Arthrospira jenneri, Arthrospira massartii, Arthrospira maxima, and Arthrospira platensis.

[0028] Fig. 1 is a diagram showing the process by which PHB-containing spirulina acts in the large intestine. Fig. 2 is a diagram explaining the release of ketone bodies by PHB. Fig. 3 is a diagram explaining the action of PHB in the large intestine. Fig. 4 is a diagram showing the chemical formulas of PHA, PHB, PHBV, and PHBH.

[0029] As shown in Figure 1, when PHB-encapsulated spirulina reaches the intestinal tract, the spirulina is digested in the small intestine, liberating PHB. When the liberated PHB reaches the large intestine, it activates, for example, butyric acid bacteria.

[0030] As shown in Figure 2, PHB is a polyester of ketone bodies. The ester bond is not hydrolyzed by mammalian digestive enzymes, but is hydrolyzed by enzymes in intestinal bacteria. Therefore, PHB is released from spirulina in the small intestine, but reaches the large intestine without being hydrolyzed there. In the large intestine, PHB is hydrolyzed by intestinal bacteria, releasing ketone bodies into the colonic lumen.

[0031] As shown in Figure 3, ketone bodies released into the colonic lumen serve as energy substrates for intestinal bacteria, activating their energy metabolism. This activates butyric acid bacteria, increasing the release of short chain fatty acids (SCFA) such as butyric acid, acetic acid, and propionic acid. SCFA are involved in inducing a butyric acid bacteria-dominated intestinal environment, and enter and circulate throughout the animal's body, where they are involved in growth promotion, infection suppression, and suppression of inflammatory bowel disease.

[0032] 1 to 3 show an example in which PHB is encapsulated in spirulina, but a PHA other than PHB may also be encapsulated in spirulina or chlorella. When an animal ingests chlorella encapsulating a PHA other than PHB, the same effect occurs in the large intestine as when the animal ingests spirulina encapsulating PHB.

[0033] Example 1 The amount of PHB contained in PHB-encapsulating spirulina was measured using high performance liquid chromatography (HPLC).

[0034] Specifically, Spirulina in the logarithmic growth phase was cultured for 3 days using the following methods: (1) control Zaroc medium, (2) Zaroc medium supplemented with trehalose at a concentration of 3 mM, and (3) nitrogen source (NaNO). ３ (4) Zaroc medium with trehalose at a concentration of 3 mM after removing the nitrogen source (NaNO ３ The spirulina was then collected and the PHB was quantified by HPLC.

[0035] Figure 5 is a diagram for explaining this experiment. Figure 5A shows the morphology of the spirulina used in this experiment. Figure 5B shows the state of spirulina cultivation.

[0036] The spirulina used was Arthrospira platensis NIES-39 obtained from the Global Environmental Forum. This spirulina was treated with NaNO as a nitrogen source. ３ Spirulina was pre-cultured in a 250 mL flask with 50 mL of Zaroc medium (pH 10.0) supplemented with 100% CO . As shown in Figure 5B, spirulina was grown in a 250 mL flask with 50 mL of Zaroc medium (pH 10.0) supplemented with 100% CO . ２The mixture was pre-incubated aerobically at 32°C under white fluorescent lighting for 7 days while bubbling with 0.1% CO₂.

[0037] After this pre-culture, the cells were harvested, and 5 mL of Spirulina was inoculated into 50 mL of each of the four media. The cells were then cultured under continuous illumination for 3 days. The Spirulina cells were centrifuged (1000 rpm for 15 minutes) to precipitate, and then dried in a dryer at 37°C to obtain four types of dried Spirulina cells.

[0038] Subsequently, the PHB content of the four dried cells was quantified. The four dried cells (0.5 g) were boiled in concentrated sulfuric acid for 1 hour to hydrolyze PHB to crotonic acid. Separation of PHB (crotonic acid) was performed using a C18 reverse-phase column (4.6 mm internal diameter, 150 mm length) packed with 5 mm diameter silica gel and equipped with an SPD-20A UV / VIS detector at 210 nm. HPLC analysis was performed using a 20 μL sample injection. The solvent was 0.1% acetic acid, 60% (v / v), and acetonitrile, 40% (v / v), at a flow rate of 0.6 mL / min. The amount of PHB in the samples was calculated using a calibration curve derived from a PHB standard solution (commercially available from Sigma-Aldrich, catalog number 363502).

[0039] Figure 6 shows the experimental results. As shown in Figure 6, only when Spirulina was cultured using "(3) nitrogen source-removed culture medium + trehalose 3 mM", 8 (=2) per dry cell weight was obtained. ３ It was confirmed that the strain accumulated 100% of PHB.

[0040] On the first day after the start of cultivation, about 2% of PHB was accumulated. Since PHB production is an enzymatic reaction and cells are in a logarithmic growth state, the enzyme product should increase exponentially. Therefore, the amount of PHB is "A Ｎ It can be expressed as (%)" (A: constant, N: number of days). Since 2% of PHB was accumulated on the first day and 8% on the third day, it can be estimated that A = 2 when N = 1, and 8% when N = 3.

[0041] From this, if the culture time is 6 days, ６It can be estimated that this method can produce PHB (=64). Therefore, it can be estimated that this method can accumulate at least 60% of PHB. In other words, it was confirmed that the PHB encapsulation rate of Spirulina can be in the range of 0% to 60%.

[0042] Similarly, chlorella accumulated 17% of PHB after 3 days of culture, suggesting that the encapsulation rate of PHB can reach at least 60% in chlorella. In other words, by adjusting the culture time of spirulina or chlorella, the encapsulation rate can be set within the range of 0% to 60%.

[0043] [Example 2] Test for inhibiting bacterial infection by feeding PHB to Artemia crustaceans

[0044] The microscopic crustacean Artemia is widely used as an experimental system for easily measuring the biological effects of PHB. When Artemia is infected with the pathogenic bacterium Vibrio, it dies within a few days. PHB inhibits this death. Artemia is easy to culture, has a digestive tract, and is fed algae such as chlorella and spirulina, making it ideal for verifying the biological activity of chlorella and spirulina containing PHB.

[0045] It is common technical knowledge that spirulina and chlorella alone cannot prevent death due to Vibrio infection. Therefore, if ingesting PHB-containing spirulina or chlorella into Artemia results in a protective effect similar to that of PHB, this would mean that PHB has been delivered to the large intestine by the spirulina or chlorella.

[0046] In this experiment, spirulina containing no PHB and spirulina containing 8% PHB were prepared, and 5% (300 mg) of dried spirulina cells were added to the culture medium, followed by cultivation with vibrio (100,000 viable cells) for three days.

[0047] The Artemia used were commercially available Brine Shrimp Eggs (Nippon Dobutsu Yakuhin Co., Ltd.). Artemia larvae were used for the test at least 48 hours after hatching. A 6-well plate was used as the test container, with 6 mL of sterilized artificial seawater (salinity 3.2%) per well (volume 15 mL). The test was carried out in an incubator maintained at 28°C, with the plate being shaken at 150 rpm.

[0048] The Vibrio solution was prepared by culturing 150 mg of a glycerol stock of Vibrio bacteria, provided by Professor Ikuo Hirono of Tokyo University of Marine Science and Technology, in 40 mL of sterilized alkaline peptone medium (salt concentration 2.0%) for 6 hours, and diluting the bacterial solution adjusted using a McFarland turbidity meter.

[0049] Prior to the start of the test, each well was filled with 6 mL of artificial seawater, and 20 Artemia larvae were pipetted into each well and allowed to acclimate to the environment at 28°C for 6 hours. ５ The number of individuals in the wells was counted using a microscope before the start of the test (0 hours) and after 48 hours. The survival rate (%) was calculated as the number of surviving individuals / total number of individuals x 100. Whether or not individuals survived was determined by visually checking the motility of the individuals.

[0050] In preliminary experiments, Vibrio ５ It was confirmed that the amount of PHB contained in Spirulina was suitable, and that adding 0.01 mg / ml of PHB could suppress the toxicity of Vibrio. Since Spirulina PHB contains 8% PHB, adding it to seawater at a concentration of 5% would be equivalent to 0.024 mg / ml of PHB, which was thought to be sufficient to suppress Vibrio infection.

[0051] The survival rate after 48 hours was examined for four samples. To conduct this experiment, four six-well plates were prepared, filled with 6 ml of seawater, and 20 Artemia were placed in each well. The six-well plates were divided into three groups: Spirulina (-), Spirulina (+), and Spirulina PHB (+) PHB. Spirulina (-) indicates that no spirulina was added, Spirulina (+) indicates that 300 mg of spirulina was added, and Spirulina PHB (+) indicates that 300 mg of spirulina containing 8% PHB was added.

[0052] One of the two well plates in each group was cultured without Vibrio, while the other was cultured with Vibrio. After culturing under these conditions for 48 hours, surviving Artemia were counted visually.

[0053] Figure 7 shows experimental results showing the survival rate of Artemia. * in Figure 7 indicates a significant difference.

[0054] The following is clear from Figure 7: (A) Spirulina (-): The addition of Vibrio reduced cell viability from 73% to 12%. (B) Spirulina (+): The addition of Vibrio reduced cell viability from 92% to 13%. This indicates that Spirulina alone did not have a protective effect against the toxicity of Vibrio.

[0055] (C) Spirulina PHB (+): The addition of Vibrio tended to decrease cell viability from 95% to 65%. This difference was not significant (P = 0.11). In other words, the addition of Vibrio did not significantly decrease cell viability. Therefore, it was confirmed that Spirulina containing PHB had the effect of protecting Artemia from the toxicity of Vibrio. In other words, it can be seen that the presence of PHB allowed Artemia to maintain their survival.

[0056] It was confirmed that the inclusion of PHB enabled Artemia to maintain survival, a phenomenon that would not occur if PHB were not present in the Artemia's intestinal tract. Therefore, it was confirmed that PHB exerted a physiological effect in the Artemia's intestinal tract. Therefore, it was confirmed that PHB-encapsulated Spirulina functions as a carrier for delivering PHB to the large intestinal lumen.

[0057] As described above, by encapsulating PHB in algae, it is possible for PHB to reach the large intestine lumen of an animal without the need for costly PHB extraction, making it easier to use PHB to suppress inflammation or infectious diseases that occur in animals.

[0058] Food and feed compositions containing PHB are described in detail below. PHB, an example of PHA, is slowly hydrolyzed by enzymes exposed in intestinal bacteria to release monomers. At least one of these monomers is 3-hydroxybutyrate (3HB). 3HB serves as an energy substrate for intestinal bacteria and can activate their energy metabolism. This activates butyric acid bacteria, increasing the release of short chain fatty acids (SCFA), such as butyric acid, acetic acid, and propionic acid. SCFA enter the animal's body, circulate throughout the body, and are thought to be involved in growth promotion, infection control, and suppression of inflammatory bowel disease.

[0059] Chlorella and spirulina are already widely recognized as human foods. The present inventors have conceived of using chlorella containing PHB to deliver PHB to the large intestine.

[0060] The proportion of chlorella or spirulina containing PHB to be used is preferably 0.01 to 50% by weight, more preferably 0.1 to 50% by weight, and even more preferably 1 to 10% by weight, based on the total weight of chlorella or spirulina containing PHB.

[0061] The ratio of PHB to the dry preparation in chlorella or spirulina containing PHB is 3% or more, preferably 5% or more, more preferably 10% or more, more preferably 20%, and even more preferably 30% or more.

[0062] PHB promotes the growth of zooplankton such as Artemia and crustaceans such as whiteleg shrimp, improving their survival rate, and providing a safe feed composition with minimal environmental impact. By feeding PHB contained in chlorella, it is possible to achieve a high growth rate (growth rate) while maintaining a high survival rate.

[0063] By providing zooplankton with PHB, it is possible to promote the rapid growth of zooplankton and crustaceans while maintaining a high survival rate. The use of chlorella containing PHB makes it possible to efficiently and stably obtain highly nutritious zooplankton. Chlorella containing PHB can also be used as a growth promoter for zooplankton and crustaceans. Chlorella containing PHB can suppress infectious diseases such as Vibrio in zooplankton such as Artemia and crustaceans such as vannamei shrimp.

[0064] PHB can be used as an immunopotentiator for fish, improving the production efficiency of farmed fish. Chlorella containing PHB can be used for fish such as yellowtail, amberjack, flounder, red sea bream, tiger pufferfish, devil stinger, flatfish, horse mackerel, mackerel, sardines, whiting, rockfish, salmon, trout, sweetfish, carp, yamame salmon, crucian carp, and goldfish. Furthermore, chlorella containing PHB can be used not only for adult fish but also for various growth stages, such as hatched seedlings and juveniles. The immunopotentiating effect for fish can be quantified using a leukocyte phagocytosis test or an envelopment index.

[0065] PHB is also useful for mammals and birds. These higher animals have intestinal microflora, and when PHB induces an intestinal environment dominated by butyric acid bacteria, it promotes growth and inhibits inflammatory bowel disease.

[0066] PHB can increase the release of short-chain fatty acids (SCFA) such as butyric acid, acetic acid, and propionic acid in pigs, which results in intestinal regulation, suppression of inflammatory bowel disease, and relief of constipation.

[0067] PHB releases ketone bodies in the large intestine, functioning as a "ketone donor." After ingestion, PHB gradually increases blood ketone body concentrations over several hours, and the increase continues for 10 hours or more. When using PHB as feed or food, a sustained increase in ketone body concentrations is necessary, and PHB is useful as a "ketone donor."

[0068] PHB activates the following intestinal bacteria: bacteria belonging to groups IV, XIVa, and XVIII of the Clostridium cluster, such as the genera Eubacterium, Roseburia, Coprococcus, Faecalibacterium, Ruminococcus, Lachnospira, Clostridium, and Butyricoccus, which are capable of producing butyric acid, acetic acid, and propionic acid, and bacteria of the genus Akkermansia.

[0069] PHB can induce an increase in butyrate concentration by proliferation of enterobacteria with high butyrate production capacity. Butyrate produced by enterobacteria in the large intestinal flora activates macrophages resident in Peyer's patches in the large intestine. Activated macrophages activate naive T cells and differentiate them into regulatory T (Treg) cells. Tregs circulate throughout the body, not just in the large intestine, and suppress many allergic and inflammatory diseases, including inflammatory bowel disease.

[0070] PHB induces the activation of Treg cells, and therefore has been shown to be effective against typical livestock diseases such as foot-and-mouth disease, anthrax, brucellosis, tuberculosis, Johne's disease, bovine spongiform encephalopathy (BSE), bluetongue, Akabane disease, Chuzan disease, bovine viral diarrhea / mucosal disease (BVD / MD), infectious bovine rhinotracheitis (IBR), bovine leukemia, Aino virus infection, Ibaraki disease, bovine ephemeral fever, black jaundice, salmonellosis, bovine adenovirus disease, bovine coronavirus disease, bovine respiratory syncytial virus disease, bovine rotavirus disease, bovine pasteurellosis, bovine piroplasmosis, bovine lungworm disease, coccidiosis, strongyloidiasis, cryptosporidiosis, neosporosis, and many more. It has an inhibitory effect on nitrate poisoning, cerebral cortical necrosis, hog cholera, Owensky's disease, porcine reproductive and respiratory syndrome (PRRS), porcine epidemic diarrhea (PED), transmissible gastrointestinal disease (TGE), swine erysipelas, toxoplasmosis, porcine pleuropneumonia, porcine colibacillosis, exudative epidermitis (soot disease), equine infectious anemia, equine influenza, equine rhinopneumonitis, contagious equine metritis, equine paratyphoid, equine heartworm disease, Newcastle disease, low pathogenic Newcastle disease, highly pathogenic avian influenza (HPAI), low pathogenic avian influenza (LPAI), avian influenza, poultry salmonellosis, avian mycoplasmosis, maggot disease, and acarinosis.

[0071] PHB releases ketone bodies in the large intestine, providing animals with ketone bodies. Ketone bodies are a collective term for 3-hydroxybutyrate (3HB), acetoacetate, and acetone, with over 80% being 3HB. They are effective in treating many age-related diseases, including cancer, heart disease, and neurological disorders such as Alzheimer's disease.

[0072] PHB can be used as a food or feed composition having preventive and / or therapeutic effects against diseases such as cancer, autoimmune diseases, etc. It can also be used for the treatment of various diseases (e.g., cancer, autoimmune diseases, etc.) in humans and animals, as well as for the purpose of improving, remission, and complete cure of the disease state.

[0073] PHB is expected to be effective in treating hepatocellular carcinoma, cholangiocarcinoma, renal cell carcinoma, squamous cell carcinoma, basal cell carcinoma, transitional cell carcinoma, adenocarcinoma, malignant gastrinoma, malignant melanoma, fibrosarcoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, malignant teratoma, angiosarcoma, Kaposi's sarcoma, osteosarcoma, chondrosarcoma, lymphangiosarcoma, malignant meningioma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, leukemia, brain tumor, epithelial cell-derived neoplasm (epithelial carcinoma), basal cell carcinoma, adenocarcinoma, gastrointestinal cancers such as lip cancer, oral cancer, esophageal cancer, small intestine cancer, and stomach cancer, colon cancer, rectal cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, cervical cancer, lung cancer, breast cancer, skin cancers such as squamous cell carcinoma and basal cell carcinoma, prostate cancer, and renal cell carcinoma.

[0074] PHB is expected to have therapeutic effects on inflammatory bowel disease (such as controlled ulcerative colitis and Crohn's disease), type 1 diabetes, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), myasthenia gravis, and the like.

[0075] PHB provides ketone bodies to butyric acid bacteria and the like in the large intestine of animals, promoting the release of short chain fatty acids (SCFA) such as butyric acid, acetic acid, and propionic acid. PHB can be used as a new prebiotic preparation. Although PHB can be applied in a wide range of industrial fields, its application as a food or feed composition is limited.

[0076] PHB can suppress colon cancer, ulcerative colitis, and obesity. PHB provides ketone bodies to intestinal bacteria, activates butyric acid bacteria, and releases butyrate into the colonic lumen. This allows PHB to activate regulatory T cells and protect the colonic epithelium.

[0077] PHB can suppress damage caused by Vibrio, a pathogenic microorganism in crustaceans. Therefore, PHB can reduce the need for antibiotics. As in mammals, PHB provides ketone bodies to the intestinal bacteria of crustaceans, activating butyric acid bacteria and releasing butyric acid into the lumen of the large intestine. Because butyric acid suppresses inflammation, it can suppress damage caused by Vibrio.

[0078] PHB promotes the growth of crustaceans such as whiteleg shrimp, improves their survival rate, and provides a safe feed composition with little environmental impact. By feeding PHB to crustaceans, a high growth rate (growth rate) can be achieved while maintaining a high survival rate.

[0079] However, despite the promise of PHB as a new prebiotic preparation, the industrial extraction of PHB has been problematic in that it is complicated and costly. To make the PHB produced in this way usable as food, it is necessary to extract it without using organic solvents, which increases costs. Therefore, a method for mass-producing PHB using bacteria has been put into practical use. Photosynthetic organisms (algae) such as chlorella and spirulina also accumulate PHB at a certain concentration within their cells depending on the culture conditions.

[0080] [Industrial Applicability] Algae containing PHB can function as a carrier for delivering PHB to the large intestine of zooplankton, crustaceans, fish, birds, and mammals (pigs, cows, horses, etc.). They can then be used as feed for these animals, promoting growth, increasing survival rates, suppressing infectious diseases, and treating inflammatory bowel disease. This method eliminates the need for PHB extraction, making it possible to produce PHB at low cost. Furthermore, PHB-containing algae can be used as a new prebiotic preparation for human food.

[0081] [Additional Note] The present invention may include the following configurations.

[0082] A method for delivering biodegradable polyesters to the large intestine of animals using chlorella encapsulating the biodegradable polyesters.

[0083] A food or feed composition containing chlorella encapsulating a biodegradable polyester.

[0084] In the feed composition described above, the chlorella is at least one species selected from the group consisting of Chlorella vulgaris, Chlorella ellipsoidea, Chlorella saccarophila, Chlorella regularis, Chlorella pyrenoidosa, Chlorella fusca, and Chlorella sorokiniana.

[0085] A food or feed composition containing chlorella encapsulating polyhydroxyalkanoate.

[0086] A food or feed composition containing chlorella encapsulating polyhydroxyalkanoate having a weight-average molecular weight of 10,000 or more.

[0087] A food or feed composition containing chlorella containing polyhydroxyalkanoate having a weight-average molecular weight of 10,000 or more in an amount of 3% or more, preferably 5% or more, more preferably 10% or more, even more preferably 20% or more, and even more preferably 30% or more, based on the dry weight of the chlorella.

[0088] The food or feed composition as described above, wherein the polyhydroxyalkanoate is at least one selected from the group consisting of polyhydroxybutyrate, polyhydroxybutyrate hexanoate, and polyhydroxybutyrate valerate.

[0089] The food or feed composition described above, wherein the form of the chlorella encapsulating the polyhydroxyalkanoate is a culture solution containing the chlorella encapsulating the polyhydroxyalkanoate, dried fungus cells, or dried fungus cells that have been crushed.

[0090] A propionic acid release agent for the large intestine, whose active ingredient is chlorella containing polyhydroxyalkanoate.

[0091] A colonic butyric acid release agent whose active ingredient is chlorella containing polyhydroxyalkanoate.

[0092] An acetic acid-releasing agent for the large intestine, whose active ingredient is chlorella containing polyhydroxyalkanoate.

[0093] An agent for lowering pH in the large intestine, the active ingredient of which is chlorella containing polyhydroxyalkanoate.

[0094] An agent for releasing butyrate in the large intestine, whose active ingredients are chlorella containing polyhydroxyalkanoates and spores of the butyric acid bacterium Clostridium Butyricum.

[0095] An acetic acid-releasing agent for the large intestine, whose active ingredients are chlorella containing polyhydroxyalkanoates and spores of the butyric acid bacterium Clostridium Butyricum.

[0096] A propionic acid release agent for the large intestine, whose active ingredients are chlorella containing polyhydroxyalkanoates and spores of the butyric acid bacterium Clostridium Butyricum.

[0097] An agent for lowering pH in the large intestine, whose active ingredients are chlorella containing polyhydroxyalkanoate and spores of Clostridium Butyricum, a butyric acid bacterium.

[0098] A feed composition containing chlorella encapsulating polyhydroxyalkanoate, which is a growth promoter or an inflammatory bowel disease inhibitor for pigs.

[0099] A feed composition containing chlorella encapsulating polyhydroxyalkanoate, which is a growth promoter or an inflammatory bowel disease inhibitor for chickens.

[0100] A feed composition containing chlorella encapsulating polyhydroxyalkanoate, which is a growth promoter or infection inhibitor for fish.

[0101] A feed composition containing chlorella encapsulating polyhydroxyalkanoate, which is a growth promoter or infection inhibitor for crustaceans.

[0102] A feed composition containing chlorella encapsulating polyhydroxyalkanoate, which is an agent for increasing the survival rate, growth promoter, or infection inhibitor of zooplankton.

[0103] A food or feed composition containing chlorella encapsulating polyhydroxyalkanoate, which is a chronic renal failure inhibitor.

[0104] A food or feed composition containing chlorella encapsulating polyhydroxyalkanoate, an inflammatory bowel disease inhibitor.

[0105] A food or feed composition containing chlorella encapsulating polyhydroxyalkanoate, an anti-obesity agent.

[0106] A food or feed composition containing chlorella encapsulating polyhydroxyalkanoate, a dermatitis inhibitor.

Claims

1. A method for delivering polyhydroxybutyrate to the large intestine of animals using algae, a photosynthetic organism that encapsulates polyhydroxybutyrate, as a carrier of polyhydroxybutyrate.

2. The method according to claim 1, wherein the algae is Chlorella, a green algae, or Spirulina, a blue-green algae.

3. The method of claim 1, wherein the algae is at least one type of chlorella selected from the group consisting of Chlorella vulgaris, Chlorella ellipsoidea, Chlorella saccarophila, Chlorella regularis, Chlorella pyrenoidosa, Chlorella fusca, and Chlorella sorokiniana.

4. The method of claim 1, wherein the algae is at least one type of spirulina selected from the group consisting of Arthrospira ardissonei, Arthrospira erdosensis, Arthrospira fusiformis, Arthrospira indica, Arthrospira innermongoliensis, Arthrospira jenneri, Arthrospira massartii, Arthrospira maxima, and Arthrospira platensis.

5. A carrier consisting of algae encapsulating polyhydroxybutyrate that delivers polyhydroxybutyrate to the large intestine of animals.

6. A food product containing a carrier consisting of algae encapsulating polyhydroxybutyrate, which delivers the polyhydroxybutyrate to the large intestine of an animal.

7. A feed composition comprising a carrier comprising algae encapsulating polyhydroxybutyrate, which delivers polyhydroxybutyrate to the large intestine of an animal.

8. An intracolonic propionate release agent comprising a carrier consisting of algae encapsulating polyhydroxybutyrate, which delivers polyhydroxybutyrate to the large intestine of an animal.

9. An intracolonic butyrate-releasing agent comprising a carrier consisting of algae encapsulating polyhydroxybutyrate, for delivering polyhydroxybutyrate to the large intestine of an animal.

10. An intracolonic acetate releasing agent comprising a carrier comprising algae encapsulating polyhydroxybutyrate for delivering polyhydroxybutyrate to the large intestine of an animal.

11. An agent for lowering pH in the large intestine, comprising a carrier consisting of algae encapsulating polyhydroxybutyrate, which delivers polyhydroxybutyrate to the large intestine of an animal.

Images