Prevention and treatment of fatty liver

Abstract

The present invention relates to Anaerobutyricum soehngenii or a related species thereof for use in the prevention and / or treatment of hepatic steatosis, in particular non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH), for increasing bile acid plasma levels to reduce hepatitis and / or reduce hepatic necroinflammatory activity scores. Said Anaerobutyricum soehngenii or a related species thereof may be combined with at least one species of Bifidobacterium genus, preferably Bifidobacterium animalis subsp. lactis or a related species thereof and / or Bifidobacterium breve or a related species thereof. Additionally or alternatively, the Anaerobutyricum soehngenii or a related species may be combined with at least one species of Akkermansia, preferably Akkermansia muciniphila or a related species. Additionally or alternatively, the Anaerobutyricum soehngenii or a related species may be combined with at least one species of Lactobacillus, preferably Lactobacillus acidophilus or a related species, Lactobacillus casei or a related species, and / or Lactobacillus reuteri or a related species.

Description

[Technical field]

[0001] The present invention relates to the field of prevention and treatment of hepatic steatosis. [Background technology]

[0002] Nonalcoholic fatty liver disease (NAFLD) is recognized as the most prevalent chronic liver disease worldwide, ranging from simple steatosis (nonalcoholic fatty liver), to nonalcoholic steatohepatitis (NASH), NASH-fibrosis, cirrhosis, and hepatocellular carcinoma. The current estimated global prevalence of NAFLD is 25%–30% in the general population and up to 80% in those with metabolic syndrome and type II diabetes. By definition, excessive alcohol intake excludes the diagnosis of NAFLD.

[0003] NAFLD refers to a group of diseases that result in the accumulation of excess fat in the liver in patients who drink little or no alcohol. The most common form of NAFLD is called nonalcoholic fatty liver disease (NAFLD). Because the development and progression of NAFLD are strongly driven by insulin resistance, several therapeutic strategies in clinical development for NAFLD aim to reduce insulin resistance.

[0004] NASH refers to hepatitis caused by lipid toxicity associated with fatty liver. NASH significantly increases the risk of developing liver cirrhosis and hepatocellular carcinoma (HCC) and is associated with increased atherosclerotic cardiovascular disease. Since the association between NAFLD / NASH and insulin resistance is well known, strategies to reduce insulin resistance may slow the progression of NAFLD / NASH disease or alleviate its symptoms.

[0005] The gut microbiota is associated with the development and prevalence of NAFLD and NASH. People who eat a plant-based diet with less animal protein have a significantly lower incidence of the disease, which is thought to be mediated by the gut microbiota. Therefore, Witjes et al. (Hepatology Communications, Vol. 4, No. 11, 2020) propose the transplantation of fecal microbiota from lean vegan donors as a potential treatment.

[0006] However, there is a need in the art for new and improved therapies in the prevention and treatment of NAFLD and NASH.

[0007] It is an object of the present disclosure, among other objects, to address the above-mentioned needs in the art and to provide new and / or improved strategies for preventing and / or treating NAFLD and NASH. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Witjes et al. Hepatology Communications, Vol. 4, No. 11, 2020 [Non-Patent Document 2] Kleiner et al., Vol. 41, No. 6 (June 2005) [Non-Patent Document 3] Shetty et al. Int J Syst Evol Microbiol. 2018 Dec;68(12):3741-3746 [Non-Patent Document 4] Collado et al. Appl Environ Microbiol 2007 Dec;73(23):7767-70 [Non-Patent Document 5] Devereux, J et al. Nucleic Acids Research (1984) 12(1):387 [Non-Patent Document 6] Atschul, SF et al. J. Molec. Biol. (1990) 215:403 [Non-Patent Document 7] Bottacini et al. 2011, J Bacteriol 193: 6387-6388 [Non-Patent Document 8] Kankainen et al. 2009 106:17193-8 [Non-Patent Document 9] Chiang Liver Res. 2017 Jun; 1(1): 3-9 Summary of the Invention

[0009] The present inventors have surprisingly found that administering Anaerobutyricum soehngenii or its relatives to subjects with fatty liver increases bile acid plasma levels, which reduces hepatitis.Therefore, the administration of Anaerobutyricum soehngenii or its relatives can be applied in the strategy for preventing and / or treating hepatic steatosis.

[0010] Furthermore, it has been found that combining Anaerobutyricum soehngenii or a closely related species thereof with Bifdobacterium, Akkermansia, and / or Lactobacillus bacteria provides a synergistic therapeutic effect in the prevention or treatment of fatty liver, particularly non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH).

[0011] The present disclosure provides new and improved strategies for preventing and / or treating fatty liver, NAFLD, and / or NASH. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present disclosure relates to Anaerobutyricum soehngenii, or a related species thereof, having a 16S rRNA gene sequence having at least 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.5, 99.9, 100% sequence identity to SEQ ID NO:1 and / or SEQ ID NO:2, in particular for use in the prevention and / or treatment of fatty liver and / or for increasing the production of propionate / propionate and / or butyrate / butyrate or derivatives thereof in the intestine.

[0013] In accordance with the above, the present disclosure relates to a method for preventing and / or treating fatty liver, e.g., in a subject in need thereof, comprising, e.g., administering to said subject said Anaerobutyricum soehngenii, or a closely related species thereof.

[0014] Fatty liver is a condition in which excess fat accumulates in the liver. There are two stages of fatty liver disease: nonalcoholic fatty liver disease (NAFLD) and alcoholic liver disease. NAFLD consists of simple fatty liver and nonalcoholic steatohepatitis (NASH).

[0015] In the present disclosure, the fatty liver may in particular be selected from non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH).

[0016] The term "nonalcoholic fatty liver disease" (NAFLD) refers to a group of conditions in which excess fat accumulates in the liver in people who drink little or no alcohol. The most common stage of NAFLD is called fatty liver. NAFLD is strongly associated with insulin resistance and type 2 diabetes, so treatment of NAFLD aims to reduce insulin resistance.

[0017] The term "nonalcoholic steatohepatitis" (NASH) refers to liver inflammation and damage caused by the accumulation of fat in the liver. NASH not only significantly increases the risk of developing cirrhosis and hepatocellular carcinoma, but is also associated with other diseases not directly related to liver damage, such as an increased risk of cardiovascular disease. The association between insulin resistance and the development of NASH ( / NAFLD) is well known, and strategies to reduce insulin resistance may reduce disease progression and symptoms of NASH ( / NAFLD).

[0018] Uses according to the present disclosure may increase plasma levels of bile acids, particularly primary bile acids (cholic acid and chenodeoxycholic acid), and / or secondary bile acids (deoxycholic acid and lithocholic acid). This, in turn, reduces hepatitis (e.g., as determined by (the sum of) a lobular inflammation score of 0-3, a microgranuloma score of 0-1, a large lipogranuloma score of 0-1, and / or a portal inflammation score of 0-1, as shown below; or as determined by a necroinflammatory activity score (NAS)). Uses according to the present disclosure may therefore reduce hepatitis (e.g., as determined by (the sum of) a lobular inflammation score of 0-3, a microgranuloma score of 0-1, a large lipogranuloma score of 0-1, and / or a portal inflammation score of 0-1, as shown below; or as determined by a necroinflammatory activity score).

[0019] Increased bile acid plasma levels as part of the present disclosure are preferably demonstrated by one or more of the following methods: thin layer chromatography, gas chromatography, high performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LC-MS), gas chromatography mass spectrometry (GC-MS) supercritical fluid chromatography, and capillary electrophoresis, immunoassays, and bioluminescence assays.

[0020] In a particularly preferred embodiment, the use according to the present disclosure is for reducing hepatic necroinflammatory activity score.

[0021] The term hepatic necroinflammatory activity score is interchangeable with the terms NAFLD score and / or NASH score.

[0022] To determine the hepatic necroinflammatory activity score, the NASH Clinical Research Network (NASH-CRN) classification can be used (e.g., use hematoxylin-eosin stained slides for steatosis, inflammation, and ballooning, and Sirius red stained slides for assessment of fibrosis) as described in Kleiner et al., Vol. 41, No. 6 (June 2005). The score is preferably an unweighted sum of the steatosis grade (0-3), lobular inflammation grade (0-3), and hepatocyte ballooning grade (0-2). See below: [Table 1A] [Table 1B] [Table 1C]

[0023] Uses according to the present disclosure also include: - a steatosis grade score, in particular as defined above (score 1, 2, 3); and / or - fibrosis stage score, specifically as defined above (score 1, 1A, 1B, 1C, 2, 3, or 4) can be reduced.

[0024] The Anaerobutyricum soehngenii or its related species according to the present disclosure is preferably selected from the genus Anaerobutyricum or the genus Eubacterium, and is preferably Anaerobutyricum soehngenii (e.g., DSM17630 / KCTC15707) and / or Anaerobutyricum hallii (DSM3353 / ATCC27751).

[0025] In a study by Shetty et al. (Int J Syst Evol Microbiol. 2018 Dec;68(12):3741-3746), the bacterial species previously known as Eubacterium hallii was reclassified into two groups: Anaerobutyricum hallii and Anaerobutyricum soehngenii. Both species, Anaerobutyricum soehngenii and / or Anaerobutyricum hallii, are considered anaerobic Gram-positive catalase-negative bacteria belonging to Clostridial cluster XIVa (also known as Lachnospiracaea) of the phylum Firmicutes.

[0026] Most preferably, the at least one Anaerobutyricum species according to the present disclosure is Anaerobutyricum soehngenii (e.g., DSM17630 / KCTC15707) or a closely related species thereof having a 16S rRNA gene sequence with at least 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.5, 99.9, 100% sequence identity to the 16S rDNA sequence of Anaerobutyricum soehngenii (SEQ ID NO: 1). Such cutoff values ​​based on 16S rDNA similarity can define bacterial species with similar properties and / or functionality.

[0027] Additionally or alternatively, the Anaerobutyricum genus according to the present disclosure is Anaerobutyricum hallii (e.g., DSM3353 / ATCC27751) or a closely related species thereof having a 16S rRNA gene sequence with at least 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.5, 99.9, 100% sequence identity to the 16S rDNA sequence of Anaerobutyricum hallii (SEQ ID NO:2). Such cutoff values ​​based on 16S rDNA similarity can define species with similar properties and / or functionality. [Table 2A] [Table 2B]

[0028] In a preferred embodiment, Anaerobutyricum soehngenii, or a related species thereof, according to the present disclosure is combined with at least one Bifdobacterium sp. This is a synergistic combination that has been found to result in an unexpected reduction in hepatic necroinflammatory activity scores.

[0029] The Bifdobacterium sp. can be administered separately, sequentially, or simultaneously with the Anaerobutyricum soehngenii or closely related species, and thus the Bifdobacterium sp. can be included in the same composition or in a separate composition with respect to the Anaerobutyricum soehngenii or closely related species.

[0030] The genus Bifidobacterium is a genus of Gram-positive, usually non-motile, often branched, anaerobic bacteria. Bifidobacteria are widespread in the digestive tract, vagina, and oral cavity of mammals, including humans. Bifidobacteria are one of the major bacterial genera that constitute the mammalian digestive tract microbiota. At least one Bifidobacterium strain according to the present disclosure is preferably capable of assimilating human milk oligosaccharides (HMOs).

[0031] The at least one Bifdobacterium genus bacteria of the present disclosure preferably comprises: - Bifidobacterium animalis sub. lactis, or a closely related species thereof, having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity with the 16S rRNA gene sequence of the type strain of Bifidobacterium animalis sub. lactis (NCBI accession code NR_040867, SEQ ID NO: 3); - Bifdobacterium infantis (capable of assimilating HMO) having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity with the 16S rRNA gene sequence of the type strain of Bifdobacterium infantis (NCBI accession code D86184, SEQ ID NO: 4), or a closely related species thereof; - Bifdobacterium longum (capable of assimilating HMO) having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity with the 16S rRNA gene sequence of the type strain of Bifdobacterium longum (NCBI accession code M58739, SEQ ID NO: 5), or a closely related species thereof; - Bifdobacterium breve (capable of assimilating HMOs) having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity with the 16S rRNA gene sequence of the type strain of Bifdobacterium breve (NCBI accession code AB006658, SEQ ID NO: 6), or a closely related species thereof; - Bifidobacterium thermophilum having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity to the 16S rRNA gene sequence of the type strain of Bifidobacterium thermophilum (NCBI accession code AB016246, SEQ ID NO: 7), or a closely related species thereof; - Bifdobacterium bifidum having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity to the 16S rRNA gene sequence of the type strain of Bifdobacterium bifidum (NCBI accession code M38018, SEQ ID NO: 8), or a closely related species thereof; - Bifidobacterium adolescentis having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity to the 16S rRNA gene sequence of the type strain of Bifidobacterium adolescentis (NCBI accession code M58729, SEQ ID NO: 9), or a closely related species thereof; - Bifidbacterium catenulatum having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity to the 16S rRNA gene sequence of the type strain of Bifidbacterium catenulatum (NCBI accession code M58732, SEQ ID NO: 10), or a close relative thereof; - Bifidobacterium pseudocatenulatum, or a closely related species thereof, having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity to the 16S rRNA gene sequence of the type strain of Bifidobacterium pseudocatenulatum (NCBI accession code D86187, SEQ ID NO: 11). Includes one or more of the following.

[0032] In a particularly preferred embodiment, the Bifdobacterium spp. - Bifdobacterium animalis sub. lactis, or a closely related species thereof, having a 16S rRNA gene sequence with at least 90, 95, 97, 99, 100% sequence identity to SEQ ID NO: 3; and / or - Bifdobacterium breve, or a closely related species thereof, having a 16S rRNA gene sequence with at least 90, 95, 97, 99, 100% sequence identity to SEQ ID NO:6. is selected from. [Table 3A] [Table 3B] [Table 3C] [Table 3D] [Table 3E] [Table 3F] [Table 3G] [Table 3H]

[0033] In another particularly preferred embodiment, Anaerobutyricum soehngenii, or a related species thereof, and / or at least one Bifdobacterium sp. according to the present disclosure is combined with at least one Akkermansia sp., preferably said at least one Akkermansia sp., which is pasteurized (i.e., heated at 55-99°C, preferably 65-80°C for 5-60 seconds, or 1-60 minutes, preferably 60-80°C for 20-40 minutes, more preferably 65-75°C for 25-35 minutes). This is yet another synergistic combination, which has been found to result in an unexpected reduction in hepatic necroinflammatory activity scores.

[0034] The at least one Akkermansia bacterium can be administered separately, sequentially, or simultaneously with the Anaerobutyricum soehngenii, or a related species thereof, and / or the at least one Bifidobacterium bacterium, and thus the Akkermansia bacterium can be included in the same composition or in a separate composition with respect to the Anaerobutyricum soehngenii, or a related species thereof, and / or the at least one Bifidobacterium bacterium.

[0035] Preferably, the at least one Akkermansia species according to the present disclosure is Akkermansia muciniphila having a 16S rRNA sequence having at least 90, 95, 97, 99, or 100% sequence identity to SEQ ID NO:12, or a closely related species thereof.

[0036] Akkermansia is a genus in the phylum Verrucomicrobia. Akkermansia has been found to improve intestinal mucosal barrier function, or intestinal barrier function, which refers to the property of the intestinal mucosa to adequately contain unwanted luminal contents in the intestinal tract while maintaining the ability to absorb nutrients. Its role in protecting mucosal tissues and the circulatory system from exposure to inflammatory molecules such as microbes, toxins, and antigens is essential for maintaining health and well-being. Thus, Akkermansia can be used to prevent or treat intestinal mucosal barrier dysfunction, which is implicated in numerous health conditions, including food allergies, microbial infections, irritable bowel syndrome, inflammatory bowel disease, celiac disease, metabolic syndrome, nonalcoholic fatty liver disease, diabetes, and septic shock. See Collado et al. 2007 (Appl Environ Microbiol 2007 Dec;73(23):7767-70). or Appl Environ Microbiol. 2020 Mar 18;86(7):e03004-19.

[0037] The at least one Akkermansia genus of the present disclosure is preferably: - Akkermansia muciniphila (capable of assimilating HMO) having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity to the 16S rRNA gene sequence of the type strain of Akkermansia muciniphila (NCBI accession code AY271254, SEQ ID NO: 12), or a closely related species thereof. - Akkermansia glycanipila, or a closely related species thereof, having a 16S rRNA gene with at least 90, 95, 97, 98, 99, or 100% sequence identity to the 16S rRNA gene sequence of the type strain of Akkermansia glycanipila (NCBI accession code NR152695, SEQ ID NO: 13). Includes one or more of the following. [Table 4A] [Table 4B] [Table 4C]

[0038] In another particularly preferred embodiment, Anaerobutyricum soehngenii or related species thereof and / or at least one Bifdobacterium sp. and / or at least one Akkermansia sp. according to the present disclosure are combined with at least one Lactobacillus sp. This is a further synergistic combination, which has been found to result in an unexpected reduction in hepatic necroinflammatory activity scores.

[0039] The at least one Lactobacillus can be administered separately, sequentially or simultaneously with the Anaerobutyricum soehngenii, or a related species thereof, and / or the at least one Bifdobacterium, and / or the at least one Akkermansia, and thus the Lactobacillus can be included in the same composition or in separate compositions with respect to the Anaerobutyricum soehngenii, or a related species thereof, and / or the at least one Bifdobacterium, and / or the at least one Akkermansia.

[0040] The Lactobacillus genus is preferably selected from the following: - Lactobacillus acidophilus having a 16S rRNA sequence with at least 90, 95, 97, 99, 100% sequence identity to SEQ ID NO: 14, or a closely related species thereof; - Lactobacillus casei having a 16S rRNA sequence with at least 90, 95, 97, 99, 100% sequence identity to SEQ ID NO: 15, or a closely related species thereof; - Lactobacillus reuteri having a 16S rRNA sequence with at least 90, 95, 97, 99, 100% sequence identity to SEQ ID NO: 16, or a closely related species thereof; and / or - Lactobacillus rhamnosus having a 16S rRNA sequence that has at least 90, 95, 97, 99, 100% sequence identity to SEQ ID NO: 17, or a closely related species thereof. [Table 5A] [Table 5B] [Table 5C] [Table 5D]

[0041] In preferred embodiments, the present disclosure excludes the use (e.g., by co-administration) of any Ruminococcus spp. (e.g., Ruminococcus flavefaciens, R. torques, or R. faecis), any Faecalibacterium spp. (e.g., Faecalibacterium prausnitzii), and / or any Prevotella spp., such as Prevotella copri. The present disclosure may include or exclude any Anaerostipes spp. (particularly Anaerostipes rhamnisovorans) or any Faecalibacterium spp. (e.g., Faecalibacterium prausnitzii) to obtain improved efficacy in prophylaxis and / or treatment by the present disclosure.

[0042] It is envisioned that Anaerobutyricum soehngenii or a closely related species thereof, Bifdobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure are contained in feces.

[0043] The Anaerobutyricum soehngenii or related species thereof, Bifdobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure may be, for example, from or derived from fecal material obtained from one or more donor subjects. As used herein, the term "donor" refers to the subject providing the feces. Fecal material according to the present disclosure may thus be derived from a donor and administered to a recipient. Optionally, after processing, the fecal material is administered to the recipient. The one or more donor subjects are preferably mammals, preferably humans. Also, the recipient is preferably a mammal, preferably humans.

[0044] Preferably, the fecal material is obtained from at least one healthy (human) donor, more preferably from at least one (human) donor who follows (or has followed) a vegetarian diet, most preferably a vegan diet. A vegetarian diet does not include any meat, poultry, or seafood, or includes at most 0.1, 0.5, 1 kg of meat, poultry, or seafood per month. A vegan diet does not include any meat, poultry, seafood, or food of animal origin, or includes at most 0.1, 0.5, 1 kg of meat, poultry, seafood, or food of animal origin per month. A healthy donor can be considered as a donor who does not have a condition, for example, as described in Table 1 of Lise Sofie et al. (2019, Transfusion and Apheresis Science, Volume 58, Issue 1, P113-116).

[0045] The selected donor subject is preferably between 18-27, preferably 20-25 kg / m 2 As used herein, the term "body mass index," or "BMI," refers to a person's mass divided by the square of their height, and is expressed as kg / m 2 The donor subjects selected are preferably aged less than 30 years, or less than 35 years. For example, at least one donor subject is aged between 18 and 30 years, for example between 20 and 25 years. Additionally or alternatively, the donor selected follows (or has followed) a diet rich in prebiotic fiber (which increases fecal butyric acid production), such as, for example, Whole Fiber, see WO2021 / 204719 (e.g., at least 0.1, 0.5, 1 kg of prebiotic fiber per month).

[0046] Additionally or alternatively, at least one donor subject has a relative abundance of Bifidobacterides order bacteria in the fecal material of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30% (compared to the abundance of bacteria in other genera). Additionally or alternatively, at least one donor subject has a relative abundance of Akkermansia genus bacteria in the fecal material of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30% (compared to the abundance of bacteria in other genera).

[0047] In a preferred embodiment, at least 10% of the Anaerobutyricum soehngenii or its related species is 8 Pieces or 10 8 At least 10 cells of the Bifdobacterium spp. are contained in the fecal material. 8 Pieces or 10 8 The fecal material contains at least 10 cells of the Akkermansia spp. 8 Pieces or 10 8 At least 10 cells of the Lactobacillus genus are contained in the fecal material. 8 Pieces or 10 8 cells are contained in the fecal material.

[0048] In other words, the Anaerobutyricum soehngenii or related species thereof, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure are preferably enriched in the fecal material, i.e. the cell count of Anaerobutyricum soehngenii or related species thereof, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. is higher than in the fecal material of the prior art, e.g. cells of Anaerobutyricum soehngenii or related species thereof, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. are added to the fecal material or the feces is enriched with said Anaerobutyricum soehngenii or related species thereof, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. When Anaerobutyricum soehngenii or its related species, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure are contained in feces, preferably, at least 10 4 , 10 5 , 2×10 5 , 3×10 5 , 4×10 5 , 5×10 5 , 6×10 5 , 7×10 5 , 8×10 5 , 9×10 5 , 10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 6×10 6 , 7×10 6 , 8×10 6 , 9×10 6 , 10 7 , 2×10 7 , 3×10 7 , 4×107 , 5×10 7 , 6×10 7 , 7×10 7 , 8×10 7 , 9×10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 Preferably, Anaerobutyricum soehngenii or a related species thereof, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. are the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, and / or tenth most predominant bacterial species in the fecal material, i.e., have the highest cell count or at least have a cell count that is in the top ten compared to other bacterial species contained in the fecal material.

[0049] Preferably, when the composition according to the present disclosure is fecal material, the fecal material can be feces, or a portion thereof, preferably a purified portion thereof. Purifying the fecal material can make it more convenient to administer. In certain embodiments, a 50-150 mg fecal material sample can be combined with 5-15 mL of isotonic saline, e.g., containing 10% glycerol, and frozen at -80°C until delivery. For example, 1 mL can be mixed with breast milk or pasteurized cow's milk to make 10 mL, and 5 mL can be administered to the recipient.

[0050] As used herein, a portion of fecal matter refers to one or more specific groups of components, including, but not limited to, enzymes, proteins, lipids, molecules, microorganisms, viruses, bacteria, fungi, yeasts, archaea, compounds, complexes, solids, liquids, particles, and fibers.

[0051] As used herein, a purified portion of fecal material indicates that undesirable components are not present in the fecal material.

[0052] Preferably, fecal material for use according to the present disclosure is contained in a liquid medium and / or does not contain solids having a diameter greater than 10, 25, 50, 75, 100, 200, 400, 600, 800, or 1000 μm, and is preferably obtained by mixing allogeneic feces with an aqueous medium followed by filtration and / or centrifugation. This significantly reduces the viscosity of the feces and increases its fluidity, facilitating administration of the fecal material to a subject. The liquid medium may consist of water or other types of liquids that may be supplemented with other components, such as, for example, salts, to provide an isotonic solution.

[0053] According to one aspect of the disclosure, the fecal material according to the disclosure is included in a composition, such as a pharmaceutical composition, more preferably a liquid dosage form, that facilitates administration of the fecal material to a recipient.

[0054] It is further envisioned that the fecal material according to the present disclosure is in a lyophilized and / or microencapsulated form (to protect from the gastric environment). Uses according to the present disclosure may include administering fecal material obtained from at least one donor subject to a recipient in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 separate doses, preferably with at least 1, 2, 3, 4, 5, 6, 7, 8 weeks between doses.

[0055] Alternatively, no Anaerobutyricum soehngenii or related species, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure are present in the feces.

[0056] At least one Anaerobutyricum soehngenii or related species thereof, at least one Bifidobacterium genus, at least one Akkermansia genus, and / or at least one Lactobacillus genus according to the present disclosure may be included in the composition.

[0057] Compositions according to the present disclosure may be administered enterally, preferably orally, nasally, or rectally, and / or via a nasoduodenal tube.

[0058] The compositions according to the present disclosure can be used as pharmaceuticals and / or with physiologically acceptable carriers, which can be any inert carrier. For example, non-limiting examples of suitable physiologically or pharma-ceutically acceptable carriers include any well-known physiological or pharmaceutical carriers, buffers, diluents, and excipients. It will be understood that the selection of a suitable physiological carrier depends on the intended mode of administration (e.g., oral) of the compositions taught herein. Those skilled in the art know how to select a physiologically acceptable carrier that is suitable or compatible with the compositions for use taught herein.

[0059] It is envisaged that compositions according to the present disclosure are contained in and / or encapsulated by an (enteric) coating, which preferably does not dissolve and / or disintegrate in the recipient's stomach environment. Such a coating can help the composition reach the intended site for delivery, e.g., the duodenum, without being degraded by the acidic environment of the stomach. Preferred (enteric) coatings function by presenting a surface that is stable at the highly acidic pH found in the stomach, but degrades more rapidly at lower pH. For example, they will not dissolve in the gastric acid of the stomach (pH about 3), but will dissolve in the alkaline (pH 7-9) environment present in the small intestine, or the duodenum.

[0060] In one embodiment, the present disclosure relates to compositions for use as probiotics. Thus, as used herein, "probiotics" refers to microorganisms, such as intestinal bacteria, that, when administered or ingested in effective amounts, provide a health benefit to a host (e.g., a human or mammal). Preferably, probiotics should be alive or viable when administered to a subject, so that they colonize the large intestine of the host. However, under certain conditions, probiotics may be dead when administered, provided that the substances produced by the probiotics still exert a probiotic beneficial effect on the host.

[0061] In one embodiment, the combination taught herein may be for use as a symbiotic. The term "symbiotic" or "symbiotic product" as used herein generally refers to a composition and / or dietary supplement that combines probiotics and one or more compounds that promote the growth and / or activity of GI microorganisms, such as prebiotics, in one product. Symbiotics beneficially affect the host by improving the survival and colonization of probiotics in the GI tract, selectively stimulating the growth of probiotics, and / or activating metabolism, thus improving the condition of the host. Those skilled in the art are familiar with symbiotics and know how to select ingredients that can be combined into symbiotics.

[0062] The inventors have further surprisingly found that microencapsulation of at least one Anaerobutyricum soehngenii, or a related species thereof, at least one Bifidobacterium sp., at least one Akkermansia sp., and / or at least one Lactobacillus sp. according to the present disclosure may provide additional synergistic therapeutic effects in the prevention or treatment of hepatic steatosis, NAFLD, and / or NASH.

[0063] The term "microencapsulation" is used to describe the encapsulation of bacteria in a matrix, coating, or membrane, typically a protective matrix or membrane. The (average) diameter of the microcapsules is between 50 nm and 2 mm, preferably between 100 nm and 1 mm. The matrix, coating, or membrane is typically comprised of milk, milk proteins, and / or polymers. The purpose of the microencapsulation may be to protect the bacteria, and its components, from destruction by the surrounding environment, such as the gastrointestinal environment, among other possible purposes. Microencapsulation of bacteria may also help improve the incorporation of bacteria into dairy products, foods, pharmaceutical preparations, and / or pharmaceutical compositions. Microencapsulation of bacteria may also support therapeutic effects.

[0064] A variety of materials can be used for microencapsulation of bacteria, including, for example, pea protein, milk, milk protein, whey protein, casein, xanthan gum, alginate, gelatin, chitosan, carboxymethylcellulose, starch, and / or carrageenan, and combinations thereof. In a preferred embodiment, the Anaerobutyricum soehngenii or related species thereof, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure are microencapsulated in one or more polymers.

[0065] The subject receiving the combination or composition taught herein may be selected from the group consisting of humans, non-human primates, mice, rats, dogs, cows, and pigs. In a preferred embodiment, the subject is a human.

[0066] According to the present disclosure, at least one species of Anaerobutyricum soehngenii or a related species thereof, at least one species of Bifidobacterium genus, at least one species of Akkermansia genus and / or at least one species of Lactobacillus genus are 4 ~10 15The combination, in amounts ranging from 0.1 to 10000 colony forming units (CFU), For example, at least one species of Anaerobutyricum soehngenii or a related species thereof, at least one species of Bifidobacterium, at least one species of Akkermansia, and / or at least one species of Lactobacillus may be present in an amount of, for example, 10% or more per dose, or per ml, or per gram of formulation or composition comprising said species. 6 CFU~10 13 CFU, preferably 10 7 CFU~10 12 CFU, preferably 10 8 CFU~10 11 CFU, more preferably 10 9 CFU~10 11 The amount of CFU may be included in the combination.

[0067] In one embodiment, the at least one Anaerobutyricum soehngenii, or related species thereof, at least one Bifidobacterium, at least one Akkermansia, and / or at least one Lactobacillus in a combination or composition taught herein may be incorporated in a lyophilized and / or microencapsulated form (e.g., as reviewed by Solanki et al., BioMed Res. Int. 2013, Article ID 620719), or other form that retains the activity and / or viability of the bacterial strain.

[0068] In one embodiment, the combination or composition taught herein may include one or more ingredients suitable for promoting the survival and / or viability of the bacteria taught herein or strains derived therefrom during storage and / or exposure to bile and / or passage through the gastrointestinal tract of a mammal (e.g., human). Non-limiting examples of such ingredients include enteric coatings and controlled release agents that allow passage through the stomach. Those skilled in the art know how to select appropriate ingredients to keep bacteria as taught herein viable and functional, i.e., capable of performing their intended functions.

[0069] For example, it may be advantageous to add one or more prebiotic ingredients to the combinations taught herein to supplement the effects of the bacteria taught herein (e.g., production of propionate / propionate and / or butyrate / butyrate or derivatives thereof). The prebiotic ingredients may also enhance the activity and / or stimulate the growth of the bacteria, or strains derived therefrom, as taught herein. As used herein, "prebiotic" generally refers to non-digestible food ingredients that promote the growth of beneficial microorganisms in the gut. Prebiotics, or prebiotic products, consist primarily of fermentable fibers, or non-digestible carbohydrates. Fermentation of these fibers by probiotics promotes the production of beneficial end-products, such as SCFAs, especially butyrate. Non-limiting examples of suitable prebiotics include fibers such as inulin, pectin, resistant starch, as well as cellobiose, maltose, mannose, salicin, trehalose, amygdalin, arabinose, melibiose, sorbitol, rhamnose, and / or xylose. Those skilled in the art are familiar with the field of prebiotics and know how to select ingredients with prebiotic activity.

[0070] In addition to or instead of preventing and / or treating hepatic steatosis, NAFLD and / or NASH, the present disclosure can be used to (enhance) butyric acid and / or butyrate production, preferably in situ, i.e. in the small intestine.Similarly, the combination according to the present disclosure can also reduce the level of lactate in the small intestine, for example in situ (lactate is known to be an undesirable compound in the intestinal tract).

[0071] As used herein, the term "butyrate" or "butyric acid" (also known by the systematic name butanoic acid) refers to a carboxylic acid having the structural formula CH3CH2CH2COOH. The term may include its derivatives, i.e., compounds derived from butyric acid, including salts and esters of butyric acid known as butyrates or butanoates. Non-limiting examples of butyrate salts include sodium butyrate, calcium butyrate, magnesium butyrate, manganese butyrate, cobalt butyrate, barium butyrate, lithium butyrate, zinc butyrate, potassium butyrate, ferrous butyrate, and the like. Non-limiting examples of butyrate esters (i.e., esters of butyric acid) include cellulose acetate butyrate, methyl butyrate, ethyl butyrate, butyl butyrate, pentyl butyrate, and the like.

[0072] Without wishing to be bound by any theory, the bacterial strains according to the present disclosure may survive and at least transiently colonize the gastrointestinal tract of a human when administered to a human or ingested in an appropriate amount by a human. This colonization may typically allow for increased butyric acid / butyrate production in situ, although other mechanisms cannot be excluded. Increased in situ production may at least partially underlie the beneficial effects of the combinations taught herein, such as the prevention and / or treatment of hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), and / or non-alcoholic steatohepatitis (NASH).

[0073] In one embodiment, at least one species of Anaerobutyricum soehngenii, or a related species thereof, at least one species of Bifidobacterium, at least one species of Akkermansia, and / or at least one species of Lactobacillus can be included in a food composition, a feed composition, a feed supplement composition, a food supplement composition, or a pharmaceutical composition. At the same time, or alternatively, at least one species of Anaerobutyricum soehngenii, or a related species thereof, at least one species of Bifidobacterium, at least one species of Akkermansia, and / or at least one species of Lactobacillus can be included in a liquid, a liquid beverage (including dairy beverages and fermented beverages), yogurt, cheese, gel, gelatin, gelatin capsule, powder, paste, tablet, or capsule.

[0074] The food or food supplement composition is preferably a dairy product, more preferably a fermented dairy product, most preferably a yoghurt or a yoghurt drink.

[0075] The pharmaceutical composition may be, for example, in liquid or solid form, more preferably in solid form, such as a solid dosage form, for example, a capsule, tablet, or powder. Preferably, the pharmaceutical composition does not involve pure water or an aqueous medium containing more than 99% water by weight.

[0076] The compositions as taught herein, including the combinations for use according to the present disclosure, may further comprise any acceptable carrier suitable for maintaining the Anaerobutyricum soehngenii or its relatives, Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. according to the present disclosure viable until consumed by a subject (e.g., a human or an animal). For example, non-limiting examples of acceptable carriers suitable for this purpose include any of the well-known physiological or pharmaceutical carriers, buffers, and excipients. It can be understood that the selection of a suitable physiological or pharmaceutical carrier depends on the intended mode of administration of the compositions as taught herein (e.g., oral) and the intended form of the compositions (e.g., beverage, yogurt, powder, capsule, etc.). Those skilled in the art know how to select suitable physiological or pharmaceutical carriers for the compositions as taught herein.

[0077] At least one species of Anaerobutyricum soehngenii or a related species thereof, at least one species of Bifidobacterium spp., at least one species of Akkermansia spp., and / or at least one species of Lactobacillus spp. taught in the present disclosure may be used in a variety of applications, including the manufacture of food products, pharmaceuticals, and pharmaceuticals that are suitable for use in food and pharmaceutical preparations. 4 ~10 15 For example, at least one species of Anaerobutyricum soehngenii or related species, at least one species of Bifidobacterium, at least one species of Akkermansia, and / or at least one species of Lactobacillus may be present in the composition in amounts ranging from 10 to 1000 mg / kg. 6 CFU~10 13 CFU, preferably 10 7CFU~10 12 CFU, preferably 10 8 CFU~10 11 CFU, more preferably 10 9 CFU~10 11 Alternatively, the amount and / or frequency of administration of at least one species of Anaerobutyricum soehngenii or related species, at least one species of Bifidobacterium, at least one species of Akkermansia, and / or at least one species of Lactobacillus may be greater than 10 CFU per day, for example, per dose, or per ml, or per g of formulation or composition comprising said species. 6 ~10 13 , preferably 10 7 ~10 12 , preferably 10 8 ~10 11 , more preferably 10 9 ~10 11 CFU.

[0078] As used herein, the terms "comprising" or "to comprise" and their conjugations refer to the context in which the term is used in an open-ended sense, meaning that items following the word are included, but items not specifically mentioned are not excluded. It also encompasses the more restrictive verbs "to consist essentially of" and "to consist of."

[0079] The reference to an element by the indefinite article "a" or "an" does not exclude the possibility that there is more than one of the element, unless the context clearly requires that there is only one of the element. Thus, the indefinite article "a" or "an" normally means "at least one".

[0080] The terms "increase" and "increased level", as well as "decrease" and "decreased level" refer to the ability to significantly increase or decrease, or to a significantly increased or decreased level. Generally, a level is increased or decreased if it is at least 5%, e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% higher or lower, respectively, than the corresponding level in a control or reference. Alternatively, a level in a sample can be increased or decreased if it is statistically significantly increased or decreased compared to the level in a control or reference.

[0081] As used herein, the term "identity" refers to a measure of the identity of nucleotide or amino acid sequences. Generally, sequences are aligned to obtain the highest sequence match. "Identity" per se has an art-recognized meaning and can be calculated using published techniques. See, for example: (COMPUTATIONAL MOLECULAR BIOLOGY, Lesk, AM, ed., Oxford University Press, New York, 1988; BIOCOMPUTING: BIOCOMPUTING: INFORMATICS AND GENOME PROJECTS, Smith, DW, ed., Academic Press, New York, 1993; COMPUTER ANALYSIS OF SEQUENCE DATA, PART I, Griffin, AM, and Griffin, HG, eds, Humana Press, New Jersey, 1994; SEQUENCE ANALYSIS IN MOLECULAR BIOLOGY, von Heinje, G., Academic Press, 1987; and SEQUENCE ANALYSIS PRIMER; Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991). Although there are many methods for measuring identity between two polynucleotide or polypeptide sequences, the term "identity" is well known to those of skill in the art (Carillo, H., and Lipton, D., SIAM J. Applied Math (1988) 48:1073).

[0082] Commonly used methods for determining identity or similarity between two sequences include, but are not limited to, those disclosed in GUIDE TO HUGE COMPUTERS, Martin J. Bishop, ed., Academic Press, San Diego, 1994, and Carillo, H., and Lipton, D., SIAM J. Applied Math (1988) 48:1073. Methods for determining identity and similarity are codified in computer programs, such as the standard settings of NCBI Nucletide Blast (blastn, https: / / blast.ncbi.nlm.nih.gov / ). Preferred computer program methods for determining identity and similarity between two sequences include the GCS program package (Devereux, J., et al., Nucleic Acids Research (1984) 12(1):387), BLASTP, BLASTN, FASTA (Atschul, SF et al., J. Molec. Biol. (1990) 215:403).

[0083] By way of example, a nucleotide sequence having at least, for example, 95% "identity" to a reference nucleotide sequence is intended to mean that the nucleotide sequence is identical to the reference sequence, except that there may be up to 5 point mutations per 100 nucleotides of the reference polypeptide sequence. In other words, to obtain a nucleotide sequence that is at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted and / or replaced with another nucleotide, and / or up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. In the sequence listing, "n" may represent a, t, g, or c.

[0084] In the event of a discrepancy between a sequence disclosed herein and a sequence disclosed in the sequence listing, the sequence disclosed herein is preferred. Alternatively, the sequence in the sequence listing may be used. [Brief description of the drawings]

[0085] [Figure 1] SCFA production in the absence or presence of Bifidobacterium animalis subsp. lactis BLC1. [Diagram 2] SCFAs produced on fucose (25 mM) in YCFA medium in the absence or presence of L. rhamnosus GG. [Diagram 3] Histological evaluation of mice. AD: inflammation grade, fibrosis grade, NAS score or global NASH score of mice. E: CRN classification.

[0086] EXAMPLES

[0087] Example 1 A. soehngenii has been shown to affect glucose metabolism in the small intestine and insulin resistance. In an in vitro model of the ileum in the presence of a synthetic microbiota, A. soehngenii contributes only to a limited extent to SCFA production. Experiments were performed to determine whether this SCFA production could be enhanced by supplementation with the commercially available probiotic Bifidobacterium animalis subsp. lactis BLC1 (Bottacini et al. 2011, J Bacteriol 193: 6387-6388).

[0088] Briefly, a bacterial synthetic consortium containing upper intestinal bacteria (Lactobacillus spp., Streptococcus spp., Enterococcus spp., Clostridium nexile, Faecalibacterium prausnitzii, Veillonella spp., Prevotella melaninogenica, and Blautia obeum) and a supporting substrate was stabilized for 14 days in the ileal mucosa-SHIME (Simulator of Human Intestinal Microbial Ecosystem) model.

[0089] 7 ml of this stabilized consortium was inoculated with A. soehngenii or a combination of A. soehngenii and B. infantis and cultured under anaerobic conditions in the presence of 3 mM bile salts at 37° C. The initial pH of the medium was 7.5.

[0090] After 24 h, samples were taken and analyzed for SCFAs (acetate, propionate, and butyrate). The results showed that in the presence of both A. soehngenii and B. infantis, all SCFAs were significantly increased compared to the SCFA levels in the presence of A. soehngenii alone (Figure 1).

[0091] This indicates metabolic synergy between A. soehngenii and B. infantis under upper intestinal conditions.

[0092] Example 2 Similarly, synergy between A. soehngenii L2-7 and various Lactobacillus species was shown in cultures with various carbon sources. The combination of A. soehngenii and the commercial probiotic strain Lactobacillus rhamnosus GG (Kankainen et al. 2009 106:17193-8) showed clear synergy in growth on fucose, a common sugar present in the intestinal tract. Although A. soehngenii does not utilize fucose, L. rhamnosus GG converted it to lactate and acetate, while the combination of both strains converted fucose to butyrate, the main metabolic end product of A. soehngenii. See Figure 2.

[0093] Example 3 Two groups of C57BL6 / J mice, 10 mice per group, were fed a Western diet (WDF) enriched with 15% fructose in drinking water for 20 weeks. A control group of 10 mice was fed a chow diet for the same period. The WDF resulted in a diet-induced obesity (DIO) mouse model of non-alcoholic steatohepatitis (NASH) (body weight increased 25% compared with control mice). Starting at week 12, DIO-NASH mice were orally challenged once weekly with 10^ CFU of A. soehngenii or a placebo. At week 20, mice were killed and blood, including portal vein samples, and liver and intestinal samples were collected. The WDF-induced DIO-NASH model was effective in inducing NASH: mean histological steatosis grade at week 20 was 3, mean NAS score was 4, and mean fibrosis grade was 1 (perihepatic or periaortic fibrosis).

[0094] A clear reduction in inflammation grade, fibrosis grade, NAS score, and global NASH score was observed with A. soehngenii compared to placebo, and the number of mice showing NASH was reduced compared to placebo (Figure 3).

[0095] Example 4 The present inventors have found that co-administration of Anaerobutyricum soehngenii or Anaerobutyricum hallii with Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp. has beneficial and synergistic effects in patients suffering from or at risk of hepatic steatosis.

[0096] method participants The study included untreated Caucasian patients with hepatic steatosis confirmed by ultrasound examination. The main inclusion criteria were age 21–69 years, males or postmenopausal females, and a body mass index (BMI) of 25 kg / m 2 Patients were required to have a fatty liver larger than 18 years of age and suspected of having NAFLD on a prior ultrasound scan (based on elevated liver enzymes, impaired glucose tolerance, and severity of steatosis on ultrasound).Exclusion criteria were a history of any of the following: cardiovascular disease, T2DM, renal disease, cholecystectomy, or immunosuppression; use of proton pump inhibitors, antibiotics, or anticoagulants in the past 3 months; current medication use; history of moderate to heavy alcohol intake (≥12 g / day); and causes of liver disease other than NAFLD (e.g., hemochromatosis, autoimmune hepatitis, cirrhosis, hepatitis B or C, hemochromatosis, alpha-1 antitrypsin deficiency, alcoholic liver disease).

[0097] intervention Subjects were treated for at least 24 weeks according to the single or combination treatment arms shown in Table 1. Necroinflammatory liver activity scores (NAFLD activity scores) were measured at baseline and after treatment. 10 The living unit was administered once daily in capsule form.

[0098] Liver biopsy Percutaneous liver biopsies were performed according to local standard procedures and based on clinical indications. All tissue specimens were scored by a hepatic pathologist blinded to other results. Hematoxylin-eosin stained slides were used to assess steatosis, inflammation, and ballooning, and Sirius red stained slides were used to assess fibrosis, according to the NASH Clinical Research Network (NASH-CRN) classification (Kleiner et al. vol. 41, no. 6, June 2005). Necroinflammatory activity scores (NAS) were determined as described herein.

[0099] plasma measurement Bile acid plasma concentrations were measured by liquid chromatography tandem mass spectrometry (LC-MS / MS).

[0100] result As shown, the inventors determined the therapeutic efficacy of Anaerobutyricum soehngenii or Anaerobutyricum hallii administered alone or in combination with Bifidobacterium spp., Akkermansia spp., and / or Lactobacillus spp.

[0101] Anaerobutyricum soehngenii or Anaerobutyricum hallii alone has limited ability to improve necroinflammatory activity scores. On the other hand, Anaerobutyricum soehngenii or Anaerobutyricum hallii alone leads to increased plasma levels of primary bile acids (cholic acid and chenodeoxycholic acid) and secondary bile acids (deoxycholic acid and lithocholic acid). Increased plasma levels of these bile acids activate the farnesoid-X-receptor (FXR) and the G protein-coupled bile acid receptor GPBAR1 (TGR5), leading to increased secretion of GLP-1, which suppresses hepatic lipogenesis and hepatic inflammation (Chiang (Liver Res. 2017 Jun; 1(1): 3-9).

[0102] The effects on bile acid plasma levels after treatment and efficacy in reducing necroinflammatory activity scores are shown in Table 6 (Table.1) according to the following ranking system: the first rank represents the lowest effect and the last rank represents the highest effect: "not measurable", "weak", "low", "low / moderate", "moderate", "high", "very high". Lower necroinflammatory activity scores can prevent the development of hepatic steatosis, NAFLD, and / or NASH in healthy individuals. It is expected that results similar to the estimated effects shown in Table 6 (Table.1) will be obtained in larger patient cohorts. [Table 6]

[0103] Example 5 Microencapsulation As shown in this experiment, we compared the effects of non-microencapsulated bacteria with the effects of microencapsulated bacteria.

[0104] The same subjects and metrics were used as described in Example 4. The same ranking system was used to demonstrate efficacy as described in Example 4. To illustrate the effect of microencapsulation of the bacteria, the dosage of the bacteria was 100 times lower compared to Example 1. The bacteria were in capsule form, administered once daily at 10 per capsule. 8 The dose was given in viable units.

[0105] result The results are shown in Table 7 (Table.2). [Table 7]

[0106] It is expected that similar benefits will be observed in larger patient cohorts, as shown in Table 7 (Table 2).

Claims

1. A composition for preventing and / or treating fatty liver, comprising Anaerobutyricum soehngenii or a closely related species thereof, having a 16S rRNA gene sequence having at least 97% sequence identity with SEQ ID NO: 1 or SEQ ID NO: 2, wherein the Anaerobutyricum soehngenii or a closely related species thereof is combined with at least one type of Bifidobacterium genus bacterium.

2. The at least one type of Bifidobacterium bacterium is - Bifidobacterium animalis subspecies lactis, or a closely related species thereof, having a 16S rRNA gene sequence with at least 97% sequence identity to SEQ ID NO: 3, and / or - Bifidobacterium breve, or a closely related species thereof, having a 16S rRNA gene sequence that has at least 97% sequence identity with SEQ ID NO:

6. The composition of claim 1 selected from:

3. The composition according to claim 1, wherein the use is further for reducing hepatic necroinflammatory activity score.

4. 2. The composition of claim 1, wherein the fatty liver is nonalcoholic fatty liver disease (NAFLD) and / or nonalcoholic steatohepatitis (NASH).

5. 2. The composition of claim 1, wherein the Anaerobutyricum soehngenii, or a related species thereof, is combined with at least one species of Bifidobacterium and at least one species of Akkermansia.

6. The composition according to claim 5 , wherein the at least one type of Akkermansia fungus is targeted for sterilization.

7. 6. The composition of claim 5, wherein the at least one Akkermansia species is Akkermansia muciniphila having a 16S rRNA sequence with at least 97% sequence identity to SEQ ID NO: 12, or a closely related species thereof.

8. 2. The composition of claim 1, wherein the Anaerobutyricum soehngenii, or a related species thereof, is combined with at least one species of Bifidobacterium and at least one species of Lactobacillus.

9. The at least one kind of Lactobacillus bacterium is - Lactobacillus acidophilus or a closely related species thereof, having a 16S rRNA sequence that has at least 97% sequence identity with SEQ ID NO: 14; - Lactobacillus casei or a closely related species thereof, having a 16S rRNA sequence that has at least 97% sequence identity with SEQ ID NO: 15; - Lactobacillus reuteri or a closely related species thereof, having a 16S rRNA sequence with at least 97% sequence identity to SEQ ID NO: 16, and / or - Lactobacillus rhamnosus or a closely related species thereof, having a 16S rRNA sequence that has at least 97% sequence identity with SEQ ID NO:

17. The composition of claim 8 selected from:

10. The composition described in claim 1, wherein the Anaerobutyricum soehngenii, or a related species thereof, is derived from fecal material, preferably the fecal material being obtained from a healthy donor.

11. 11. The composition of claim 10, wherein the fecal material is obtained from a donor following a vegan diet.

12. The fecal material comprises at least 10 8 The composition of claim 10 comprising cells of Anaerobutyricum soehngenii, or a closely related species thereof.

13. The composition described in claim 1, wherein the Anaerobutyricum soehngenii, or a related species thereof, is in a microencapsulated or freeze-dried form.

14. The composition described in claim 1, wherein the Anaerobutyricum soehngenii, or a related species thereof, is contained in a composition preferably including a physiologically acceptable carrier.

15. The Anaerobutyricum soehngenii or its related species is 10 14 ~10 15 15. The composition of claim 14, wherein the composition is present in an amount in the range of colony forming units (CFU).

16. The composition of claim 14, - a pharmaceutical composition, preferably in solid dosage form such as a capsule, tablet or powder, and / or a food composition, which is preferably a dairy product, more preferably a fermented dairy product, most preferably a yogurt or a yogurt drink The composition.

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