Media supplements and methods to culture human gastrointestinal anaerobic microorganisms

A media supplement for culturing anaerobic bacteria, the media supplement comprising a filtrate of effluent from a chemostat vessel in which a target bacterial ecosystem has been cultured, wherein the target bacterial ecosystem has been cultured in culture media which is Media 1, Media 1 comprising:

• 0.4% w/v Peptone;
• 0.4% w/v Yeast extract;
• 0.4% w/v NaHCO3;
• 0.4% w/v Pectin;
• 0.4% w/v Xylan;
• 0.4% w/v Arabinogalactan;
• 0.6% w/v Casein;
• 1% w/v unmodified wheat starch;
• 0.2% w/v inulin;
• 0.1% w/v bile salts;
• 0.1% w/v L cysteine HCl;
• 0.0002% w/v CaCl2;
• 0.0002% w/v NaCl;
• 0.0008% w/v K2HPO4;
• 0.0008% w/v KH2PO4;
• 0.0002% w/v MgSO4;
• 0.0001% w/v Hemin;
• and 0.00002% w/v menadione.

A method for preparing a media supplement for culturing anaerobic bacteria, wherein the method comprising the steps of:

• a) culturing a target bacterial ecosystem in culture media in a single-stage chemostat under conditions for a system retention time of about 20 to 70 hours, a temperature of about 37° C., a pH of about 6.9 to 7 and maintenance of anaerobic conditions to the chemostat;
• b) collecting effluent from the chemostat;
• and
• c) filtering the effluent through a 0.2 μm filter to remove bacterial cells, in order to produce the media supplement;
• wherein the culture media and the target bacterial ecosystem comprises:
• (1) Acidaminococcus intestinalis, Bacteroides ovatus, Bifidobacterium adolescentis, Bifidobacterium longum, Blautia product, Clostridium cocleatum, Collinsella aerofaciens, Dorea longicatena, Escherichia coli, Eubacterium desmolans Eubacterium eligens, Eubacterium limosum, Eubacterium rectale, Eubacterium ventriosum, Faecalibacterium prausnitzii, Lachnospira pectinoshiza, Lactobacillus casei/paracasei, Lactobacillus casei, Parabacteroides distasonis, Raoultella sp., Roseburia faecalis Roseburia intestinalis, Ruminococcus torques Ruminococcus obeurn, and Streptococcus mills;
• (2) Acetobacterimn sp., Acidaminococcits intestinalis, Anaerostipes hadrus, Atopobimn minntum. Bacteroides fragilis, Bacteroides ovatus, Bacteroides vulgatus, Bildobacterium adolescentis Bitidobacterium longum, Blautia coccoides, Blautia producta, Clostridium aldenense. Clostridium citroniae, Clostridium cocleatum, Clostridium hathewayi, Clostridium lacialifermentans. Clostridium orbiscindens. Collinsella aerofaciens Dorea longicatena Escherichia coli Eubacterimn desmolans Eubacterium eligens, Eubacterium fissicatena Eubacterium limosum. Eubacterium rectale, Ellbacterium sp., Eubacterium yentriosum Faecalibacterium prausnitzii, Lachnospira pectinoshiza, Lactobacillus casei, Lactobacillus paracasei. Parabacteroides distasonis, Raoultella sp., Roseburia faecalis, Roseburia hominis Roseburia intestinalis. Roseburia inulinivorans Ruminococcus torques, Ruminococcus obeum, and Streptococcus mitis;
• (3) Adlercreutzia equolifaciens, Akkermansia muciniphila, Alistipes finegoldii, Alistipes utredinis, Alistipes shahii, Alistipes sp., Bacteroides capillosus, Bacteroids cellulosilyticus, Bacteroides eggerthii, Bacteroides ovatus. Bacteroides thetaiotaomicron, Bacteroides uniformis. Bacteroides vulgatus, Bacillus circulans, Bacillus, simplex. Bifidobacterium longum Blautia hydrogenotrophica, Blautia sp., Blautia/Clostridum coccoides, Brevibacillus parabrivis, Catabacter hongkongensis, Catabactei sp., Catenibacterium mitsuokai, Clostridium aldenense 1, Clostridium aldenense 2, Clostridium asparagiforme, Clostridium bolteae, Clostridium celerecrescens. Clostridium hathewayi 4, Clostridium hathewayi 2, Clostridium hathewayi 3, Clostridium hathewayi 4, Clostridium hvlemonae 1 Clostridium hylemonae 2 Clostridium inocuum, Clostridium lavalense, Clostridium leptum, Clostridium orbiscindens Clostridium ramosum, Clostridium scindens, Clostridium staminisolvens. Clostridium sultatireducens, Clostridium symbiosum. Clostridium thermocellum, Clostridium sp. 1, Clostridium sp. 2, Clostridium sp. 3, Clostridium sp 4, Clostridium sp 5, Clostridium sp 6, Collinsella aerofaciens, Coprococcus catus, Coprococcus comes, Coprococcus eutactus, Dorea formicigenerans, Dorea longicatena, Escherichia coli, Eubacterium biforme Eubacterium callanderi, Eubacterium dolichum, Euhacteriwn eligens, Eubacterium fissicatena, Eitbacterium limosum, Eubacterium rectale, Eubacterium siraeum, Eubacterium ventriosum, Eubacteria xylanophilum 1, Eubacteria xylanophilum 2, Eubacterium sp, Faecalibacterium prausnitzii Gemmiger/Subdoligramulum formicilis/variabile 1, Gemmiger/Subdoligramulum formicilis/variabile 2, Holdemania filiformis, Micro bacterium schleiferi, Micrococcus luteus, Odoribacter splanchnicus. Oscillibactei-valericigenes, Oscillibacter sp., Parabacteroides gordonii, Parabacteroides merdae. Parasutterella excrementihontints, Phascolardobacterium sp., Roseburiafaecalis. 1, Roseburiafaecalis 2, Roseburis hominis. Roseburia intestinalis, Roseburia sp., Ruminococcus albus, Ruminococcus bromii 1, Ruminococcus bromii 2, Ruminococcus lactaris, Ruminococcus luti, Ruminorocells obeum, Ruminococcus torques 1, Ruminococcus torques 2, Ruminococcus torques 3, Ruminococcus torques 4, Ruminococcus torques 5, Ruminococcus sp. 1, Ruminococcus sp. 2, Ruminococcus sp 3, Staphylococcus epidermidis, Streptococcus mitis, Streptococcus thermophiles, Synergistes sp., and Turicibacter sanguinis;
• or
• (4) a community of bacterial strains representing an enterotype of human gut, wherein the enterotype is the Bacteroides, the Prevotella or the Ruminococcus enterotype.