Claims for Patent: 10,314,864

Summary for Patent: 10,314,864

Title:	Media supplements and methods to culture human gastrointestinal anaerobic microorganisms
Abstract:	A media supplement for culturing anaerobic bacteria is provided which comprises a filtrate of eilluent from a chemostat vessel in which a target bacterial ecosystem has been culnn-ed. Methods of using the supplement for culturing or isolating anaerobic microbial strains or communities, particularly anaerobic bacteria from the human gut, are also provided.
Inventor(s):	Allen-Vercoe; Emma (Guelph, CA), McDonald; Julie (Guelph, CA)
Assignee:	Nubiyota LLC (Teaneck, NJ)
Application Number:	15/440,906
Patent Claims:	1. method of isolating anaerobic bacteria from human gut, comprising: (a) culturing a fecal sample comprising a microbial community of the human gut in culture media in a single-stage chemostat under conditions replicating normal human colonic gastrointestinal tract, until equilibrium is reached; (b) diluting the culture and plating onto anaerobe agar supplemented with a media supplement, and optionally supplemented with defibrinated sheep blood, wherein the media supplement comprises 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 media comprising: 0.2% w/v Peptone: 0.2% w/v Yeast extract 0.2% w/v NaHCO.sub.3: 0.2% w/v Pectin; 0.2% w/v Arabinogalactan; 0,3% w/v Casein; 0,5% w/v unmodified wheat starch: 0.1% w/v inulin; 0,05% w/v bile salts; 0.05% w/v L cysteine HCl; 0.0001% w/v CaCl.sub.2; 0.0001% w/v NaCI; 0.0004% w/v K.sub.2HPO.sub.4: 0.0004% w/v KH.sub.2PO.sub.4; 0.0001% w/v MgSO.sub.4; 0.00005% w/v Hemin; and 0.00001% w/v menadione; and wherein the target bacterial ecosystem comprises Akkermansia muciniphila, Bacteroides ovatus, Bifidobacterium longum, Collinsella aerofaciens, Dorea longicatena, Eubacterium eligens, Eubacterium rectale, Eubacterium ventriosum, Faecalibacterium prausnitzii, Flavonifractor plautii, which is formerly Clostridium orbiscindens, Roseburia intestinalis, and Ruminococcus obeum; (c) incubating plates in an anaerobe chamber; (d) purifying individual anaerobic bacterial colonies grown in step (c) to isolate anaerobic bacteria from the human gut; and optionally, culturing the purified individual anaerobic bacterial colonies from step (d) in liquid culture in a single-stage chemostat under conditions replicating normal human colonic gastrointestinal tract, optionally wherein the media supplement is used to supplement culture media at about 1% v/v to about 10% v/v. 2. The method of claim 1, wherein the anaerobe chamber contains an atmosphere of N.sub.2, CO.sub.2 or H.sub.2, or a mixture thereof. 3. The method of claim 1, wherein the target bacterial ecosystem comprises a human fecal sample. 4. The method of claim 3, wherein the human fecal sample which is cultured is a 10% w/v fecal slurry supernatant or a 20% w/v fecal slurry supernatant. 5. The method of claim 1, wherein the culture media of step a) further comprises 0.2% w/v Xylan. 6. The method of claim 1, wherein the anaerobic bacteria obtained is Faecalibacterium prausnitzii or Ruminococcus callidus (ATCC27760). 7. The method of claim 1, wherein obtaining at least one of the anaerobic bacteria of Faecalibacterium prausnitzii, Ruminococcus callidus (ATCC27760), or a Roseburia species serves as a positive indicator of effectiveness of the method.

Details for Patent 10,314,864

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	PANHEMATIN	hemin for injection	For Injection	101246	07/20/1983		2031-09-14