Polyvalent culture medium for anaerobic bacteria under aerobic conditions

Patent metadata

Field	Value
Publication number	US-11091735-B2
Application number	US-201816636014-A
Country	US
Kind code	B2
Filing date	Aug 1, 2018
Priority date	Aug 8, 2017
Publication date	Aug 17, 2021
Grant date	Aug 17, 2021

Abstract

The present invention relates to a polyvalent culture medium for anaerobic bacteria under aerobic conditions in blood culture bottles.The present invention provides a polyvalent culture medium for culture in aerobic atmosphere of anaerobic bacteria or aerobic bacteria comprising a basal culture medium for bacteria characterized in that it further comprises a mixture of the following antioxidant compounds: sodium hydrosulphide (Na2S), L-cysteine, ascorbic acid, glutathione, catalase, ubiquinol and lipoic acid.

First claim

The invention claimed is:

1. A polyvalent liquid culture medium for culture in aerobic atmosphere of anaerobic bacteria or aerobic bacteria comprising:
a basal culture medium for aerobic and anaerobic bacteria;
wherein the basal culture medium comprises a mixture of the following antioxidant compounds: sodium hydrosulphide (Na 2 S), L-cysteine, ascorbic acid, glutathione, catalase, ubiquinol and lipoic acid.

2. The culture medium according to claim 1 , wherein said antioxidant compounds comprise the following quantities and weight proportions per 1 L:
sodium hydrosulphide: at least 0.25 g (0.025%);
L-cysteine: at least 0.25 g (0.025%);
ascorbic acid: at least 0.5 g (0.05%);
glutathione: at least 0.1 g (0.01%);
catalase: at least 0.06 g (0.006%);
ubiquinol: at least 0.06 g (0.006%); and
lipoic acid: at least 0.01 g (0.001%).

3. The culture medium according to claim 1 , further comprising the following nutrient components in said basal culture medium:
a source of carbon and nitrogen selected from the group consisting of a yeast extract, an acetate salt, and a tryptic peptone,
a source of phosphorus,
at least one sugar,
at least one salt of metals selected from K, Mg, Na, and Ca,
at least one pH-regulating buffer substance for adjusting the pH from 7 to 8, or NaHCO 3 for adjusting the pH to 7.5, and
at least one vitamin or growth factor.

4. The culture medium according to claim 1 , wherein said basal culture medium is a conventional acellular bacterial medium comprising components selected from an extract of ground material, or lysate of multicellular tissue, an enzyme digestate, soybean and/or animal tissue, a peptone, a yeast extract, a sugar, a NaCl and/or Na 2 PO 4 salt.

5. The culture medium according to claim 1 , wherein said basal culture medium is a liquid culture medium for culturing anaerobic bacteria in a blood, stool, sputum, or vaginal secretion sample.

6. The culture medium according to claim 1 , wherein said basal culture medium comprises the following nutrient components:
casein hydrolysate;
proteose peptone;
yeast extract;
glucose;
sodium chloride (NaCl); and
a growth factor: α-ketoglutarate.

7. The culture medium according to claim 6 , comprising the following components in the following quantities and weight proportions per 1 L:
casein hydrolysate: 15 g (1.5%),
proteose peptone: 15 g (1.5%),
yeast extract: 10 g (1%),
α-ketoglutarate: 2 g (0.2%),
sodium chloride (NaCl): 5 g (0.5%),
glucose: 10 g (1%),
dipotassium phosphate (K 2 HPO 4 ): 0.83 g (0.083%),
sodium hydrosulphide (Na 2 S): 0.5 g (0.05%),
L-cysteine: 0.5 g (0.05%),
ascorbic acid: 1 g (0.1%),
glutathione: 0.1 g (0.01%),
catalase: 0.16 g (0.016%),
ubiquinol: 0.16 g (0.016%),
lipoic acid: 0.010 g (0.001%),

wherein the pH of the solution is adjusted to 7.5±0.2 with 10 M KOH.

8. A process for the in vitro culture of an aerobic or anaerobic bacterium under aerobic atmosphere with a culture medium as claimed in claim 1 .

9. The process according to claim 8 , wherein said bacterium is cultured in a blood, stool, sputum, or vaginal secretion sample.

10. The process according to claim 8 , wherein said anaerobic bacterium is selected from the group consisting of Akkermansia muciniphila, Bacteroides fragilis, Bacteroides nordii, Bacteroides thetaiotaomicron, Clostridium beijeirinckii, Clostridium butyricum, Clostridium massilioamazoniensis, Clostridium irregulare, Finegoldia magna, Propionibacterium acnes, Propionibacterium avidum, Prevotella denticola , and Prevotella histicola.

11. The culture medium according to claim 1 , wherein said antioxidant compounds comprise the following quantities and weight proportions per 1 L:
sodium hydrosulphide: from 0.25 g to 0.5 g (0.025 to 0.05%);
L-cysteine: from 0.25 g to 0.5 g (0.025 to 0.05%);
ascorbic acid: from 0.5 to 1 g (0.05 to 0.1%);
glutathione: from 0.1 to 0.5 g (0.01 to 0.05%);
catalase: from 0.06 to 0.16 g (0.006 to 0.016%);
ubiquinol: from 0.06 to 0.16 g (0.006 to 0.016%); and
lipoic acid: from 0.01 to 0.015 g (0.001 to 0.0015%).

12. The culture medium according to claim 3 , wherein the at least one salt is NaCl.

13. The culture medium according to claim 3 , wherein the at least one pH-regulating buffer substance for adjusting the pH from 7 to 8 is K 2 HPO 4 .

14. The culture medium according to claim 4 , wherein the enzyme digestate is an enzyme digestate of casein.

15. The culture medium according to claim 4 , wherein the sugar is dextrose or glucose.

Assignees

• Fond Mediterranee Infection
• Univ Aix Marseille
• Assist Publique—Hopitaux De Marseille
• Centre Nat Rech Scient
• Inst Nat Sante Rech Med
• Fond Mediterranee Infection Faculte De Medecine

Inventors

• Raoult Didier
• Khelaifia Saber
• Bonnet Marion

Classifications

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title