1. M17 Broth: 0.05% ascorbic acid, 0.5% lactose, 0.025% magnesium sulfate, 0.5% meat extract, 0.25% meat peptone (peptic),
1.9% sodium glycerophosphate, 0.5% soya peptone (papainic),
0.25% tryptone, 0.25% yeast extract.
2. M17 agar: 0.05% ascorbic acid, 0.5% lactose, 0.025%magnesium sulfate, 0.5% meat extract, 0.25% meat peptone (peptic),
1.9% sodium glycerophosphate, 0.5% soya peptone (papainic),
0.25% tryptone, 0.25% yeast extract, 1.5% agar.
3. Glucose solution 50%.
4. PBS 0.01 M (1Â): 137 mM NaCl, 2.7 mM KCl, 10 mM
Na 2 HPO 4 , 2 mM KH 2 PO 4.
5. Bacterial samples (L. lactis).
6. Xylazine hydrochloride.
7. Ketamine.
8. Forceps.
9. Micropipette and tips.
10. 0.22 μm syringe filter.
11. Petri dishes.
12. Syringe.
13. Gavage needle (20–22 G).
14. Autoclave.
15. Spectrophotometer.
16. pH meter.
17. 30
C incubator.
18. Refrigerated centrifuge.
19. Laminar flow cabinet.
20. Magnetic bar.
21. Magnetic stirrer.
3 Methods
3.1 M17 Broth
Preparation (1 L)
1. Suspend 42.25 g of the mixture in 900 mL distilled water (the
quantity can vary according to the manufacturer).
2. Add water q.s. (Quantum satis, “quantity required”).
3. Heat, if necessary, to dissolve the medium completely.
4. Sterilize by autoclaving.
5. Add glucose to 0.5% (i.e., for 1 L of broth add 10 mL of
glucose 50% (stock solution)) (see Note 2).
6. Store in a tightened container at 2–8
C (see Note 3).
Lactic Acid Bacteria for Therapeutic Applications
449
1.9% sodium glycerophosphate, 0.5% soya peptone (papainic),
0.25% tryptone, 0.25% yeast extract.
2. M17 agar: 0.05% ascorbic acid, 0.5% lactose, 0.025%magnesium sulfate, 0.5% meat extract, 0.25% meat peptone (peptic),
1.9% sodium glycerophosphate, 0.5% soya peptone (papainic),
0.25% tryptone, 0.25% yeast extract, 1.5% agar.
3. Glucose solution 50%.
4. PBS 0.01 M (1Â): 137 mM NaCl, 2.7 mM KCl, 10 mM
Na 2 HPO 4 , 2 mM KH 2 PO 4.
5. Bacterial samples (L. lactis).
6. Xylazine hydrochloride.
7. Ketamine.
8. Forceps.
9. Micropipette and tips.
10. 0.22 μm syringe filter.
11. Petri dishes.
12. Syringe.
13. Gavage needle (20–22 G).
14. Autoclave.
15. Spectrophotometer.
16. pH meter.
17. 30
C incubator.
18. Refrigerated centrifuge.
19. Laminar flow cabinet.
20. Magnetic bar.
21. Magnetic stirrer.
3 Methods
3.1 M17 Broth
Preparation (1 L)
1. Suspend 42.25 g of the mixture in 900 mL distilled water (the
quantity can vary according to the manufacturer).
2. Add water q.s. (Quantum satis, “quantity required”).
3. Heat, if necessary, to dissolve the medium completely.
4. Sterilize by autoclaving.
5. Add glucose to 0.5% (i.e., for 1 L of broth add 10 mL of
glucose 50% (stock solution)) (see Note 2).
6. Store in a tightened container at 2–8
C (see Note 3).
Lactic Acid Bacteria for Therapeutic Applications
449