63using a 0.2 μm filter. Store the medium between 2 °C and 8 °C,
in the dark and use within 2 months of the production date. The
composition of CMYCO solution 1 per liter is CaCl 2 ·2H 2 O
(0.055 g), MgSO 4 ·7H 2 O (21.40 g), ZnSO 4 ·7H 2 O (2.88 g), and
distilled water (1.0 L). To prepare CMYCO solution 1, add the
ingredients to the water and stir to dissolve. Store it at 2–8 °C. Use
it within a 6 month period from the date of production.
The composition of CMYCO solution 2 per liter is
CoCl 2 ·6H 2 O (0.048 g), CuSO 4 ·5H 2 O (0.0025 g),
MnCl·4H 2 O (0.002 g), concentrated HCL (0.5 mL), distilled
water (1.0 L). To prepare CMYCO solution 2, add the ingre-
dients to the water and stir to dissolve. Store it at 2–8 °C. Use
it within a 6 month period from the date of production. The
composition of CMYCO solution 3 per liter is l-serine (1.0 g),
l-alanine (1.0 g), l-arginine (1.0 g) l-asparagine (20.0 g),
l-aspartic acid (1.0 g), l-glycine (1.0 g), l-glutamic acid
(1.0 g), l-isoleucine (1.0 g), l-leucine (1.0 g), distilled water
(1.0 L). To prepare CMYCO solution 3, add the ingredients
to the water and stir to dissolve. Store it at 2–8 °C. Use it
within a week of production. The composition of CMYCO
solution 4 per liter is pyruvic acid sodium salt (100.0 g) and
distilled water (1.0 L). Prepare fresh CMYCO solution 4 every
time CMM is prepared. Add the pyruvic acid to the water and
stir to dissolve the ingredients. The composition of CMYCO
solution 5 per liter is FeSO 4 ·7H 2 O (1.0 g), concentrated HCL
(0.5 mL), and distilled water (1.0 L). Prepare fresh CMYCO
solution 5 every time CMM is made. Add the ingredients to
the water and stir to dissolve the ingredients.- Mycobacterium tuberculosis strain H37Rv.
- Chemostat (set up as in Subheading 3.1).
- CMM Mod2 medium (see Subheading 2.1 for the recipe).
- Chemostat (set up as in Subheading 3.1).
- Spare 1 L Glass Vessel: (DASGIP) (Eppendorf, Stevenage,
UK). - CMM medium (see Subheading 2.1 for the recipe).
- Antibiotics.
- CMM Mod2 medium (See Subheading 2.1 for the recipe).
- Minisart syringe filters (0.2 μM pore; Sigma-Aldrich, Poole,
UK). - Sterile water.
- Mityvac handheld vacuum pump (mityvac.com).
2.2 Establishing
Steady-State Growth
2.3 Alteration
of Growth Rates
in Chemostat Culture
2.4 Addition
of Antibiotics
to the Culture System
Application of Continuous Culture for Assessing Antibiotic Activity Against...