NUTRIENT MEDIA
For reactivation of lyophilized cultures 0.3 - 0.5 ml of sterile 0.5%
NaCl or an appropriate nutrient medium is added into an ampule opened
under sterile conditions. Bacterial cell suspension is kept for 30 min
at room temperature. For primary inoculations it is recommended to use
a medium indicated in the published Catalogue for a certain strain.
Best for utilization are the cultures after second and third
inoculations.
1. BEEF-EXTRACT BROTH
Peptone 10.0 g
NaCl 5.0 g
Beef water 1000.0 ml
pH = 7.2 - 7.4. Sterilized at 121 C for 30 min.
Preparation of meat water: 500.0 g of meat without bones, fat and
tendons is minced, 1000.0 ml of tap water is added, and this is kept at
room temperature for 12 h or in a thermostate at 37 C for 2 h. Further
meat is wrung out using cheese-cloth or linen, and the filtrate is
boiled for 30 min. The cooled substance is filtrated through a cotton
wool filter, and water is added to obtain the initial volume.
2. BEEF-EXTRACT AGAR
Peptone 10.0 g
NaCl 5.0 g
Agar 20.0 g
Beef water
(see medium 1) 1000.0 ml
pH = 7.2 - 7.4. Sterilized at 121 C for 30 min.
3. WORT-AGAR 6 B
Malt extract 6 B 1000.0 ml
Agar 20.0 g
Sterilized at 105 C for 30 min.
250.0 g of ground malt are mixed with 1000.0 ml of tap water, heated up
to 55 - 60 C and kept at this temperature for 1.5 - 2.0 h while mixing
regularly. Then the temperature is elevated up to 80 C, and the mixture
is kept for more 10 min. The wort is cooled, wrung out through a linen
bag, the concentration of sugars 6 B is adjusted, and agar is added.
Sterilization at 105 C for 30 min.
4. GORBENKO MEDIUM
Dry nutrient agar 5.0 g
Agar 13.5 g
Lake water 1000.0 ml
Reference: Gorbenko Yu.A. About the most favourable quantity of dry
nutrient agar in a medium for cultivation of sea heterotrophic
bacteria. - Mikrobiologiya. Vol. 30. N. 1. P. 168-172.
5. MEDIUM KIEVSKAYA (MEDIUM K)
KNO3 1.0 g
KH2PO4 1.0 g
K2HPO4 1.0 g
NaCl 1.0 g
MgSO4 0.2 g
CaCl2 0.02 g
FeCl3 0.001 g
pH = 6.8 - 7.0. Sterilize liquid media at 121 C for 30 min, agarized -
at 105 C for 20 min. Gas mixture composition: propane(n-butane) - air
1:5.
Reference: Malashenko Yu.A., Romanovskaya V.A., Bogachenko V.N.,
Khotian L.V., Voloshin N.V. ªËaracteristics nutrition of microorganisms
growing on natural gas. - Mikrobiologiya. 1973. Vol. 42. N. 3. P.
403-408.
6. RAYMOND'S MEDIUM
NH4NO3 2.0 g
KH2PO4 2.0 g
Na2HPO4 3.0 g
NaCl 1.0-5.0 g
MgSO4 0.2 g
Na2CO3 0.1 g
ªÄCl2 0.01 g
MnSO4 0.02 g
FeSO4 0.01 g
pH = 6.8 - 7.0. n-Hexadecane is used as the only carbon source at a
concentration of 1.0 vol.%. Sterilized at 121 ©áfor 20 min.
Reference: Raymond R.L. Microbial oxidation of n-paraffinic
hydrocarbons. - Develop. Indust. Microbiol. 1961. Vol. 2. P. 23-32.
7. FAYBITCH'S SUCROSE-GELATIN-AGAR MEDIUM
Sucrose 100.0 g
Gelatine 15.0 g
Agar 10.0 g
Water 1000.0 ml
Reference: Faybich M.M. Liophilic dry preparations of living vaccines
and collection cultures. - Bulletin of practice exchange of the
institutes of epidemiology and microbiology, health protection
ministry. 1947. N. 4/22. P. 1-17.
8. MEDIUM KIEVSKAYA
KNO3 1.0 g
KH2PO4 1.0 g
K2HPO4 1.0 g
NaCl 1.0 g
MgSO4 0.2 g
CaCl2 0.02 g
FeCl3 0.001 g
pH = 6.8 - 7.0. n-Hexadecane is used as the sole carbon source at a
concentration of 1.0 vol.%.
Reference: Malashenko Yu.A., Romanovskaya V.A., Bogachenko V.N.,
Khotian L.V., Voloshin N.V. ªËaracteristics nutrition of microorganisms
growing on natural gas. - Mikrobiologiya. 1973. Vol. 42. N. 3. P.
403-408.