Working with Lactobacillus _"Firm-5"
The
Lactobacillus "Firm 5" clade are gram positive bacterial symbionts of the honey bee (
Apis mellifera) gut core microbiota that can be cultivated
in vitro. Their isolation and characterization were recently carried out by
Waldan Kwong in the laboratory of
Dr. Nancy Moran and are described in depth
here. Genomes for two Firm-5 strains, wkB8 and wkB10 are described
here
In vitro culture conditions
Plate Growth
Firm 5 strains wkB8 and wkB10 are generally cultured at 35°C and 5% CO
2. They grow robustly on agar plates after 48-72 hours on the following media:
- Columbia Agar supplement with 5% Sheep's Blood
- MRS agar
Colonies will appear tiny on CBA agar but will have a slight clearing of blood (alpha hemolysis), along the streaks/path where they are growing. The hemolysis/clearing will increase over time if left in the incubator.
Liquid Media
Firm 5 strains wkB8 and wkB10 can be cultivated in the following liquid media (same general culture conditions as above, without shaking):
Antibiotic Concentrations
This table lists some of the antibiotics tested on
Lactobacillus Firm-5 strains. These concentrations apply to
Lactobacillus (wkB8 and wkB10). Antibiotic selection in other organisms may use different concentrations, so check the primary literature for more info.
Reagent |
Abb |
Stock Conc. |
Working Conc. |
Dilution |
Solvent |
MSDS |
Notes |
Kanamycin |
Kan |
50 mg/mL |
50 µg/mL |
2,500× |
ddH2O |
link |
Lactobacillus Firm-5 wkB8 and wkB10 are both naturally resistant to Kan. Use 50 µg/mL. Plates start to lose sensitivity after 2 weeks; recommended Kanamycin plates only last a few weeks at 4°C; best to make fresh Kan plates every 2 - 4 weeks. |
Erythromycin |
Ery |
250 mg/mL |
250 µg/mL |
1,000× |
EtOH |
link |
May crystallize when added to liquid culture. Still works. Selection strength may vary in non-model bacteria. |
Tetracycline |
Tet |
10 mg/mL |
10 µg/mL |
1,000× |
70% EtOH |
link |
Light sensitive, stability < 2 weeks. |
-- Main.SeanLeonard - 23 Feb 2017
Contributors to this topic

SeanLeonard, ZuberiAshraf
Topic revision: r2 - 2025-03-20 - 22:41:16 - Main.ZuberiAshraf