Assessment of Salmonella Typhimurium load within infected nematodes
Instrument: Precellys® 24 Dual
Sample: C.elegans with S. Typhimurium strain L1019.
Protocol:
- Precellys kits: 03961-1-004 (0.5 mm glass beads)
- Sample: C.elegans (Six animals at L4 stage) infected with S. Typhimurium strain L1019.
- Buffer: 200 μl M9 with 25 mM levamisole hydrochloride.
- 10 replicates / 5 time points (5 days).
- Run the Precellys at 6500rpm, 1x10s.
- For Analysis Lysates were serially diluted in M9 and plated onto LB plates containing kanamycin (30 μg/ml) to select for L1019. Colonies were counted by eye and scaled to the original concentration per nematode.
Results:
Infection load was assessed through viable counts for 5 days / each 24 hours. lys-7 and abl-1 mutant animals are tolerant of S. Typhimurium infection. There was no difference between any of the strains (p.0.2) showing that lys-7 and abl-1 single mutants are tolerant to S. Typhimurium infection for at least six days.
Lysing nematodes infected with Salmonella using the Precellys does not affect the bacteria viability, allowing infection load to be accurately assessed.
