There is a continuous need for the development of simple animal models for the study of host-pathogen interactions. There is also a need for the identification and study of virulence mechanisms as well. Diverse bacteria are pathogens of C.elegans and may constitute a useful model for the study of both pathogens and symbionts.
Using WMicrotracker, it is possible to easily study bacterial infection in C.elegans (and other small organisms) in order to discover novel antibiotics, natural killers, or genes that improve immune resistance. Below we present an example of application.
Kinetics of killing of Pseudomonas fluorescens CHA0 in C.elegans
In this experiment we can observe a long-term kinetic and dose response effect of Pseudomonas CHA0 infection in C.elegans. Paralytic killing is reported to depend on bacterial hydrogen cyanide production.
(1) Grow synchronized populations of worms in NGM plates. (2)Remove worms from plates using buffer. (3)Count #worms in 10ul, and adjust volume to get [5 worms/10ul]. (4)Transfer 90ul of worm solution to 96 well microplates using multichannel pipette. (1′)Prepare a 96 deep well plate containing the bacterial library to test and growth the bacteria overnight at 37? C. (2′)Pipette 10 ul of bacteria to worm microplate. Reach a final OD600<1.0 in order to prevent reading interference and oxygen depletion. (optional :)It is possible to include chemicals to test at this point. (6)Register the plate with WMicrotracker.