Modeling Aeromonas Pathogenesis in C. elegans Using a Survival Assay

Samenvatting

Source: Chen, Y., et al. Evaluating Virulence and Pathogenesis of Aeromonas Infection in a Caenorhabditis elegans Model. J. Vis. Exp. (2018)

This video demonstrates a survival assay using C. elegans to model Aeromonas pathogenesis, where oral ingestion of the bacteria leads to intestinal infection and mortality, enabling quantitative assessment of bacterial virulence in a live host system.

Protocol

1. C. elegans Survival Assay with Aeromonas

1. On the day of seeding L1 worms on the ENGM (enriched nematode growth medium) spreading with E. coli OP50, pick a single colony of each of the four Aeromonas strains or E. coli OP50 and culture with 2 mL LB broth, respectively, at 37 °C for 16 h.
   NOTE: To avoid contamination, this step should be performed in a laminar flow hood. Here, the following bacterial strains were used: E. coli OP50, the normal food source of C. elegans. A. dhakensis AAK1, the first fully sequenced pathogenic clinical A. dhakensis isolate. A. hydrophila A2-066, A. veroni....

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Shaker incubator	YIH DER	LM-570R	Bacteria incubation
NaCl	SIGMA	31434	Culture medium preparation
MgSO₄	SIGMA	M7506	Culture medium preparation
Agar	Difco	214530	Culture medium preparation
CaCl₂	SIGMA	C1016	Culture medium preparation
Cholesterol	SIGMA	C8503	Culture medium preparation
6 cm Petri plate	ALPHA PLUS	46	Agar plate preparation
Bacterial peptone	Affymetrix/USB	AAJ20048P2	Culture medium preparation
FudR	SIGMA	1271008	Culture medium preparation
Microscope system	Nikon	Eclipase Ti inverted	Microscope imaging