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Evaluation of Burkholderia cepacia Complex Bacteria Pathogenicity Using Caenorhabditis elegans   

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Abstract

This protocol describes two biological assays to evaluate pathogenicity of Burkholderia cepacia complex (Bcc) strains against the nematode Caenorhabditis elegans. Specifically, these two assays allow one to identify if the under-investigated Bcc strains are able to kill the nematodes by intestinal colonization (slow killing assay, SKA) or by toxins production (fast killing assay, FKA). The principal differences between the two assays rely on the different killing kinetics for worms.

Keywords: Burkholderia cepacia complex strains, Caenorhabditis elegans, Animal model

Background

The Burkholderia cepacia complex (Bcc) occupies a critical position among Gram-negative multi-drug resistant bacteria. It consists of at least 20 closely related species. Many Bcc strains are multi drug and pandrug-resistant opportunistic human pathogens caused problematic lung infections in immune-compromised individuals, including cystic fibrosis (CF) patients. The use of non-vertebrate host model can be useful for dissecting virulence and pathogenicity determinants as well as identifying novel therapeutic targets (Kothe et al., 2003).

There are a good number of assays for detecting Bcc virulence against a large panel of host models, in liquid or in solid surface. However, some of those are mostly focused on phenotypic observations, which are difficult to detect and have a low reproducibility (Cardona et al., 2005). Herein, we developed two assays based on the analysis of surviving worms, which is a more reproducible and allows easy and fast comparison among the Bcc strains tested. In addition, these assays permit the detection of death mechanisms of Bcc towards nematode.

These killing assays allow us to identify bacterial strains that are able to colonize the nematode intestine and produce diffusible toxins capable of killing the host.

Copyright: © 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite:  Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
  1. Tedesco, P., Di Schiavi, E., Palma Esposito, F. and de Pascale, D. (2016). Evaluation of Burkholderia cepacia Complex Bacteria Pathogenicity Using Caenorhabditis elegans. Bio-protocol 6(20): e1964. DOI: 10.21769/BioProtoc.1964.
  2. Tedesco, P., Visone, M., Parrilli, E., Tutino, M. L., Perrin, E., Maida, I., Fani, R., Ballestriero, F., Santos, R., Pinilla, C., Di Schiavi, E., Tegos, G. and de Pascale, D. (2015). Investigating the role of the host multidrug resistance associated protein transporter family in Burkholderia cepacia complex pathogenicity using a Caenorhabditis elegans infection model. PLoS One 10(11): e0142883.
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