Effects of sequential infections of Caenorhabditis elegans with Staphylococcus aureus and Proteus mirabilis


K. Balamurugan, Department of Biotechnology, Alagappa University, Karaikudi-630 003, India. Tel: 91 4565 225215; fax: 91 4565 225202; e-mail: bsuryar@yahoo.com


Caenorhabditis elegans can be used to study the dynamics of polymicrobial infections, specifically those between Gram-positive and Gram-negative bacteria. With C. elegans, Proteus mirabilis acts as an opportunistic pathogen and does not kill this host. Hence, in the present study, C. elegans was immunochallenged by pre-infecting it with the pathogen Staphylococcus aureus in order to study the subsequent effect of P. mirabilis on the host. It was found that 12 hrs of S. aureus and 80 hrs of subsequent P. mirabilis infection significantly reduced the life span of exposed C. elegans by 80%. However, preinfection with S. aureus for 8 and 4 hrs reduced the life span of C. elegans by only 60 and 30%, respectively. Further, there was greater production of reactive oxygen species in the sequentially infected samples than in the S. aureus and P. mirabilis controls. Real time PCR analysis indicated regulation of candidate immune regulatory genes, lysozyme (lys-7), CUB-like proteins (F08G5.6), neuropeptide-like factors (nlp-29), transcription factors of mitogen-activated protein kinase (ATF-7) and daf-2–daf-16 (daf-16), insulin-like signaling pathways and C-type lectin (clec-60 and clec-87) family members during S. aureus and subsequent P. mirabilis-mediated infections, indicating possible roles of, and contributions by, the above factors during host immune responses against these sequential infections. The present findings demonstrate that S. aureus infections increase the vulnerability of the C. elegans host by subverting its immune system, which then permits the opportunistic pathogen P. mirabilis to be pathogenic to this host