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Open Access Research article

A zebrafish high throughput screening system used for Staphylococcus epidermidis infection marker discovery

Wouter J Veneman1*, Oliver W Stockhammer2, Leonie de Boer2, Sebastian A J Zaat2, Annemarie H Meijer1 and Herman P Spaink1*

Author Affiliations

1 Institute of Biology, Leiden University, PO Box 9502, Leiden RA, 2300, the Netherlands

2 Department of Medical Microbiology, Center of Infection and Immunity Amsterdam, (CINIMA), Academic Medical Center, PO Box 22660, Amsterdam, 1100 DD, the Netherlands

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BMC Genomics 2013, 14:255  doi:10.1186/1471-2164-14-255

Published: 15 April 2013

Abstract

Background

Staphylococcus epidermidis bacteria are a major cause of biomaterial-associated infections in modern medicine. Yet there is little known about the host responses against this normally innocent bacterium in the context of infection of biomaterials. In order to better understand the factors involved in this process, a whole animal model with high throughput screening possibilities and markers for studying the host response to S. epidermidis infection are required.

Results

We have used a zebrafish yolk injection system to study bacterial proliferation and the host response in a time course experiment of S. epidermidis infection. By combining an automated microinjection system with complex object parametric analysis and sorting (COPAS) technology we have quantified bacterial proliferation. This system was used together with transcriptome analysis at several time points during the infection period. We show that bacterial colony forming unit (CFU) counting can be replaced by high throughput flow-based fluorescence analysis of embryos enabling high throughput readout. Comparison of the host transcriptome response to S. epidermidis and Mycobacterium marinum infection in the same system showed that M. marinum has a far stronger effect on host gene regulation than S. epidermidis. However, multiple genes responded differently to S. epidermidis infection than to M. marinum, including a cell adhesion gene linked to specific infection by staphylococci in mammals.

Conclusions

Our zebrafish embryo infection model allowed (i) quantitative assessment of bacterial proliferation, (ii) identification of zebrafish genes serving as markers for infection with the opportunistic pathogen S. epidermidis, and (iii) comparison of the transcriptome response of infection with S. epidermidis and with the pathogen M. marinum. As a result we have identified markers that can be used to distinguish common and specific responses to S. epidermidis. These markers enable the future integration of our high throughput screening technology with functional analyses of immune response genes and immune modulating factors.

Keywords:
Zebrafish; Staphylococcus epidermidis; High throughput screening; Transcriptome analysis; RNA deep sequencing; COPAS; Host pathogen interaction