Live cell imaging of Salmonella Typhimurium interaction with zebrafish larvae after injection and immersion delivery methods

Laboratorio de Microbiología de Sistemas, Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile
Department of Biology, Universidad de Chile, Santiago, Chile
Laboratorio de Biología Estructural y Molecular, Universidad de Chile, Santiago, Chile
Centro FONDAP de Regulación del Genoma, Facultad de Ciencias, Universidad de Chile, Santiago, Chile
Laboratorio de Microbiología, Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
DOI
10.7287/peerj.preprints.2319v1
Subject Areas
Aquaculture, Fisheries and Fish Science, Microbiology, Infectious Diseases
Keywords
Danio rerio, zebrafish, infection, live-cell imaging, immune response., Host-pathogen interaction, Salmonella
Copyright
© 2016 Varas et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Varas MA, Fariña A, Díaz-Pascual F, Ortíz-Severín J, Marcoleta AE, Allende ML, Santiviago CA, Chávez FP. 2016. Live cell imaging of Salmonella Typhimurium interaction with zebrafish larvae after injection and immersion delivery methods. PeerJ Preprints 4:e2319v1

Abstract

Surrogate host models have been employed to study bacterial virulence mechanisms of important human pathogens. Particularly, zebrafish (Danio rerio) has been used to determine the role of vertebrate innate immunity during bacterial infections. The easy-to-obtain large number of embryos and optical transparency of larvae allow live cell imaging of the infection progress and the major cellular types of the innate immune system that develop during the first days of embryogenesis. In zebrafish model, microinjecting bacteria into embryos and/or larvae can cause infection. Alternatively, an infection can be generated by static immersion of larvae on a microbial suspension. Both methods differ in the mode and time of infection, inoculum size and host response. In this work, we compare the in vivo immune response induced by Salmonella enterica serovar Typhimurium (S. Typhimurium) inoculated by immersion and microinjection in zebrafish larvae. To this end, an immersion protocol using transgenic zebrafish larvae was developed for in vivo monitoring of GFP-tagged S. Typhimurium infection progress and immune response during 72 h. The infection progress was compared to that of zebrafish larvae inoculated by microinjection. Our results in zebrafish corroborate previous Salmonella virulence studies in murine models and reveal that host-pathogen interaction not only depends on the virulence of the strain, but also on the inoculation method and host conditions.

Author Comment

This is a submission to PeerJ for review.

Supplemental Information

Raw data for zebrafish mortality by injection method

Raw data for zebrafish mortality caused by microinjection of S. Typhimurium in 3dpf transgenic line.

DOI: 10.7287/peerj.preprints.2319v1/supp-1

Raw data for Salmonella colonization in zebrafish larvae

GFP-tagged S. Typhimurium colonization in 9 dpf zebrafish transgenic line

DOI: 10.7287/peerj.preprints.2319v1/supp-2