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Evaluation of the pathogenesis and treatment of Mycobacterium marinum infection in zebrafish
Journal article   Peer reviewed

Evaluation of the pathogenesis and treatment of Mycobacterium marinum infection in zebrafish

Kevin Takaki, J Muse Davis, Kathryn Winglee and Lalita Ramakrishnan
Nature protocols, Vol.8(6), pp.1114-1124
06/2013
DOI: 10.1038/nprot.2013.068
PMCID: PMC3919459
PMID: 23680983

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Abstract

Mycobacterium marinum-infected zebrafish are used to study tuberculosis pathogenesis, as well as for antitubercular drug discovery. The small size of zebrafish larvae coupled with their optical transparency allows for rapid analysis of bacterial burdens and host survival in response to genetic and pharmacological manipulations of both mycobacteria and host. Automated fluorescence microscopy and automated plate fluorimetry (APF) are coupled with facile husbandry to facilitate large-scale, repeated analysis of individual infected fish. Both methods allow for in vivo screening of chemical libraries, requiring only 0.1 μmol of drug per fish to assess efficacy; they also permit a more detailed evaluation of the individual stages of tuberculosis pathogenesis. Here we describe a 16-h protocol spanning 22 d, in which zebrafish larvae are infected via the two primary injection sites, the hindbrain ventricle and caudal vein; this is followed by the high-throughput evaluation of pathogenesis and antimicrobial efficacy.
 Animal Husbandry - methods Antitubercular Agents - pharmacology Phagocytosis - physiology Zebrafish Fluorometry Mycobacterium Infections, Nontuberculous - physiopathology Animals Larva - microbiology Mycobacterium Infections, Nontuberculous - immunology Macrophages - immunology Microscopy, Fluorescence Mycobacterium Infections, Nontuberculous - drug therapy Antitubercular Agents - therapeutic use Disease Models, Animal

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