PUBLICATION
Detection of Aeromonas salmonicida subsp. salmonicida infection in zebrafish by labelling bacteria with GFP and a fluorescent probe based on the siderophore amonabactin
- Authors
- Rodríguez-Pedrouzo, A., Cisneros-Sureda, J., Martínez-Matamoros, D., Rey-Varela, D., Balado, M., Rodríguez, J., Lemos, M.L., Folgueira, M., Jiménez, C.
- ID
- ZDB-PUB-231020-56
- Date
- 2023
- Source
- Microbial pathogenesis 185: 106394 (Journal)
- Registered Authors
- Folgueira Otero, Monica
- Keywords
- A. salmonicida, Fluorescence microscopy, Fluorescent probes, Green fluorescent protein (GFP), Siderophore amonabactin, Sulforhodamine B conjugate (AMB-SRB), Zebrafish
- MeSH Terms
-
- Aeromonas salmonicida*/genetics
- Animals
- Fish Diseases*/microbiology
- Fluorescent Dyes/metabolism
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Siderophores/metabolism
- Zebrafish
- PubMed
- 37858632 Full text @ Microb. Pathog.
Citation
Rodríguez-Pedrouzo, A., Cisneros-Sureda, J., Martínez-Matamoros, D., Rey-Varela, D., Balado, M., Rodríguez, J., Lemos, M.L., Folgueira, M., Jiménez, C. (2023) Detection of Aeromonas salmonicida subsp. salmonicida infection in zebrafish by labelling bacteria with GFP and a fluorescent probe based on the siderophore amonabactin. Microbial pathogenesis. 185:106394.
Abstract
Zebrafish (Danio rerio) is an excellent model to study bacterial infections in fish and their treatment. We used zebrafish as a model of infection for Aeromonas salmonicida subsp. salmonicida (hereinafter A. salmonicida), the causative agent of fish furunculosis. In this work, we studied the infection process of A. salmonicida by immersion of zebrafish larvae in 2 different doses of the bacteria and the fish mortality was monitored for three days. The bacterium caused a high mortality (65 %) in zebrafish larvae only when they were exposed to a high bacterial concentration (107 bacterial cells/mL). To evaluate the use of fluorescence microscopy to follow A. salmonicida infection in vivo, two different fluorescent strains generated by labeling an A. salmonicida strain with either, the green fluorescent protein (GFP), or with a previously reported siderophore amonabactin-sulforhodamine B conjugate (AMB-SRB), were used. The distribution of both labeled bacterial strains in the larvae tissues was evaluated by conventional and confocal fluorescence microscopy. The fluorescent signal showed a greater intensity with the GFP-labeled bacteria, so it could be observed using conventional fluorescence microscopy. Since the AMB-SRB labeled bacteria showed a weaker signal, the larvae were imaged using a laser scanning confocal microscope after 48 h of exposure to the bacteria. Both fluorescent signals were mainly observed in the larvae digestive tract, suggesting that this is the main colonization route of zebrafish for waterborne A. salmonicida. This is the first report of the use of a siderophore-fluorophore conjugate to study a bacterial infection in fish. The use of a siderophore-fluorophore conjugate to study a bacterial infection in fish has the advantage that it is a specific marker and that does no require genetic manipulation of the bacteria.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping