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Updated: Dec 20, 2025
Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions
Published on: May 16, 2020
Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions
Savini Thrikawala1, Emily E Rosowski2
1Department of Biological Sciences, Clemson University.
Abstract:
Invasive aspergillosis (IA) is one of the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal drugs, IA can cause >50% mortality in infected immunocompromised patients. It is crucial to determine both host and pathogen factors that contribute to infection susceptibility and low survival rates in infected patients in order to develop novel therapeutics. Innate immune responses play a pivotal role in recognition and clearance of Aspergillus spores, though little is known about the exact cellular and molecular mechanisms. Reliable models are required to investigate detailed mechanistic interactions between the host and pathogen. The optical clarity and genetic tractability of zebrafish larvae make them an intriguing model to study host-pathogen interactions of multiple human bacterial and fungal infections in a live and intact host. This protocol describes a larval zebrafish Aspergillus infection model. First, Aspergillus spores are isolated and injected into the zebrafish hindbrain ventricle via microinjection. Then, chemical inhibitors such as immunosuppressive drugs are added directly to the larval water. Two methods to monitor the infection in injected larvae are described, including the 1) homogenization of larvae for colony forming unit (CFU) enumeration and 2) a repeated, daily live imaging setup. Overall, these techniques can be used to mechanistically analyze the progression of Aspergillus infection in vivo and can be applied to different host backgrounds and Aspergillus strains to interrogate host-pathogen interactions.
Insights
This study introduces a zebrafish model to study invasive aspergillosis (IA), a deadly fungal infection. This model helps investigate host-pathogen interactions to develop new treatments for immunocompromised patients.
Area of Science:
- Mycology
- Immunology
- Zebrafish models
Background:
- Invasive aspergillosis (IA) is a severe fungal infection in immunocompromised individuals, with high mortality rates despite antifungal treatments.
- Understanding host and pathogen factors is critical for developing novel therapeutics against IA.
- Current knowledge of innate immune mechanisms in IA is limited, necessitating better experimental models.
Purpose of the Study:
- To establish and validate a larval zebrafish model for studying invasive aspergillosis (IA).
- To investigate host-pathogen interactions during Aspergillus infection in a live, intact organism.
- To provide a platform for mechanistic analysis and drug screening for IA.
Main Methods:
- Development of a protocol for Aspergillus spore isolation and microinjection into zebrafish larvae hindbrain ventricle.
- Application of chemical inhibitors (e.g., immunosuppressive drugs) in larval water to modulate host response.
- Monitoring infection progression using two methods: homogenization for colony-forming unit (CFU) enumeration and daily live imaging.
Main Results:
- Successfully established a reproducible larval zebrafish model for invasive aspergillosis.
- Demonstrated the utility of both CFU enumeration and live imaging for assessing infection.
- The model allows for in vivo analysis of Aspergillus infection dynamics and host responses.
Conclusions:
- The larval zebrafish model provides a powerful tool for studying the mechanisms of invasive aspergillosis.
- This model can be adapted for various host backgrounds and Aspergillus strains to explore host-pathogen interactions.
- The established techniques facilitate the development of new therapeutic strategies against IA.
