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Quantitative Detection of Active Vibrios Associated with White Plague Disease in Mussismilia braziliensis Corals
Luciane A Chimetto Tonon1,2,3, Janelle R Thompson2, Ana P B Moreira3
1Laboratory of Organic Chemistry of Biological Systems, Chemical Institute of São Carlos, University of São Paulo, São Carlos, Brazil.
Abstract:
Over recent decades several coral diseases have been reported as a significant threat to coral reef ecosystems causing the decline of corals cover and diversity around the world. The development of techniques that improve the ability to detect and quantify microbial agents involved in coral disease will aid in the elucidation of disease cause, facilitating coral disease detection and diagnosis, identification and pathogen monitoring, pathogen sources, vectors, and reservoirs. The genus Vibrio is known to harbor pathogenic strains to marine organisms. One of the best-characterized coral pathogens is Vibrio coralliilyticus, an aetilogic agent of White Plague Disease (WPD). We used Mussismilia coral tissue (healthy and diseased specimens) to develop a rapid reproducible detection system for vibrios based on RT-QPCR and SYBR chemistry. We were able to detect total vibrios in expressed RNA targeting the 16S rRNA gene at 5.23 × 106 copies/μg RNA and V. coralliilyticus targeting the pyrH gene at 5.10 × 103 copies/μg RNA in coral tissue. Detection of V. coralliilyticus in diseased and in healthy samples suggests that WPD in the Abrolhos Bank may be caused by a consortium of microorganism and not only a single pathogen. We developed a more practical and economic system compared with probe uses for the real-time detection and quantification of vibrios from coral tissues by using the 16S rRNA and pyrH gene. This qPCR assay is a reliable tool for the monitoring of coral pathogens, and can be useful to prevent, control, or reduce impacts in this ecosystem.
Insights
A new RT-qPCR method efficiently detects and quantifies Vibrio bacteria, including the coral pathogen Vibrio coralliilyticus, in coral tissues. This tool aids in understanding coral diseases like White Plague Disease (WPD) and monitoring reef health.
Area of Science:
- Marine Biology
- Microbial Pathogenesis
- Molecular Diagnostics
Background:
- Coral diseases pose a significant threat to global coral reef ecosystems, leading to reduced coral cover and biodiversity.
- Accurate detection and quantification of microbial agents are crucial for understanding coral disease etiology and management.
- The genus Vibrio includes known marine pathogens, with Vibrio coralliilyticus identified as a causative agent of White Plague Disease (WPD).
Purpose of the Study:
- To develop a rapid, reproducible, and cost-effective detection system for vibrios in coral tissues using RT-qPCR.
- To quantify total Vibrio and specifically Vibrio coralliilyticus in healthy and diseased Mussismilia coral specimens.
- To provide a reliable tool for monitoring coral pathogens and aiding in the prevention and control of coral diseases.
Main Methods:
- Development of a SYBR-based RT-qPCR assay targeting the 16S rRNA gene for total Vibrio detection.
- Design of a specific RT-qPCR assay targeting the pyrH gene for Vibrio coralliilyticus quantification.
- Application of the developed assays to RNA extracted from healthy and diseased Mussismilia coral tissues.
Main Results:
- The assay successfully detected total vibrios at 5.23 × 10^6 copies/μg RNA and Vibrio coralliilyticus at 5.10 × 10^3 copies/μg RNA in coral tissue.
- Vibrio coralliilyticus was detected in both diseased and healthy coral samples, suggesting a potential role of microbial consortia in WPD.
- The developed RT-qPCR system offers a practical and economical alternative to probe-based methods for real-time pathogen quantification.
Conclusions:
- The established RT-qPCR assay is a reliable tool for the detection and quantification of Vibrio species in coral tissues.
- Findings suggest that White Plague Disease may involve a complex interplay of microorganisms rather than a single pathogen.
- This diagnostic tool can significantly contribute to the monitoring, prevention, and management of coral diseases in vulnerable reef ecosystems.
