Development and validation of the first species-specific MALDI-TOF MS main spectra profile database for rapid identification of Aeromonas schubertii associated with white nodular disease in freshwater fish.
7 affiliations
- 1Veterinary Paramedic Study Program, Faculty of Vocational Studies, Hasanuddin University, Makassar 90245, South Sulawesi, Indonesia.
- 2Veterinary Study Program, Faculty of Medicine, Hasanuddin University, Makassar 90245, South Sulawesi, Indonesia.
- 3The International Graduate Course of Veterinary Science and Technology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
- 4Department of Aquatic Animal Medicine, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt.
- 5Department of Veterinary Preventive Medicine, College of Veterinary Medicine, Qassim University, Buraydah 52571, Saudi Arabia.
- 6Center of Excellence in Fish Infectious Diseases Research Unit, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
- 7Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
Abstract
Background and aim
Aeromonas schubertii is an emerging aquatic bacterial pathogen associated with distinctive white nodular disease in freshwater fish and is increasingly relevant to aquaculture health and disease surveillance. Accurate species-level identification remains challenging because commercial matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) databases have limited species-specific spectral representation of A. schubertii. This study aimed to develop and validate the first species-specific main spectra profile (MSP) database for rapid and reliable identification of A. schubertii.Materials and methods
Six representative A. schubertii isolates were used to construct MSPs following ethanol-formic acid-acetonitrile protein extraction. A minimum of 20 high-quality spectra per isolate were retained for MSP generation. The customized database was evaluated using 19 A. schubertii isolates, including the six isolates used for database construction and 13 additional isolates. Identification reliability and repeatability were evaluated, and discriminatory performance was assessed against four closely related non-target Aeromonas species. Principal component analysis (PCA), spectral profiling, and dendrogram analyses were used to evaluate spectral relatedness and species differentiation.Results
Using the commercial database alone, 15/19 (78.95%) isolates achieved reliable species-level identification (log scores of 2.01-2.22), whereas 4/19 (21.05%) were identified only at the genus level (scores of 1.95-1.98). After incorporating the customized MSP database, all 19 isolates (100%) were correctly identified as A. schubertii, with highly reliable log scores ranging from 2.49 to 2.68. Technical replicates showed high spectral reproducibility, with Pearson correlation coefficients of 0.95-0.97. PCA and dendrogram analyses clearly separated A. schubertii from the tested non-target Aeromonas species, supporting the discriminatory performance of the customized database. Molecular analysis of 16S rRNA and gyrB sequences further supported the taxonomic identification of the isolates.Conclusion
The species-specific MSP database substantially improved MALDI-TOF MS identification of A. schubertii, increasing species-level identification from 78.95% to 100%. The database provides a rapid and reliable framework for improving diagnosis and surveillance of this emerging aquaculture pathogen. Further expansion with geographically diverse isolates and multicenter validation is warranted to strengthen database transferability and broader diagnostic application.
Related
- Identification of candidate selenium-associated transcriptomic biomarkers in chronic obstructive pulmonary disease: an integrated bioinformatics and expression validation study
- Identification and validation of T cell proliferation-associated transcriptomic biomarkers in ischemic stroke
- Identification and validation of early vital sign trajectory subphenotypes for risk stratification in sepsis-associated liver injury: a multicenter retrospective cohort study
- A novel multiplex assay for species-level identification of non-tuberculous mycobacteria in culture-positive respiratory specimens based on capillary electrophoresis: development and clinical validation
- Electronic Health Record-Based Machine-Learning for Cardiac Arrhythmia Identification in Adults with High-Risk Chronic Disease: Model Development and Validation Study
BibTeX
@article{mursalim2026development,
title = {{Development and validation of the first species-specific MALDI-TOF MS main spectra profile database for rapid identification of Aeromonas schubertii associated with white nodular disease in freshwater fish.}},
author = {Muhammad Fadhlullah Mursalim and Hendri Budiyansah and Mahmoud Mabrok and Sulaiman F Aljasir and Rian Hari Suharto and Channarong Rodkhum},
journal = {Veterinary world},
year = {2026},
month = sep,
doi = {10.14202/vetworld.2026.4175-4190},
url = {https://europepmc.org/article/MED/42928764},
note = {CC BY},
}
RIS
TY - JOUR
TI - Development and validation of the first species-specific MALDI-TOF MS main spectra profile database for rapid identification of Aeromonas schubertii associated with white nodular disease in freshwater fish.
AU - Muhammad Fadhlullah Mursalim
AU - Hendri Budiyansah
AU - Mahmoud Mabrok
AU - Sulaiman F Aljasir
AU - Rian Hari Suharto
AU - Channarong Rodkhum
PY - 2026
DA - 2026/09/22
JO - Veterinary world
DO - 10.14202/vetworld.2026.4175-4190
UR - https://europepmc.org/article/MED/42928764
AB - Background and aim Aeromonas schubertii is an emerging aquatic bacterial pathogen associated with distinctive white nodular disease in freshwater fish and is increasingly relevant to aquaculture health and disease surveillance. Accurate species-level identification remains challenging because commercial matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) databases have limited species-specific spectral representation of A. schubertii. This study aimed to develop and validate the first species-specific main spectra profile (MSP) database for rapid and reliable identification of A. schubertii.Materials and methods Six representative A. schubertii isolates were used to construct MSPs following ethanol-formic acid-acetonitrile protein extraction. A minimum of 20 high-quality spectra per isolate were retained for MSP generation. The customized database was evaluated using 19 A. schubertii isolates, including the six isolates used for database construction and 13 additional isolates. Identification reliability and repeatability were evaluated, and discriminatory performance was assessed against four closely related non-target Aeromonas species. Principal component analysis (PCA), spectral profiling, and dendrogram analyses were used to evaluate spectral relatedness and species differentiation.Results Using the commercial database alone, 15/19 (78.95%) isolates achieved reliable species-level identification (log scores of 2.01-2.22), whereas 4/19 (21.05%) were identified only at the genus level (scores of 1.95-1.98). After incorporating the customized MSP database, all 19 isolates (100%) were correctly identified as A. schubertii, with highly reliable log scores ranging from 2.49 to 2.68. Technical replicates showed high spectral reproducibility, with Pearson correlation coefficients of 0.95-0.97. PCA and dendrogram analyses clearly separated A. schubertii from the tested non-target Aeromonas species, supporting the discriminatory performance of the customized database. Molecular analysis of 16S rRNA and gyrB sequences further supported the taxonomic identification of the isolates.Conclusion The species-specific MSP database substantially improved MALDI-TOF MS identification of A. schubertii, increasing species-level identification from 78.95% to 100%. The database provides a rapid and reliable framework for improving diagnosis and surveillance of this emerging aquaculture pathogen. Further expansion with geographically diverse isolates and multicenter validation is warranted to strengthen database transferability and broader diagnostic application.
SN - 0972-8988
SN - 2231-0916
ER -