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Intraspecific geographical variability of Phlebotomus perniciosus assessed by MALDI-TOF MS protein profiling
Vit Dvorak1, Carla Maia2, Rafael Marmé2
1Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
Background:
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein profiling has emerged over the last decade as a method of choice for species identification of many medically important arthropods. However, the influence of intraspecific variability on the performance of this popular technique has seldom been tested. This study provides the first standardized comparison of different geographical populations of Phlebotomus perniciosus, a vector of Leishmania infantum and Toscana virus in the western Mediterranean, by MALDI-TOF MS protein profiling.
Methods:
Phlebotomus perniciosus males were collected in five countries (Portugal, Spain, France, Italy, Croatia) that represent most of its distribution in Europe. All samples were trapped, stored and processed according to a highly standardized protocol to avoid effects other than geographical origin on their protein spectra acquired by MALDI-TOF MS protein profiling. The obtained protein spectra were compared with laboratory-reared specimens of Ph. perniciosus.
Results:
Twenty-two analysed specimens from five geographical populations provided protein spectra that were highly similar, species-specific and clustering according to their quality. No grouping according to geographical origin was observed, and the protein spectra of field-collected specimens showed similar composition and complexity to spectra from Ph. perniciosus laboratory colony-reared in captivity for several decades.
Conclusions:
Our findings demonstrate that in samples of a same sex, with the same collection method and storage time, MALDI-TOF MS protein profiling does not reflect the geographical origin of analysed specimens, confirming the value of this technique for high-fidelity and reproducible species identification of sand flies regardless of their geographical origin.
Insights
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein profiling reliably identifies sand fly species. Geographical origin does not impact MALDI-TOF MS results for Phlebotomus perniciosus, confirming its value for reproducible species identification.
Area of Science:
- Entomology
- Medical Entomology
- Molecular Biology
Background:
- Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein profiling is a key technique for identifying medically important arthropods.
- Intraspecific variability's impact on MALDI-TOF MS performance is understudied.
- Phlebotomus perniciosus is a vector for Leishmania infantum and Toscana virus in the western Mediterranean.
Purpose of the Study:
- To assess the influence of geographical origin on MALDI-TOF MS protein profiles of Phlebotomus perniciosus.
- To provide the first standardized comparison of different European populations of Phlebotomus perniciosus using MALDI-TOF MS.
- To evaluate the reliability of MALDI-TOF MS for species identification across diverse geographical locations.
Main Methods:
- Collected male Phlebotomus perniciosus from five European countries (Portugal, Spain, France, Italy, Croatia).
- Employed a highly standardized protocol for trapping, storage, and processing to minimize external variables.
- Compared protein spectra from field-collected specimens with laboratory-reared Ph. perniciosus using MALDI-TOF MS.
Main Results:
- Protein spectra from 22 specimens across five geographical populations were highly similar and species-specific.
- No geographical clustering was observed in the protein spectra.
- Spectra from field-collected sand flies matched those from long-term laboratory-reared colonies.
Conclusions:
- MALDI-TOF MS protein profiling is not influenced by the geographical origin of Phlebotomus perniciosus samples when sex, collection, and storage methods are standardized.
- The technique offers high-fidelity and reproducible species identification for sand flies.
- MALDI-TOF MS is a robust tool for identifying Phlebotomus perniciosus irrespective of its geographical origin.
