20.500.12556/RUP-23186 Intraspecific geographical variability of Phlebotomus perniciosus assessed by MALDI-TOF MS protein profiling Background Matrix‑assisted laser desorption/ionization time‑of‑flight mass spectrometry (MALDI‑TOF MS) pro‑ tein 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 popula‑ tions 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, Croa‑ tia) 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. Ozadje Proteinsko profiliranje z matrično podprto lasersko desorpcijo/ionizacijo in masno spektrometrijo s časom preleta (MALDI-TOF MS) se je v zadnjem desetletju uveljavilo kot ena izmed najprimernejših metod za identifikacijo vrst številnih medicinsko pomembnih členonožcev. Vendar pa je bil vpliv znotrajvrstne variabilnosti na učinkovitost te priljubljene tehnike le redko preučen. Ta raziskava predstavlja prvo standardizirano primerjavo različnih geografskih populacij vrste Phlebotomus perniciosus, prenašalca Leishmania infantum in virusa Toscana v zahodnem Sredozemlju, z uporabo proteinskega profiliranja MALDI-TOF MS. Metode Samci vrste Phlebotomus perniciosus so bili zbrani v petih državah (Portugalska, Španija, Francija, Italija in Hrvaška), ki zajemajo večino območja razširjenosti te vrste v Evropi. Vsi vzorci so bili ulovljeni, shranjeni in obdelani po strogo standardiziranem protokolu, da bi se izognili vplivom drugih dejavnikov kot geografski izvor na njihove proteinske spektre, pridobljene z metodo MALDI-TOF MS. Dobljeni proteinski spektri so bili primerjani z osebki vrste Ph. perniciosus, vzrejenimi v laboratoriju. Rezultati Dvaindvajset analiziranih osebkov iz petih geografskih populacij je zagotovilo proteinske spektre, ki so bili med seboj zelo podobni, vrstno specifični in so se združevali glede na njihovo kakovost. Združevanja glede na geografski izvor ni bilo opaziti. Proteinski spektri osebkov, zbranih na terenu, so pokazali podobno sestavo in kompleksnost kot spektri osebkov iz laboratorijske kolonije vrste Ph. perniciosus, ki je bila vzrejana v ujetništvu več desetletij. Sklepi Naše ugotovitve kažejo, da pri vzorcih istega spola, zbranih z enako metodo in shranjenih enako dolgo, proteinsko profiliranje MALDI-TOF MS ne odraža geografskega izvora analiziranih osebkov. To potrjuje vrednost te tehnike za zanesljivo, natančno in ponovljivo identifikacijo vrst peščenih muh ne glede na njihov geografski izvor. intraspecific geographical variability Phlebotomus perniciosus MALDI-TOF intraspecifična geografska variabilnost Phlebotomus perniciosus MALDI-TOF true false true Angleški jezik Slovenski jezik Članek v reviji 2026-06-29 12:04:23 2026-06-29 12:04:25 2026-06-30 03:04:55 0000-00-00 00:00:00 2026 0 0 str. 1-8 article no. ǂ197 Vol. 19 2026 0000-00-00 Zaloznikova NiDoloceno NiDoloceno 0000-00-00 0000-00-00 2026-03-23 595.772.1 1756-3305 282921731 RAZ_Dvořak_Vit_2026.pdf RAZ_Dvořak_Vit_2026.pdf 1 ED916D712F4FC8451C6C3EFACF8CAE47 60d0ec09e7f8930927861f02e16b2f9cf1ea9fe637f11d54e201dc8d68d0e4f8 e9bf9b1f-73a1-11f1-9e8d-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=33895 https://link.springer.com/article/10.1186/s13071-026-07354-9 1 bf14c23b-73a1-11f1-9e8d-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=33894 Fakulteta za matematiko, naravoslovje in informacijske tehnologije 0 0 0