Naslov
EXPLORING BALKAN SAND FLY FAUNA USING MALDI-TOF MASS SPECTROMETRY

Autori
Hlavackova, Kristyna, Dvorak, Vit, Volf, Petr, Halada, P., Alten, Bulent, Ivović, Vladimir, Omeragić, Jasmin, Pajović, Igor, Martinković, Franjo

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, ostalo

Izvornik
3rd Conference on Neglected Vectors and Vector-Borne Diseases (EurNegVec): with MC and WG Meeting of the COST Action TD1303 Abstract Book / - , 2016, 49-50

Skup
3rd Conference on Neglected Vectors and Vector-Borne Diseases (EurNegVec): with MC and WG Meeting of the COST Action TD1303

Mjesto i datum
Zaragoza, Španjolska, 24.05.2016. - 26.05.2016

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
BALKAN ; SAND FLY FAUNA ; MALDI-TOF MASS SPECTROMETRY

Sažetak
Bloodsucking females of phlebotomine sand-flies are the only proven vectors of Leishmania parasites, causative agents of leishmaniases. Their conclusive species identification in endemic areas, where morphologically similar species with different vectorial capacity and ecology occur, is profound. Traditional morphological approach relies on minute and sometime dubious characters ; therefore several molecular methods were applied recently. Protein profiling by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry is a promising technique for identification of various organisms including insects. It is simple, accurate, and requires minimal sample preparation. The obtained mass spectrum represents a characteristic and unique protein fingerprint, which allows rapid and effective species identification. We optimized methods of specimen capture, storage, sample preparation, tested suitability of different body parts of sand fly for protein identification, and developed a multi-approach protocol that utilizes a single insect body for several identification methods (morphological, DNA and protein based). Further on, the influence of blood meal at several time intervals after feeding on the quality of protein profile of females was investigated. Also spectra of larvae (L4) and pupae were measured and compared with profiles of adult sand flies. A reference spectra database comprising 16 sand fly species was established using MALDI Biotyper software. It includes species maintained in laboratory colonies as well as field-collected specimens identified by morphological analysis and DNA sequencing. During two field trips to Balkan countries (Bosnia and Hercegovina, Bulgaria, Croatia, Hungary, Macedonia, Montenegro, Serbia and Slovenia) in summer 2015, 680 specimens (Larroussius, Adlerius, Paraphlebotomus, Phlebotomus, Sergentomyia) out of 7257 totally captured were identified by protein profiling which proved as reliable. It was useful for female specimens of the subgenus Larroussius (492 analyzed specimens) and differentiation between its closely related species (P. neglectus, P. perfiliewi, P. tobbi). Also two species of the subgenus Adlerius (P. balcanicus, P. simici), hardly distinguishable by morphology, were unambiguously differentiated by protein spectra. All dubious samples (difference between morphological and protein based ID) were clarified by a sequencing analysis, which confirmed the identification according to MALDI-TOF MS in all cases. In summary, MALDI-TOF protein profiling is a suitable, time- and cost- effective method for species identification of large sets of field-caught sand flies if these are collected, stored and analyzed using a standard, optimized protocol.

Izvorni jezik
Engleski

Znanstvena područja
Veterinarska medicina

POVEZANOST RADA