First Report of Molecular-Genetic Description and Barcoding of Hard Ticks (Akari:Ixodidae) Distributed in Georgia

Author: Mariam Zakalashvili
Co-authors: Mariam Zakalashvili, Yvonne-Marie Linton , Gvantsa Chanturia, Nino Abashidze , Giorgi Chakhunashvili, Julieta Manveliani, Giorgi Tomashvili , Roena Sukhiashvili, David Tsereteli, Paata mnadze
Keywords: (Acari: Ixodidae), COI region, primers HCO/LCO and ChelF1/ChelR2PCR amplified, species rhipicephalus annulatus
Annotation:

Abstract Molecular methods are increasingly being utilized for the accurate identification of ticks (Acari: Ixodidae), especially in cases of morphologically highly similar species. In this study, we performed molecular research of the tick fauna in Georgia using DNA barcoding method. Ticks were sampled in three (shida qartli, samtskhe Javakheti, kakheti) bio geographical regions and five species were recorded: Five different species were identified Georgian samples, Riphicepalus anulatus, Riphicepalus bursa, Hyaloma scupenze, Hyaloma marginatum, Dermacentor marginatus. All of the listed species pose a threat to humans (including Crimean-Congo hemorrhagic fever), except for the species rhipicephalus annulatus, which mostly infects cattle The phylogenetic tree of hard ticks was built according to the sequencing results. For this purpose, magnetic bead DNA extraction and PCR amplification protocols were selected and optimized. Samples of hard ticks were collected from different regions of Georgia; gDNA was extracted from 91 samples and COI region was PCR amplified with primers HCO/LCO and ChelF1/ChelR2. The best amplifications were done by ChelF1/ChelR2 primers for further research as they showed more specificity in amplification reactions. PCR amplicons were sequenced with Sanger sequencing method and results were analyzed in Geneious Prime. All sample species were identified with high reliability (Query coverage 99.3 %). sequences were analyzed by species delimitation methods together with the sequences of conspecific and congeneric species from the public BOLD database. Our specimens of H. punctata represent a new, genetically distinct MOTU