MORPHOLOGCAL AND MOLECULAR STUDY OF HARD TICKS SPECIES THAT INFESTED SMALL RUMINANTS IN DUHOK GOVERNORATE, KURDISTAN REGION, IRAQ

Abstract

Ticks are harmful ectoparasite that feed on human and animal blood and causing many diseases through the world. They infested many hosts including: mammals, reptiles and birds. Ticks are important vector and they have the ability to transmit a variety of pathogenic agent to humans and animals. Ticks are divided into two major groups which are hard tick (Ixodidae) and soft tick (Argasidae). Because there was no such study done on identification of tick species by PCR technique in Kurdistan and particularly in Duhok Governorate, therefore present study was done to identify tick species by using molecular study by using of 16S rRNA and DNA sequencing. About 1000 ticks were collected from both sheep and goat respectively (500 and 500), form Duhok Governorate including: Barwaria, Zakho, Sumeil, Mangeshik, Sersin, Shekhan and Akre, Iraqi Kurdistan, between May and June 2016, between April and June 2017. The results of present study three genera of tick were detected in small ruminants by microscopic identification including: Rhipicephalus spp., Hyalomma spp. and Boophilus spp. Distribution of tick among sheep and goat according to the gender, the rate of infection in female was higher than in male in both species Ewe and Doe was 32.6% and 31.11% respectively as compared to male in both species (Ram and Buck) was 21.15% and 15.11% respectively. The distribution of gender of tick in was higher in male ticks than female tick with ratio 2:1. Distribution of identified ticks in present study including (Rhipicephalus, Hyalomma and Boophilus) respectively, in Barwaria were (82.6%, 13.3%, and 4.1) respectively, in Zaxo were (48.3%, 42.5% and 10.3%), in Sumel were (47%, 42.7% and 10.3%), in Mangeshik were (73,2%, 26.8%, and 0%), in Sersink were (61.5%, 38.5% and 0%), in Shekhan were (78.8%, 11.8% and 9.4%) and in Akre were (60%, 34% and 6%). On molecular study, 60 samples from 150 were positive with size 460 bp after 16S rRNA amplification and have got clear bands on agarose gel 1% and electrophoresis and 20 PCR positive products were sent to Humanizing Genomics, Macrogen Company (Korea) using primer 16S_rRNA gene for sequencing both forward and Reverse. Six species of tick under two genera were founded including: Rhipicephalus and Hyalomma were identified which including: Hyalomma anatolicum, H. marginatum, R. annulatus, R. sanguineus and R. turanicus. H. asiaticum asiaticum for the first time was recorded in Kurdistan, and especially in Duhok city. Moreover, all sequences were submitted to NCBI using BankIt software and we obtained accession number. Phylogenetic tree was constructed based on 16S rRNA for both samples: 16S rRNA (MN594483) and (MN594490) were identical 100% to reference sequences respectively: (KU664367.1 and HM176656.1) and other sequences were identical 99% to the references sequence. In conclusion the present study is the first study for identification of tick species among sheep and goats in Duhok Governorate, Iraqi Kurdistan by sequencing analysis.