DOI: 10.5061/dryad.7h44j1073
title: Data from: Identification and genetic diversity of ticks in local sheep in Samarra city, northern of Baghdad, Iraq

description: This study reported hard tick (Ixodida: Ixodidae) infestation in sheep in
 Samara city, north of Baghdad, from June to December 2023. A total of 480
 ticks were manually pulled out from the sheep with an ethanol-soaked
 tissue and preserved in plastic containers containing 70% ethyl alcohol.
 All samples were counted and examined individually under a light
 microscope to identify the genus. Preliminary microscopic examination
 revealed that the majority of the collected ticks were Hyalomma, with
 91.4%, followed by Rhipicephalus (6.8%) and Ixodes (1.8%). Furthermore,
 Hyalomma was dominant across all months, with a peak relative abundance in
 October (100%). The peak relative abundances for Rhipicephlus (16%) and
 Ixodes (4.3%) occurred in July. Under further microscopic evaluation,
 morphological features demonstrated three species, namely, Hyalomma
 detritum (Koch), Hy. anatolicum (Koch) and Rhipicephalus sanguineus sensu
 lato (Latreille). To confirm genera and species, molecular diagnosis was
 performed, based on the COX1 gene, resulting in the identification of five
 species, namely, Hy. detritum , Hy. anatolicum , Hy. excavatum (Koch,
 1844), Rh. sanguineus sensu lato and an unidentified Ixodes sp. 
 While the Ixodes scapularis isolation unexpectedly revealed genetic
 resemblance to North American samples, indicating a wider geographical
 spread, the Hyalomma and Rhipicephalus isolates shared ancestry with other
 Middle Eastern strains, demonstrating regional genetic stability. These
 results showed the comprehensive diversity of tick species in sheep,
 including Hyalomma, Rhipicephalus, and Ixodes, and also provided good
 epidemiological data to support the preventative procedures of tick-borne
 infections in Samarra city. 
description: <strong>Ethics</strong>. This project was approved by
 the ethical committee (ACUC) at the University of Baghdad (No.1667 on
 22/03/2023).           <strong>Study
 area.</strong> Samarra is a city in Salah Adin province, 120 km
 north of Baghdad. The city is bordered in the North-East by Diyala
 province, which shares borders with Iran (Figure 1). This study was
 conducted mainly in open farm areas in Samarra villages, where these areas
 are open to domestic and wild animals. Most of the farms lie at the border
 with Diyala province. The weather in this city is a typical desert
 climate, where it is extremely hot-dry in summer, and cold and dry to
 moderately wet in winter. The temperature ranges from 0°C on winter nights
 to over 50°C on summer days. <strong>Parasite
 samples.</strong> Four hundred and eighty tick samples were obtained
 from more than 150 naturally infested, al-Awassi breed of sheep, from
 multiple farms in Samarra city (Figure 1). Sheep were thoroughly
 investigated for ticks during the period from June to December 2023. Hard
 ticks were collected from parts of the animals body which were devoid of
 wool, such as eyes, ears, udders, external genitalia and under tail areas.
 Samples were collected by a veterinarian twice per week in cooperation
 with the owners. After collection, hard ticks were transported to the
 parasitology laboratory of the College of Veterinary Medicine, University
 of Baghdad for microscopic examination then preserved in 10 ml of 70%
 ethyl alcohol for further processing.
 <strong>Morphological identification of
 ticks.</strong> Ticks’ specimens were placed on a clean slide using
 double sided tape. Suitable forceps were used to manipulate ticks dorsally
 or ventrally and for spreading legs to examine internal structures. A
 small drop of glycerol was used to stabilize the tick on the slide. Tick
 samples were morphologically categorized under dissecting microscope into
 genera using taxonomic keys (<strong>Makwarela et al.,
 2024)</strong>. Dorsal and ventral sides of adult ticks were
 precisely examined for distinguishing characteristics of structures under
 light microscopy. Subsequently, identified adults’ ticks were attached
 dorsally on the slides using the tape to photograph the specimens using a
 digital camera. <strong>Extraction of DNA and
 PCR.</strong> Ticks were subjected to molecular analysis based on
 the mitochondrial cytochrome c oxidase subunit 1 gene
 (<em>COX</em>1). DNA extraction was performed on hard ticks,
 according to manufacturer’s guidelines using PCR Premix Kit (Bioneer,
 South Korea). Amplification of 680 bp of <em>COX</em>1 locus
 was done for molecular identification of tick species using one sets of
 primers. The primers were <em>COX</em>1-F 5’ATCATAAAKAYHTTGG
 3’ and<em>COX</em>1 -R 5’ GGGTGACCRAARAAHCA 3’
 (<strong>Lv et al., 2014)</strong>. PCR was done using a
 Veriti thermal cycler (ThermoFisher Scientific, USA) using the following
 parameters: initial denaturation step of 95 ◦C for 5 min, 30 cycles of
 denaturation at 95 ◦C for 45 s, annealing at 54–58 ◦C for 45 s and
 extension at 72 ◦C for 1 min, and a final extension of 72 ◦C for 5 min.
 All the PCR products were analysed by electrophoresis on agarose gel,
 stained with RedSafe (Intron, Korea), and visualized on a UV
 transilluminator. Amplicons were sent for sequencing (Macrogen, South
 Korea) and sequences were submitted to Basic Local Alignment Search Tool
 (BLAST) at the National Centre for Biotechnology information (NCBI).
 Multiple sequence alignment of the partial <em>COX</em>1 locus
 sequences were made by MUSCLE in SnapGene (Version 5.0.5. Iowa university,
 USA). A phylogenetic analysis was conducted using neighbor joining
 analyses with 500 bootstrap replicates in MEGA X (<strong>Kumar, et
 al., 2012</strong>).
description: # Data from: Identification and genetic diversity of ticks in local sheep
 in Samarra city, north of Baghdad, Iraq Dataset DOI:
 [10.5061/dryad.7h44j1073](10.5061/dryad.7h44j1073) **Description of the
 data and file structure** #### File Structure #### Input Files **File:**
 **TicksRawData2.xlsx** **Description:** Input file used for the number,
 genera, and species of ticks and sheep, and which tick infects which
 sheep. * Sheep ID: This is the unique identifier assigned to the sheep
 sampled. - Tick ID: ticks IDs that were collected from this sheep. *
 Tick_Species: The ticks collected belong to the respective genus. -
 Date_Collected: The samples were collected. * Location: The sampling took
 place in a location referred. - Sheep_Age: The sheep is either young or
 adult. * Sheep_Sex: Male or Female. - Infection_Status: whether the sheep
 was found to be infested with ticks. * Notes: Additional note stating that
 the sheep belongs to which local breed. The following files are provided
 in the under`.doc` format and named according to the figure or table they
 support in the publication. Each `doc` file contains the figure or table
 and describes its internal files and purpose. **File:** **Datainput_Figure
 1.doc** **Description:** Input file used for a map of Iraq, showing
 Samarra city and the study area, which is located within Salah Adin
 province, 120 km north of Baghdad. **File:** **Datainput_Figure 2.doc**
 **Description:** Input file used for the morphological characterisations
 of three genera of female ticks: A, B. Hyalomma species. C, Rhipicephalus
 species. D, Ixodes species collected from sheep in Samarra, Iraq.
 **File:** **Datainput_Figure 3.doc** **Description:** Input file used for
 the Phylogenetic relationship of Hyalomma (A), Rhipicephalus (B), and
 Ixodes (C) species of ticks isolated from sheep in Samarra, Iraq, based on
 the partial COX1 gene sequence. The analysis was generated using the
 maximum likelihood approach. The proportion of isolates is grouped based
 on the bootstrap test (100 repetitions), and posterior probabilities of
 nucleotide substitutions are displayed next to each branch and reflected
 in the scale bar. The GenBank accession number for each isolate is
 displayed next to the scientific names, and samples from this study are
 labelled with red dots. **File:** **Datainput_Table 1.doc**
 **Description:** Input file used for the Relative abundance of different
 tick genera as determined by microscopic identification for the seven
 months of the study period in Samarra, Iraq. Data input gene accession
 numbers links Description: Input files used links of accession numbers
 deposited to the GenBank:
 [https://www.ncbi.nlm.nih.gov/nuccore/PP857589.1?report=GenBank](https://www.ncbi.nlm.nih.gov/nuccore/PP857589.1?report=GenBank) [https://www.ncbi.nlm.nih.gov/nuccore/PP857588.1?report=GenBank](https://www.ncbi.nlm.nih.gov/nuccore/PP857588.1?report=GenBank) [https://www.ncbi.nlm.nih.gov/nuccore/PP857590.1?report=GenBank](https://www.ncbi.nlm.nih.gov/nuccore/PP857590.1?report=GenBank) [https://blast.ncbi.nlm.nih.gov/Blast.cgi](https://blast.ncbi.nlm.nih.gov/Blast.cgi) [https://www.ncbi.nlm.nih.gov/nuccore/PP857593.1?report=GenBank](https://www.ncbi.nlm.nih.gov/nuccore/PP857593.1?report=GenBank) [https://www.ncbi.nlm.nih.gov/nuccore/PP857587.1?report=GenBank](https://www.ncbi.nlm.nih.gov/nuccore/PP857587.1?report=GenBank)