Molecular characterization of Rhipicephalus (Boophilus) annulatus from Iran by sequences of cytochrome c oxidase subunit I (COI) and the second internal transcribed spacer (ITS2)

Document Type : Pathology - Clinical Pathology

Authors

1 Graduated from the Faculty of Veterinary Medicine, University of Tehran, Iran

2 Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Iran Iranian Center of Ticks and Tick-born Diseases, Faculty of Veterinary Medicine, Tehran, Iran

3 Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Iran Iranian Center of Ticks and Tick-born Diseases, Faculty of Veterinary Medicine, Tehran, Iran Abstract:

Abstract

Background: Traditionally, morphological features of Rhipicephalus (Boophilus) annulatus from closely-related ticks have been considered for their identification and differentiation. However, it is difficult and requires expertise in order to accurately identify and differentiate engorged female ticks and some developmental stages such as larva and nymph from other similar ticks. Hence, molecular markers may be a suitable alternative. OBJECTIVES: Mitochondrial cytochrome c oxidase subunit I (COI) gene and the second internal transcribed spacer (ITS2) fragments of Rh. (Bo.) annulatus were sequenced to assess the use of molecular techniques for identifications and phylogenetic studies of these ticks. METHODS: Polymerase chain reaction (PCR) technique was performed based on the analyses of COI and ITS2 sequences of ticks collected from two different regions in Iran (Golestan and Mazandaran). RESULTS: The length of COI and ITS2 sequences were 1539 and 1158bp, respectively. The nucleotide similarity of COI gene was 91.3% between the ticks examined from the two different regions. The deduced amino acid sequences from COI showed 98.6% similarity between the ticks studied and showed 98.2 and 99.6% similarity with the only complete sequence of Rh. (Bo.) annulatus (AGH19677) registered in GenBank. The obtained complete nucleotide sequences of ITS2 from Rh. (Bo.) annulatus from Golestan and Mazandaran revealed 99.9% similarity, while the other ticks registered in GenBank 95 to 99% similarity (KC503267, AF271270, AF271272, JQ412126). CONCLUSIONS: It seems that COI and ITS2 sequences could provide suitable genetic markers for discrimination and genetic characterization of Rhipicephalus (Boophilus) annulatus.
 

Keywords


Barker, S., Murrell, A. (2004) Systematics and evolution of ticks with a list of valid genus and species names. J Parasitol. 129: S15-S36.
Barker, S.C. (1998) Distinguishing species and populations of rhipicephaline ticks with its 2 ribosomal RNA. J Parasitol. 84: 887-892.
Brahma, R.K., Dixit, V., Sangwan, A.K., Doley, R. (2014) Identification and characterization of Rhipicephalus (Boophilus) microplus and Haemaphysalis bispinosa ticks (Acari: Ixodidae) of North East India by ITS2 and 16S rDNA sequences and morphological analysis. Exp Appl Acarol. 62: 253-265.
Cruickshank, R.H. (2002) Molecular markers for the phylogenetics of mites and ticks. Sys  Appl Acarol. 7: 3-14.
Dantas-Torres, F., Latrofa, M.S., Annoscia, G., Giannelli, A., Parisi, A., Otranto, D. (2013) Morphological and genetic diversity of Rhipicephalus sanguineus sensu lato from the new and old worlds. Parasit Vectors. 6: 213.
Hwang, U.-W., Kim, W. (1999) General properties and phylogenetic utilities of nuclear ribosomal DNA and mitochondrial DNA commonly used in molecular systematics. Korean J Parasitol. 37: 215-228.
Jongejan, F., Uilenberg, G. (2004) The global importance of ticks. Parasitology. 129: S3-S14.
Latrofa, M.S., Dantas-Torres, F., Annoscia, G., Cantacessi, C., Otranto, D. (2013) Comparative analyses of mitochondrial and nuclear genetic markers for the molecular identification of Rhipicephalus spp. Infection, Genet Evol. 20: 422-427.
Liu, G.H., Chen, F., Chen, Y.Z., Song, H.Q., Lin, R.Q., Zhou, D.H., Zhu X.Q. (2013) Complete mitochondrial genome sequence data provides genetic evidence that the brown dog tick Rhipicephalus sanguineus (Acari: Ixodidae) represents a species complex. Int J Biol Sci. 9: 361.
Murrell, A., Barker, S.C. (2003) Synonymy of Boophilus Curtice, 1891 with Rhipicephalus Koch, 1844 (Acari: Ixodidae). Sys Parasitol. 56: 169-172.
Nava, S., Guglielmone, A. A., Mangold, A. J. (2008) An overview of systematics and evolution of ticks. Front Biosci (Landmark edition). 14: 2857-2877.
Navajas, M. (1998) Host plant associations in the spider mite Tetranychus urticae (Acari: Tetranychidae): insights from molecular phylogeography. Exp Appl Acarol. 22: 201-214.
Rahbari, S., Nabian, S., Shayan, P. (2007) Primary report on distribution of tick fauna in Iran. Parasitol Res. 101: 175-177.
Razmi, G.R., Glinsharifodini, M., Sarvi, S. (2007) Prevalence of ixodid ticks on cattle in Mazandaran province, Iran. Korean  J Parasitol. 45: 307-310.
Razmi, G.R., Hosseini, M., Aslani, M. (2003) Identification of tick vectors of ovine theileriosis in an endemic rion of Iran. Vet Parasitol. 116: 1-6.
Spitalska, E., Namavari, M.M., Hosseini, M.H., Shad-del, F., Amrabadi, O. R., Sparagano, O. A. (2005) Molecular surveillance of tick-borne diseases in Iranian small ruminants. Small Rum Res. 57: 245-248.
Szabó, M.P., Mangold, A.J., João, C.F., Bechara G.H., Guglielmone, A