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      Genetic characterization of Rhipicephalus sanguineus (sensu lato) ticks from dogs in Portugal

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          The taxonomic status of the brown dog tick Rhipicephalus sanguineus ( sensu stricto) is a subject of on-going debate; there is a consensus that populations of this tick species should be referred to as R. sanguineus ( sensu lato) until its taxonomic status is resolved. Recent genetic studies revealed the existence of more than one lineage of R. sanguineus ( s.l.) in temperate countries. In this study, we assessed the genetic identity of ticks collected from rural dogs living in several areas located in all major geographical regions of Portugal.


          A total of 347 ticks were collected from rural dogs living in different regions of Portugal. These ticks were morphologically identified and partial mitochondrial 16S rRNA gene sequences (~300 bp) were obtained from representative specimens.


          The ticks were morphologically identified as Ixodes ricinus (seven males and 27 females), Rhipicephalus bursa (one male), Rhipicephalus pusillus (one female) and R. sanguineus ( s.l.) (two larvae, 101 nymphs, 108 males and 100 females). Partial mitochondrial 16S rRNA gene sequences were obtained from 58 R. sanguineus ( s.l.) specimens, and all of them were genetically identified as belonging to the so-called temperate lineage of R. sanguineus ( s.l.)


          These results strongly suggest that the temperate species of R. sanguineus ( s.l.) is the only representative of this tick group found on dogs in Portugal. It also adds weight to the hypothesis that Rhipicephalus turanicus is not present in this country, although further investigations are necessary to confirm this.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13071-017-2072-1) contains supplementary material, which is available to authorized users.

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          Most cited references 29

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          MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.

          We announce the release of an advanced version of the Molecular Evolutionary Genetics Analysis (MEGA) software, which currently contains facilities for building sequence alignments, inferring phylogenetic histories, and conducting molecular evolutionary analysis. In version 6.0, MEGA now enables the inference of timetrees, as it implements the RelTime method for estimating divergence times for all branching points in a phylogeny. A new Timetree Wizard in MEGA6 facilitates this timetree inference by providing a graphical user interface (GUI) to specify the phylogeny and calibration constraints step-by-step. This version also contains enhanced algorithms to search for the optimal trees under evolutionary criteria and implements a more advanced memory management that can double the size of sequence data sets to which MEGA can be applied. Both GUI and command-line versions of MEGA6 can be downloaded from free of charge.
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            Clustal W and Clustal X version 2.0.

            The Clustal W and Clustal X multiple sequence alignment programs have been completely rewritten in C++. This will facilitate the further development of the alignment algorithms in the future and has allowed proper porting of the programs to the latest versions of Linux, Macintosh and Windows operating systems. The programs can be run on-line from the EBI web server: The source code and executables for Windows, Linux and Macintosh computers are available from the EBI ftp site
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              A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences.

               Motoo Kimura (1980)
              Some simple formulae were obtained which enable us to estimate evolutionary distances in terms of the number of nucleotide substitutions (and, also, the evolutionary rates when the divergence times are known). In comparing a pair of nucleotide sequences, we distinguish two types of differences; if homologous sites are occupied by different nucleotide bases but both are purines or both pyrimidines, the difference is called type I (or "transition" type), while, if one of the two is a purine and the other is a pyrimidine, the difference is called type II (or "transversion" type). Letting P and Q be respectively the fractions of nucleotide sites showing type I and type II differences between two sequences compared, then the evolutionary distance per site is K = -(1/2) ln [(1-2P-Q) square root of 1-2Q]. The evolutionary rate per year is then given by k = K/(2T), where T is the time since the divergence of the two sequences. If only the third codon positions are compared, the synonymous component of the evolutionary base substitutions per site is estimated by K'S = -(1/2) ln (1-2P-Q). Also, formulae for standard errors were obtained. Some examples were worked out using reported globin sequences to show that synonymous substitutions occur at much higher rates than amino acid-altering substitutions in evolution.

                Author and article information

                [1 ]Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation (Fiocruz), Recife, Pernambuco 50740-465 Brazil
                [2 ]ISNI 0000 0001 0120 3326, GRID grid.7644.1, Department of Veterinary Medicine, , University of Bari, ; 70010 Valenzano Bari, Italy
                [3 ]ISNI 0000000121511713, GRID grid.10772.33, Current address: Global Health and Tropical Medicine, GHTM, , Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNL, ; Rua de Junqueira 100, 1349-008 Lisboa, Portugal
                [4 ]ISNI 0000 0000 8484 6281, GRID grid.164242.7, Faculty of Veterinary Medicine, , Universidade Lusófona de Humanidades e Tecnologias, ; Campo Grande, 376, 1749-024 Lisboa, Portugal
                [5 ]ISNI 0000000121821287, GRID grid.12341.35, Department of Veterinary Sciences, School of Agrarian and Veterinary Sciences, , University of Trás-os-Montes e Alto Douro (UTAD), ; Vila Real, Portugal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                13 March 2017
                13 March 2017
                : 10
                28285602 5346838 2072 10.1186/s13071-017-2072-1
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.

                Funded by: Global Health and Tropical Medicine
                Award ID: UID/Multi/04413/2013
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                © The Author(s) 2017


                brown dog ticks, rhipicephalus, dogs, genetics, morphology, portugal


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