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      Genetic and phylogenetic analysis of the ticks from the Sinai Massif, Egypt, and their possible role in the transmission of Babesia behnkei

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          Abstract

          Following the description of Babesia behnkei in the region of St. Katherine, Sinai, the present study was undertaken to determine the role of local tick species as vectors of piroplasms. First we assessed the local fauna of ticks, especially species occurring on rodents, camels and encountered in the environment, and then we compared genotypes of ticks from isolated wadis. Finally, we assessed the role of local tick species as potential vectors of Babesia spp. During our expedition to the Sinai Massif in a 4-week period in August–September 2012, 393 ticks were collected, including 235 adult questing ticks collected from the environment (ground level in the wadis) and 158 engorging ticks from camels and rodents. Amplification and sequencing of a 600 bp fragment of the conservative 18S rDNA and a 440 bp fragment of the more variable mitochondrial (mt) 16S rDNA were carried out to enable the identification of 54 ticks and to assess the genetic variability of ticks collected from two distant isolated wadis. The camel tick Hyalomma dromedarii constituted the majority (80–90%) of adult ticks collected from three wadis in the Sinai Mountains near St. Katherine. Among juvenile ticks collected from rodents, three genotypes were identified: H. dromedarii; Hyalomma sp. showing low homology with H. dromedarii, H. lusitanicum or H. aegyptium; and Rhipicephalus sp. A new genotype of Hyalomma was identified in an isolated montane valley, W. Gebal. Babesia/Theileria DNA was not detected in any of the ticks, which is likely due to the low infection rate in the limited number of ticks that were examined.

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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 www.megasoftware.net free of charge.
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            Phylogeny of hard- and soft-tick taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences.

            Ticks are parasitiform mites that are obligate hematophagous ectoparasites of amphibians, reptiles, birds, and mammals. A phylogeny for tick families, subfamilies, and genera has been described based on morphological characters, life histories, and host associations. To test the existing phylogeny, we sequenced approximately 460 bp from the 3' end of the mitochondrial 16S rRNA gene (rDNA) in 36 hard- and soft-tick species; a mesostigmatid mite, Dermanyssus gallinae, was used as an outgroup. Phylogenies derived using distance, maximum-parsimony, or maximum-likelihood methods were congruent. The existing phylogeny was largely supported with four exceptions. In hard ticks (Ixodidae), members of Haemaphysalinae were monophyletic with the primitive Amblyomminae and members of Hyalomminae grouped within the Rhipicephalinae. In soft ticks (Argasidae), the derived phylogeny failed to support a monophyletic relationship among members of Ornithodorinae and supported placement of Argasinae as basal to the Ixodidae, suggesting that hard ticks may have originated from an Argas-like ancestor. Because most Argas species are obligate bird octoparasites, this result supports earlier suggestions that hard ticks did not evolve until the late Cretaceous.
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              Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus ticks fed on infected blood in a new skin-feeding technique.

              Although Babesia divergens is the the principal confirmed zoonotic Babesia sp. in Europe, there are gaps in our knowledge of its biology and transmission by the tick Ixodes ricinus. In order to reproduce the part of the parasite cycle that occurs in the vector, an in vitro animal skin feeding technique on blood containing in vitro cultivated B. divergens was developed. Parasite DNA was detected in all samples of salivary glands of nymphs and adults that had fed on parasitized blood as larvae and nymphs, respectively, indicating acquisition as well as a transtadial persistence of B. divergens. PCR performed on eggs and larvae produced by females that had fed on parasitized blood demonstrated the existence of a transovarial transmission of the parasite. Gorging B. divergens infected larvae on non-infected gerbils showed persistance of the parasite over moulting into the resulting nymphs. These results indicate that the parasitic stages infective for the vector (i.e. the sexual stages) can be produced in vitro. To our knowledge, this is the first report of artificial feeding of I. ricinus via membrane as well as in vitro transmission of B. divergens to its vector. The opportunities offered by the use of such a transmission model of a pathogen by I. ricinus are discussed.
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                Author and article information

                Contributors
                muha@biol.uw.edu.pl
                e.mierzejewska@biol.uw.edu.pl
                emanmohallal@yahoo.com
                jerzy.behnke@nottingham.ac.uk
                anabena@biol.uw.edu.pl
                Journal
                Exp Appl Acarol
                Exp. Appl. Acarol
                Experimental & Applied Acarology
                Springer International Publishing (Cham )
                0168-8162
                1572-9702
                28 August 2017
                28 August 2017
                2017
                : 72
                : 4
                : 415-427
                Affiliations
                [1 ]ISNI 0000 0004 1937 1290, GRID grid.12847.38, Department of Parasitology, Institute of Zoology, Faculty of Biology, , University of Warsaw, ; 1 Miecznikowa Street, 02-096 Warsaw, Poland
                [2 ]ISNI 0000 0004 5373 9159, GRID grid.466634.5, Desert Research Center, ; Cairo, Egypt
                [3 ]ISNI 0000 0004 1936 8868, GRID grid.4563.4, School of Life Sciences, Faculty of Medicine and Health Sciences, , University of Nottingham, ; Nottingham, NG7 2RD UK
                Article
                164
                10.1007/s10493-017-0164-4
                5583268
                28849399
                2ee040a2-2576-40a8-8a5f-06957dcea36c
                © The Author(s) 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), 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.

                History
                : 8 March 2017
                : 17 July 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100004281, Narodowe Centrum Nauki;
                Award ID: OPUS 2011/03/B/NZ6/02090 (2012-2015)
                Award Recipient :
                Categories
                Article
                Custom metadata
                © Springer International Publishing AG 2017

                Entomology
                hyalomma dromedarii,rhipicephalus,babesia,genotyping,rodents,sinai
                Entomology
                hyalomma dromedarii, rhipicephalus, babesia, genotyping, rodents, sinai

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