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      Isolation and partial characterisation of a novel Trypanosoma from the tick Ixodes ricinus

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          Abstract

          Trypanosomes have long been recognised as being amongst the most important protozoan parasites of vertebrates, from both medical and veterinary perspectives. Whilst numerous insect species have been identified as vectors, the role of ticks is less well understood. Here we report the isolation and partial molecular characterisation of a novel trypanosome from questing Ixodes ricinus ticks collected in Slovakia. The trypanosome was isolated in tick cell culture and then partially characterised by microscopy and amplification of fragments of the 18S rRNA and 24Sα rDNA genes. Analysis of the resultant sequences suggests that the trypanosome designated as Trypanosoma sp. Bratislava1 may be a new species closely related to several species or strains of trypanosomes isolated from, or detected in, ticks in South America and Asia, and to Trypanosoma caninum isolated from dogs in Brazil. This study highlights the potential involvement of ixodid ticks in the epidemiology of trypanosomes, as well as the use of tick cell lines for isolation of such tick-borne protozoa. Further studies are required to investigate the epidemiology, transmission and life cycle of this putative novel species.

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          Ixodes ricinus and Its Transmitted Pathogens in Urban and Peri-Urban Areas in Europe: New Hazards and Relevance for Public Health

          Tick-borne diseases represent major public and animal health issues worldwide. Ixodes ricinus, primarily associated with deciduous and mixed forests, is the principal vector of causative agents of viral, bacterial, and protozoan zoonotic diseases in Europe. Recently, abundant tick populations have been observed in European urban green areas, which are of public health relevance due to the exposure of humans and domesticated animals to potentially infected ticks. In urban habitats, small and medium-sized mammals, birds, companion animals (dogs and cats), and larger mammals (roe deer and wild boar) play a role in maintenance of tick populations and as reservoirs of tick-borne pathogens. Presence of ticks infected with tick-borne encephalitis virus and high prevalence of ticks infected with Borrelia burgdorferi s.l., causing Lyme borreliosis, have been reported from urbanized areas in Europe. Emerging pathogens, including bacteria of the order Rickettsiales (Anaplasma phagocytophilum, “Candidatus Neoehrlichia mikurensis,” Rickettsia helvetica, and R. monacensis), Borrelia miyamotoi, and protozoans (Babesia divergens, B. venatorum, and B. microti) have also been detected in urban tick populations. Understanding the ecology of ticks and their associations with hosts in a European urbanized environment is crucial to quantify parameters necessary for risk pre-assessment and identification of public health strategies for control and prevention of tick-borne diseases.
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            Tick cell lines: tools for tick and tick-borne disease research.

            Over 40 cell lines are currently available from 13 ixodid and one argasid tick species. The successful isolation and propagation of several economically important tick-borne pathogens in tick cell lines has created a useful model to study interactions between tick cells and these viral and bacterial disease agents. Tick cell lines have already proved to be a useful tool in helping to define the complex nature of the host-vector-pathogen relationship. With the availability of genomics tools, tick cell lines will become increasingly important as a complement to tick and tick-borne disease research in vivo once genetic transformation and gene silencing using RNA interference become routine.
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              Rickettsia monacensis sp. nov., a spotted fever group Rickettsia, from ticks (Ixodes ricinus) collected in a European city park.

              We describe the isolation and characterization of Rickettsia monacensis sp. nov. (type strain, IrR/Munich(T)) from an Ixodes ricinus tick collected in a city park, the English Garden in Munich, Germany. Rickettsiae were propagated in vitro with Ixodes scapularis cell line ISE6. BLAST analysis of the 16S rRNA, the citrate synthase, and the partial 190-kDa rickettsial outer membrane protein A (rOmpA) gene sequences demonstrated that the isolate was a spotted fever group (SFG) rickettsia closely related to several yet-to-be-cultivated rickettsiae associated with I. ricinus. Phylogenetic analysis of partial rompA sequences demonstrated that the isolate was genotypically different from other validated species of SFG rickettsiae. R. monacensis also replicated in cell lines derived from the ticks I. ricinus (IRE11) and Dermacentor andersoni (DAE100) and in the mammalian cell lines L-929 and Vero, causing cell lysis. Transmission electron microscopy of infected ISE6 and Vero cells showed rickettsiae within the cytoplasm, pseudopodia, nuclei, and vacuoles. Hamsters inoculated with R. monacensis had immunoglobulin G antibody titers as high as 1:16,384, as determined by indirect immunofluorescence assay. Western blot analyses demonstrated that the hamster sera cross-reacted with peptides from other phylogenetically distinct rickettsiae, including rOmpA. R. monacensis induced actin tails in both tick and mammalian cells similar to those reported for R. rickettsii. R. monacensis joins a growing list of SFG rickettsiae that colonize ticks but whose infectivity and pathogenicity for vertebrates are unknown.
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                Author and article information

                Contributors
                Journal
                Ticks Tick Borne Dis
                Ticks Tick Borne Dis
                Ticks and Tick-Borne Diseases
                Elsevier
                1877-959X
                1877-9603
                1 September 2020
                September 2020
                : 11
                : 5
                : 101501
                Affiliations
                [a ]Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UK
                [b ]School of Science, Engineering and Environment, G32 Peel Building, University of Salford, Salford M5 4WT, UK
                [c ]Centre of Rickettsiosis and Arthropod-borne Diseases, CIBIR, C/ Piqueras, 98, Logroño 26006, La Rioja, Spain
                [d ]Institute of Zoology, Slovak Academy of Sciences, Dubravska cesta 9, SK-84506 Bratislava, Slovakia
                [e ]The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK
                Author notes
                [* ]Corresponding author at: Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UK. L.Bell-Sakyi@ 123456liverpool.ac.uk
                Article
                S1877-959X(20)30270-3 101501
                10.1016/j.ttbdis.2020.101501
                7397511
                32723658
                71e0d727-bd59-45a9-9546-3b6f60e5d6e4
                © 2020 The Authors

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

                History
                : 24 May 2020
                : 14 June 2020
                : 21 June 2020
                Categories
                Article

                Infectious disease & Microbiology
                ixodes ricinus,tick cell line,trypanosome,slovakia
                Infectious disease & Microbiology
                ixodes ricinus, tick cell line, trypanosome, slovakia

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