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      Anaplasma phagocytophilum, Bartonella spp ., haemoplasma species and Hepatozoon spp . in ticks infesting cats: a large-scale survey

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          Abstract

          Background

          Ticks derived from cats have rarely been evaluated for the presence of pathogens. The aim of this study was to determine the prevalence of Anaplasma phagocytophilum, Bartonella spp ., haemoplasma species and Hepatozoon spp. in ticks collected from cats in the UK.

          Methods

          Five hundred and forty DNA samples extracted from 540 ticks collected from cats presenting to veterinarians in UK practices were used. Samples underwent a conventional generic PCR assay for detection of Hepatozoon spp. and real-time quantitative PCR assays for detection of Anaplasma phagocytophilum and three feline haemoplasma species and a generic qPCR for detection of Bartonella spp. Feline 28S rDNA served as an endogenous internal PCR control and was assessed within the haemoplasma qPCR assays. Samples positive on the conventional and quantitative generic PCRs were submitted for DNA sequencing for species identification.

          Results

          Feline 28S rDNA was amplified from 475 of the 540 (88.0%) ticks. No evidence of PCR inhibition was found using an internal amplification control. Of 540 ticks, 19 (3.5%) contained DNA from one of the tick-borne pathogens evaluated. Pathogens detected were: A. phagocytophilum ( n = 5; 0.9%), Bartonella spp. ( n = 7; 1.3%) [including Bartonella henselae ( n = 3; 0.6%) and Bartonella clarridgeiae ( n = 1; 0.2%)], haemoplasma species ( n = 5; 0.9%), “ Candidatus Mycoplasma haemominutum” ( n = 3; 0.6%), Mycoplasma haemofelis ( n = 1; 0.2%), “ Candidatus Mycoplasma turicensis” ( n = 1; 0.2%), Hepatozoon spp. ( n = 2; 0.4%), Hepatozoon felis ( n = 1; 0.2%) and Hepatozoon silvestris ( n = 1; 0.2%).

          Conclusion

          These data provide important information on the prevalence of tick-borne pathogens in ticks infesting cats, with the identification of haemoplasma species, A. phagocytophilum, H. felis and Bartonella spp. (including B. henselae and B. clarridgeiae). This study also documents the first report of H. silvestris in ticks collected from domestic cats.

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          Most cited references21

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          Multiplex real-time PCR for detection of anaplasma phagocytophilum and Borrelia burgdorferi.

          A multiplex real-time PCR assay was developed for the simultaneous detection of Anaplasma phagocytophilum and Borrelia burgdorferi. The assay was tested on various Anaplasma, Borrelia, Erhlichia, and Rickettsia species, as well as on Bartonella henselae and Escherichia coli, and the assay was found to be highly specific for A. phagocytophilum and the Borrelia species tested (B. burgdorferi, B. parkeri, B. andersonii, and B. bissettii). The analytical sensitivity of the assay is comparable to that of previously described nested PCR assays (A. phagocytophilum, 16S rRNA; B. burgdorferi, fla gene), amplifying the equivalent of one-eighth of an A. phagocytophilum-infected cell and 50 borrelia spirochetes. The dynamic range of the assay for both A. phagocytophilum and B. burgdorferi was >/=4 logs of magnitude. Purified DNA from A. phagocytophilum and B. burgdorferi was spiked into DNA extracted from uninfected ticks and from negative control mouse and human bloods, and these background DNAs were shown to have no significant effect on sensitivity or specificity of the assay. The assay was tested on field-collected Ixodes scapularis ticks and shown to have 100% concordance compared to previously described non-probe-based PCR assays. To our knowledge, this is the first report of a real-time multiplex PCR assay that can be used for the simultaneous and rapid screening of samples for A. phagocytophilum and Borrelia species, two of the most common tick-borne infectious agents in the United States.
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            Development of a novel genus-specific real-time PCR assay for detection and differentiation of Bartonella species and genotypes.

            The genus Bartonella includes numerous species with varied host associations, including several that infect humans. Development of a molecular diagnostic method capable of detecting the diverse repertoire of Bartonella species while maintaining genus specificity has been a challenge. We developed a novel real-time PCR assay targeting a 301-bp region of the ssrA gene of Bartonella and demonstrated specific amplification in over 30 Bartonella species, subspecies, and strains. Subsequent analysis of ssrA sequences was sufficient to discriminate Bartonella species and provided phylogenetic data consistent with that of gltA, a commonly used gene for differentiating Bartonella genotypes. Using this assay, we identified Bartonella DNA in 29% and 47% of blood specimens from elk in Wyoming and cattle in the Republic of Georgia, respectively. Sequence analysis of a subset of genotypes from elk specimens revealed a cluster most closely related to Bartonella capreoli, and genotypes from cattle were identified as Bartonella bovis, both Bartonella species commonly found in wild and domestic ruminants. Considering the widespread geographic distribution and infectivity potential to a variety of hosts, this assay may be an effective diagnostic method for identification of Bartonella infections in humans and have utility in Bartonella surveillance studies.
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              Tick modulation of host immunity: an important factor in pathogen transmission.

              S K Wikel (1999)
              Immunological interactions at the tick host interface involve innate and specific acquired host immune defenses and immunomodulatory countermeasures by the tick. Tick feeding stimulates host immune response pathways involving antigen-presenting cells, cytokines, B-cells, T-cells, circulating and homocytotropic antibodies, granulocytes, and an array of biologically active molecules. In response to host immune defenses, tick-mediated host immunosuppressive countermeasures inhibit: host antibody responses; complement activation; T-cell proliferation; and cytokine elaboration by macrophages and Th1-lymphocytes. Immunosuppressive proteins identified in tick salivary glands and saliva have been partially characterised. Tick-induced host immunosuppression facilitates blood meal acquisition and is an important factor in the transmission/establishment of the tick-borne disease-causing agent, Borrelia burgdorferi. A novel strategy for control of tick-borne pathogens is proposed.
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                Author and article information

                Contributors
                fd16816@bristol.ac.uk
                sd13044@bristol.ac.uk
                serina.filler@bristol.ac.uk
                swaid.abdullah@bristol.ac.uk
                sophie.keyte@bristol.ac.uk
                hannah.newbury@merck.com
                c.r.helps@bristol.ac.uk
                richard.wall@bristol.ac.uk
                s.tasker@bristol.ac.uk
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                20 March 2018
                20 March 2018
                2018
                : 11
                : 201
                Affiliations
                [1 ]ISNI 0000 0004 1936 7603, GRID grid.5337.2, Small Animal Hospital, Langford Vets, , University of Bristol, ; Langford, United Kingdom
                [2 ]ISNI 0000 0004 1936 7603, GRID grid.5337.2, Veterinary Parasitology and Ecology Group, School of Biological Sciences, , University of Bristol, ; Bristol, United Kingdom
                [3 ]ISNI 0000 0004 1936 7603, GRID grid.5337.2, Bristol Veterinary School, , University of Bristol, ; Langford, United Kingdom
                [4 ]MSD Animal Health, Walton Manor, Walton, Milton Keynes, United Kingdom
                [5 ]ISNI 0000 0004 1936 7603, GRID grid.5337.2, Molecular Diagnostic Unit, Diagnostic Laboratories, Langford Vets, , University of Bristol, ; Langford, United Kingdom
                Article
                2789
                10.1186/s13071-018-2789-5
                5861659
                29558992
                5aea5fe0-32fc-4e0e-8bc7-99fa3716c636
                © The Author(s). 2018

                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. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 4 January 2018
                : 7 March 2018
                Funding
                Funded by: Zutshi-Smith Studentship
                Funded by: MSD Animal Health
                Categories
                Research
                Custom metadata
                © The Author(s) 2018

                Parasitology
                feline,tick-borne pathogens,anaplasma phagocytophilum,bartonella henselae,bartonella clarridgeiae,haemoplasma,mycoplasma haemofelis,“candidatus mycoplasma haemominutum”,“candidatus mycoplasma turicensis”,hepatozoon felis,hepatozoon silvestris

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