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Experimental infection of horses with Rickettsia rickettsii

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      Abstract

      Background

      Rickettsia rickettsii is vectored by ticks, and some vertebrate hosts can be sources of infection to ticks during bacteremic periods. In Brazil, the main vector for R. rickettsii is the tick Amblyomma sculptum, a member of the A. cajennense complex. Horses, in turn, are one of the major hosts for A. sculptum. In this study, horses experimentally infected with R. rickettsii were assessed for clinical changes and their capability to transmit the infection to A. sculptum ticks.

      Methods

      Four horses were infected with R. rickettsii through either intraperitoneal injection or infestation with R. rickettsii-infected A. sculptum ticks. Simultaneously, the animals were infested with non-infected A. sculptum ticks. The horses were monitored for 30 days by clinical examination, hematological and biochemical tests, real-time PCR of blood for the detection of Rickettsia, and inoculation of blood in guinea pigs. IgG antibody titers were followed until the horses have shown seronegativity or until the end of the experiment. Uninfected ticks that fed on horses were subjected to real-time PCR and/or were fed on susceptible rabbits.

      Results

      The horses showed no clinical, hematological or blood biochemical alterations, and bacteremia was not detected by real-time PCR or by inoculation of horse blood into guinea pigs. Anti- R. rickettsii antibodies were detected in horses from 10 days to 2 years after infection. Uninfected ticks, after feeding on infected horses, showed 2.1 % positivity in real-time PCR, but failed to transmit the infection to rabbits at a next feeding stage.

      Conclusions

      Rickettsia rickettsii-infected horses did not manifest illness and are not competent amplifier hosts of R. rickettsii for A. sculptum ticks.

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      Most cited references 37

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      Essentals of Veterinary hematology

       NC JAIN,  N.C JAIN,  NC Jain (1993)
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        Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes.

        DNA sequences from specific genes, amplified by the polymerase chain reaction technique, were used as substrata for nonisotopic restriction endonuclease fragment length polymorphism differentiation of rickettsial species and genotypes. The products amplified using a single pair of oligonucleotide primers (derived from a rickettsial citrate synthase gene sequence) and cleaved with restriction endonucleases were used to differentiate almost all recognized species of rickettsiae. A second set of primers was used for differentiation of all recognized species of closely related spotted fever group rickettsiae. The procedure circumvents many technical obstacles previously associated with identification of rickettsial species. Multiple amplified DNA digest patterns were used to estimate the intraspecies nucleotide sequence divergence for the genes coding for rickettsial citrate synthase and a large antigen-coding gene of the spotted fever group rickettsiae. The estimated relationships deduced from these genotypic data correlate reasonably well with established rickettsial taxonomic schemes.
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          Phylogeny of hard- and soft-tick taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences.

           W Black,  J Piesman (1994)
          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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            Author and article information

            Affiliations
            [1 ]Agribusiness Technology Agency of São Paulo State, São José do Rio Preto, Brazil
            [2 ]Faculty of Veterinary Medicine, University of São Paulo, São Paulo, Brazil
            Contributors
            tatianaueno@apta.sp.gov.br
            franc.borges@yahoo.com.br
            jonasmfilho@hotmail.com
            wca@usp.br
            wilsonrf@usp.br
            labruna@usp.br
            Journal
            Parasit Vectors
            Parasit Vectors
            Parasites & Vectors
            BioMed Central (London )
            1756-3305
            13 September 2016
            13 September 2016
            2016
            : 9
            : 1
            27624315
            5022194
            1784
            10.1186/s13071-016-1784-y
            © The Author(s). 2016

            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.

            Funding
            Funded by: FundRef http://dx.doi.org/10.13039/501100002322, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior;
            Award ID: CAPES/PROEX 2327/2015
            Award Recipient :
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            © The Author(s) 2016

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