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      Hepatozoon silvestris sp. nov.: morphological and molecular characterization of a new species of Hepatozoon (Adeleorina: Hepatozoidae) from the European wild cat ( Felis silvestris silvestris)

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          SUMMARY

          Based on morphological and genetic characteristics, we describe a new species of Hepatozoon in the European wild cat ( Felis silvestris silvestris), herein named Hepatozoon silvestris sp. nov. The study also provides the first data on the occurrence of H. felis in this wild felid. Hepatozoon meronts were observed in multiple cross-sections of different organs of four (44%) cats. Additionally, extracellular forms, resembling mature gamonts of Hepatozoon, were found in the spleen and myocardium of two cats. Furthermore, tissues of six animals (67%) were positive by PCR. Hepatozoon felis was identified infecting one cat (11%), whereas the 18S rRNA sequences of the remaining five cats (56%) were identical, but distinct from the sequences of H. felis. Phylogenetic analyses revealed that those sequences form a highly supported clade distant from other Hepatozoon spp. Future studies should include domestic cats from the areas where the wild cats positive for H. silvestris sp. nov. were found, in order to investigate their potential role to serve as intermediate hosts of this newly described species. Identification of its definitive host(s) and experimental transmission studies are required for elucidating the full life cycle of this parasite and the possible alternative routes of its transmission.

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          Precision Farming: Technologies and Information as Risk-Reduction Tools

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            Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR.

            We have identified a novel 529bp fragment that is repeated 200- to 300-fold in the genome of Toxoplasma gondii. This 529bp fragment was utilised for the development of a very sensitive and specific PCR for diagnostic purposes, and a quantitative competitive-PCR for the evaluation of cyst numbers in the brains of chronically infected mice. The 529bp fragment was found in all 60 strains of T. gondii tested, and it discriminates DNA of T. gondii from that of other parasites. Toxoplasma gondii DNA was detected in amniotic fluid of patients, as well as in various tissues from infected mice. Polymerase chain reaction with the 529bp fragment was more sensitive than with the 35-copy B1 gene. For the quantitative competitive-PCR, a 410-bp competitor molecule was co-amplified with similar efficiency as the 529bp fragment. Quantitative competitive-PCR produced a linear relationship between the relative amounts of PCR product and the number of tachyzoites in the range of 10(2)-10(4) tachyzoites and 100-3000 tissue cysts. A highly significant correlation between visual counting of brain cysts and quantitative competitive-PCR was obtained in mice chronically infected with Toxoplasma. Thus, quantitative competitive-PCR with the 529bp fragment can be used as an alternative for the tedious visual counting of brain cysts in experimental animals. With the quantitative competitive-PCR, furthermore, we could confirm the copy number of the 529bp fragment in tachyzoites and estimate the number of bradyzoites per cyst.
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              The genus Hepatozoon (Apicomplexa: Adeleina).

              Hemogregarines of the genus Hepatozoon are intraerythrocytic apicomplexan parasites that have been described from all groups of tetrapod vertebrates. Gametogenesis, fertilization, and sporogonic development, which culminates in the formation of polysporocystic oocysts, occur in the gut or hemocoel of a hematophagous arthropod definitive host. Merogonic development occurs in the internal organs of vertebrate hosts after they ingest these infected arthropods. The presence of cystic stages, observed for many Hepatozoon species, increases life cycle complexity and exploits the feeding behavior of vertebrate hosts. The inconsistency of morphological characteristics of these parasites, especially those associated with gamont structure, coupled with low host specificity of the parasites for their invertebrate and vertebrate hosts, have rendered species differentiation difficult. A systematic review of the hemogregarine complex has resulted in the expansion of the genus Hepatozoon to include all members of the genus Haemogregarina that infect amphibians, snakes, lizards, crocodilians, birds, and mammals.
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                Author and article information

                Journal
                Parasitology
                Parasitology
                PAR
                Parasitology
                Cambridge University Press (Cambridge, UK )
                0031-1820
                1469-8161
                April 2017
                12 December 2016
                : 144
                : 5
                : 650-661
                Affiliations
                [1 ]Department of Pathobiology, Institute of Parasitology, University of Veterinary Medicine Vienna , Veterinaerplatz 1, 1210 Vienna, Austria
                [2 ]Department of Pathology, Faculty of Veterinary Medicine, University of Sarajevo , Zmaja od Bosne 90, 71000 Sarajevo,Bosnia and Herzegovina
                [3 ]Department of Veterinary Medicine, University of Bari , Str. prov. per Casamassima km 3, 70010 Valenzano, Italy
                [4 ]Koret School of Veterinary Medicine, Hebrew University of Jerusalem , P.O. Box 12, 76100 Rehovot, Israel
                Author notes
                [* ]Corresponding author: Department of Pathobiology, Institute of Parasitology, University of Veterinary Medicine Vienna , Veterinaerplatz 1, 1210 Vienna, Austria. E-mail: georg.duscher@ 123456vetmeduni.ac.at
                Article
                S0031182016002316 00231
                10.1017/S0031182016002316
                5426326
                27938443
                3be6b44b-756e-45ff-9726-264648169768
                © Cambridge University Press 2016

                This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 26 August 2016
                : 04 November 2016
                : 04 November 2016
                Page count
                Figures: 5, Tables: 2, References: 47, Pages: 12
                Categories
                Research Article

                Parasitology
                hepatozoon silvestris sp. nov.,hepatozoon felis,european wild cat,felis silvestris silvestris,morphology,molecular characterization,bosnia and herzegovina

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