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      Establishment of Babesia vulpes n. sp. (Apicomplexa: Babesiidae), a piroplasmid species pathogenic for domestic dogs

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          Abstract

          Background

          Canine babesiosis is a severe disease caused by several Babesia spp. A number of names have been proposed for the canine-infecting piroplasmid pathogen initially named Theileria annae Zahler, Rinder, Schein & Gothe, 2000. It was shown to be a member of the Babesia ( sensu lato) group infecting carnivores and is also closely related to the Babesia microti group. Subsequently, the same parasite species was reclassified as a member of the genus Babesia and the name Babesia vulpes Baneth, Florin-Christensen, Cardoso & Schnittger, 2015 was proposed for it. However, both names do not meet the requirements of the International Code of Zoological Nomenclature (no accompanying descriptions, no deposition of type-specimens) and cannot be recognized as available names from the nomenclatural point of view. The purpose of this study was to further characterize this parasite in order to confirm its validity, to provide its description and to introduce zoological nomenclature for it with the name Babesia vulpes n. sp.

          Results

          Morphological description of the parasite in canine erythrocytes demonstrated that it takes the shape of small (1.33 × 0.98 µm), round to oval forms reminiscent of the pyriform and ring shapes of other small canine Babesia spp., such as Babesia gibsoni Patton, 1910 and Babesia conradae Kjemtrup, Wainwright, Miller, Penzhorn & Carreno, 2006. However, these parasite forms were overall smaller than those measured for the latter two species and no tetrad (Maltese cross) form was reported. Furthermore, phylogenetic analysis using the cytochrome c oxidase subunit 1 (COX1) amino acid sequences substantiates the species identity of this parasite as previously demonstrated based on phylogenetic analysis of the 18S rRNA and β-tubulin genes. The holotype of the parasite species was designated and deposited in an accessible public collection.

          Conclusions

          This study ratifies the name Babesia vulpes n. sp. proposed for the parasite previously referred to as Theileria annae Zahler, Rinder, Schein & Gothe, 2000, Babesia annae (Zahler, Rinder, Schein & Gothe, 2000) or Babesia vulpes Baneth, Florin-Christensen, Cardoso & Schnittger, 2015, or mentioned as “ Babesia microti-like piroplasm”, “ Babesia Spanish dog isolate” and Babesia cf. microti.

          Electronic supplementary material

          The online version of this article (10.1186/s13071-019-3385-z) contains supplementary material, which is available to authorized users.

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          Most cited references42

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          Amendment of Articles 8, 9, 10, 21 and 78 of the International Code of Zoological Nomenclature to expand and refine methods of publication

          (2012)
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            Reclassification of Theileria annae as Babesia vulpes sp. nov.

            Background Theileria annae is a tick-transmitted small piroplasmid that infects dogs and foxes in North America and Europe. Due to disagreement on its placement in the Theileria or Babesia genera, several synonyms have been used for this parasite, including Babesia Spanish dog isolate, Babesia microti-like, Babesia (Theileria) annae, and Babesia cf. microti. Infections by this parasite cause anemia, thrombocytopenia, and azotemia in dogs but are mostly subclinical in red foxes (Vulpes vulpes). Furthermore, high infection rates have been detected among red fox populations in distant regions strongly suggesting that these canines act as the parasite’s natural host. This study aims to reassess and harmonize the phylogenetic placement and binomen of T. annae within the order Piroplasmida. Methods Four molecular phylogenetic trees were constructed using a maximum likelihood algorithm based on DNA alignments of: (i) near-complete 18S rRNA gene sequences (n = 76 and n = 93), (ii) near-complete and incomplete 18S rRNA gene sequences (n = 92), and (iii) tubulin-beta gene sequences (n = 32) from B. microti and B. microti-related parasites including those detected in dogs and foxes. Results All phylogenetic trees demonstrate that T. annae and its synonyms are not Theileria parasites but are most closely related with B. microti. The phylogenetic tree based on the 18S rRNA gene forms two separate branches with high bootstrap value, of which one branch corresponds to Babesia species infecting rodents, humans, and macaques, while the other corresponds to species exclusively infecting carnivores. Within the carnivore group, T. annae and its synonyms from distant regions segregate into a single clade with a highly significant bootstrap value corroborating their separate species identity. Conclusion Phylogenetic analysis clearly shows that T. annae and its synonyms do not pertain to Theileria and can be clearly defined as a separate species. Based on the facts that T. annae and its synonyms have not been shown to have a leukocyte stage, as expected in Theileria, do not infect humans and rodents as B. microti, and cluster phylogenetically as a separate species, this study proposes to name this parasite Babesia vulpes sp. nov., after its natural host, the red fox V. vulpes. Electronic supplementary material The online version of this article (doi:10.1186/s13071-015-0830-5) contains supplementary material, which is available to authorized users.
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              Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe. Part I. Epizootiological aspects.

              Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced. Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following: Piroplasmid infections: Approximately from 50 to 70% of wild or domestic mammals (symptomatic) were infected. Piroplasmids detected in ruminants were:COW: B. bovis, T. annulata and Theileria sp. (type C). Sheep and goat: B. ovis. Piroplasmids present in canids were: Babesia canis vogeli, Babesia canis canis, Theileria annae and B. equi. The only piroplasmid found in asymptomatic dogs was B. equi. Piroplasmids found in horse were: B. equi and B. canis canis.H. canis infections in canids: H. canis was absent of domestic dog samples, whereas all foxes studied were infected by this protozoa. Genetic analysis showed that most of piroplasmid and Hepatozoon isolates from southern Europe matched unambigously with previously described species, as demonstrated by the high level sequence identity between them, usually between 99 and 100%. Minor differences, usually detected in hypervariable regions of 18s rRNA gene are probably due to strain variations or rare genetic polymorphisms. A possible exception was B. bovis, which shows a relatively lower degree of homology (94%) with regard to other B. bovis isolates from several countries. The same is true for B. ovis, that showed a 94% identity with regard to Babesia sp. from South African cow and a 92% with rapport to B. bovis from Portugal.
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                Author and article information

                Contributors
                gad.baneth@mail.huji.ac.il
                lcardoso@utad.pt
                paulabrilhante@inno.pt
                schnittger.leonhard@inta.gob.ar
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                26 March 2019
                26 March 2019
                2019
                : 12
                : 129
                Affiliations
                [1 ]ISNI 0000 0004 1937 0538, GRID grid.9619.7, Koret School of Veterinary Medicine, , Hebrew University, ; P.O. Box 12, 76100 Rehovot, Israel
                [2 ]ISNI 0000000121821287, GRID grid.12341.35, Department of Veterinary Sciences, and Animal and Veterinary Research Centre (CECAV), , University of Trás-os-Montes e Alto Douro (UTAD), ; Vila Real, Portugal
                [3 ]Inno – Serviços Especializados em Veterinária, Braga, Portugal
                [4 ]ISNI 0000 0001 2167 7174, GRID grid.419231.c, Instituto de Patobiología Veterinaria, (CICVyA), , Instituto Nacional de Tecnología Agropecuaria (INTA), ; 1686 Hurlingham, Argentina
                [5 ]ISNI 0000 0001 1945 2152, GRID grid.423606.5, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), ; Buenos Aires, Argentina
                Author information
                http://orcid.org/0000-0002-7549-1305
                Article
                3385
                10.1186/s13071-019-3385-z
                6434798
                30909951
                a359f35e-4c29-4a6c-98ab-153597c5f0b4
                © The Author(s) 2019

                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
                : 27 November 2018
                : 7 March 2019
                Categories
                Research
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                © The Author(s) 2019

                Parasitology
                babesia vulpes n. sp.,babesia microti,babesia cf. microti,babesia microti-like piroplasm,theileria annae,“babesia (theileria) annae”,babesia annae,“babesia spanish dog isolate”,red fox,dog

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