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      Diversity of Anaplasma and Ehrlichia/Neoehrlichia Agents in Terrestrial Wild Carnivores Worldwide: Implications for Human and Domestic Animal Health and Wildlife Conservation

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          Abstract

          Recently, the incidence and awareness of tick-borne diseases in humans and animals have increased due to several factors, which in association favor the chances of contact among wild animals and their ectoparasites, domestic animals and humans. Wild and domestic carnivores are considered the primary source of tick-borne zoonotic agents to humans. Among emergent tick-borne pathogens, agents belonging to family Anaplasmataceae (Order Rickettsiales) agents stand out due their worldwide distribution and zoonotic potential. In this review we aimed to review the genetic diversity of the tick-transmitted genera Ehrlichia, Anaplasma and “Candidatus Neoehrlichia sp.” in wild carnivores Caniformia (Canidae, Mustelidae and Ursidae) and Feliformia (Felidae, Hyanidae, Procyonidae and Viverridae) worldwide, discussing the implications for human and domestic animal health and wildlife conservation. Red foxes ( Vulpes vulpes) have been identified as hosts for Anaplasma spp. ( A. phagocytophilum, Anaplasma ovis, A. platys), Ehrlichia canis and “ Candidatus Neoehrlichia sp.” (FU98 strain) and may contribute to the maintaenance of A. phagocytophilum in Europe. Raccoons ( Procyon lotor) have been reported as hosts for E. canis, A. bovis, “ Candidatus Neoehrlichia lotoris” and A. phagocytophilum, and play a role in the maintenance of A. phagocytophilum in the USA. Raccoon dogs ( Nyctereutes procyonoides) may play a role as hosts for A. bovis and A. phagocytophilum. New Ehrlichia and/or Anaplasma genotypes circulate in wild canids and felids from South America and Africa. While Ehrlichia sp. closely related to E. canis has been reported in wild felids from Brazil and Japan, Anaplasma sp. closely related to A. phagocytophilum has been detected in wild felids from Brazil and Africa. Red foxes and mustelids (otters) are exposed to E. canis in countries located in the Mediaterranean basin, probably as a consequence of spillover from domestic dogs. Similarly, E. canis occurs in procyonids in North (raccoons in USA, Spain) and South ( Nasua nasua in Brazil) Hemispheres, in areas where E. canis is frequent in dogs. While “ Candidatus Neoehrlichia lotoris” seems to be a common and specific agent of raccoons in the USA, “ Candidatus Neoehrlichia sp.” (FU98 strain) seems to show a broader range of hosts, since it has been detected in red fox, golden jackal ( Canis aureus) and badger ( Meles meles) in Europe so far. Brown ( Ursus arctos) and black ( Ursus americanus) bears seem to play a role as hosts for A. phagocytophilum in the North Hemisphere. Anaplasma bovis has been detected in wild Procyonidae, Canidae and Felidae in Asia and Brazil. In order to assess the real identity of the involved agents, future works should benefit from the application of MLST (Multi Locus Sequence Typing), WGS (Whole Genome Sequencing) and NGS (Next Generation Sequencing) technologies aiming at shedding some light on the role of wild carnivores in the epidemiology of Anaplasmataceae agents.

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          Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and 'HGE agent' as subjective synonyms of Ehrlichia phagocytophila.

          The genera Anaplasma, Ehrlichia, Cowdria, Neorickettsia and Wolbachia encompass a group of obligate intracellular bacteria that reside in vacuoles of eukaryotic cells and were previously placed in taxa based upon morphological, ecological, epidemiological and clinical characteristics. Recent genetic analyses of 16S rRNA genes, groESL and surface protein genes have indicated that the existing taxa designations are flawed. All 16S rRNA gene and groESL sequences deposited in GenBank prior to 2000 and selected sequences deposited thereafter were aligned and phylogenetic trees and bootstrap values were calculated using the neighbour-joining method and compared with trees generated with maximum-probability, maximum-likelihood, majority-rule consensus and parsimony methods. Supported by bootstrap probabilities of at least 54%, 16S rRNA gene comparisons consistently clustered to yield four distinct clades characterized roughly as Anaplasma (including the Ehrlichia phagocytophila group, Ehrlichia platys and Ehrlichia bovis) with a minimum of 96.1% similarity, Ehrlichia (including Cowdria ruminantium) with a minimum of 97.7% similarity, Wolbachia with a minimum of 95.6% similarity and Neorickettsia (including Ehrlichia sennetsu and Ehrlichia risticii) with a minimum of 94.9% similarity. Maximum similarity between clades ranged from 87.1 to 94.9%. Insufficient differences existed among E. phagocytophila, Ehrlichia equi and the human granulocytic ehrlichiosis (HGE) agent to support separate species designations, and this group was at least 98.2% similar to any Anaplasma species. These 16S rRNA gene analyses are strongly supported by similar groESL clades, as well as biological and antigenic characteristics. It is proposed that all members of the tribes Ehrlichieae and Wolbachieae be transferred to the family Anaplasmataceae and that the tribe structure of the family Rickettsiaceae be eliminated. The genus Anaplasma should be emended to include Anaplasma (Ehrlichia) phagocytophila comb. nov. (which also encompasses the former E. equi and the HGE agent), Anaplasma (Ehrlichia) bovis comb. nov. and Anaplasma (Ehrlichia) platys comb. nov., the genus Ehrlichia should be emended to include Ehrlichia (Cowdria) ruminantium comb. nov. and the genus Neorickettsia should be emended to include Neorickettsia (Ehrlichia) risticii comb. nov. and Neorickettsia (Ehrlichia) sennetsu comb. nov.
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            Circulation of four Anaplasma phagocytophilum ecotypes in Europe

            Background Anaplasma phagocytophilum is the etiological agent of granulocytic anaplasmosis in humans and animals. Wild animals and ticks play key roles in the enzootic cycles of the pathogen. Potential ecotypes of A. phagocytophilum have been characterized genetically, but their host range, zoonotic potential and transmission dynamics has only incompletely been resolved. Methods The presence of A. phagocytophilum DNA was determined in more than 6000 ixodid ticks collected from the vegetation and wildlife, in 289 tissue samples from wild and domestic animals, and 69 keds collected from deer, originating from various geographic locations in The Netherlands and Belgium. From the qPCR-positive lysates, a fragment of the groEL-gene was amplified and sequenced. Additional groEL sequences from ticks and animals from Europe were obtained from GenBank, and sequences from human cases were obtained through literature searches. Statistical analyses were performed to identify A. phagocytophilum ecotypes, to assess their host range and their zoonotic potential. The population dynamics of A. phagocytophilum ecotypes was investigated using population genetic analyses. Results DNA of A. phagocytophilum was present in all stages of questing and feeding Ixodes ricinus, feeding I. hexagonus, I. frontalis, I. trianguliceps, and deer keds, but was absent in questing I. arboricola and Dermacentor reticulatus. DNA of A. phagocytophilum was present in feeding ticks and tissues from many vertebrates, including roe deer, mouflon, red foxes, wild boar, sheep and hedgehogs but was rarely found in rodents and birds and was absent in badgers and lizards. Four geographically dispersed A. phagocytophilum ecotypes were identified, that had significantly different host ranges. All sequences from human cases belonged to only one of these ecotypes. Based on population genetic parameters, the potentially zoonotic ecotype showed significant expansion. Conclusion Four ecotypes of A. phagocytophilum with differential enzootic cycles were identified. So far, all human cases clustered in only one of these ecotypes. The zoonotic ecotype has the broadest range of wildlife hosts. The expansion of the zoonotic A. phagocytophilum ecotype indicates a recent increase of the acarological risk of exposure of humans and animals. Electronic supplementary material The online version of this article (doi:10.1186/1756-3305-7-365) contains supplementary material, which is available to authorized users.
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              The natural history of Anaplasma phagocytophilum.

              Anaplasma phagocytophilum is the recently designated name replacing three species of granulocytic bacteria, Ehrlichia phagocytophila, Ehrlichia equi and the agent of human granulocytic ehrlichiosis, after the recent reorganization of the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales. Tick-borne fever (TBF), which is caused by the prototype of A. phagocytophilum, was first described in 1932 in Scotland. A similar disease caused by a related granulocytic agent was first described in horses in the USA in 1969; this was followed by the description of two distinct granulocytic agents causing similar diseases in dogs in the USA in 1971 and 1982. Until the discovery of human granulocytic anaplasmosis (HGA) in the USA in 1994, these organisms were thought to be distinct species of bacteria infecting specific domestic animals and free-living reservoirs. It is now widely accepted that the agents affecting different animal hosts are variants of the same Gram-negative obligatory intracellular bacterium, which is transmitted by hard ticks belonging to the Ixodes persulcatus complex. One of its fascinating features is that it infects and actively grows in neutrophils by employing an array of mechanisms to subvert their bactericidal activity. It is also able to survive within an apparently immune host by employing a complex mechanism of antigenic variation. Ruminants with TBF and humans with HGA develop severe febrile reaction, bacteraemia and leukopenia due to neutropenia, lymphocytopenia and thrombocytopenia within a week of exposure to a tick bite. Because of the severe haematological disorders lasting for several days and other adverse effects on the host's immune functions, infected animals and humans are more susceptible to other infections.
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                Author and article information

                Contributors
                Journal
                Front Vet Sci
                Front Vet Sci
                Front. Vet. Sci.
                Frontiers in Veterinary Science
                Frontiers Media S.A.
                2297-1769
                23 November 2018
                2018
                : 5
                : 293
                Affiliations
                Laboratório de Imunoparasitologia, Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista (FCAV/UNESP) , Jaboticabal, Brazil
                Author notes

                Edited by: Michael Kosoy, Centers for Disease Control and Prevention (CDC), United States

                Reviewed by: Allen L. Richards, Naval Medical Research Center, United States; Irina Goodrich, Centers for Disease Control and Prevention (CDC), United States

                *Correspondence: Marcos Rogério André marcosandre.fcav@ 123456gmail.com

                This article was submitted to Parasitology, a section of the journal Frontiers in Veterinary Science

                Article
                10.3389/fvets.2018.00293
                6265506
                30533417
                5c2a5ed3-403e-499f-9443-fdd5b8a37897
                Copyright © 2018 André.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 23 August 2018
                : 01 November 2018
                Page count
                Figures: 0, Tables: 2, Equations: 0, References: 127, Pages: 24, Words: 18577
                Categories
                Veterinary Science
                Review

                anaplasma,ehrlichia,“candidatus neoehrlichia sp.”,carnivora,genetic diversity,ticks

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