Article

Molecular characterization of Hepatozoon sp from Brazilian dogs and its phylogenetic relationship with other Hepatozoon spp

Oklahoma State University - Stillwater, SWO, Oklahoma, United States
Veterinary Parasitology (Impact Factor: 2.55). 05/2007; 145(1-2):21-30. DOI: 10.1016/j.vetpar.2006.10.023
Source: PubMed

ABSTRACT To characterize phylogenetically the species which causes canine hepatozoonosis at two rural areas of Rio de Janeiro State, Brazil, we used universal or Hepatozoon spp. primer sets for the 18S SSU rRNA coding region. DNA extracts were obtained from blood samples of thirteen dogs naturally infected, from four experimentally infected, and from five puppies infected by vertical transmission from a dam, that was experimentally infected. DNA of sporozoites of Hepatozoon americanum was used as positive control. The amplification of DNA extracts from blood of dogs infected with sporozoites of Hepatozoon spp. was observed in the presence of primers to 18S SSU rRNA gene of Hepatozoon spp., whereas DNA of H. americanum sporozoites was amplified in the presence of either universal or Hepatozoon spp.-specific primer sets; the amplified products were approximately 600bp in size. Cloned PCR products obtained from DNA extracts of blood from two dogs experimentally infected with Hepatozoon sp. were sequenced. The consensus sequence, derived from six sequence data sets, were blasted against sequences of 18S SSU rRNA of Hepatozoon spp. available at GenBank and aligned to homologous sequences to perform the phylogenetic analysis. This analysis clearly showed that our sequence clustered, independently of H. americanum sequences, within a group comprising other Hepatozoon canis sequences. Our results confirmed the hypothesis that the agent causing hepatozoonosis in the areas studied in Brazil is H. canis, supporting previous reports that were based on morphological and morphometric analyses.

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    • "Many recent studies have shown that canine babesiosis (caused by Babesia vogeli), hepatozoonosis (caused by Hepatozoon canis), monocytic ehrlichiosis (caused by Ehrlichia canis), and anaplasmosis (caused by Anaplasma platys) are endemic in many parts of the country (RAMOS et al., 2010; SPOLIDORIO et al., 2011; VIEIRA et al., 2011; COSTA-JÚNIOR et al., 2012; SILVA et al., 2012; DEMONER et al., 2013). While H. canis seems to be primarily associated with ticks of the genus Amblyomma in Brazil (FORLANO et al., 2007; DEMONER et al., 2013), the other agents mentioned above have been associated with Rhipicephalus sanguineus sensu lato ticks (DANTAS-TORRES, 2008; VIEIRA et al., 2011). A recent study described clinical illness in domestic dogs due to Rickettsia rickettsii, the agent of Rocky Mountain spotted fever (RMSF), for the first time in Brazil (LABRUNA et al., 2009). "
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    ABSTRACT: Blood samples were collected from 99 domestic dogs from the urban and rural areas of the Lábrea municipality, state of Amazonas, Brazil. Canine serum samples were tested by immunofluorescence assay against Rickettsia spp., which revealed that only 3.0% (1/33) and 7.6% (5/66) of the dogs from urban and rural areas, respectively, reacted positively to at least one Rickettsia species. DNA was extracted from canine blood and tested by a battery of PCR assays targeting protozoa of the genera Babesia and Hepatozoon, and bacteria of the genera Rickettsia and Ehrlichia and family Anaplasmataceae. All samples were negative in the PCR assays targeting the genera Babesia, Hepatozoon, Ehrlichia and Rickettsia. For Anaplasmataceae, 3% (1/33) and 39.4% (26/66) of the urban and rural dogs, respectively, yielded amplicons that generated DNA sequences 100% identical to the corresponding sequence of Wolbachia endosymbiont of Dirofilaria immitis. Because of these results, all canine DNA samples were further tested in a PCR assay targeting filarial nematodes, which was positive for 18.2% (6/33) and 57.6% (38/66) urban and rural dogs, respectively. Filarial-PCR products generated DNA sequences 100% identical to D. immitis. While tick-borne infections were rare in Lábrea, D. immitis infection rates were among the highest reported in South America.
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    • "During the cycle in arthropod vectors , oocysts containing sporocysts and sporozoites are formed in the hemocoel and are transmitted when infected ticks are ingested by intermediate vertebrate hosts. Several tick species have been incriminated as potential vectors (Forlano et al., 2007); this paper reports on the presence of H. canis oocysts in Rhipicephalus (Boophilus) microplus collected from an infected dog. "
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    • "In the genus Hepatozoon, two species have been described infecting dogs, H. canis and Hepatozoon americanum (Baneth et al. 2000). However, only H. canis has been described in Brazil, based on molecular studies (Rubini et al. 2005; Forlano et al. 2007). E. canis and A. platys are obligatory intracellular rickettsia with tropism for leukocytes and platelets, respectively. "
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