Description of three new species of Hepatozoon (Apicomplexa, Hepatozoidae) from Rattlesnakes (Crotalus durissus terrificus) based on molecular, morphometric and morphologic characters

Hepatozoon spp. are commonly found infecting snakes. Since the latter are parasitized by diverse forms and data in the literature show divergence, we studied Hepatozoon spp. diversity on Crotalus durissus terrificus snakes using both molecular and morphological approaches. Naturally infected animals were employed. Blood was collected, blood smears were prepared and an aliquot was stored at -20. °C for DNA extraction. Five specimens of C. durissus terrificus were selected, each of them infected with one gamont type. Morphological and morphometric analyses of the found gamonts led to their grouping into three populations. For molecular characterization, seven oligonucleotide pairs that amplify distinct regions of rDNA gene were tested by adopting the PCR technique. Only the oligonucleotide pairs HepF300/Hep900 and HEMO1/HEMO2 were efficient in amplifying and distinguishing different isolates of Hepatozoon spp. from snakes. The better results were obtained when both oligonucleotide pairs were used in association. Based on the molecular and morphologic differences, three new species were proposed: Hepatozoon cuestensis sp. nov.; Hepatozoon cevapii sp. nov. and Hepatozoon massardii sp. nov. This is the first description of new Hepatozoon species from snakes, based on molecular characterization and morphological data, in South America. © 2013 Elsevier Inc.

Hepatozoon griseisciuri in grey squirrels (Sciurus carolinensis): changes of blood leucocyte numbers resulting from infection

Numbers of leucocytes in squirrels with gametocytes of Hepatozoon in their blood (infected) were compared with animals without gametocytes (uninfected). Typical values for leucocytes/mm(3) blood in uninfected squirrels were: leucocytes 5-7 x 10(3), granulocytes 3-4 x 10(3), lymphocytes 2-0 x 10(3) and monocytes 0-3 x 10(3) cells. Infection caused an increase in monocytes, lymphocytes and granulocytes, and there was a significant positive association between parasitaemia level and numbers of both total leucocytes and monocytes. Infected animals had more uninfected monocytes/mm(3) blood than did uninfected animals. The proportions of monocytes were more variable over time in infected animals, but no shift between infected and uninfected status was detected. Transfer of serum from infected squirrels to mice resulted in elevated counts of total blood leucocytes, monocytes and granulocytes, but not of lymphocytes, as compared with controls. Serum from squirrels with high parasitaemias had a more marked effect than serum from squirrels with low parasitaemias. Results indicate an infection - related monocytosis, possibly controlled by cytokines, that increases the number of cells available for invasion by gametocytes, thus enhancing the chances of parasite transmission.

Morphologic and morphometric analysis of Hepatozoon spp. (Apicomplexa, Hepatozoidae) of snakes

Hepatozoon species are the most abundant hemoparasites of snakes. Its identification has been based mainly on the morphologic characterization of the gamonts in the peripheral blood of the vertebrate host and also of the cysts found in the internal organs of the vertebrate and invertebrate hosts. Using a computerized image analysis system, we studied five species of Hepatozoon from recently captured snakes in Botucatu, State of São Paulo, Brazil, to evaluate the importance of the morphology and morphometry of the gamonts for the characterization of Hepatozoon species and to analyze the morphologic changes induced in the erythrocytes by the parasite. The studied species were H. terzii of Boa constrictor amarali, Hepatozoon sp. of Crotalus durissusterrificus, H. philodryasi of Philodryas patagoniensis, and H. migonei and H. cyclagrasi of Hydrodynastes gigas. We observed three different groups, one of them including the species H. terzii, H. philodryasi and Hepatozoon sp. of C. durissus terrificus; and the other two consisting of H. migonei and H. cyclagrasi. Degree of alterations in the erythrocytes was variable and it may be useful for characterization of Hepatozoon species.

Molecular characterization of Hepatozoon sp in cats from Sao Luis Island, Maranhao, Northeastern Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological, morphometric, and molecular characterization of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from naturally infected Caudisona durissa terrifica (Serpentes, Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Description of gamontogonic and sporogonic stages of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from Caudisoma durissa terrifica (Serpentes, Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hepatozoon canis infection associated with dog ticks of rural areas of Rio de Janeiro State, Brazil

Hepatozoon canis is a tick-borne protozoan that infects dogs and has been reported throughout the world. Manifestation of H. canis infection varies from being sub-clinical in apparently healthy dogs to severe illness. The main vector of the infection is the dog tick, Rhipicephalus sanguineus although other species may also transmit this agent. H. canis has been reported previously in Brazil, but mostly as an occasional finding during laboratory exams and always associated with other diseases. The prevalence of H. canis in dogs of rural areas of Brazil has been little studied. For this study, 250 dogs from seven counties of Rio de Janeiro state were examined. All the dogs were from rural areas, near forest. of the dogs examined, 26 dogs were from Seropedica, 82 from Itaguai, 41 from Paracambi, 26 from Mangaratiba, 32 from Barra do Pirai, 32 from Pirai and 11 from Miguel Pereira. Blood smears from the peripheral blood of the ear were taken and ticks found on the dogs were collected for identification in the laboratory. Using blood smear evaluation, H. canis was identified in 39.2% of the animals examined. Other hemoparasites identified were Babesia canis (5.2%) and Ehrlichia canis (4.8%). Four tick species were found parasitizing the dogs: Amblyomma cajennense (23.6%), R. sanguineus (12.4%), Amblyomma aureolatum (2.8%) and Amblyomma ovale (2.0%). There was a positive correlation between the presence of A. cajennense and H. canis infection. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Seasonal variation of Hepatozoon spp. (Apicomplexa, Hepatozoidae) parasitemia from Boa constrictor amarali (Serpentes, Boidae) and Hydrodynastes gigas (Serpentes, Colubridae)

The aim of this study was to evaluate the parasitemia variation of three Hepatozoon species in Brazilian snakes. This study was conducted between 2001 and 2003 and included Hepatozoon terzii from Boa constrictor amarali, and Hepatozoon migonei and Hepatozoon cyclagrasi from Hydrodynastes gigas. It was observed that the parasitemia tended to decrease in all three Hepatozoon species but the parasites were not eliminated. This data suggest that Hepatozoon infection may be similar to Toxoplasma gondii infection, in that it persists throughout host life.

Molecular identification and characterization of canine Hepatozoon species from Brazil

Canine Hepatozoon species from Brazil was molecular identified and characterized for the first time. From 31 dogs, 7 were positive for blood smear examination and 21 positive for PCR. Partial sequences of the 18S rRNA gene from eight naturally infected dogs were analyzed. Sequences revealed that Brazilian Hepatozoon is closely related with the Japanese Hepatozoon, that has 99% nucleotide identity with Hepatozoon canis from Israel, and different from Hepatozoon americanum. These results indicate that the canine Hepatozoon species from Brazil is H. canis.

Detection of Hepatozoon spp in naturally infected Brazilian dogs by polymerase chain reaction

Hepatozoon canis was molecularly identified in Rio de Janeiro State, Brazil. Twelve dogs from urban areas were studied by blood smear examination and the polymerase chain reaction (PCR) assay. From these dogs, only 1 was positive in both blood smears and PCR.

Prevalence of Hepatozoon spp. (Apicomplexa, Hepatozoidae) among recently captured Brazilian snakes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Oocysts of Hepatozoon canis in Rhipicephalus (Boophilus) microplus collected from a naturally infected dog

Canine hepatozoonosis is a tick-borne disease caused by protozoans of the genus Hepatozoon. Several tick species have been implicated as potential vectors. Therefore, extensive studies are needed to determine the 'natural' endemic cycle of this parasite. This paper presents the first report of the presence of Hepatozoon canis oocysts in Rhipicephalus (Boophilus) microplus collected from an infected dog. (C) 2011 Elsevier B.V. All rights reserved.

Prevalence, hematology and serum biochemistry in stray dogs naturally infected by Hepatozoon canis in São Paulo

Cães capturados nas ruas de quatro municípios do estado de São Paulo foram examinados para estudar a prevalência de Hepatozoon canis. Dos 222 animais, 13 (5.9%) encontravam-se infectados. Exames hematológicos realizados em nove animais evidenciaram três com anemia e um com leucocitose. A única alteração encontrada nos exames bioquímicos foi hiperglobulinemia.

Description of the gamonts of a small species of Hepatozoon sp (Apicomplexa, Hepatozoidae) found in Crotalus durissus terrificus (Serpentes, Viperidae)

A small species of the genus Hepatozoon found in a specimen of Crotalus durissus terrificus from the Botucatu region, São Paulo State, Brazil is described. The morphologic alterations induced in the snake's erythrocytes by the presence of this parasite are described. Morphology and morphometric analyses were performed using the Qwin Lite 2.5 computerized image analysis system (Leica). The Hepatozoon possessed a small and short body (8.1+/-0.5 mum long and 3.8+/-0.4 mum wide), with round extremities. The cytoplasm varied from pale blue to basophilic and had no granulations. Its nucleus was large, occupied a large area of the cytoplasm, and was irregular in shape and not condensed. Despite its small size, this parasite induced important changes in the host cell. Total parasitemia observed was 56.6%.

Molecular characterization of feline Hepatozoon species from Brazil

Feline Hepatozoon species from Brazil was molecular identified and characterized for the first time in São Paulo state, Brazil. Partial sequences of the 18S rRNA gene from the Hepatozoon from three naturally infected cats were analyzed. Sequences revealed that feline Hepatozoon was closely related to the canine Hepatozoon canis from Brazil. (C) 2005 Elsevier B.V All rights reserved.