title: Description of three new species of Hepatozoon (Apicomplexa, Hepatozoidae) from Rattlesnakes (Crotalus durissus terrificus) based on molecular, morphometric and morphologic characters.
abstract: 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.
authors.affiliation: Instituto de Biociências de Botucatu, Unesp - Univ Estadual Paulista, Campus de Botucatu, Departamento de Parasitologia, Distrito de Rubião Júnior, Botucatu, SP 18618-970, Brazil. Electronic address: odwyer@ibb.unesp.br.
authors.forename: Lucia Helena
authors.initials: LH
authors.lastname: O'Dwyer
authors.forename: Tatiana Cristina
authors.initials: TC
authors.lastname: Moço
authors.forename: Karina dos Santos
authors.initials: Kdos S
authors.lastname: Paduan
authors.forename: Carine
authors.initials: C
authors.lastname: Spenassatto
authors.forename: Reinaldo José
authors.initials: RJ
authors.lastname: da Silva
authors.forename: Paulo Eduardo Martins
authors.initials: PE
authors.lastname: Ribolla
mesh_terms: D000818:Animals; D001938:Brazil / Q000453:epidemiology; D003044:Coccidia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D003048:Coccidiosis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D017839:Crotalus / Q000469:parasitology*; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics; D004912:Erythrocytes / Q000469:parasitology; D005260:Female; D007091:Image Processing, Computer-Assisted; D008297:Male; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D015995:Prevalence; D016415:Sequence Alignment
chemical_list: D016054:DNA, Protozoan; D004275:DNA, Ribosomal
keywords: Hepatozoon; Molecular characterization; Morphology; Rattlesnakes