title: Plasmodium (Novyella) nucleophilum from an Egyptian Goose in São Paulo Zoo, Brazil: microscopic confirmation and molecular characterization.
abstract: Plasmodium (Novyella) nucleophilum was identified using microscopy and PCR, in an Egyptian Goose (Alopochen aegyptiacus) that died in São Paulo Zoo, Brazil. This parasite is characterized by elongated gametocytes, small meronts with scant cytoplasm, less than eight merozoites and mainly for having all the stages appressed to the nuclei of infected erythrocytes. Additionally, Plasmodium (Haemamoeba) sp. was identified by microscopy in the same blood sample. The latter parasite lacks nucleophilic blood stages and is characterized by large roundish trophozoites, each with a large prominent centrally collated vacuole. This co-infection was not confirmed by PCR amplification of the mitochondrial cytochrome b (cytb) gene and sequencing; only one Plasmodium sp. cytb sequence was detected in the blood sample. Since parasitemia of P. nucleophilum (2.4%) was much higher than that of P. (Haemamoeba) sp. (0.2%), PCR may have favored the amplification of the cytb sequence of the former. Phylogenetic analysis is in agreement with this conclusion because the reported cytb sequence was positioned in the same branch of sequences of several Novyella species. This is the first assignment of the mitochondrial cytb gene sequence to P. nucleophilum. The P. (Haemamoeba) parasite is particularly similar to Plasmodium (Haemamoeba) tejerai, because its advanced trophozoites and young erythrocytic meronts possess a large vacuole with prominent pigment granules arranged around it, the characteristic features of development in this species. For definitive identification of P. (Haemamoeba) species, mature meronts and gametocytes are required; however, these were absent from the thin blood smear. Representative images of the blood stages of P. nucleophilum and P. (Haemamoeba) sp. are provided. Together with microscopy data, the P. nucleophilum cytb sequence will assist in molecular identification (barcoding) of this Plasmodium species in other birds.
authors.affiliation: Zoological Park Foundation, São Paulo, Brazil.
authors.forename: Carolina Romeiro Fernandes
authors.initials: CR
authors.lastname: Chagas
authors.affiliation: Institute of Ecology, Nature Research Centre, Lithuania.
authors.forename: Gediminas
authors.initials: G
authors.lastname: Valkiūnas
authors.affiliation: Zoological Park Foundation, São Paulo, Brazil.
authors.forename: Carolina Vaz Cabral
authors.initials: CV
authors.lastname: Nery
authors.affiliation: Zoological Park Foundation, São Paulo, Brazil.
authors.forename: Paloma Canedo
authors.initials: PC
authors.lastname: Henrique
authors.affiliation: Zoological Park Foundation, São Paulo, Brazil.
authors.forename: Irys Hany Lima
authors.initials: IH
authors.lastname: Gonzalez
authors.affiliation: Malaria Research Center, Superintendence for Endemic Disease Control, São Paulo Institute of Tropical Medicine, University of São Paulo, Brazil.
authors.forename: Eliana Ferreira
authors.initials: EF
authors.lastname: Monteiro
authors.affiliation: Malaria Research Center, Superintendence for Endemic Disease Control, São Paulo Institute of Tropical Medicine, University of São Paulo, Brazil.
authors.forename: Lilian de Oliveira
authors.initials: Lde O
authors.lastname: Guimarães
authors.affiliation: Laboratory of Virology, São Paulo Institute of Tropical Medicine, University of São Paulo, Brazil.
authors.forename: Camila Malta
authors.initials: CM
authors.lastname: Romano
authors.affiliation: Malaria Research Center, Superintendence for Endemic Disease Control, São Paulo Institute of Tropical Medicine, University of São Paulo, Brazil.
authors.forename: Karin
authors.initials: K
authors.lastname: Kirchgatter
keywords: Alopochen aegyptiacus; Avian malaria diagnosis; Brazil; Goose; P. (Haemamoeba) sp.; Plasmodium nucleophilum