The Australian skink Egernia stokesii had been recognised as a host of two species of Plasmodium, Plasmodium mackerrasae and P. circularis; nevertheless, molecular data are available for only a single haemosporidian species of this host. Its sequences are labelled as "Plasmodium sp." or "Plasmodium mackerrasae", but morphological characteristics of this isolate are unavailable. Phylogenetic analyses of these sequences placed them into the clade of the genus Haemocystidium. In this study, blood samples of six E. stokesii were analysed by both, molecular and microscopic methods to clarify the haemosporidia of this lizard. Application of these approaches offered discordant results. Whereas sequence analysis clustered our isolates with lizard species of Haemocystidium, morphology of blood stages is more akin to Plasmodium than Haemocystidium. However, limited sampling, indistinguishable nuclei/merozoites and risk of possible hidden presence of mixed infection prevent reliable species identification of detected parasites or their description as new species of Haemocystidium.

CEITEC Central European Institute of Technology University of Veterinary Sciences Brno Palackého 1946 1 612 42 Brno Czech Republic

College of Science and Engineering Flinders University Adelaide South Australia 5001 Australia

Department of Biology and Wildlife Diseases Faculty of Veterinary Hygiene and Ecology University of Veterinary Sciences Brno Palackého 1946 1 Brno 612 42 Czech Republic

Zobrazit více v PubMed

Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990;215:403–410. doi: 10.1016/S0022-2836(05)80360-2. PubMed DOI

Bensch S, Stjernman M, Hasselquist D, Ústman Ú, Hansson B, Westerdahl H, Pinheiro RT. Host specificity in avian blood parasites: a study of Plasmodium and Haemoproteus mitochondrial DNA amplified from birds. Proc R Soc B. 2000;267:1583–1589. doi: 10.1098/rspb.2000.1181. PubMed DOI PMC

Boysen KE, Perkins SL, Hunjan S, Oliver P, Gardner MG, Balasubramaniam S, Melville J. Diversity and phylogenetic relationships of haemosporidian and hemogregarine parasites in Australian lizards. Mol Phylogenet Evol. 2022;167:107358. doi: 10.1016/j.ympev.2021.107358. PubMed DOI

de Oliveira JP, André MR, Alves Júnior JRF, Lustosa APG, Werther K. Molecular detection of hemogregarines and haemosporidians in Brazilian free-living testudines. Int J Parasitol Parasites Wildl. 2018;7:75–84. doi: 10.1016/j.ijppaw.2018.01.008. PubMed DOI PMC

Galen SC, Borner J, Martinsen ES, Schaer J, Austin CC, West J, Perkins SL. The polyphyly of Plasmodium: comprehensive phylogenetic analyses of the malaria parasites (order Haemosporida) reveal widespread taxonomic conflict. R Soc Open Sci. 2018;5:171780. doi: 10.1098/rsos.171780. PubMed DOI PMC

Guindon S, Dufayard J-F, Lefort V, Anisimova M, Hordijk W, Gascuel O. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol. 2010;59:307–321. doi: 10.1093/sysbio/syq010. PubMed DOI

Hellgren O, Waldenström J, Bensch S. A new PCR assay for simultaneous studies of Leucocytozoon, Plasmodium, and Haemoproteus from avian blood. J Parasitol. 2004;90:797–802. doi: 10.1645/GE-184R1. PubMed DOI

Huelsenbeck JP, Ronquist F. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics. 2001;17:754–755. doi: 10.1093/bioinformatics/17.8.754. PubMed DOI

Javanbakht H, Kvičerová J, Dvořáková N, Mikulíček P, Sharifi M, Kautman M, Maršíková A, Široký P. Phylogeny, diversity, distribution, and host specificity of Haemoproteus spp. (Apicomplexa: Haemosporida: Haemoproteidae) of palaearctic tortoises. J Euk Microbiol. 2015;62:670–678. doi: 10.1111/jeu.12227. PubMed DOI

Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012;28:1647–1649. doi: 10.1093/bioinformatics/bts199. PubMed DOI PMC

Kvičerová J, Pakandl M, Hypša V. Phylogenetic relationships among Eimeria spp. (Apicomplexa, Eimeriidae) infecting rabbits: evolutionary significance of biological and morphological features. Parasitology. 2008;135:443–452. doi: 10.1017/S0031182007004106. PubMed DOI

Kvičerová J, Hypša V, Dvořáková N, Mikulíček P, Jandzik D, Gardner MG, Javanbakht H, Tiar G, Široký P. Hemolivia and Hepatozoon: haemogregarines with tangled evolutionary relationships. Protist. 2014;165:688–700. doi: 10.1016/j.protis.2014.06.001. PubMed DOI

Maia JP, Harris DJ, Carranza S. Reconstruction of the evolutionary history of Haemosporida (Apicomplexa) based on the cyt b gene with characterization of Haemocystidium in geckos (Squamata: Gekkota) from Oman. Parasitol Int. 2016;65:5–11. doi: 10.1016/j.parint.2015.09.003. PubMed DOI

Martinsen ES, Perkins SL, Schall JJ. A three-genome phylogeny of malaria parasites (Plasmodium and closely related genera): evolution of life-history traits and host switches. Mol Phylogenet Evol. 2008;47:261–273. doi: 10.1016/j.ympev.2007.11.012. PubMed DOI

O’Donoghue P. Haemoprotozoa: making biological sense of molecular phylogenies. Int J Parasitol Parasites Wildl. 2017;6:241–256. doi: 10.1016/j.ijppaw.2017.08.007. PubMed DOI PMC

Pacheco MA, Cepeda AS, Bernotiene R, Lotta IA, Matta NE, Valkiunas G, Escalante AA. Primers targeting mitochondrial genes of avian haemosporidians: PCR detection and differential DNA amplification of parasites belonging to different genera. Int J Parasitol. 2018;48:657–670. doi: 10.1016/j.ijpara.2018.02.003. PubMed DOI PMC

Paperna I, Landau I. Haemoproteus (Haemosporidia) of lizards. Bull Mus Nat Hist Natur Paris. 1991;13:309–349.

Perkins SL, Schall JJ. A molecular phylogeny of malarial parasites recovered from cytochrome b gene sequences. J Parasitol. 2002;88:972–978. doi: 10.1645/0022-3395(2002)088[0972:AMPOMP]2.0.CO;2. PubMed DOI

Pineda-Catalan O, Perkins SL, Peirce MA, Engstrand R, Garcia-Davila C, Pinedo-Vasquez M, Aguirre AA. Revision of hemoproteid genera and description and redescription of two species of chelonian hemoproteid parasites. J Parasitol. 2013;99:1089–1098. doi: 10.1645/13-296.1. PubMed DOI

Stöver BC, Müller KF. TreeGraph 2: combining and visualizing evidence from different phylogenetic analyses. BMC Bioinforma. 2010;11:7. doi: 10.1186/1471-2105-11-7. PubMed DOI PMC

Telford SR. A malarial parasite of Australian skinks, Plasmodium mackerrasae sp. n. J Parasitol. 1979;65:409–413. doi: 10.2307/3280285. PubMed DOI

Telford SR. Re-discovery of Haemoproteus kopki (De Mello, 1916) in two geckoes from the Afghan border of Pakistan. Parasitology. 1982;84:527–536. doi: 10.1017/S0031182000052811. DOI

Telford SR, Stein J. Two malaria parasites (Apicomplexa: Plasmodiidae) of the Australian skink Egernia stokesii. J Parasitol. 2000;86:395–406. doi: 10.1645/0022-3395(2000)086[0395:TMPAPO]2.0.CO;2. PubMed DOI

Telford SR, Peirce MA, Samour J. A new Haemocystidium (Apicomplexa: Plasmodiidae) species of the Dhub lizard, Uromastyx aegyptia microlepis, in Abu Dhabi, distinguished by the absence of pigment. J Parasitol. 2012;98:654–656. doi: 10.1645/JP-GE-3004R1. PubMed DOI

Telford SR (2009) Hemoparasites of the Reptilia: Color Atlas and Text. CRC Press, Boca Raton.10.1201/9781420080414