Abstract
PCR-based methods have been increasingly used in diagnosis of parasitic diseases. Over 40 morphospecies of avian malaria parasites of the genus Plasmodium have been described. However, only nine of them have been identified on the level of their mitochondrial cytochrome b (cyt b) gene lineages, which are frequently used in molecular biology studies of avian blood haemosporidian parasites. In this study, we linked two common mitochondrial cyt b lineages P-SGS1 and P-TURDUS1 with their morphospecies. Light infections with two species of malaria parasites of the subgenera Haemamoeba and Giovannolaia were isolated from naturally infected adult Hawfinches, Coccothraustes coccothraustes Linnaeus, on the Curonian Spit in the Baltic Sea. These parasites were inoculated to naive juveniles of the Common Crossbill, Loxia curvirostra Linnaeus, and House Sparrow, Passer domesticus Linnaeus. Heavy parasitemia of Plasmodium (Haemamoeba) relictum Grassi & Feletti, 1891 (lineage P-SGS1) and Plasmodium (Giovannolaia) circumflexum Kikuth, 1931 (P-TURDUS1) developed in the subinoculated Common Crossbills and House Sparrows, respectively, which enabled the detailed illustration of all main blood stages of these parasites and the deposition of their voucher specimens. The parasites of both lineages are actively transmitted in Europe and inhabit a broad range of avian hosts. Lineages closely related to P. relictum and P. circumflexum were identified. This study contributes to establishing of combined PCR-based and morphological identification of avian malaria parasites.References
Atkinson, C.T. & van Riper, C. III. (1991) Pathogenicity and epizootiology of avian haematozoa: Plasmodium, Leucocytozoon, and Haemoproteus. In: Loye, J.E. & Zuk, M. (Eds), Bird-parasite interactions: ecology, evolution, and behavior. Oxford University Press, Oxford, UK, pp. 19–48.
Beadell, J.S., Gering, E., Austin, J., Dumbacher, J.P., Peirce, M., Pratt, T.K., Atkinson, C.T. & Fleischer, R.C. (2004) Prevalence and differential host-specificity of two avian blood parasite genera in the Australo-Papuan region. Molecular Ecology, 13, 3829–3844.
Beadell, J.S., Ishtiaq, F., Covas, R., Melo, M., Warren, B.H., Atkinson, C.T., Bensch, S., Graves, G.R., Jhala, Y.V., Peirce, M.A., Rahmani, A.R., Fonseca, D.M. & Fleischer, R.C. (2006) Global phylogeographic limits of Hawaii’s avian malaria. Proceedings of the Royal Society of London Series B, Biological Sciences B, 273, 2935–2944.
Bensch, S., Stjernman, M., Hasselquist, D., Ostman, O., Hansson, B., Westerdahl, H. & Pinheiro, R.T. (2000) Host specificity in avian blood parasites: a study of Plasmodium and Haemoproteus mitochondrial DNA amplified from birds. Proceedings of the Royal Society of London Series B, Biological Sciences B, 267, 1583 – 1589.
Corradetti, A., Garnham, P.C.C. & Laird, M. (1963) New classification of the avian malaria parasites. Parasitologia, 5, 1 –4.
Fallon, S.M., Bermingham, E. & Ricklefs, R.E. (2005) Host specialization and geographic localization of avian malaria parasites: A regional analysis in the Lesser Antilles. American Naturalist, 165, 466–480.
Feldman, R.A., Freed, L.A. & Cann, R.L. (1995) A PCR test for avian malaria in Hawaiian birds. Molecular Ecology, 4, 663–673.
Garnham, P.C.C. (1966) Malaria parasites and other Haemosporidia. Blackwell Scientific Publications, Oxford, UK, 1144 pp.
Godfrey, R.D., Fedynich, A.M. & Pence, D.B. (1987) Quantification of hematozoa in blood smears. Journal of Wildlife Diseases, 23, 558–565.
Grassi, B. & Feletti, R. (1891) Malariaparasiten in den Vögeln. Zentralblatt für Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene, 9, 403–409.
Hall, T.A. (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows95/98/NT. Nucleic Acids Symposium Series, 41, 95–98.
Hellgren, O., Waldenström, J. & Bensch, S. (2004) A new PCR assay for simultaneous studies of Leucocytozoon, Plasmodium and Haemoproteus from avian blood. Journal of Parasitology, 90, 797 – 802.
Hellgren, O. (2005) The occurrence of haemosporidian parasites in the Fennoscandian bluethroat (Luscinia svecica) population. Journal of Ornithology, 146, 55–60.
Hellgren, O., Waldenström, J., Pérez-Tris, J., Szöll, Ö.E., Hasselquist, D., Križanauskienè-Iezhova, T.A., Valki„nas, G. & Bairlein, F. (2005) Vertebrate host specificity of two avian malaria parasites of the subgenus Novyella: Plasmodium nucleophilum and Plasmodium vaughani. Journal of Parasitology, 91, 472–474.
Kikuth, W. (1931) Immunbiologische und chemotherapeutische Studien an verschiedenen Stämmen von Vogelmalaria. Zentralblatt für Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene, 121, 401–409.
Križanauskienè, A., Hellgren, O., Kosarev, V., Sokolov, L., Bensch, S. & Valki„nas, G. (2006) Variation in host specificity between species of avian haemosporidian parasites: evidence from parasite morphology and cytochrome b gene sequences. Journal of Parasitology, 92, 1319–1324.
Kumar, S., Tamura, K. & Nei, M. (2004) MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Briefings in Bioinformatics, 5, 150–163.
Li, J., Wirtz, R.A., McConkey, G.A., Sattabongkot, J., Waters, A.P., Rogers, M.J. & McCutchan, T.F. (1995) Plasmodium: genus – conserved primers for species identification and quantitation. Experimental Parasitology, 81, 182–190.
Martinsen, E.S., Paperna, I. & Schall, J.J. (2006) Morphological versus molecular identification of avian Haemosporidia: an exploration of three species concepts. Parasitology, 133, 279–288.
Martinsen, E.S., Waite, J.L. & Schall, J.J. (2007) Morphologically defined subgenera of Plasmodium from avian hosts: test of monophyly by phylogenetic analysis of two mitochondrial genes. Parasitology, 134, 483–490.
Pérez-Tris, J. & Bensch, S. (2005) Diagnosing genetically diverse avian malaria infections using mixed-sequence analysis and TA-cloning. Parasitology, 131, 1–9.
Perkins, S.L. & Schall, J.J. (2002) A molecular phylogeny of malarial parasites recovered from cytochrome b gene sequences. Journal of Parasitology, 88, 972 – 978.
Reullier, J., Pérez-Tris, J., Bensch, S. & Secondi, J. (2006) Diversity, distribution and exchange of blood parasites meeting at an avian moving contact zone. Molecular Ecology, 15, 753–763.
Ricklefs, R.E. & Fallon, S.M. (2002) Diversification and host switching in avian malaria parasites. Proceedings of the Royal Society of London Series B, Biological Sciences B, 269, 885 – 892.
Szymanski, M.M. & Lovette, I.J. (2005) High lineage diversity and host sharing of malarial parasites in a local avian assemblage. Journal of Parasitology, 91, 768–774.
Valki„nas, G. (2005) Avian malaria parasites and other haemosporidia. CRC Press, Boca Raton, Florida, USA, 946 pp.
Valki„nas, G., Bensch, S., Iezhova, T.A., Križanauskienè, A., Hellgren, O. & Bolshakov, C.V. (2006) Nested cytochrome b PCR diagnostics underestimate mixed infections of avian blood hemosporidian parasites: microscopy is still essential. Journal of Parasitology, 92, 418–422.
Valki„nas, G., Zehtindjiev, P., Hellgren, O., Ilieva, M., Iezhova, T.A. & Bensch, S. (2007) Linkage between mitochondrial cytochrome b lineages and morphospecies of two avian malaria parasites, with a description of Plasmodium (Novyella) ashfordi sp. nov. Parasitology Research, 100, 1311–1322.
Waldenström, J., Bensch, S., Kiboi, D., Hasselquist, D. & Ottosson, U. (2002) Cross-species infection of blood parasites between resident and migratory songbirds in Africa. Molecular Ecology, 11, 1545–1554.
Waldenström, J., Bensch, S., Hasselquist, D. & Östman, Ö. (2004) A new nested polymerase chain reaction method very efficient in detecting
Wiersch, S.C., Maier, W.A. & Kampen, H. (2005) Plasmodium (Haemamoeba) cathemerium gene sequences for phylogenetic analysis of malaria parasites. Parasitology Research, 96, 90–94.