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Genetic diversity of avian blood parasites in SE Europe: Cytochrome b lineages of the genera Plasmodium and Haemoproteus (Haemosporida) from Bulgaria

1Institute of Zoology, Bulgarian Academy of Sciences, 1 Tzar Osvoboditel Blvd., 1000, Sofia, Bulgaria

2Central Laboratory of General Ecology, Bulgarian Academy of Sciences, 2 Gagarin Street, 1113, Sofia, Bulgaria

3Department of Ecology, Lund University, Ecology Building, SE-22362, Lund, Sweden

© 2010 W. Stefański Institute of Parasitology, PAS. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. (CC BY-NC-ND 3.0)

Citation Information: Acta Parasitologica. Volume 55, Issue 3, Pages 201–209, ISSN (Online) 1896-1851, ISSN (Print) 1230-2821, DOI: 10.2478/s11686-010-0029-z, August 2010

Publication History

Published Online:


We used a nested PCR protocol to examine the genetic diversity of cytochrome b (cyt b) lineages from blood parasites of the genera Plasmodium and Haemoproteus in birds in Bulgaria. In total, 460 birds of 43 species and 14 families (mostly passerines) were examined for the presence of infections. Of them, 267 were recognised as infected with haemosporidian parasites. Mixed infections were recorded in 24 individuals (9%). Besides the 24 individuals with mix infections, 114 (43%) were positive for Plasmodium spp. and 129 (48%) for Haemoproteus spp. We identified 52 genetic lineages of haemosporidian parasites: 38 of Haemoproteus and 14 of Plasmodium. Twelve new cyt b lineages of Haemoproteus were recorded; they occurred in the following hosts: grey-faced woodpecker (Picus canus), golden oriole (Oriolus oriolus), jay (Garrulus glandarius), barred warbler (Sylvia nisoria), song thrush (Turdus philomelos), spotted flycatcher (Muscicapa striata), spanish sparrow (Passer hispaniolensis), hawfinch (Coccothraustes coccothraustes), and cirl bunting (Emberiza cirlus). We also detected 22 new host records for previously known lineages. The most common lineage was SGS1 (Plasmodium relictum), which had a total prevalence of 14% and occurred in 8 host species belonging to 5 families. Three of the cyt b lineages of genus Haemoproteus (DURB1, DURB2 and SYNIS2) showed more than 5% divergence from all described morphologically lineages. These lineages probably represent at least 2 different morphospecies which remains to be identified.

Keywords: Cytochrome b lineages; Plasmodium; Haemoproteus; specificity

  • [1] Bairlein F. 1995. Manual of field methods. European-African songbird migration network. Institut für Vogelforschung, Wilhelmshaven.

  • [2] 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. DOI: 10.1111/j.1365-294X.2004.02363.x. http://dx.doi.org/10.1111/j.1365-294X.2004.02363.x [CrossRef]

  • [3] 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. DOI: 10.1098/rspb.2006.3671. http://dx.doi.org/10.1098/rspb.2006.3671 [CrossRef]

  • [4] Bensch S., Hellgren O., Pérez-Tris J. 2009. MalAvi: a public database of malaria parasites and related haemosporidians in avian hosts based on mitochondrial cytochrome b lineages. Molecular Ecology Resources, 9, 1353–1358. DOI: 10.1111/j.1755-0998.2009.02692.x. http://dx.doi.org/10.1111/j.1755-0998.2009.02692.x [Web of Science] [CrossRef]

  • [5] Bensch S., Pérez-Tris J., Waldenstrom J., Hellgren O. 2004. Linkage between nuclear and mitochondrial DNA sequences in avian malaria parasites: multiple cases of cryptic speciation? Evolution, 58, 1617–1621. DOI: 10.1111/j.0014-3820.2004.tb01742.x. [CrossRef]

  • [6] 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. DOI: 10.1098/rspb.2000.1181. http://dx.doi.org/10.1098/rspb.2000.1181 [CrossRef]

  • [7] Bensch S., Waldenström J., Jonzén N., Westerdahl H., Hansson B., Sejberg D., Hasselquist D. 2007. Temporal dynamics and diversity of avian malaria parasites in a single host species. Journal of Animal Ecology, 76, 112–122. DOI: 10.1111/j.1365-2656.2006.01176.x. http://dx.doi.org/10.1111/j.1365-2656.2006.01176.x [Web of Science] [CrossRef]

  • [8] BWPi 2006. The birds of the western Palearctic interactive, 2006 Upgrade. DVD Birdguides, Shrewsbury.

  • [9] 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. DOI: 10.1086/428430. http://dx.doi.org/10.1086/428430 [CrossRef]

  • [10] 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.

  • [11] Hellgren O., Križanauskienė A., Valkiūnas G., Bensch S. 2007a. Diversity and phylogeny of mitochondrial cytochrome b lineages from six morphospecies of avian Haemoproteus (Haemosporida: Haemoproteidae). Journal of Parasitology, 93, 889–896. DOI: 10.1645/GE-1051R1.1. http://dx.doi.org/10.1645/GE-1051R1.1 [CrossRef] [Web of Science]

  • [12] 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. DOI: 10.1645/GE-184R1. http://dx.doi.org/10.1645/GE-184R1 [CrossRef]

  • [13] Hellgren O., Waldenström J., Pérez-Tris J., Szöll Ö.E., Hasselquist D., Križanauskienė A., Ottosson U., Bensch S. 2007b. Detecting shifts of transmission area in avian blood parasites — a phylogenetic approach. Molecular Ecology, 16, 1281–1290. DOI: 10.1111/j.1365-294X.2007.03227.x. http://dx.doi.org/10.1111/j.1365-294X.2007.03227.x [Web of Science] [CrossRef]

  • [14] Howard R., Moore A. 2008. Complete checklist of the birds of the world. 3rd edition (corrigenda 8). Avibase — the world bird database, Denis Lepage, Bird Studies Canada. Available from: www.bsc-eoc.org/avibase/avibase.jsp (accessed 12 March 2010).

  • [15] Križanauskienė A., Hellgren O., Kosarev V., Sokolov L., Bensch S., Valkiūnas 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. DOI: 10.1645/GE-873R.1. http://dx.doi.org/10.1645/GE-873R.1 [CrossRef]

  • [16] Križanauskienė A., Pérez-Tris J., Palinauskas V., Bensch S., Valkiūnas G. 2010. Molecular phylogenetic and morphological analysis of haemosporidian parasites (Haemosporida) in a naturally infected European songbird, the blackcap Sylvia atricapilla, with description of Haemoproteus pallidulus sp. nov. Parasitology, 137, 217–227. DOI: 10.1017/S0031182009991235. http://dx.doi.org/10.1017/S0031182009991235 [Web of Science] [CrossRef]

  • [17] Marzal A., Bensch S., Reviriego M., Balbontin J., De Lope F. 2008. Effects of malaria double infection in birds: one plus one is not two. Journal of Evolutionary Biology, 21, 979–987. DOI: 10.1111/j.1420-9101.2008.01545.x. http://dx.doi.org/10.1111/j.1420-9101.2008.01545.x [CrossRef] [Web of Science]

  • [18] Palinauskas V., Kosarev V., Shapoval A., Bensch S., Valkiūnas G. 2007. Comparison of mitochondrial cytochrome b lineages and morphospecies of two avian malaria parasites of the subgenera Haemamoeba and Giovannolaia (Haemosporida: Plasmodiidae). Zootaxa, 1626, 39–50.

  • [19] Pérez-Tris J., Bensch S. 2005. Diagnosing genetically diverse avian malaria infections using mixed-sequence analysis and TA-cloning. Parasitology, 131, 1–9. DOI: 10.1017/S003118200500733X. http://dx.doi.org/10.1017/S003118200500733X [CrossRef]

  • [20] 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. DOI: 10.1645/0022-3395(2002)088[0972:AMPOMP]2.0.CO;2. http://dx.doi.org/10.1645/0022-3395(2002)088[0972:AMPOMP]2.0.CO;2 [CrossRef]

  • [21] 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. DOI: 10.1098/rspb.2001.1940. http://dx.doi.org/10.1098/rspb.2001.1940 [Web of Science] [CrossRef]

  • [22] Sambrook J., Fritsch E.F., Maniatis T. 1989. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.

  • [23] Sehgal R.N.M., Jones H.I., Smith T.B. 2005. Blood parasites of some West African birds. Journal of Veterinary Medical Science, 67, 295–301. http://dx.doi.org/10.1292/jvms.67.295

  • [24] Shurulinkov P., Chakarov N. 2005. Prevalence of blood parasites in different local populations of reed warbler (Acrocephalus scirpaceus) and great reed warbler (Acrocephalus arundinaceus). Parasitology Research, 99, 588–592. DOI: 10.1007/s00436-006-0202-3. http://dx.doi.org/10.1007/s00436-006-0202-3 [CrossRef]

  • [25] Shurulinkov P., Golemansky V. 2002. Haemoproteids (Haemosporida: Haemoproteidae) of wild birds in Bulgaria. Acta Protozoologica, 41, 359–374.

  • [26] Shurulinkov P., Golemansky V. 2003. Plasmodium and Leucocytozoon (Sporozoa: Haemosporida) of wild birds in Bulgaria. Acta Protozoologica, 41, 205–214.

  • [27] Svensson L. 1992. Identification Guide to European Passerines. British Trust for Ornithology, 4th ed., Stockholm.

  • [28] 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. DOI: 10.1645/GE-417R1.1. http://dx.doi.org/10.1645/GE-417R1.1 [CrossRef]

  • [29] Tamura K., Dudley J., Nei M., Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24, 1596–1599 (Publication PDF at http://www.kumarlab.net/publications). http://dx.doi.org/10.1093/molbev/msm092

  • [30] Valkiūnas G. 2005. Avian malaria parasites and other haemosporidia. CRC Press, Boca Raton, Florida, USA, 946 pp.

  • [31] Valkiūnas G., Bensch S., Iezhova T.A., Križanauskienė A., Hellgren O., Bolshakov C.V. 2006. Nested cytochrome b PCR diagnostics underestimate mixed infections of avian blood haemosporidian parasites: microscopy is still essential. Journal of Parasitology, 92, 418–422. DOI: 10.1645/GE-3547RN.1. http://dx.doi.org/10.1645/GE-3547RN.1 [CrossRef]

  • [32] Valkiūnas G., Iezhova T., Golemansky V., Pilarska D., Zehtindjiev P. 1999. Blood protozoan parasites (Protozoa: Kinetoplastida and Haemosporida) in wild birds from Bulgaria. Acta Zoologica Bulgarica, 51, 127–129.

  • [33] Valkiūnas G., Iezhova T.A., Loiseau C., Chasar A., Smith T.B., Sehgal R.N.M. 2008a. New species of haemosporidian parasites (Haemosporida) from African rainforest birds, with remarks on their classification. Parasitology Research, 103, 1213–1228. DOI: 10.1007/s00436-008-1118-x. http://dx.doi.org/10.1007/s00436-008-1118-x [CrossRef] [Web of Science]

  • [34] Valkiūnas G., Križanauskienė A., Iezhova T.A., Hellgren O., Bensch S. 2007a. Molecular phylogenetic analysis of circumnuclear hemoproteids (Haemosporida: Haemoproteidae) of sylviid birds, with a description of Haemoproteus parabelopolskyi sp. nov. Journal of Parasitology, 93, 680–687. DOI: 10.1645/GE-1102R.1. http://dx.doi.org/10.1645/GE-1102R.1 [Web of Science] [CrossRef]

  • [35] Valkiūnas G., Liutkevičius G., Iezhova T. 2002. Complete development of three species of Haemoproteus (Haemosporida, Haemoproteidae) in the biting midge Culicoides impunctatus (Diptera: Ceratopogonidae). Journal of Parasitology, 88, 864–868. DOI: 10.1645/0022-3395(2002)088[0864:CDOTSO]2.0.CO;2. http://dx.doi.org/10.1645/0022-3395(2002)088[0864:CDOTSO]2.0.CO;2 [CrossRef]

  • [36] Valkiūnas G., Sehgal R.N.M., Iezhova T.A., Smith T.B. 2005. Further observations on the blood parasites of birds in Uganda. Journal of Wildlife Diseases, 41, 580–587. [CrossRef]

  • [37] Valkiūnas G., Zehtindjiev P., Dimitrov D., Križanauskienė A., Iezhova T.A., Bensch S. 2008b. Polymerase chain reactionbased identification of Plasmodium (Huffia) elongatum, with remarks on species identity of haemosporidian lineages deposited in GenBank. Parasitology Research, 102, 1185–1193. DOI: 10.1007/s00436-008-0892-9. http://dx.doi.org/10.1007/s00436-008-0892-9 [CrossRef] [Web of Science]

  • [38] Valkiūnas G., Zehtindjiev P., Hellgren O., Ilieva M., Iezhova T.A., Bensch S. 2007b. 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. DOI: 10.1007/s00436-006-0409-3. http://dx.doi.org/10.1007/s00436-006-0409-3 [Web of Science] [CrossRef]

  • [39] Waldenström J., Bensch S., Hasselquist D., östman Ö. 2004. A new nested polymerase chain reaction method very efficient in detecting Plasmodium and Haemoproteus infections from avian blood. Journal of Parasitology, 90, 191–194. DOI: 10.1645/GE-3221RN. http://dx.doi.org/10.1645/GE-3221RN [CrossRef]

  • [40] 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. DOI: 10.1046/j.1365-294X.2002.01523.x. http://dx.doi.org/10.1046/j.1365-294X.2002.01523.x [CrossRef]

  • [41] Zehtindjiev P., Ilieva M., Križanauskienė A., Oparina O., Oparin M., Bensch S. 2009. Occurrence of haemosporidian parasites in the paddyfield warbler, Acrocephalus agricola (Passeriformes, Sylviidae). Acta Parasitologica, 54, 295–300. DOI: 10.2478/s11686-009-0052-0. http://dx.doi.org/10.2478/s11686-009-0052-0 [CrossRef] [Web of Science]

  • [42] Zehtindjiev P., Ilieva M., Westerdahl H., Hansson B., Valkiūnas G., Bensch S. 2008. Dynamics of parasitemia of malaria parasites in a naturally and experimentally infected migratory songbird, the great read warbler Acrocephalus arundinaceus. Experimental Parasitology, 119, 99–110. DOI:10.1016/j.exppara.2007.12.018. http://dx.doi.org/10.1016/j.exppara.2007.12.018 [Web of Science] [CrossRef]

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