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Complete sporogony of Plasmodium relictum (lineage pGRW4) in mosquitoes Culex pipiens pipiens, with implications on avian malaria epidemiology

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Abstract

Plasmodium relictum (lineage pGRW4) causes malaria in birds and is actively transmitted in countries with warm climates and also temperate regions of the New World. In Europe, the lineage pGRW4 has been frequently reported in many species of Afrotropical migrants after their arrival from wintering grounds, but is rare in European resident birds. Obstacles for transmission of this parasite in Europe have not been identified. Culex quinquefasciatus is an effective vector of pGRW4 malaria, but this mosquito is absent from temperate regions of Eurasia. It remains unclear if the lineage pGRW4 completes sporogony in European species of mosquitoes. Here we compare the sporogonic development of P. relictum (pGRW4) in experimentally infected mosquitoes Culex pipiens pipiens form molestus, C. quinquefasciatus, and Ochlerotatus cantans. The pGRW4 parasite was isolated from a garden warbler Sylvia borin, multiplied, and used to infect laboratory-reared Culex spp. and wild-caught Ochlerotatus mosquitoes by allowing them to take blood meals on infected birds. The exposed females were maintained at a mean laboratory temperature of 19 °C, which ranged between 14 °C at night and 24 °C during daytime. They were dissected on intervals to study the development of sporogonic stages. Only ookinetes developed in O. cantans; sporogonic development was abortive. The parasite completed sporogony in both Culex species, with similar patterns of development, and sporozoites were reported in the salivary glands 16 days after infection. The presence of sporogonic stages of the lineage pGRW4 in mosquitoes was confirmed by PCR-based testing of (1) the sporozoites present in salivary glands and (2) the single oocysts, which were obtained by laser microdissection from infected mosquito midguts. This study shows that P. relictum (pGRW4) completes sporogony in C. p. pipiens at relatively low temperatures. We conclude that there are no restrictions for spreading this bird infection in Europe from the point of view of vector availability and temperature necessary for sporogony. Other factors should be considered and were discussed for the explanation of rare reports of this malaria parasite in Europe.

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Acknowledgments

We would like to thank the director of the Biological Station “Rybachy,” Casimir V. Bolshakov, for generously providing the facilities for the experimental research and the staff of the Biological Station “Rybachy” for assistance in the field. The authors are grateful to Dr. Roland Kuhn and Dr. Ana Rivero for providing samples of the C. p. pipiens f. molestus and C. quinquefasciatus mosquitoes, respectively, and Dr. Arūnas Bukantis for consultations on climatology. Data about long-term mean air degrees in Europe were provided by the Physical Science Division, Earth System Research Laboratory, NOAA, Boulder, Colorado, from their Web site at http://www.esrl.noaa.gov/psd/. Care and handling of experimental animals was in accordance with the current laws of Lithuania and Russia. This research was supported by the Open Access to research infrastructure of the Nature Research Centre under Lithuanian open access network initiative. Our sincere thanks to the Department of Molecular and Regenerative Medicine, Hematology, Oncology and Transfusion Medicine Centre of the Vilnius University Hospital Santariškių Klinikos for opportunities to use the laser microdissection microscope. This study was partly supported by the Lithuanian Science Foundation (award no. MIP-15022).

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  1. Nature Research Centre, Akademijos 2, LT-08412, Vilnius 2100, Lithuania

    Gediminas Valkiūnas, Rita Žiegytė, Vaidas Palinauskas, Rasa Bernotienė, Dovilė Bukauskaitė, Mikas Ilgūnas, Dimitar Dimitrov & Tatjana A. Iezhova

  2. Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Gagarin Street, 1113, Sofia, Bulgaria

    Dimitar Dimitrov

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  1. Gediminas Valkiūnas
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  2. Rita Žiegytė
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Valkiūnas, G., Žiegytė, R., Palinauskas, V. et al. Complete sporogony of Plasmodium relictum (lineage pGRW4) in mosquitoes Culex pipiens pipiens, with implications on avian malaria epidemiology. Parasitol Res 114, 3075–3085 (2015). https://doi.org/10.1007/s00436-015-4510-3

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