Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter March 25, 2015

PCR identification of five species from the Anopheles maculipennis complex (Diptera: Culicidae) in North-Eastern Romania

Maria-Larisa Ivanescu, Dumitru Acatrinei, Ionuţ Pavel, Tatiana Sulesco and Liviu Miron
From the journal Acta Parasitologica


The members of the Anopheles maculipennis complex have been incriminated for the transmission of the malaria in Europe, which was endemic until the middle of the century. The global warming and the intensification of the intercontinental travel constitute a risk of the re-emergence of the malaria in Europe, given the presence of the Anopheles vectors. The study has attempted the identification by using the PCR (Polymerase Chain Reaction) of the members of the Anopheles maculipennis complex from the North-eastern area of Romania from the city of Iaşi. In total there have been identified by using the PCR amplifying the ITS2 sequence of the ribosomal DNA, 217 specimens belonging to the complex of A. maculipennis among which: 58 A. atroparvus, 18 A. melanoon, 2 A. labranchiae, 52 A. maculipennis and 87 A. messeae. The ITS2 sequences of the ribosomal DNA have been compared to those of the species belonging to the A. maculipennis available in GenBank. The Species A. labranchiae is reported for the first time in Romania, being identified in the larval stage IV. The adaptation of a new species to the climatic conditions present in the North-eastern Romania, confirms the phenomenon of global warming and also the intensification of the travelling. As a result of the analysis of the A. labranchiae sequence, this one corresponds to the extent of 96% to the species from Italy, registered in GenBank, given the fact that a high number of the inhabitants of the municipality of Iaşi are working in this country.


Alten B., Kampen H., Fontenille D. 2007. Malaria in Southern Europe: resurgence from the past?. In: (Eds. W. Takken, B. G.Knols) , Emerging Pests and Vector-Borne Diseases in Europe Vol.1. Wageningen University Press, 35-57. DOI: 10.3920/978-90-8686-626-7Search in Google Scholar

Becker N., Petric D., Boase C., Zgomba M., Dahl C., Lane J., Kaiser A. 2003. Mosquitoes and their Control. Kluwer Academic/ Plenum Publishers, New York. 163-186Search in Google Scholar

Bietolini S., Candura F., Coluzzi M. 2006. Spatial and long term temporal distribution of the Anopheles maculipennis complex species in Italy. Parassitologia, 48, 581-608. Cote INIST: 26177, 35400014676523.0060Search in Google Scholar

Calleri G., Behrens R.H., Schmid M.L., Gobbi F., Grobusch M.P., Castelli F., Gascon J., Bisoffi Z., Jelinek T., Caramello P. 2011. Malaria chemoprophylaxis recommendations for immigrants to Europe, visiting relatives and friends - a Delphi method study. Malaria Journal, 10, 137. DOI: in Google Scholar

Cobo F. 2014. Current status of malaria, Imported Infectious Diseases, The Impact in Developed Countries, 1st Edition, Elsevier, 61-90Search in Google Scholar

Di Luca M., Boccolini D., Severini F., Toma L., Mancini-Barbieri F., Massa A., Romi R. 2009. A 2-Year Entomological Study of Potential Malaria Vectors in Central Italy. Vector-Borne and Zoonotic Diseases, 9, 703-711. DOI: 10.1089/vbz.2008.0129Search in Google Scholar

Djadid N.D., Gholizadeh S., Tafsiri E., Romi R., Gordeev M., Zakeri S., 2007. Molecular identification of Palearctic members of Anopheles maculipennis in northern Iran. Malar Journal, 6, 6. DOI: 10.1186/1475-2875-6-6Search in Google Scholar

Kronefeld M., Dittmann M., Zielke D., Werner D., Kampen H., 2012. Molecular confirmation of the occurrence in Germany of Anopheles daciae (Diptera, Culicidae). Parasites & Vectors, 5, 250. DOI: 10.1186/1756-3305-5-250Search in Google Scholar

Mouchet J., Carnevale P., Coosemans M., Julvez J., Manguin S., Richard-Lenoble D., Sircoulon J., 2004. Biodiversité du paludisme dans le monde, Paris: John Libbey Eurotext, 425 pp.Search in Google Scholar

Nitzulescu V. and Gherman I., 1990. Entomologie medicală, Editura Academiei Romane, 117-129Search in Google Scholar

Nicolescu G. and Purcărea-Ciulacu V. 2012. Modelul de stratificare a zonelor de risc de re-emergenţă şi al strategiei de supraveghere ş i control al malariei în România. Edit. Ars Docendi- Universitatea din Bucuresti Search in Google Scholar

Nicolescu G., Linton Y.M., Vladimirescu A., Howard T.M., Harbach R.E. 2004. Mosquitoes of the Anopheles maculipennis group (Diptera: Culicidae) in Romania, with the discovery and formal recognition of a new species based on molecular and morphological evidence. Bulletin of Entomological Research, 94, 525-35. DOI: 10.1079/BER2004330Search in Google Scholar

Odolini S., Gautret P., Parola P. 2012. Epidemiology of Imported Malaria in the Mediterranean Region. Mediterranean Journal of Hematology and Infections Diseases, 4, e2012031. DOI: 10.4084/MJHID.2012.031Search in Google Scholar

Proft J., Maier A.M., Kampen H., 1999. Identifcation of six sibling species of the Anopheles maculipennis complex (Diptera: Culicidae) by a polymerase chain reaction assay. Parasitol Research (1999) 85, 837-843. DOI: 10.1007/s004360050642Search in Google Scholar

Pages F., Orlandi-Pradines E., Corbel V. 2007. Vecteurs du paludisme :biologie, diversite, controle et protection individuelle. Médecine et maladies infectieuses 37, 153-161. DOI: 10. 1016/j.medmal.2006.10.009Search in Google Scholar

Patz A.J., Olson H.S. 2006. Malaria risk and temperature: Influences from global climate change and local land use practices. National Academy of Sciences of the United States of America, PNAS, vol. 103 no. 15. 5635-5636. DOI: 10.1073/pnas.060 1493103Search in Google Scholar

Romi R., Boccolini D., Menegon M., Rezza G. 2012. Probable autochthonous introduced malaria cases in Italy in 2009-2011 and the risk of local vector-borne transmission. Eurosurveillance. Volume 17, Issue 48, 29 November 17, 20325. 20325Search in Google Scholar

Romi R., Sabatinelli G., Majori G. 2001. Could malaria reappear in Italy? Emerging Infections Diseases. Nov-Dec, 7, 915-919. DOI: 10.3201/eid0706.010601Search in Google Scholar

Romi R., Boccolini D., D’Amato S., Corrado C., Peragallo M., D’Ancona F., Pompa M.G., Majori G. 2010. Incidence of malaria and risk factors in Italian travelers to malaria endemic countries. Travel Medicine and Infectious Disease, May, 8, 144-154. DOI: 2010.02.001Search in Google Scholar

Shkurti K., Vyshka G., Velo E., Bocari A., Kokici M., Kraja D., 2013. Imported malaria in Albania and the risk factors that could allow its reappearance. Malar Journal. 2013, 12, 197. DOI: 10.1186/1475-2875-12-197Search in Google Scholar

Sedaghat M.M., Linton Y.M., Oshaghi M.A., Vatandoost H., Harbach R.E. 2003. The Anopheles maculipennis complex (Diptera: Culicidae) in Iran: molecular characterization and recognition of a new species. Bulletin of Entomological Research. Dec, 93, 527-535. DOI: http// BER2003272Search in Google Scholar

Sinka M. E., Bangs M.J., Manguin S., Coetzee M., Mbogo C.M., Hemingway J., Patil A.P., Temperley W.H., Gething P.W., Kabaria C.W., Okara R.M., Van Boeckel T., Godfray C.J., Harbach R.E., Hay S.I. 2010. The dominant Anopheles vectors of human malaria in Africa, Europe and the Middle East: occurrence data, distribution maps and bionomic precis. Parasites & Vectors, 3, 117. DOI: 10.1186/1756-3305-3-117Search in Google Scholar

Sinka M. E. et al . 2012. A global map of dominant malaria vectors Parasites & Vectors, 5, 69. DOI: 10.1186/1756-3305-5-69Search in Google Scholar

Tolle M. A. 2009. Mosquito-borne Diseases. in Current Problem in Pediatric and Adolescent Health Care, 39, 97-140. DOI: 10.1016/j.cppeds.2009.01.001Search in Google Scholar

Vicente L., Sousa C. A., Alten B., Caglar S.S., Falcuta E., Latorre J.M., Toty C., Barre H., Demirci B., Di Luca M., Toma L., Alves R., SalgueiroP., SilvaT. L., BarguesM. D., Mas-ComaS., Boccolini D., Romi R., Nicolescu G., RosarioV. E., Ozer N., Fontenille D., Pinto J. 2011. Genetic and phenotypic variation of the malaria vector Anopheles atroparvus in southern Europe. Malaria Journal, 10, 5. DOI: 10.1186/1475-2875-10-5Search in Google Scholar

Zahar A.R., 1990. Vector bionomics in the epidemiology and control of malaria. Part II. The WHO Europeanregion and the WHO Eastern Mediterranean region. Volume II: Applied field studies. Section III: Vector bionomics, malaria epidemiology and control by geographical areas. (A): The Mediterranean Basin. WHO; 226 pp Search in Google Scholar

Received: 2014-1-24
Revised: 2014-12-9
Accepted: 2015-1-9
Published Online: 2015-3-25
Published in Print: 2015-6-1

© 2015 W. Stefański Institute of Parasitology, PAS