Skip to main content Accesibility Help

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

  • G. Nicolescu (a1), Y.-M. Linton (a2), A. Vladimirescu (a1), T.M. Howard (a2) and R.E. Harbach (a2)...

Mosquitoes of the Anopheles maculipennis group were collected in five districts of Romania (Constanta, Giurgiu, Ilfov, Mehedinti and Suceava) between March 2000 and June 2003. Two hundred and ninety-seven specimens were identified by molecular methods. Nuclear rDNA ITS2 sequences of 178 specimens were compared with GenBank sequences for nine known Palaearctic species of the group, and 119 specimens were identified using an ITS2 PCR–RFLP assay developed during the study. Five genetically distinct species of the group were identified: A. atroparvus van Thiel, A. maculipennis Meigen, A. melanoon Hackett and A. messeae Falleroni and a previously unrecognized species. The new species, herein formally described and named A. daciaesp. n., was collected in the Black Sea coastal region and plains adjacent to the Danube River in southern Romania. Anopheles daciae is most similar to and sympatric with A. messeae. It is contrasted with A. messeae and characterized on the basis of unique nuclear ITS2 and mitochondrial COI DNA sequences and morphological characters of the eggs. The larval, pupal and adults stages of the two species were also compared, but no reliable characters were found to distinguish them. It seems likely that A. daciae is more widespread in eastern Europe and the Balkan States, and could be responsible for malaria transmission in these regions that is currently attributed to A. messeae. Anopheles melanoon is reported from Romania for the first time.

Corresponding author
*Fax: 00442079425229 E-mail:
Hide All
Alten, B., Caglar, S.S., Özel, O. (2000) Malaria and its vectors in Turkey. European Mosquito Bulletin 7, 2733.
Boccolini, D., Di Luca, M., Marinucci, .M, Romi, R. (2003) Further molecular and morphological support for the formal synonymy of Anopheles subalpinus Hackett & Lewis with A. melanoon Hackett. European Mosquito Bulletin 16, 15.
Bruce-Chwatt, L.J., de Zulueta, J. (1980) The rise and fall of malaria in Europe. London: Butler and Tanner.
Cianchi, R., Sabatini, A., Boccolini, D., Bullini, L., Coluzzi, M. (1987) Electrophoretic evidence of reproductive isolation between sympatric populations of Anopheles melanoon and A. subalpinus. p. 1560 in Third International Congress on Malaria and Babesiosis International Laveran Foundation, Annecy France.
Clary, D.O. & Wolstenholme, D.R. (1985) The mitochondrial DNA molecule of Drosophila yakuba: nucleotide sequence, gene organisation and genetic code. Journal of Molecular Evolution 22, 252271.
Collins, F.H. & Paskewitz, S.M. (1996) A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species. Insect Molecular Biology 5, 19.
Di Luca, M., Boccolini, D., Marinucci, M., Romi, R. (2004) Intrapopulation polymorpism in Anophyelas messeae (An. masulipennis complex) inferred by molecular analysis. Journal of Medical Entomology 41, 582586.
Falleroni, D. (1926) Fauna anofelica italiana e suo “habitat” (paludi, risaie, canali). Metodi di lotta contro la malaria. Rivista di Malariologia 5, 553593.
Frizzi, G. (1953) Etude cytogenetique d' Anopheles maculipennis en Italie. Bulletin of the World Health Organization 9, 335344.
Gordeev, M., Goriacheva, I., Shaikevitch, E., Ejov, M. (2004) Intraspecific variability of the second internal transcribed spacer of the ribosomal DNA among five Palaearctic species of anopheline mosquitoes. European Mosquito Bulletin 17, 1419.
Hackett, L.W. (1934) The present status of our knowledge of the sub-species of Anopheles maculipennis. Transactions of the Royal Society of Tropical Medicine and Hygiene 28, 109128.
Hackett, L.W. & Lewis, D.J. (1935) A new variety of Anopheles maculipennis in southern Europe. Rivista di Malariologia 14, 377383.
Harbach, R.E. & Knight, K.L. (1980) Taxonomists' glossary of mosquito anatomy Plexus Publishing Inc., Marlton New Jersey.
International Commission on Zoological Nomenclature. (1999) International code of zoological nomenclature 4th edn. The International Trust for Zoological Nomenclature London.
Jetten, T.H., Takken, W. (1994) Anophelism without malaria in Europe. A review of the ecology and distribution of the genus Anopheles in Europe. Wageningen Agricultural University Papers, 94–5. Agricultural University Wageningen, The Netherlands.
Kitzmiller, J.B., Frizzi, G., Baker, R. (1967) Evolution and speciation within the maculipennis complex of the genus Anopheles. Genetics of insect vectors of disease. pp. 151210 in Wright, J.W., Pal, R. (Eds) LondonElsevier Publishing.
Kumar, S., Tamura, K., Jakobsen, I.B., Nei, M. (2001) MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17, 12441245.
Linton, Y.-M., Samanidou-Voyadjoglou, A., Smith, L. & Harbach, R.E. (2001a) New occurrence records for Anopheles maculipennis and An. messeae in northern Greece based on DNA sequence data. European Mosquito Bulletin 11, 3136.
Linton, Y.-M., Harbach, R.E., Chang, M.S., Anthony, T.G., Matusop, A. (2001b) Morphological and molecular identity of Anopheles (Cellia) sundaicus (Diptera: Culicidae), the nominotypical member of a malaria vector species complex in Southeast Asia. Systematic Entomology 26, 357366.
Linton, Y.-M., Samanidou-Voyadjoglou, A. & Harbach, R.E. (2002a) Ribosomal ITS2 sequence data for Anopheles maculipennis and An. messeae in northern Greece, with a critical assessment of previously published sequences. Insect Molecular Biology 11, 379383.
Linton, Y.-M., Smith, L. & Harbach, R.E. (2002b) Molecular confirmation of sympatric populations of Anopheles messeae and Anopheles atroparvus overwintering in Kent, southeast England. European Mosquito Bulletin 13, 816.
Linton, Y.-M., Smith, L. & Harbach, R.E. (2002c) Observations on the taxonomic status of Anopheles subalpinus Hackett & Lewis and An. melanoon Hackett. European Mosquito Bulletin 13, 17.
Linton, Y.-M., Smith, L., Koliopoulos, G., Samanidou-Voyadjoglou, A., Zounos, A.K. & Harbach, R.E. (2003) Morphological and molecular characterization of Anopheles (Anopheles) maculipennis Meigen, type species of the genus and nominotypical member of the Maculipennis Complex. Systematic Entomology 28, 3656.
Linton, Y.-M., Smith, L., Koliopoulos, G., Zounos, A.K., Samanidou-Voyadjoglou, A., Patsoula, E. & Harbach, R.E. (2004) The Anopheles maculipennis complex (Diptera: Culicidae) in Greece. Medical and Veterinary Entomology 18 (in press).
Marinucci, M., Romi, R., Mancini, P., Di Luca, M., Severini, C. (1999) Phylogenetic relationships of seven Palaearctic members of the maculipennis complex inferred from ITS2 sequence data. Insect Molecular Biology 8, 469480.
Martini, E., Zotta, G. (1934) Rapport sur un voyage d'études effectué à travers la Roumanie pendant les mois d'Août et de Septembre 1933. Races d' A. maculipennis en Roumanie. Societé des Nations, C.H. (Malaria) 218, 384.
Missiroli, A. (1939) The varieties of Anopheles maculipennis and the malaria problem in Italy. Proceedings of the 7th International Congress of Entomology 3, 16191640.
Nicolescu, G. (1995) The mosquitoes of Romania: an annotated checklist and bibliography. Romanian Archives of Microbiology and Immunology 54, 75109.
Nicolescu, G. (1996) George Zotta (1886–1942): an early concept of malaria stratification. Romanian Archives of Microbiology and Immunology 2, 173179.
Nikolaeva, N. (1996) Resurgence of malaria in the former Soviet Union (FSU). Society of Vector Ecology Newsletter 27, 1011.
Proft, J., Maier, W.A., Kampen, H. (1999) Identification of six sibling species of the Anopheles maculipennis complex (Diptera: Culicidae) by a polymerase chain reaction assay. Parasitology Research 85, 837843.
Romi, R., Boccolini, D., Hovanesyan, I., Grigoryan, G., Di Luca, M., Sabatinelli, G. (2002) Anopheles sacharovi (Diptera: Culicidae): a re-emerging malaria vector in the Ararat Valley of Armenia. Journal of Medical Entomology 39, 446450.
Sedaghat, M.M., Linton, Y.-M., Nicolescu, G., Smith, L., Koliopoulos, G., Zounos, A.K., Oshaghi, M.A., Vatandoost, H. & Harbach, R.E. (2003a) Morphological and molecular characterization of Anopheles (Anopheles) sacharovi Favre, a primary vector of malaria in the Middle East. Systematic Entomology 28, 241256.
Sedaghat, M.M., Linton, Y.-M., Oshaghi, M.A., Vatandoost, H. & Harbach, R.E. (2003b) The Anopheles maculipennis complex (Diptera: Culicidae) in Iran: molecular characterization and recognition of a new species. Bulletin of Entomological Research 93, 527535.
Simon, C., Frati, F., Beckenbach, A., Crespi, B., Lui, H., Flook, P. (1994) Evolution, weighting and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Annals of the Entomological Society of America 87, 651701.
Stegnii, V.N. (1982) Genetic adaptation and speciation in sibling species of the Eurasian Maculipennis Complex. pp. 454465 in Steiner, W.M., Tabachnick, W.J., Rai, K.S., Narang, S. (Eds) Recent developments in the genetics of insect vectors, Champaign: Illinois Stipes Publishing.
Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F. & Higgins, D.G. (1997) The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 24, 48764882.
White, G.B. (1978) Systematic reappraisal of the Anopheles maculipennis complex. Mosquito Systematics 10, 1344.
Zotta, G. (1938) Contribution à l'étude de la distribution des races d' A. maculipennis en rapport avec les grandes lignes de repartition du paludisme en Roumanie. Archives Roumaines de Pathologie Expérimentale et de Microbiologie 2, 209246.
Zotta, G., Georgescu, M., Ionescu, V., Lupascu, G., Mardare, I. & Teodorescu, A.M. (1940) Nouvelle carte de la disribution des races d' Anopheles en Roumanie. Bulletin de la Section Scientifique de l'Académie Roumaine 2, 7387.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Bulletin of Entomological Research
  • ISSN: 0007-4853
  • EISSN: 1475-2670
  • URL: /core/journals/bulletin-of-entomological-research
Please enter your name
Please enter a valid email address
Who would you like to send this to? *


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed