Insect Molecular Biology (2002) 11(4), 379 – 383 © 2002 The Royal Entomological Society 379 Blackwell Science, Ltd SHORT NOTE Ribosomal ITS2 sequence data for Anopheles maculipennis and An. messeae in northern Greece, with a critical assessment of previously published sequences Y.-M. Linton*, A. Samanidou-Voyadjoglou† and R. E. Harbach* *Department of Entomology and Biomedical Sciences Theme, The Natural History Museum, Cromwell Road, London, UK; †Department of Parasitology, Entomology and Tropical Diseases, National School of Public Health, 196 Alexandras Avenue, Athens, Greece Abstract DNA sequences were generated for eight specimens of the Anopheles maculipennis complex from Florina in NW Greece, and identified to species on the basis of comparison with ITS2 sequences for members of the complex already in GenBank. The sequences revealed the presence of An. maculipennis and An. messeae in Florina. Problems with sequence reliability and acces- sibility of sequences generated in earlier studies of Palaearctic members of the complex are discussed. Keywords: Anopheles maculipennis, Anopheles messeae, ITS2. Introduction Anopheles maculipennis, the historical malaria vector in Europe, was exposed as the first sibling species complex of mosquitoes more than 70 years ago (Falleroni, 1926; van Thiel, 1927). The complex comprises nine Palaearctic mem- bers: An. atroparvus van Thiel, An. beklemishevi Stegnii & Kabanova, An. labranchiae Falleroni, An. maculipennis Meigen, An. martinius Shingarev, An. melanoon Hackett, An. messeae Falleroni, An. sacharovi Favre and An. subalpinus Hackett & Lewis (White, 1978; de Zulueta et al., 1983; Cianchi et al., 1987; Ribeiro et al., 1988). Early records indicated the presence of An. maculipennis, An. messeae, An. subalpinus and An. sacharovi in North- ern areas of Greece (Hackett & Lewis, 1935; Hackett & Missiroli, 1935; Pandazis, 1935; Shannon, 1935; Livadas & Sphangos, 1940; Shannon & Hadjinicolaou, 1941), but little is known about their current distribution (see Ramsdale & Snow, 2000). Although An. maculipennis, An. messeae and An. sacharovi are capable of transmitting malaria, they exhibit different vector capacities (Kasap, 1990; Nikolaeva, 1996; Alten et al., 2000). Given this and the proximity of Greece to areas of resurgent malaria transmission, correct vector identification is essential to assess the potential risk of malaria in the border regions of Greece, and devise appropriate control or monitoring strategies. In this study, DNA sequences of the nuclear internal trans- cribed spacer (ITS2) were generated and the specimens identified by comparison to ITS2 sequences in GenBank. Problems with sequence reliability and accessibility of sequences generated in earlier studies of Palaearctic members of the complex are discussed. Results and discussion ITS2 sequence data DNA sequences were generated for eight individuals (Table 1) and compared to those in GenBank. Two sequences shared 97.6% identity with An. messeae (Z50105; Marinucci et al., 1999), and the other six shared 97.7% similarity with An. maculipennis (Z50104; Marinucci et al., 1999) (Table 1). Positive identification of An. maculipennis and An. messeae in Florina, NW Greece constitutes new distribution records for these species (Lin- ton et al., 2001b). Percentage AT content was 46.9% in An. maculipennis (24.5% A, 22.4% T, 24.8% C, 28.3% G) and 46.8% in An. messeae (24.4% A, 22.4% T, 25.4% C, 27.8% G). These values are concordant with 40–50% AT values reported for other mosquitoes of subgenus Anopheles, including members of the Quadrimaculatus, Maculipennis and Hyrcanus groups (Cornel et al., 1996; Received 17 January 2002; accepted after revision 5 March 2002. Corre- spondence: Yvonne-Marie Linton, Department of Entomology, The Natural History Museum, Cromwell Road, London SW7 5BD, UK. Tel.: +44 (0)207 9425819; fax: +44 (0)207 9425229; e-mail: yvol@nhm.ac.uk