The second internal transcribed spacer of nuclear ribosomal DNA as a tool for Latin American anopheline taxonomy: a critical review

Among the molecular markers commonly used for mosquito taxonomy, the internal transcribed spacer 2 (ITS2) of the ribosomal DNA is useful for distinguishing among closely-related species. Here we review 178 GenBank accession numbers matching ITS2 sequences of Latin American anophelines. Among those, we found 105 unique sequences corresponding to 35 species. Overall the ITS2 sequences distinguish anopheline species, however, information on intraspecific and geographic variations is scarce. Intraspecific variations ranged from 0.2 percent to 19 percent and our analysis indicates that misidentification and/or sequencing errors could be responsible for some of the high values of divergence. Research in Latin American malaria vector taxonomy profited from molecular data provided by single or few field capture mosquitoes. However we propose that caution should be taken and minimum requirements considered in the design of additional studies. Future studies in this field should consider that: (1) voucher specimens, assigned to the DNA sequences, need to be deposited in collections, (2) intraspecific variations should be thoroughly evaluated, (3) ITS2 and other molecular markers, considered as a group, will provide more reliable information, (4) biological data about vector populations are missing and should be prioritized, (5) the molecular markers are most powerful when coupled with traditional taxonomic tools.

Evaluation of insectide resistence and biochemical mechanisms in a population of Culex quinquefasciatus (Diptera: Culicidae) from Säo Paulo, Brazil

To establish an insectidical resistance surveillance program, Culex quinquefasciatus mosquitoes from Säo Paulo, Brazil, were colonized (PIN95 strain) and analyzed for levels of resistance. The PIN95 strain showed low levels of resistance to organophosphates [malathion (3.3-fold), fenitrothion (11.2-fold)] and a carbamate [propoxur (3.0-fold)]. We also observed an increase of 7.4 and 9.9 in alpha and ß esterase activities, respectively, when compared with the reference IAL strain. An alteration in the sensitivity of acetycholinesterase to insectide inhibition was also found in the PIN95 mosquitoes. The resistant allele (Ace.1r), however, was found at low frequencies (0.12) and does not play an important role in the described insecticide resistance. One year later, Cx quinquefasciatus mosquitoes were collected (PIN96 strain) at the same site and compared to the PIN95 strain. The esterase activity patterns observed for the PIN96 strain were similar to those of the PIN95 mosquitoes. However, the occurence of the Ace.1r allele was statistically higher in the PIN96 strain. The results show that esterase-based insecticide resistance was established in the PIN95 Cx. quinquefasciatus population and that an acethylcholinesterase based resistant mechanism has been selected for. A continuous monitoring of this phenomenon is fundamental for rational mosquito control and insecticide application programs.

Resistência a inseticidas organofosforados e carbamatos em populaçäo de Culex quinquefasciatus / Organophosphorous and carbamate resistence in a population of Culex quinquefasciatus

Relata-se a ocorrência, na populaçäo de Culex quinquefasciatus, de resistência a pelo menos dois organofosforados e a um carbamato (malathion, fenitrothion e propoxur, respectivamente), mostrando a necessidade de haver programa de manejo de inseticidas. A suscetibilidade dessa populaçäo ao piretróide permetrina indica essa classe como de escolha numa eventual necessidade de substituiçäo dos organofosforados.