Abstract Drosophila butantan sp. nov., a species belonging to the guarani group and closely related to Drosophila nigrifemur from Bolivia, is described based on a female, and some of its offspring, collected at the forest reserve of the Instituto de Biociências da Universidade de São Paulo, Cidade Universitária "Armando de Salles Oliveira", São Paulo City, state of São Paulo, Brazil. Although externally similar, the two apparently forest-dwelling species can be told apart by having distinct oviscapt valves and spermathecal introverts and tips. Accordingly, a proposal is made to also include D. nigrifemur, a previously unassigned species, in the guarani group. The two species seem to be also related to Drosophila alexandrei and Drosophila guaraja as indicated by their external morphology, their elongate spermathecae and the not so sharply pointed oviscapt valves. The karyotypes of the new species differ from those described for D. alexandrei and D. guaraja, while those of D. nigrifemur remain still unknown. Photomicrographs of the male and female imagines, in addition to drawings and photos of their terminalia, are also included.

Drosophila piratininga sp. nov. e Drosophila sampa sp. nov., espécies neotropicais pertencentes ao grupo canalinea, são descritas no presente artigo. Elas podem ser facilmente separadas com base no padrão das asas, sendo que as duas nervuras transversais são nitidamente esfumaçadas na primeira e hialinas na segunda. As coletas foram realizadas na Reserva Florestal da Cidade Universitária "Armando de Salles Oliveira", um remanescente da Mata Atlântica de planalto localizado na área urbana da zona oeste do município de São Paulo, Estado de São Paulo, Brasil. Ilustrações detalhadas das terminálias são fornecidas.

Drosophila piratininga sp. nov. and Drosophila sampa sp. nov., belonging to the canalinea group of the subgenus Drosophila, are described based mostly on wild-caught specimens collected at a forest reserve of the Cidade Universitária "Armando de Salles Oliveira", an urban remnant of the montane Atlantic Forest located at west São Paulo city, state of São Paulo, Brazil. The two species are readily distinguished from each other mostly by having different wing patterns: the main crossveins being remarkably clouded in the first one and unclouded in the latter. Illustrations of terminalia are also provided.

Drosophila pauliceia sp. nov., espécie com tórax pintado e muito semelhante à Drosophila boraceia Vilela & Val, 2004, é descrita a partir de exemplares procedentes de um fragmento urbano da Mata Atlântica de planalto, localizado na reserva da Cidade Universitária "Armando de Salles Oliveira", cidade de São Paulo, estado de São Paulo, Brasil. As duas espécies podem ser separadas principalmente com base no padrão das pintas do tórax e, juntamente com Drosophila peruensis Wheeler, 1959 e Drosophila atalaia Vilela & Sene, 1982, são incluídas em um novo grupo proposto no presente artigo sob o nome peruensis. Além das ilustrações detalhadas das terminálias masculina e feminina da nova espécie, desenhos inéditos dessas estruturas foram preparados também para Drosophila atalaia. Adicionalmente é proposto o grupo caponei que inclui D. caponei Pavan & Cunha, 1947 e D. neochracea Wheeler, 1959, duas espécies com tórax não pintado embora previamente consideradas próximas à D. atalaia.

Drosophila pauliceia sp. nov., a spot-thoraxed species closely related to Drosophila boraceia Vilela & Val, 2004, is described from flies collected in an urban remnant of the montane Atlantic Forest located at the Cidade Universitária "Armando de Salles Oliveira", São Paulo city, state of São Paulo, Brazil. The two forest-dwelling species can be externally distinguished mainly by having a different pattern of thoracic spots. A new group of spotted flies (peruensis group) is proposed to include them, in addition to Drosophila peruensis Wheeler, 1959, from Urubamba River, Peru, and Drosophila atalaia Vilela & Sene, 1982, from Brazil and Argentina. To facilitate comparisons, illustrations of the male and female terminalia of the new species and of some unpublished views of those of D. atalaia are included. Additionally, a new group of unspotted species (named caponei group) is proposed to include Drosophila caponei Pavan & Cunha, 1947 and D. neochracea Wheeler, 1959, both previously considered to be related to D. atalaia.