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High morphological homogeneity and cryptic speciation may cause the diversity within Simuliidae to be underestimated. Recent molecular studies on population genetics and phylogeography have contributed to reveal which factors influenced the diversity within this group. This study aimed at examining the genetic diversity of Simulium hirtipupa Lutz, 1910 in populations from the biomes Caatinga, Cerrado, and Atlantic Forest. In this study, we carried out phylogeographic and population genetic analyses using a fragment of the mitochondrial gene COI. The 19 populations studied were clustered into seven groups, most of which are associated with geography indicating certain genetic structure. The northern region of the state of Minas Gerais is most likely the center of origin of this species. The average intergroup genetic distance was 3.7%, indicating the presence of cryptic species. The species tree as well as the haplotype network recovered all groups forming two major groups: the first comprises groups Gr-Bahia (in which the São Francisco river has not acted as geographical barrier), Gr-Pernambuco, and Gr-Mato Grosso do Sul. The second included groups comprising populations of the states of Goiás, Tocantins, Minas Gerais, Bahia, São Paulo, and Espírito Santo. The mismatch distribution for groups was consistent with the model of demographic expansion, except for the Gr-Central-East_1 group. The diversification in this group occurred about 1.19 Mya during the Pleistocene, influenced by paleoclimatic oscillations during the Quaternary glacial cycles.

The black fly Simulium (Trichodagmia) hirtipupa Lutz (Diptera: Simuliidae) is widely distributed in southern Brazil, with one report from Amapá state in the northern region of Brazilian Amazonia. Morphological comparison of northern and southern populations revealed differences in all life stages, corroborated by chromosomal and molecular analyses, and indicated that the population previously identified as S. hirtipupa from Amapá state represents an undescribed species. This new species is described based on all life stages above the egg, and its chromosomal and molecular divergence from S. hirtipupa is highlighted. Simulium criniferum n. sp. can be diagnosed by the deeply concave male ventral plate with a prominent median projection bearing a ventral keel; female anal lobe in lateral view with a broadly rounded, distal membranous area about as long as wide; pupa with a boot-shaped cocoon bearing a minutely bubbled surface, cephalic plate and thorax with abundant hair-like tubercles, and gill of 12 translucent filaments with darkly sclerotized, acuminate tips; and larva with the body cuticle bearing spiniform setae, abdomen truncated posteriorly, and gill histoblast in situ with the filament tips directed ventrally. Chromosomally, the new species has five unique fixed inversions and uniquely shares three additional fixed inversions with its nearest relative, S. hirtipupa. Partial COI sequences indicate a genetic distance of ∼9% between the new species and S. hirtipupa. Females of the new species are anthropophilic.