The taxonomic status of Phlebotomus oliverioi ( Diptera , Psychodidae ) and a lectotype designation for Psathyromyia brasiliensis

Phlebotomus oliverioi Barretto & Coutinho, 1941 was originally described based only on the male holotype and has since been considered junior-synonym of Psathyromyia brasiliensis (Costa Lima, 1932). The study of the holotype of Ph. oliverioi allowed us to conclude that the head of this specimen belongs to the genus Psychodopygus and the wings, thorax and abdomen belong to a different species of Psathyromyia. Thus, Ph. oliverioi is a not valid species, and must be removed from the synonym and excluded from the provisions of the International Code of Zoological Nomenclature (Articles 17.2 and 23.8). The specimen was measured, drawn, photographed and the morphological characters are discussed. Lectotype and paralectotypes to Pa. brasiliensis are designated.

Phlebotomine sand flies play an important role in the transmission of Leishmania spp., the causative agents of leishmaniases.Approximately 267 species of phlebotomine sand flies have been recorded in Brazil (AndrAde et al., 2013), and a revision of Brazilian species of sand flies recently undertaken by one of the authors (AJA) has found that Phlebotomus oliverioi Barretto & Coutinho, 1941 has since been considered a junior-synonym of Psathyromyia brasiliensis (Costa Lima, 1932) is not a valid species.
Psathyromyia Barretto, 1962, was described as a subgenus of Lutzomyia França, 1924, and it was elevated to genus level by GAlAti (1995), with three subgenera within the subtribe Psychodopygina: Forattiniella Vargas, 1978, Xyphomyia Artemiev, 1991, and Psathyromyia sensu strictu.In accordance to BArretto (1962), the males of Psathyromyia are characterized by ascoids with posterior spurs, the fifth palpus segment longer than the third, the femur without spines, the genitalia longer than head, the gonocoxite without setae, the gonostyli with four spines and without pre-apical seta, simple paramere, and the lateral lobes (surstyli) simple.GAlAti (2003) also considered as characters of this genus the posterior legs with the metatarsomere I to be longer than the sum of the tarsomeres II+III+IV+V, the absence of tergal papillae on all abdominal tergites, and the setae on the anterior margin of the katepisternum; 43 species, including one fossil and three unclassified species (incertae sedis), were included in this genus, 26 of which have been recorded in Brazil, and 10 in the state of São Paulo.
Phlebotomus oliverioi was described in 1941 by BArretto & Coutinho based on one male collected in forested areas in the municipality of Osasco (state of São Paulo, Brazil).In the description, Ph. oliverioi is included among species presenting "the fifth palpus shorter than the third palpus".Based on this character, they compared the male of Ph. oliverioi to other phlebotomine sand fly species, which are today included in the genera Psychodopygus and Nyssomyia, according to the more recent classification of GAlAti (1995GAlAti ( , 2003)), and also provided a taxonomic discussion based on genitalia characters.In the key to American sand flies, BArretto (1950) did not include or provide any comments on Ph. oliverioi.In his thesis, reported Ph. oliverioi as an aberrant species.Once again, the author did not include Ph. oliverioi in the proposed species key and he termed Ph. oliverioi "insufficiently known" (author's pers.obs.).
Further taxonomic studies included Ph. oliverioi in the subgenus Nyssomyia (intermedia group) based on the absence of setae on the gonocoxite and the presence of four spines on the gonostyli (MArtins et al., 1978;reAdy & FrAihA, 1981), while theodor (1965) considered it as "inquirenda species".ForAttini (1973) examined the holotype and concluded that it was conspecific to Psathyromyia brasiliensis (Costa Lima, 1932), but Iheringia, Série Zoologia, Porto Alegre, 104(4):426-431, 31 de dezembro de 2014 MArtins et al. (1978) andreAdy &FrAihA (1981) did not accept the synonym proposed by ForAttini (1973), and the latter authors claimed that Ph. oliverioi did not have the antennal ascoids with proximal spurs, and that the scutum was darker than the pleura, although they did not examine the holotype and could not have observed these characters.One of us (EABG) studied the holotype and observed that the characters of the head were similar to those of Psychodopygus lloydi (Antunes, 1937) and the characteristics of genitalia similar to those of Pa.brasiliensis, suggesting a probable mistake in mounting the specimen, leading to the misidentification of Ph. oliverioi.
Here we discuss the taxonomic status of Ph. oliverioi, only known from the morphological characters of its male holotype and, as a revision of Pa.brasiliensis was performed, its lectotype and paralectotypes were designated.

MATERIAL AND METHODS
The male holotype of Ph. oliverioi was measured with a Zeiss TM eye-piece calibrated according to a standard Zeiss TM scale, and drawings were made using an Olympus TM camera lucida.All measurements are given in micrometres (µm).The photomicrographs were captured using a Leica DFC295 digital camera attached to a Leica DM5000 B optical microscope, and were taken using the Leica Application Suite (LAS) software version 4.1.0.Terminology and nomenclature follows GAlAti (1995GAlAti ( , 2003)) (Fonseca, 1936).
The holotype of Pa.oliverioi is deposited in the entomological collection of the Departamento de Epidemiologia da Faculdade de Saúde Pública da Universidade de São Paulo (FSP).The specimens of Ps. lloydi and Ps.arthuri are deposited in the Coleção Entomológica do Laboratório de Entomologia em FSP/ Phlebotominae (LESP/Phlebotominae).
The male holotype of Ph. oliverioi was originally mounted in Canada balsam, and the specimen body parts were separated and covered by three coverslips: (1) one head, (2) one wing and (3), one wing, thorax and abdomen.

DISCUSSION
The holotype of Ph. oliverioi presents the head with both maxillary palpi and antenna complete, and is mounted ventrally in Canada balsam between slide and cover slip.The fifth palpus is much shorter than the third, with the sum of the fourth and fifth palpi being equal to the length of the third palpus (both palpi were measured); the ascoids do not present proximal spur, and there are two or three papillae on the first flagellomere.These characteristics are compatible  with the genus Psychodopygus in accordance to GAlAti (2003).On the other hand, in the subgenus Psathyromyia as proposed by BArretto (1962), the fifth palpus is longer than the third and the ascoids present proximal spur.reAdy & FrAihA (1981) mentioned that the antennal ascoids lack proximal spurs and the scutum is darker than the pleura.The presence of proximal spurs on the ascoids was not mentioned in the original description, and examination of the holotype revealed that the proximal spurs of the ascoids are absent on all flagellomeres.Also, the scutum is not dark, but slightly brown.BArretto & Coutinho (1941) present the measurements of the fifth and fourth palpus segment as being longer than the third in Ph. oliverioi, but this is incorrect, as we showed by measuring the holotype.Probably, the head belongs either to the species Ps. lloydi or Ps.arthuri recorded in Osasco municipality (MArtins et al., 1978;GAlAti, 1981;AndrAde Filho et al., 1997), but the characters are not enough to distinguish between them.
Both wings were well-preserved and well-mounted on the slide.The analysis of wing venation showed that delta is longer than gamma (ratio delta/gamma = 2.0:1.0) and that delta reaches the middle of alpha.These measurements are consistent with the wing measurements in the description of Pa.brasiliensis (CostA liMA, 1932).
The thorax, with one wing, and the abdomen with one leg, were not separated, and thus they belong to the same individual.Examination of the terminalia revealed that the specimen belongs to Psathromyia sp.We also observed the absence of abdominal papillae in the tergites I-VII.In Pa.brasiliensis, the ejaculatory duct is long, approximately 10x the length of the ejaculatory apodeme (younG & dunCAn, 1994;GAlAti, 2003), and in Ph. oliverioi it is 6x shorter than the ejaculatory apodeme, considering the length of 972 µm, which is also found in the Amazonian sister species Pa.abunaensis (Martins, Falcão & Silva, 1965), known to occur in Brazil, in the states of Rondônia (GAlAti, 2003) and Amazonas (Alves et al., 2012).Phlebotomus oliverioi presents the basiphallus (aedeagus sensu GAlAti) reaching beyond the middle of the paramere, and in Pa.abunaensis the basiphallus does not reach the middle of the structure.Others characters are very similar between the males of Pa.brasiliensis and Pa.abunaensis, but the latter is not recorded in the southeast region of Brazil.
younG & dunCAn (1994) suggested that Ph. oliverioi belongs to the aragaoi group, based solely on the original description and drawings of the species.These characters include the broad wing, paramere with proximal hump and external superior spine of the gonostylus on a well marked tubercle, and fifth palpus segment shorter than the third.In fact, these characters can be observed in the original description, and were probably used by ForAttini (1973) to propose the synonym (the wing, thorax and abdomen are similar to the male of Pa.brasiliensis), after examining the holotype.Even though ForAttini (1973) did not explicitly mention the observation of the holotype of Ph. oliverioi, he commented on the similarity of Ph. oliverioi and Pa.brasiliensis.However, MArtins et al. (1978) and reAdy & FrAihA (1981) did not consider Ph. oliverioi as a junior synonymy of Pa.brasiliensis, and included it in the Nyssomyia subgenus, as proposed by theodor (1965).
After the studying the sand fly species mentioned above, as well as the species description of Ph. oliverioi, we conclude that this species is a not valid, and that is constituted by the head of a species of the genus Psychodopygus sp. and the wing, thorax and abdomen of a Psathyromyia sp.Recently, a similar taxonomic study showed that Phlebotomus breviductus Barretto & Coutinho, 1950 was also not a valid species (Andrade et al., 2013).