Description of evandromyia spelunca, a new phlebotomine species of the cortelezzii complex, from a cave in Minas Gerais State, Brazil (Diptera: Psychodidae: Phlebotominae)

Background The cave fauna of the Brazil is poorly documented, and among the insects those live or frequent caves and their adjacent environments phlebotomine sand flies call for special attention because several species are vectors of pathogens among vertebrates hosts. A new species of sand fly from Minas Gerais is described based in females and males collected in a cave of the municipality of Lassance. Results The morphological characters of the new species permit to include in the Evandromyia genus, cortelezzii complex. This complex consists of three species: Evandromyia corumbaensis (Galati, Nunes, Oshiro & Rego, 1989), Evandromyia cortelezzii (Brethes, 1923) and Evandromyia sallesi (Galvao & Coutinho, 1940). Conclusions The new species can be separate from the others of the cortelezzii complex through morphological characters of the male terminalia and female spermathecae.


Background
Sand flies are responsible for transmission of the genus Leishmania among vertebrates hosts and study of this insect group is of great importance in attempts to control of leishmaniasis [1]. In Brazil, the cave fauna of insects is poorly documented, and among the insects that live or frequent caves and their adjacent environments, phlebotomine sand flies call for special attention because several species are vectors of arboviruses and protozoa, among other parasites.
The geographical distribution of the sand flies species depends on their ability to adapt to different ecological niches. Thus, by its development in the immature stages and their feeding habits when adults, the species of sand flies are found where both larvae and adults can find appropriate environments for their development, including blood supply for females. In this way, the geographical distribution of species can be restricted to their access to a specific environment and vertebrate host. Already the occurrence of leishmaniasis is basically determined by the presence of both a susceptible vector and a host/reservoir equally susceptible to the infection [2].
The growing of tourism, involving a search for natural attractions such as cave exploration, demands a better knowledge of the threats to health that people may face in those new areas.
In the present investigation a new species of sand fly from Minas Gerais is described based on both genders collected in a cave in the Lassance municipality.

Methods
Sand flies were collected using CDC light traps (HP model), conducted between June 2008 to May 2010, in the cave named "Gruta Rebenta Bombas" city of Lassance, Minas Gerais state. Sand flies were mounted in Canada balsam (males) and Berlese liquid (females), measured with a binocular Olympus CH-2 microscope with the aid of a micrometer ocular and the drawings were done with the help of a camera lucida. All the measurements presented in this paper are given in micrometers. The nomenclature and classification is that proposed by Galati [3] and the abbreviation of the names for phlebotomine genera by Marcondes [4]. The new species is described based on 10 females and 10 males, which were associated by their capture in the same place at the same time as well as by the association of the morphological characteristics (antennal and palpal formulas; length association of the genital filaments; pigmentation of the thorax; and other structures) that include all the specimens of both sexes in the same genus and group.
Allotype female sand fly of medium size, measuring ca. 2,567 (2,574 ± 260.6; n = 10) in length. Coloration as in the males holotype and paratypes.

Etymology
The name Evandromyia spelunca sp. nov. has been given in regards to the meaning of cave in Latin.
The males of the genus Evandromyia present the gonocoxite with a tuft of bristles, basal and compact; the paramere is simple or branched; the lateral lobe apex is thin and the females present the relationship between the lengths: clypeus/head wider than 1/3; eye/ head less than 1/2; and spermathecae, with variable body. The cortelezzii complex belongs to the cortelezzi series of the Barretomyia subgenus. This series presents as main morphological characteristics: males, with the paramere and aedeagus simple; the gonostyle with upper external spine implanted in his middle, and lower spine, inserted in the basal third. Females with the globular spermathecae's body and the individual ducts are tubular [3]. Evandromyia edwardsi (Mangabeira, 1941) also belong to this series.
The new species can be separated from others of the cortelezzii complex through morphological characters. All the males of the complex are very close being separated by the following characteristics: paramere, which provides more similar aspect to Ev. cortelezzii and Ev. sallesi, however, with the internal suture, the curvature and the shape of the paramere, being enough to separate the new species from the other two closer (see Figures 3,9 and 10). Ev. sallesi presents the paramere with the most consistent shape compared to all other species, so being the most robust ( Figure 9). The suture that divides the inside of the paramere is more central and follows almost the entire length of paramere in the new species. This is more evident in the parameres of the Ev.   cortelezzii and Ev. corumbaensis, the latter being the thinnest and convex paramere of all species of the complex ( Figure 11). The lateral lobe is slightly longer than all other species of the complex. The base of the tuft presents a small pigmented area, being this characteristic common to the three other species of the cortelezzii complex. But with the tuft of the gonocoxite in the new species being slightly different from the others three species of the complex, which provides a more similar aspect to Ev. corumbaensis, both separated by the aspect of the bristles, more thinner in Ev. corumbaensis, and the tubercle that presents less setae in the new species.
The separation of the females from the others of the complex is more confused, and was not always possible, usually when the spermathecae are not present or totally visible. However, the new species can be separated from the Ev. cortelezzii and Ev. sallesi ( Figure 12) by spermathecae and characteristics of their ducts: individual and common, which are noticeably longer in the new species ( Figure 8) and more similar to Ev. corumbaensis. Only characteristics of the cibarium were not enough to separate the females of the complex. We can utilize characteristics of the cibarium to separate the species of the complex, observing the distance between the vertical teeth implantation, and position of the arch that form the horizontal teeth, this distance being greater in the new species. Added to this, it is important to check the layout of vertical teeth in the new species, which appears in the form of an arc, starting at the sides of cibarium, from top to bottom, which usually causes the distance between the deployments mentioned above, higher in Ev. spelunca sp. nov.
Due to the difficulty of identifying the species of this complex, it became routine, with rare exceptions [6], to use geographic distribution for identify them in Brazilian regions. However, it was recently demonstrated that the three species of the complex occur in all states of the Center-west Region, and only Ev. corumbaensis is restricted to that region. The species of this complex that present the greater distribution in the country is Ev. cortelezzii, demonstrating its omission from the lists of species in several states due Ev. sallesi, which has more restricted distribution than the one quoted in the literature [7]. Thus, we do not suggest the use of geographic distribution to identify the species of this complex, but we can suggest the observation of its occurrence in caves, beyond the morphological characteristics, since the new species had their captures practically restricted to inside the cave studied, showing preference for this environment.
It is still important to emphasize that the new species is sympatric with Ev. sallesi inside or outside the cave, however the latter one are always captured inside the cave in less number than outside the cave (data not shown).

Conclusions
The present study adds a new species, namely Ev. spelunca sp. nov., to the Brazilian phlebotomine sand fly fauna, presenting characters for its separation from members of the cortelezzii complex, to which it belongs With the description of the new species, now the