Description and molecular characterization of a new species of tarantula , Pamphobeteus verdolaga , from Colombia ( Araneae : Mygalomorphae : Theraphosidae )

A new species of Pamphobeteus Pocock, 1901 is described from the Colombian Andes, Medellín, Antioquia. The biochemistry and molecular characteristics of the venom of this new species (previously identified as Pamphobeteus aff. nigricolor) has been already described. A detailed taxonomic study, comparing this species with holotypes and additional material of Pamphobeteus species, allowed us to recognize it as new, and to describe it here as Pamphobeteus verdolaga sp. nov. The male of P. verdolaga sp. nov. is distinguished by the palpal bulb with broad embolus, poorly developed apical keel (A), prolateral inferior keel (PI) and prolateral accessory keel (PAc) present but poorly developed, and retrolateral keel (R) of similar length as A. Females are distinguished by the morphology of spermatheca with a wide base and very short oval seminal receptacles, which are curved toward the center. This is the thirteenth species described in Pamphobeteus and the sixth species reported from Colombia. The species description is complemented by a molecular characterization of a partial

Pamphobeteus Pocock, 1901 comprises 12 species of tarantulas, distributed in northern South America (World Spider Catalog 2016).They are mainly distributed on the Andean region, with high diversity in Colombia, although they are also found in the Amazon rainforest (Bertani et al. 2008).Pamphobeteus includes large terrestrial species, usually dark colored with violet shades.They inhabit burrows or natural cavities.
Males of Pamphobeteus can be distinguished morphologically by the presence of a palpal bulb with a concave-convex embolus, spoon-shaped, with a well developed apical keel (A) that extends up to the edge of the embolus, a well developed retrolateral keel (R), and a prolateral superior keel (PS) (Bertani et al. 2008).Females can be distinguished by the spermatheca with two seminal receptacles largely fused at the base and apically elongated (pérez-MileS et al. 1996).Pamphobeteus males and females resemble Xenesthis Simon, 1891 but they can be distinguished from those by having the scopulae on metatarsi IV restricted to the apical portion (fully extended on Xenesthis) (Bertani et al. 2008).
The biochemistry and molecular characteristics of the venom Pamphobeteus were described by eStrada-goMez et al. (2013) based on specimens from Medellín, Colombia, which had been identified as Pamphobeteus aff.nigricolor.A detailed taxonomic study, and the comparison of this specimens with holotypes, additional specimens, and the original descriptions of Pamphobeteus species, allowed us to recognize that the males from Medellín do not belong to Pamphobeteus nigricolor (Ausserer, 1875) and differ from all other species of the genus in the morphology of the palpal bulb.Based on this comparison we have determined these specimens as a new species, which is diagnosed and described as Pamphobeteus verdolaga sp.nov.The species description is complemented by a molecular characterization of a partial CO1 sequence, the first to be sequenced for the genus.

MATERIAL AND METHODS
All measurements are given in millimeters (mm).Body and legs measurements were taken with a vernier caliper; legs and palp measurements were taken in dorsal view along the central axis of the left-side limbs.Others measurements were taken using an ocular micrometer.The terminology used to described the palpal organ keel follows Bertani (2000) and for the urticating setae follows Cooke et al. (1972).The number and disposition of spines were enumerated from the anterior third to the posterior third, modified from petrunkevitCh (1925).The geographic coordinates and altitude data (meters above sea level: m asl) were referenced by GPS, Datum WGS84, or determined using the Instituto Geográfico Agustín Codazzi Gazetter (http:// www.igac.gov.co/digeo/app/index.html).The coordinates are given in UTM notation.The distribution map was produced using SimpleMappr (ShorthouSe 2010).
Holotype material is deposited at the ICN Arachnological Collection, Bogotá, Colombia and paratypes are in the Entomological Collection and Serpentarium of the Universidad de Antioquia, Medellín, Colombia, stored in 70% ethanol.Diagnosis.Male differs from known Pamphobeteus species by having palpal bulb with broad embolus, poorly developed apical keel (A) and prolateral inferior keel (PI), retrolateral keel (R) of similar length to A, and a prolateral accessory keel (PAc).Prolateral superior keel (PS) and R well developed (Figs.1-7).Females have spermathecae with a wide base and very short seminal oval receptacles, curved toward the center (Fig. 9).The palpal bulb of Pamphobeteus verdolaga sp.nov.resembles that of P. antinous Pocock, 1903;P. crassifemur Bertani, Fukushima & Silva, 2008;P. grandis Bertani, Fukushima & Silva, 2008;P. ultramarinus Schmidt, 1995;and P. vespertinus (Simon, 1889) by the broad embolus.But, all of them present palpal bulb with R shorter than A (R similar to A on P. verdolaga sp.nov.; Figs. 2, 6).
Color (alive): cephalotorax and legs brown, violet overtones on the anterior region of cephalotorax, femora greenish dark brown, abdomen dark brown.
Molecular characterization: mitochondrial CO1 partial sequence ( 414  apical keel, and retrolateral keel length similar to apical keel.Pamphobeteus augusti, P. crassifemur, P. ferox and P. ornatus have prolateral inferior keel; it is poorly developed in the first two as in P. verdolaga sp.nov., and the last two have PI well developed.In the description of P. grandis (Bertani et al. 2008: fig. 13) the apical keel was erroneously indicated as prolateral inferior keel.
Pamphobeteus females have similar spermathecae and are difficult to differentiate.They can be distinguished from other genera, except Xenesthis, by the presence of two elongated seminal receptacles, largely fused at the base.Pamphobeteus verdolaga sp.nov. is characterized by the very short oval seminal receptacles, which are curved toward the center.
Pamphobeteus petersi Schmidt, 2002 is not included in this discussion because this species is misplaced within Pamphobeteus.The copulatory organs of P. petersi resemble those of Megaphobema Pocock, 1901, thereby P. petersi must be transferred to Megaphobema.This transfer will be implemented and discussed within the context of a revision of the genus Pamphobeteus (in prep.).

Figure 12 .
Figure 12.Geographic distribution of Pamphobeteus verdolaga sp.nov.Green circle shows distribution area of the species.Arrow points to Medellín city, Antioquia, Colombia.