Taxonomic revision of Parampheres ( Arachnida : Opiliones : Gonyleptidae )

Parampheres Roewer, 1913 is a relatively common genus of South American harvestmen. This genus is easily diagnosed by the remarkable yellow patches on the prosoma. Nonetheless, species determination within this group is challenging due the convoluted taxonomic history of the group and lack of a recent revision. In this study we revise Parampheres and describe a new species, Parampheres tenebris sp. nov., from Parque Nacional da Serra Geral, Rio Grande do Sul, Brazil. The new species can be distinguished from the other species of the genus by having dorsal scutum dark, and apophysis of coxa IV of male elongated. Furthermore, we propose the following new synonymies: Callampheres boliviensis Roewer, 1913, Pertyana ronae Mello-Leitao, 1927 and Parampheres tibialis Roewer, 1917 with Parampheres pectinatus Roewer, 1913. Parampheres now includes four species distributed from southern Brazil to adjacent areas in Argentina and Uruguay. In addition, we present a phylogenetic hypothesis based on morphological characters that supports the transfer of Parampheres from Gonyleptinae to Caelopyginae.

When Caelopyginae was revised (PINTO-DA-ROCHA 2002), it did not include Parampheres (Roewer, 1913) (PINTO-DA-ROCHA et al. 2014).The species of this genus are relatively common in southern Brazil, Uruguay and northeastern Argentina.They are easily recognized by the presence of two large yellowish orange patches on the prosoma, lateral to the ocularium.
The remarkable similarity between some species of Parampheres raises questions about their validity, particularly when the troubled taxonomic history of the group is taken into account (see subfamily-level revisions cited above).For instance, KURY (2003: 135) argued that the differences between species are so subtle that they might correspond to mere intraspecific variations of a single, widespread species.
Although there have been recent studies on the distribution, behavior and physiology of Parampheres (GONZÁLES et al. 2004, HARA & GNASPINI 2003, STANLEY 2011), identification of the included species is not an easy task, since descriptions and illustration are not very informative.The last revision of the genus, which only included redescriptions of some species, was published by SOARES & SOARES (1985).
The subfamily placement of Parampheres is controversial, which can be explained by the few number of characters used during the last century to distinguish Caelopyginae from Gonyleptinae.SØRENSEN (1884) first proposed the use of pedipalp characters to separate the two subfamilies.Subsequently, he argued that, except for specific cases, the chelate shape of the pedipalp was not consistent enough to separate the two groups (HENRIKSEN 1932: 279).In the identification keys constructed by ROEWER (1913), the claws of tarsi III and IV, which are smooth in Gonyleptinae and pectinated in Caelopyginae, were used to distinguish between the two subfamilies.Based on this dichotomy, P. pectinatus was placed in Caelopyginae.MELLO-LEITÃO ZOOLOGIA 31 (6): 541-556, December, 2014541-556, December, (1949) ) advocated the use of pedipalp morphology to ascertain subfamily affinities, although he described different species of Parampheres in Caelopyginae, Gonyleptinae, and Pachylinae.SOARES & SOARES (1985) questioned the use of smooth or pectinated claws to separate among Caelopyginae and Gonyleptinae, arguing that it is common to find both states within the same genus or even among conspecifics.They also rejected the morphology of the pedipalp as a source of diagnostic features.However, they proposed the use of the tarsal counts: the fewest number of articles is found in Gonyleptinae, whereas the greatest is found in Caelopyginae.Based on this criterion, they transferred Parampheres from Caelopyginae to Gonyleptinae.However, a molecular-based phylogenetic analysis (PINTO-DA-ROCHA et al. 2014) recovered Parampheres bimaculatus Roewer, 1943 as a member of Caelopyginae, supporting ROEWER's (1913) original proposal.
In this paper we revise Parampheres, including updated redescriptions of species, new distribution records, an identification key and the description of a new species.Additionally, we present a new phylogenetic hypothesis, based on morphological data, to test the subfamily placement of the genus within Gonyleptidae.
Specimens were examined in 70% ethanol or dried.Drawings were made with a camera lucida coupled with a stereomicroscope Leica MZ75.Photographs of live animals were taken with a digital camera Cannon EOS Digital Rebel XS.Photomontage images were obtained with a Leica DFC 290 coupled with a stereomicroscope Leica M125, using the Leica Suite 3.3.0software application.The penises of specimens were observed under an optical microscope Axioskop 2 plus.Scanning electron micrographs were obtained with a Zeiss DSM 940 scanning electron microscope (IBUSP) and prepared according to PINTO-DA-ROCHA (2002).
The data matrix (42 taxa, 63 characters, Table I) was built using NDE 0.5.0 software (PAGE 2011) and the cladistic analysis was performed using TNT 1.0 (GOLOBOFF et al. 2008).The analysis was conducted using 10,000 random addition sequences with 100 trees saved per replicate.The swapping algorithm was tree bisection reconnection (TBR).The consensus cladogram was obtained using Winclada version 1.00.08 (NIXON 1999).Characters were equally weighted and treated as unordered or non-additive, and 12 of them are multistate.

TAXONOMY
Diagnosis.Parampheres resembles some Caelopyginae genera, for example Ampheres Koch, 1839, Caelopygus Koch, 1839, and Proampheres Roewer, 1913, by having rows of tubercles on femur IV (Figs 5-8), the outer apophysis of coxa IV and coloration pattern .It resembles Ampheres and Proampheres in the swollen male basitarsus I.It also shares with Proampheres the ocularium armed with spines (or higher tubercles).Parampheres can be easily distinguished from those genera cited above by the presence of two large yellow patches on the sides of the ocularium, the rounded shape of the ocularium (medially depressed in most Caelopyginae) and the presence of the fourth area of the dorsal scutum (Figs 1-4), which is absent in other Caelopyginae.
Redescription.Male.Dorsum.Anterior margin of prosoma with two or three pointed tubercles on angles and a median frontal hump with two or more pointed tubercles.Dorsal scutum densely granulated, with five transversal grooves delimiting four scutal areas (fourth groove is conspicuous only in the middle), scutal area I divided by longitudinal median groove.All areas with a pair of median larger tubercles.Maximal width of dorsal scutum reaching groove IV.Lateral margin of dorsal scutum with two rows of tubercles, from prosoma to posterior margin, generally larger and sharper near groove IV.Posterior margin and free tergites with a row of tubercles, central one larger (Figs 1-4).Chelicerae.Isomorphic in males and females, with 1-3 small tubercles on bulla, fixed finger with five teeth, movable finger with three teeth.Pedipalps.Coxa with two ventral tubercles, trochanter with three ventral tubercles, femur with row of 4-5 ventral tubercles, tarsus biconvex.Legs.Coxa I-III with dorsal tubercles, coxa IV with numerous tubercles on lateral region, a large retrolateral tubercle followed by a row of smaller tubercles and one large prolateral apophysis perpendicular to body axis.Trochanter I-IV tuberculated, trochanter IV with a retrolateral row of three tubercles (apical larger), one dorsoapical tubercle and one robust prolateral tubercle.Femur with prolateral and dorsal rows of robust tubercles.Tibia III and IV with two ventral rows of tubercles .Distitarsus II 3-segmented.Basitarsus I swollen.Penis.Ventral plate with short and wide cleft on distal margin; lateral margin with three subdistal pairs of large setae, two pairs of reduced setae placed more ventrally; one pair of very small setae of intermediate position; and four pairs of basal (basalmost very reduced) setae.Stylus with cleft, sinuous, thin, long, with subapical trichomes on the apical third of lateral and ventral sides.Presence of a subapical process in P. lucidus and P. tenebris sp.nov.Ventral process of glans with lateral prominences and longitudinal cleft .
Female.similar to male but with much shorter, reduced and conical apophysis on coxa IV.Dorsal scutum narrower, spines on lateral margins, posterior margin and free tergites higher and sharper.Femur IV armature very reduced.

Key to the males of Parampheres
1. Dorsal apophysis of coxa IV conical and short (Figs 2,26).
Femur IV with a retrolateral row of 3-4 large tubercles increasing in size from base to middle of segment (Fig. 6 Diagnosis.Parampheres bimaculatus resembles P. pectinatus, but it can be distinguished from the latter by the shorter and thicker femur IV with fewer and more agglomerated tubercles, especially on dorsal portion (Figs 5,8).The claws on tarsi III-IV are smooth or undulated but not pectinated (Figs 21,22).There are fewer tubercles on the dorsal areas and they are arranged in a more orderly pattern.The posterior margin of P. bimaculatus has a median tubercle about the same size as the others in the same row.In the others species this median tubercle is clearly larger than the others (Figs 1-4).The coloration pattern of P. bimaculatus is usually in shades of brown, brighter than in P. tenebris sp.nov.and P. lucidus, but is considerably darker than in P. pectinatus.The morphology of the male genitalia is similar to P. pectinatus, but in P. bimaculatus the trunk of the penis has a distinct process (Fig. 13), which is absent in P. pectinatus (Fig. 16).
Diagnosis.Parampheres lucidus resembles P. tenebris sp.nov. in the coloration pattern (Figs 30,31) and morphology of the penis.Both species have a subapical process on the stylus.This process is absent in the remaining species of the genus, which only have setae on it .Other unique features, ob- served only in males, are the curved tibia III and relatively short and conical apophysis of coxa IV.P. lucidus also has a characteristic femur IV with a retrolateral row of three or four tubercles, increasing in size apically.This row is limited to the first half of the femur whereas in the other species the retrolateral row is present on the entire segment (this distribution is conspicuous in male but also recognizable in females) (Figs 5-8).
Diagnosis.The external morphology of P. pectinatus is similar to P. bimaculatus, but there are some discrete differences.The body coloration pattern of P. pectinatus is predominantly yellow, with darker areas in the posterior margin and area IV, in contrast with the more uniform and darker brown pattern of P. bimaculatus.The yellow patches in P. pectinatus are elongated and blend with the body coloration, whereas in P. bimaculatus they are rounded and well delimited .The male femur IV resembles that of P. tenebris sp.nov., but it is thinner, longer, and has more tubercles that are less clustered than in P. bimaculatus .The claws in tarsi III-IV are clearly pectinated, a distinctive characteristic within the genus that is apomorfic for Caelopyginae (Figs 21,22).
Redescription.Male (MCNRS1364): Measurements.Dorsal scutum: length 8.1; maximum width 9.3.Prosoma length: 2.9; width: 3.9.Femur IV length: 12.1.Dorsum.Anterior margin of prosoma with two groups of three pointed tubercles on each side.Elevated and rounded ocularium with four anterior, three posterior tubercles, and two slightly divergent spines pointing upwards.Two large and elongated yellow patches, parallel to the ocularium.Tubercles small and sparsely distributed, mostly found behind ocularium (17), some (6) anterior to it.Yellow patches almost smooth.Four areas densely covered with randomly dispersed tubercles; each area also has a pair of higher tubercles in the center, aligned in a median row in the first three areas.Lateral margin with two irregular rows of tubercles, two larger on outer face, near area III.Posterior margin and free tergites with a row of tubercles and a larger medium one.Anal operculum with three rows of tubercles (Figs 4,28).Venter.Coxae I-IV uniformly covered with numerous ZOOLOGIA 31 (6): 541-556, December, 2014 tubercles.Stigmatic area with a few scattered small tubercles.
Posterior margin and free sternites with one row of small tubercles.Anal operculum with eight small tubercles in two rows.Chelicerae.Three granules on bulla of the first segment, second segment with 5 teeth, third with 3 teeth.Pedipalps.Coxa with two ventral tubercles.Trochanter with three ventral (one larger) and three dorsal tubercles.Femur with four tubercles in a longitudinal anterior ventral row and six small dorsal tubercles.Patella unarmed.Tibia setation: mesal IiIi, ectal iiiIi.Tarsal setation: mesal IiIii, ectal IiIii.Legs.Coxa I with one high and one smaller tubercle; II with one high anterior to ozopore, one smaller, one posterior fused to other of coxa III.
Coxa III with one fused with other of coxa II.Coxa IV with a large retrolateral tubercle followed by a row of smaller tubercles dorsally disposed, one long prolateral apophysis, perpendicular to body axis, first half curved frontward with the apical third curved backwards and pointed down.Trochanter I with seven ventral, two retrolateral and three retrodorsal tubercles Trochanter II with seven ventral, three retrolateral and two retrolateral tubercles.Trochanter III with eleven ventral, five retrodorsal and three retrolateral tubercles.Trochanter IV with several ventral tubercles, two large apical and one small basal tubercles on a retrolateral row, two retrodorsal and one robust pro-dorsal tubercles.Femora I-II tuberculated; III with three rows of tubercles slightly larger than the others.Femur IV with a retrolateral row of robust tubercles (larger in middle), a prolateral row of tubercles (larger and clustered in basal region) throughout all length and a dorsal row of high tubercles occupying the first half of the segment (Fig. 8).Tibiae III-IV with two ventral rows of tubercles (posterior larger).Tarsal formula: 6, 10(3), 7, 8. Claws pectinated (Fig. 22).Penis.Ventral plate with short and wide cleft; lateral margin with three subdistal pairs of large setae, two pairs of reduced setae placed more ventrally; one pair of very small setae, intermediate in position; and four pairs of basal setae (basalmost very reduced).
Parampheres tenebris sp.nov.Figs 3,11,15,19,23,24,27,31 Diagnosis.Parampheres tenebris sp.nov. is similar to P. lucidus in the dark color pattern (Figs 30,31).It can be easily distinguished from P. lucidus by having an elongated (more than twice trochanter IV length) and sinuous apophysis on coxa IV of the male, contrasting with the short (length less than trochanter IV length) and conical apophysis of P. lucidus (Figs 26,27).Other striking difference is the male femur IV with a retrolateral row of tubercles in P. tenebris sp.nov.and three large and spaced tubercles in P. lucidus .
Description.Male (MCNRS0044): Measurements.Dorsal scutum: length: 8.1; maximum width: 9.0.Prosoma length: 2.9; width: 4.2; Femur IV length: 11.4.Dorsum.Anterior margin of prosoma with three tubercles on each side.Frontal hump with three pointed tubercles.Ocularium elevated and rounded with two anterior, three posterior tubercles and two larger median tubercles (slightly larger than eye diameter).Two large and round yellow patches, parallel to the ocularium to groove I. Seventeen sparse granules in front and thirteen behind ocularium, yellow patches smooth.Four areas, I to III with a row of tubercles (central pair larger), also some randomly dispersed tubercles and area IV with only small scattered tubercles.Tubercles count: area I-47; area II-58; area III-45; area IV-27.Lateral margin with two irregular rows of tubercles extending from posterior margin to ozopore, tubercles are larger on outer face, near area III.Posterior margin and free tergites with a row of tubercles, with one much larger median.Anal operculum with thirteen small tubercles in three irregular rows (Figs 3,27).Venter.Coxae I-IV densely covered with tubercles.Stigmatic area with a few scattered small tubercles.Posterior margin and free sternites with one row of small tubercles.Chelicerae.Two dorsal granules on bulla.Pedipalps.Coxa with two ventral tubercles.Trochanter with three ventral and one dorsal tubercle.Femur with four tubercles in longitudinal ven-tral row, five small dorsal tubercles.Patella unarmed.Tibia setation: mesal IiIi ectal iiIi.Tarsal setation: mesal IIiii ectal: IiIiii.Legs.Coxa I with two dorsal tubercles, II with three dorsal tubercles, III with two dorsal tubercles.Coxa IV with numerous tubercles on lateral region, with a large retrolateral tubercle followed by a row of smaller tubercles until apophysis and one long prolateral apophysis perpendicular to body axis, with posterior end curved and pointing backwards.Trochanter I with seven small ventral tubercles and six small dorsal tubercles.Trochanter II with thirteen ventral tubercles and two dorsal ones.Trochanter III with nine ventral tubercles and a row of four dorsal tubercles.Trochanter IV with sixteen ventral tubercles, a retrolateral row of three tubercles, apical larger than the others, dorsoapical tubercle and one robust prolateral tubercle.Femora I-II with scattered small tubercles and a retrolateral row of small tubercles, femur III with two rows of ventral tubercles.Femur IV with a retrolateral row of robust and curved tubercles (larger in the middle), dorsal row of tubercles (much larger in the basal third), prodorsal row of tubercles (larger in the basal third) and a prolateral row of tubercles (posterior ones acute and longer), with dorsoapical reduced tubercles (Fig. 7).Tibiae III and IV with two ventral rows of tubercles (longer near apex).Tarsal formula: 6, 11(3), 7, 9. First article of tarsus I inflated.Claws slightly corrugated.Penis.Ventral plate with short and wide cleft; lateral margin with three subdistal pairs of large setae, two pairs of reduced setae more ventrally placed; one pair of very small setae on intermediate position; four pairs of basal setae (basalmost very reduced).Stylus with cleft, sinuous, thin, long, with a subapical process and lateral trichomes.Ventral process of the glans with lateral prominences and a longitudinal cleft (Figs 11,15,19).Coloration (in ethanol, Fig. 27).Most of body and legs light brown with darker apophysis on coxa IV and femur IV.Tubercles slightly darker.One rounded yellow patch occupying each lateral side of ocularium.In vivo (Fig. 31): very dark brown (almost black) body and legs, patches on the side of ocularium deep orange.Size variation of males (n = 3).Dorsal scutum length: 7.4-8.7;maximum width: 7.3-9.5.Prosoma length: 3.0-3.2;width: 3.9-4.3.Femur IV length: 9.5-11.9.
The subfamily placement of Parampheres has been controversial (see introduction), bouncing from Caelopyginae to Gonyleptinae and even Pachylinae.A close relationship between the genera of Caelopyginae was recently recovered in an analysis using four molecular markers (three mitochondrial and one nuclear) and over 100 gonyleptids by PINTO-DA-ROCHA et al. (2014).The authors included Parampheres bimaculatus and four other Caelopyginae genera and obtained the following relationships for this subfamily: (Ampheres (Parampheres (Arthrodes (Metarthrodes + Pristocnemis))))).According to their hypothesis, the pectinate claws were lost in some species of Parampheres.
The cladistic analysis of morphological characters, under equal weighting, supported the hypothesis that the subfamily is monophyletic and is sister to Gonyleptes saprophilus (Gonyleptinae).The number of synapomorphies supporting Caelopgyinae that are reversed in Parampheres is remarkable.In our analysis, Parampheres is sister to Ampheres.According to this hypothesis, the pectinate claws arose in Caelopyginae and were lost in some species of Parampheres.The shape of the ocularium, widened and with a median depression, was considered synapomorphic for the subfamily by PINTO-DA-ROCHA (2002).Nevertheless, a rounded and medially high ocularium is present in Parampheres and a similar shape occurs in related groups such as Gonyleptinae and Hernandariinae.Other features, such as the three-segmented distitarsus I (and also tarsal count) and lack of white spots on the anal operculum also show reversals in this genus.As a result, the generic placement of species is a somewhat difficult task for a taxonomist who is not familiar with the group.Among the K92 members, only Progonyleptoidellinae and other Caelopyginae genera (including Parampheres) share the moderately long pedipalp with a biconcave tarsus.The Progonyleptoidellinae genera share a short male coxa IV, which is only visible dorsally at the distal portion.On the other hand, the species of Parampheres have a wide male coxa IV.
Both analyses (molecular and morphological) supported a close relationship of Parampheres with other genera of Caelopyginae.However, these results need to be looked at with caution, since taxon coverage was different in both analyses.

Table I .
Data matrix for the species of Caelopyginae and outgroups.