Zootaxa 1067: 1–36 (2005) ISSN 1175-5326 (print edition) www.mapress.com/zootaxa/ ZOOTAXA 1067 Copyright © 2005 Magnolia Press ISSN 1175-5334 (online edition)

The Neotropical annual killifish Garman (Teleostei: : ): phylogenetic relationships, descriptive morphology, and taxonomic revision

WILSON J. E. M. COSTA * * Laboratório de Ictiologia Geral e Aplicada, Departamento de Zoologia, Universidade Federal do Rio de Janeiro, Caixa Postal 68049, CEP 21944-970, Rio de Janeiro, Brasil. [email protected]

Abstract

Pterolebias is revised on the basis of osteology, external morphology, latero-sensory system, and color patterns. Two are recognized as valid: P. longipinnis, from the eastern and southern Amazon River basin and the ParanáParaguay River system, in Brazil, Bolivia, Paraguay and Argentina; and P. phasianus, from the Paraguay River basin, in Brazil and Bolivia. Pterolebias bokermanni and P. luelingi are considered synonyms of P. longipinnis. Monophyly of Pterolebias is corroborated by the morphology of the angulo-articular, second pharyngobranchial, maxilla, metapterygoid, quadrate, basihyal, and two derived color patterns. In contrast to molecular studies, monophyly of an assemblage including Pterolebias and Gnatholebias is herein strongly supported by reduction of the interarcual cartilage, morphology of the anterior proximal radials of the anal fin and pelvic girdle, presence of scale rows on the anal-fin base, derived jaw dentition, numerous pelvic-fin rays, and long pelvic fins. A recent proposal to unite Pterolebias and Gnatholebias in a single genus is rejected.

Key words: Killifishes, Cyprinodontiformes, Rivulidae, Pterolebias, Neotropical, Amazon, Pantanal, systematics, phylogenetic relationships

Resumo

Pterolebias é revisado com base em osteologia, morfologia externa, sistema látero-sensorial e padrões de colorido. Duas espécies são reconhecidas como válidas: P. longipinnis, do leste e sul da bacia amazônica e do sistema fluvial ParanáParaguai, no Brasil, Bolívia, Paraguai e Argentina, e P. phasianus, da bacia do rio Paraguai, no Brasil e Bolívia; P. bokermanni e P. luelingi são consideradas sinônimos de P. longipinnis. Monofiletismo de Pterolebias é corroborado pela morfologia do ângulo-articular, segundo faringo-branquial, maxila, metapterigóide, quadrado, basial e dois padrões de colorido derivados. Ao contrário de estudos moleculares, monofiletismo de um agrupamento incluindo Pterolebias e Gnatholebias é aqui altamente sustentado pela redução de cartilagem interarcual, morfologia dos radiais proximais anteriores da nadadeira anal e da

Accepted by C. Gilbert: 16 Sept. 2005; published: 21 Oct. 2005 1 ZOOTAXA cintura pélvica, presença de escamas na base da nadadeira anal, dentição derivada em maxilas, 1067 numerosos raios na nadadeira pélvica e longa nadadeira pélvica. Uma recente proposta para unir Pterolebias e Gnatholebias num único gênero é rejeitada.

Introduction

Pterolebias Garman, 1895, occurs over a vast area of South America, including the eastern and southern Amazon River basin and the ParanáParaguay River system (Costa, 1998a). It is a well defined clade of annual killifishes, supported both by morphological (Costa, 1998a) and molecular studies (Murphy et al., 1999; Hrbek & Larson, 1999). However, its is still poorly known. It includes four nominal species: P. longipinnis Garman, 1895; P. bokermanni Travassos, 1969; P. luelingi (Meinken, 1969); and P. phasianus Costa, 1988. Pterolebias longipinnis has been recorded over the entire geographic range of the genus, from Marajó Island in northern Brazil (about 1º S) to Corrientes in Argentina (about 28° S). Two nominal species from the Madeira River basin, P. bokermanni and P. luelingi, are insufficiently diagnosed and not distinguishable from P. longipinnis (Thomerson, 1984; Costa, 1988). Pterolebias phasianus is a distinctively slender species with a unique color pattern. Relationships of Pterolebias to other rivulid genera are also controversial. The hypothesis generated by morphological phylogenetic analysis indicates that Pterolebias is the sister group to Gnatholebias Costa, 1998, an annual killifish genus occurring in the Orinoco River basin and adjacent coastal river basins in Venezuela and Colombia (Costa, 1998a). However, hypotheses derived from molecular studies support Gnatholebias as more closely related to other annual rivulid genera endemic to northern South America (i.e., Renova Thomerson & Taphorn, 1995; Terranatos Taphorn & Thomerson, 1978; Micromoema Costa, 1998; Myers, 1927; and Myers, 1932) than to Pterolebias (Murphy et al., 1999; Hrbek & Larson, 1999). The objectives of the present study are: 1) to provide a detailed description of morphological traits of Pterolebias, which in turn allows better morphological character analyses; 2) to identify informative characters useful for the diagnoses of species, based on large collections totaling over 400 specimens over the entire geographic range of the genus; and 3) to recognize and redescribe the valid species.

Taxonomic History

Pterolebias was first described to include a single species, P. longipinnis Garman, collected in the eastern Brazilian Amazon during the Thayer Expedition (18651866) (Garman, 1895). In Garman’s key to identification of cyprinodontiform genera, Pterolebias was distinguished from the other two rivulid genera known at that time, Rivulus Poey, 1860, and Cynolebias Steindachner, 1876, by having “body sharp-edged

2 © 2005 Magnolia Press COSTA behind vent”. This condition was repeatedly listed as diagnostic for Pterolebias (e. g., ZOOTAXA Regan, 1912; Myers, 1927), but it was considered to be a possible artifact of preservation 1067 by Myers (in Weitzman & Wourms, 1967). In the 20th century, nine new species over a wide area of South America, including the Orinoco, Amazonas and Paraguay river basins, were assigned to the genus Pterolebias: P. peruensis Myers, P. wischmanni Seegers, and P. rubrocaudatus Seegers, from the Peruvian Amazon (Myers, 1954; Seegers, 1983, 1984), P. bokermanni from the Madeira River drainage of Brazil (Travassos, 1955), P. zonatus Myers, P. maculipinnis Radda, and P. hoignei Thomerson from the Orinoco basin of Venezuela (Myers, 1935; Radda, 1964; Thomerson, 1974), P. staecki Seegers from the Brazilian central Amazon basin (Seegers, 1987), P. phasianus Costa from the Paraguay River basin, Brazil (Costa, 1988), and P. obliquus Costa, Sarmiento & Barrera from the Bolivian Amazon (Costa et al., 1996). When new collections of P. longipinnis were made in regions distant from the locality, including the Paraná-Paraguay River basin of Argentina and Paraguay (Alonso- de-Aramburu, 1961), this species became the most geographically widespread South American annual known. During the 1970s, a new annual fish species of unknown South American origin was reported in the aquarium literature as Pterolebias NSC-1 (e. g., Terceira, 1973). Costa (1989) recognized it as an undescribed genus and species, which he described as piriana Costa from the eastern Brazilian Amazon. Thomerson (1984), following comparison of the original description of Rivulichthys luelingi Meinken, from the Chapare River floodplains, Madeira River drainage, Bolivian Amazon, with data obtained from the examination of type specimens of P. longipinnis, concluded that the former is a synonym of the latter. Costa (1988) examined the type specimens of P. bokermanni and did not find characters useful for distinguishing it from P. longipinnis, but preferred not to synonymize these species, since data on live color patterns were not available at that time. However, Huber (1995) identified Pterolebias sp. aff. longipinnis from the Paraguay River basin, thus suggesting it to be a distinct species, but he provided no justification for his opinion. Schindler & Staeck (1993a, b) provided data on recent collections of Pterolebias, and, in addition, Schindler (2004) recognized Pterolebias luelingi as a valid species, and reported diagnostic features for P. bokermanni and P. longipinnis. For almost 60 years after Garmans paper, no additional diagnostic data were provided for the genus Pterolebias and no hypothesis of relationships was erected. Myers (1954) suggested Pterolebias as being closely related to Rachovia, due to the common occurrence of a short pelvic fin in Rachovia and in P. peruensis. However, a short pelvic fin is the primitive condition for rivulids, occurring in most aplocheiloid taxa, and does not, by itself, indicate a close relationship between Pterolebias and Rachovia. Weitzman & Wourms (1967) hypothesized that Rachovia, Pterolebias and Austrofundulus (including Terranatos) form a natural assemblage, based on their common

PTEROLEBIAS © 2005 Magnolia Press 3 ZOOTAXA geographic distribution in northern South America and presence of scales on the caudal 1067 fin. However, although both Rachovia and Austrofundulus have scales extensively covering the caudal fin, this condition does not occur in Pterolebias, but is present in other rivulid genera (i. e., Neofundulus Myers and Trigonectes Myers); therefore, it does not support monophyly of the group including Rachovia, Pterolebias and Austrofundulus. Weitzman & Wourms (1967) also proposed a group including Cynolebias, Cynopoecilus Regan, Pterolebias, Rachovia, Austrofundulus, Leptolebias Myers, and Simpsonichthys de Carvalho, based on the possession of thickened rays on the anterior half of the anal fin in females. According to Parenti (1981), however, this derived condition is not present in Pterolebias. Thomerson (1974) suggested that Pterolebias is an artificial assemblage, but provided no justification for this conclusion. In contrast, Parenti (1981) considered Pterolebias to be a monophyletic group diagnosed by the absence of an interarcual cartilage, based on examination of cleared and stained specimens of P. longipinnis and P. zonatus. However, Costa (1990) noted that the interarcual cartilage is present in Pterolebias, although very reduced, a condition later considered a synapomorphy for the genera Pterolebias and Gnatholebias (Costa, 1998a). Parenti (1981) recognized Pterolebias and Trigonectes as sister to a clade including Neofundulus, Rachovia, Austrofundulus, and Cynolebias, the latter including Terranatos, Leptolebias, Cynopoecilus, and Simpsonichthys, which were considered synonyms of Cynolebias. Costa (1989, 1990) hypothesized Pterolebias to be the sister group of Pituna Costa based on a reduced interarcual cartilage and widened proximal radials of the anal fin, and Pterolebias was diagnosed by the possession of eight pelvic-fin rays and a lengthened caudal fin in males. Thomerson & Taphorn (1992) criticized the employment of eight pelvic-fin rays to diagnose Pterolebias, since Rivulichthys luelingi (then considered as a synonym of P. longipinnis) and P. hoignei may sometimes have fewer pelvic-fin rays. However, Costa (1998a) asserted the non-variability of this character state in all material examined, which is in accordance with the present study. Costa (1998a) analyzed morphological traits, including osteology, laterosensory system, fin morphology and color patterns, and some sexual behavior patterns. This analysis indicated that Pterolebias was a paraphyletic assemblage, and consequently it was divided into four genera: Pterolebias, including P. longipinnis and P. phasianus; Gnatholebias, including G. zonatus and G. hoignei; Costa, including A. peruensis, A. wischmanni, A. rubrocaudatus, and A. obliquus; and Micromoema Costa, including M. xiphophora (also see Table 1, which shows the present generic placement of all species previously assigned to Pterolebias). Pterolebias was accordingly diagnosed by an anteriorly expanded ventral tip of the autopalatine, constricted dorsal portion of metapterygoid, long process of quadrate, narrow and long basihyal, reduced number of vertebrae, rounded pectoral fin, metallic orange humeral spot, and black bars on the pectoral fin in males. A sister group relationship between Pterolebias and Gnatholebias

4 © 2005 Magnolia Press COSTA was supported by large teeth on premaxilla and dentary laterally directed, long pelvic fins, ZOOTAXA and enlarged caudal fin with filaments on posterior margin in males. Pterolebias and 1067 Gnatholebias were placed in a clade that also includes Aphyolebias, Micromoema, Moema, Renova, Trigonectes and Neofundulus.

TABLE 1. Nominal species placed in Pterolebias and their present assignment according to Costa (1998a).

Nominal species Reference Assignment according to Costa (1998a) Pterolebias longipinnis Garman (1895) Pterolebias longipinnis Pterolebias zonatus Myers (1935) Gnatholebias zonatus Pterolebias peruensis Myers (1954) Aphyolebias peruensis Pterolebias bokermanni Travassos (1955) Pterolebias longipinnis Pterolebias maculipinnis Radda (1964) Rachovia maculipinnis Pterolebias NSC-1 Terceira (1973) Moema piriana Pterolebias hoignei Thomerson (1974) Gnatholebias hoignei Pterolebias wischmanni Seegers (1983) Aphyolebias wischmanni Pterolebias rubrocaudatus Seegers (1984) Aphyolebias rubrocaudatus Pterolebias staecki Seegers (1987) Moema staecki Pterolebias phasianus Costa (1988) Pterolebias phasianus Pterolebias xiphophorus Thomerson & Taphorn (1992) Micromoema xiphophora Pterolebias obliquus Costa et al. (1996) Aphyolebias obliquus

Subsequently, monophyly of both Pterolebias (sensu Costa, 1998a) and Gnatholebias were supported by molecular data published in two independent papers (Murphy et al., 1999; Hrbek & Larson, 1999). However, these studies contained unexpectedly different hypotheses of relationships involving Pterolebias and Gnatholebias. Murphy et al. (1999) analyzed four segments of the mitochondrial genome, generating an hypothesis based on parsimony analysis, in which Pterolebias would be the sister group to a clade including Renova, Terranatos, Gnatholebias, Micromoema, Rachovia, and Austrofundulus. Hrbek & Larson (1999) also analyzed mitochondrial DNA and found a most parsimonious tree, in which Pterolebias would be the sister group to Renova, and the clade Pterolebias plus Renova, the sister group to a clade including Terranatos, Gnatholebias, Micromoema, Rachovia, and Austrofundulus.

Material and methods

Material used in the taxonomic revision of Pterolebias is deposited in CBF, Collección Boliviana de Fauna, Museo Nacional de Historia Natural, La Paz; MNRJ, Museu

PTEROLEBIAS © 2005 Magnolia Press 5 ZOOTAXA Nacional, Universidade Federal do Rio de Janeiro, Rio de Janeiro; MZUSP, Museu de 1067 Zoologia, Universidade de São Paulo, São Paulo; UFRJ, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro; and USNM, National Museum of Natural History (former United States National Museum), Smithsonian Institution, Washington. All comparative material used in the phylogenetic analysis is listed in the Appendix 1. Measurements and counts follow Costa (1995). Measurements are presented as percentages of standard length (SL), except for those related to head morphology, expressed as percentages of head length. Fin-ray counts include all elements. Number of vertebrae, gill-rakers, and caudal-fin rays were recorded only from cleared and stained specimens. The compound caudal centrum was counted as a single element. Osteological preparations were made according to Taylor and Van Dyke (1985). Terminology for frontal squamation follows Hoedeman (1958), and for cephalic neuromast series follows Costa (2001). The phylogenetic analysis followed the cladistic methodology according to recent approaches discussed in Kitching et al. (1998) and Wiens (2000). With regard to testing monophyly and hypotheses of relationships of Pterolebias with other rivulid lineages, terminal taxa were species of Pterolebias (P. longipinnis and P. phasianus) and members of genera considered closely related to Pterolebias in recent phylogenetic studies (Costa, 1998a; Murphy et al., 1999; Hrbek & Larson, 1999): Aphyolebias boticarioi Costa, Austrofundulus transilis Myers, Gnatholebias hoignei (Thomerson), Gnatholebias zonatus (Myers), Micromoema xiphophora (Thomerson & Taphorn), Moema apurinan Costa, Neofundulus paraguayensis ((Eigenmann & Kennedy), Pituna compacta (Myers), Rachovia maculipinnis (Radda), Rachovia stellifer (Thomerson & Turner), Renova oscari Thomerson & Taphorn, Terranatos dolichopterus (Taphorn & Thomerson), and Trigonectes rubromarginatus Costa. The four levels of outgroups included: Rivulus janeiroensis Costa, an non annual rivulid; Kryptolebias brasiliensis (Valenciennes), a basal species of the most basal lineage of the Rivulidae (Costa, 2004); Scriptaphyosemion guignardi (Romand) of the Nothobranchidae, the sister group of Rivulidae (Murphy & Collier, 1997; Costa, 2004); and, Aplocheilus panchax (Hamilton-Buchanan) of the Aplocheilidae, the sister group of Nothobranchidae plus Rivulidae (Murphy & Collier, 1997; Costa, 2004). Character states of multi-state characters were treated as ordered whenever possible. Boundaries between states of quantitative characters were determined according to variations found among terminal taxa, in order to accommodate the real range observed for each terminal species included in that category (i. e., character state). Since these characters may involve some degree of subjectivity, they were excluded from a second analysis, which produced a single most parsimonious cladogram (tree length = 186, CI = 0.60, RI = 0.69), identical to the first of the two most parsimonious cladograms found in the first analysis (see Results below). All characters and character states used in the

6 © 2005 Magnolia Press COSTA phylogenetic analysis are listed and coded in the Appendix 2 and are organized in the data ZOOTAXA matrix of Appendix 3. Most parsimonious cladograms, consistency indices (CI) and 1067 retention indices (RI) were obtained using the algorithm mhennig*;bb* of the program Hennig86 (Farris, 1988). TreeGardener 2.2.1 (Ramos, 1996) was used for optimization of character state changes in the strict consensus tree of most parsimonious cladograms, using ACCTRAN. Bootstrap analysis (Felsenstein, 1985) was used to establish nodal support through the simple heuristic algorithm of PAUP 4.0 (Swoford, 1998) with 1000 replicates.

Results

Phylogenetic analysis. The present phylogenetic analysis is based on both a re- examination of previously reported morphological characters, plus several not previously used, for a total of 93 characters (see Appendix 2) that are included in the data matrix (Appendix 3). Two most parsimonious cladograms were found (tree length = 211, CI = 0.57, RI = 0.67), differing only in the position of the clade {T. rubromarginatus + N. paraguayensis}, which is the sister group of the clade {R. stellifer {P. compacta {A. transilis {R. maculipinnis + T. dolichopterus}}}} plus {{{A. boticarioi + M. apurinan} {M. xiphophora + R. oscari}} {{G. hoignei + G. zonatus} {P. longipinnis + P. phasianus}}} or the sister group only of {{{A. boticarioi + M. apurinan} {M. xiphophora + R. oscari}} {{G. hoignei + G. zonatus} {P. longipinnis + P. phasianus}}}. Figure 1 illustrates the strict consensus tree (three length = 211, CI = 0.57, RI = 0.67).

FIGURE 1. Strict consensus of two equally parsimonious shortest phylogenies among 17 species of the Rivulidae (L = 211, CI = 0.57, RI = 0.67). Numbers left of branches are bootstrap values.

PTEROLEBIAS © 2005 Magnolia Press 7 ZOOTAXA Taxonomic accounts 1067 Pterolebias Garman

Pterolebias Garman, 1895: 141 (type species: Pterolebias longipinnis Garman, 1895; type by monotypy).

Diagnosis: Pterolebias differs from all other genera of Rivulidae in possessing a narrow and pointed ventral process of angulo-articular, an expanded medial flap on the second pharyngobranchial, small humeral metallic orange spots in males, and dark gray bars on the pectoral fin in males. Other derived characters shared by the species of Pterolebias but hypothesized to be independently acquired in other rivulid lineages are: distal portion of maxilla strongly twisted, long posterior process of quadrate, and a narrow and long basihyal. Osteology. Superficial dermal bones and neurocranium (Figs. 2a–d): Nasal thin, scale- like. Lacrymal thin and twisted, almost cylindrical, without posterior extensions. Dermosphenotic thin and ovoid, its outer surface gently concave. Neurocranium depressed. Dorsal surface with thin ossification, bones with unclear limits. Frontal approximately rectangular, with short lateral borders. Parietal subtriangular. Supraoccipital with narrow, paired posterior process, its tip near neural spine of first vertebra. Lateral process of sphenotic narrow. Limits of bones along ventral surface of neurocranium conspicuous. Vomer trapezoidal with elongated, pointed posterior process. Two to ten conical teeth on anteromedian portion of vomer. Lateral ethmoid with distinctive anterior retrorse process. Anteromedian portion of lateral ethmoid slightly dorsally overlapping lateral portion of vomer, but never abutting lateral surface of anterior process of parasphenoid. Parasphenoid cross-shaped, with two lateral processes. Anterior portion of parasphenoid wide and pointed, overlapping dorsally posterior process of vomer; posterior portion wide and truncate, firmly attached to basioccipital; anteriormost lateral process connected to pterosphenoid; posterior lateral process attached to the prootic. Jaws, jaw suspensorium, and opercular apparatus (Figs. 2a–c): Jaw bones relatively short. Premaxilla with teeth along medial half of its anterior face; ascending process narrow, approximately rectangular, anterodistal process about trapezoidal. Maxilla rod- shaped, strongly twisted in its distal portion, bifid in its medial portion, with subtriangular dorsal process and ventral process bent. Rostral cartilage longer than wide. Dentary with teeth on its distal half. Angulo-articular triangular, with short and pointed ventral process. Retro-articular small. Meckels cartilage elongate. Premaxillary and dentary teeth conical, arranged in irregular rows; most teeth small, some larger ones forming outer row, in which, on middle of row, one or two teeth are strongly laterally displaced.

8 © 2005 Magnolia Press COSTA ZOOTAXA 1067

FIGURE 2. Dermal bones of head, neurocranium, and axial and caudal skeleton of Pterolebias longipinnis, UFRJ 6140, male, 48.3 mm SL: a, infraorbital series, left lateral view; b, neurocranium, general ventral view; c, left posterior edge of neurocranium, dorsal view, showing relative position between parietal, sphenotic and pterotic; d, posterior portion of neurocranium and first three vertebrae, left lateral view; e, fifth caudal vertebrae, left lateral view; f, last caudal vertebrae and caudal skeleton, left lateral view. BO = basioccipital; CR = caudal-fin rays; D1 = first dorsal-fin ray; DE = dermosphenotic; E1–3 = epipleural ribs of vertebrae 1–3; EO = exoccipital; EP = epural; FR = frontal; HP = hypural plate; LA = lachrymal; LE = lateral ethmoid; NA = nasal; NS = neural spine; P2–3 = pleural ribs of vertebrae 2–3; PA = parietal; PH = parhypural; PL = pleural rib; PP = pterosphenoid; PR = prootic; PS = parasphenoid; PT = pterotic; SO = supraoccipital; SP = sphenotic; U2–3 = preural centra 2–3; V1–3 = vertebrae 1–3; VO = vomer. Larger stippling indicates cartilage. Scale bar 1 mm.

PTEROLEBIAS © 2005 Magnolia Press 9 ZOOTAXA Autopalatine and ectopterygoid completely fused, forming single structure, in which 1067 ventral tip slightly abuts quadrate. Mesopterygoid thin, posteriorly contacting metapterygoid, ventrally overlapping dorsal portion of quadrate, and anterodorsally overlapping autopalatine. Quadrate approximately triangular, with long posterior process, longer that quadrate without posterior process. Sympletic elongate. Metapterygoid wide in its ventral portion, becoming abruptly narrower in its dorsal portion. All opercular apparatus bones thin. Dorsal extremity of opercle and subopercle pointed. Preopercle curved, dorsal portion forming a distinct canal. Hyoid and branchial arches (Figs. 3d–g): Basihyal subtriangular, longer than space occupied by basibranchials, and narrow, its width about 35–45% of its length; basihyal cartilage short, about 20–25% of basihyal length. Dorsal and ventral hypohyals small, always ossified. Anterior and posterior ceratohyals separated by wide cartilage. Six branchiostegal rays. Interhyal cartilaginous and minute. Urohyal with long anterodorsal process and short anterior process. Second pharyngobranchial approximately triangular, longer than wide, with prominent medial flap; two to four small conical teeth on proximal border. Third pharyngobranchial with conical teeth. Interarcual cartilage minute. Second pharyngobranchial without subdistal process. Third epibranchial with short uncinate process. Proximal edge of first hypobranchial bifid; cartilage along entire lateral margin of first hypobranchial. Fourth ceratobranchial with small conical teeth along most dorsal surface. Fifth ceratobranchial subtriangular, wide in P. longipinnis and narrow in P. phasianus, with robust conical teeth. Gill-rakers of first branchial arch 2 + 9–10. Vertebrae and caudal skeleton (Figs. 2d–f): Neural spines of first five vertebrae wider than neural spines of posterior vertebrae. Paired process on anterior subdorsal portion of first vertebra. Base of second epipleural rib wide, often with short dorsal process. Caudal vertebrae with long neural prezygapophyses. Total vertebrae 30–34.

FIGURE 3. Jaw suspensorium, opercular apparatus, and hyoid and branchial arches of Pterolebias longipinnis, UFRJ 6140, male, 48.3 mm SL: a, jaws, jaw suspensorium and opercular apparatus, left side, lateral view; b, left upper jaw, dorsal view; c, left lower jaw, ventrolateral view; d, hyoid and branchial arches, left and middle region, dorsal view of ventral part and ventral view of dorsal part; e, left hyoid bar, lateral view; f, urohyal, left lateral view; g, detailed ventral view of second pharyngobranchial and interarcual cartilage. AA = angulo-articular; ACH = anterior ceratohyal; B1–3 = basibranchials 1–3; BB = basibranchial cartilage; BC = basihyal cartilage; BH = basihyal; C1–5 = ceratobranchials 1–5; DE = dentary; DHH = dorsal hypohyal; E1–4 = epibranchials 1–4; H1–3 = hypobranchials 1–3; HY = hyomandibula; IC = interarcual cartilage; IH = interhyal; IO = interopercle; MC = Meckels cartilage; MS = mesopterygoid; MT = metapterygoid; MX = maxilla; OP = opercle; P2–3 = pharyngobranchials 2–3; PCH = posterior ceratohyal; PL = autopalatine; PM = premaxilla; PO = preopercle; QU = quadrate; RA = retro-articular; R1–6 = branchiostegal rays 1–6; RC = rostral cartilage; SO = subopercle; SY = sympletic; T4 = tooth plate pharyngobranchial 4; VHH = ventral hypohyal. Larger stippling indicates cartilage. Scale bar 1 mm.

10 © 2005 Magnolia Press COSTA ZOOTAXA 1067

PTEROLEBIAS © 2005 Magnolia Press 11 ZOOTAXA Epural and parhypural with similar shape, proximal portion respectively near neural 1067 and hemal spine of preural centrum. Hypurals fused, forming two plates separated by median gap. Accessory caudal cartilages absent. Dorsal and anal fins (Figs. 4a–b): Males and females with same number of dorsal and anal-fin rays. Most rays branched. Dorsal-fin origin on vertical between 18th and 21st vertebra, anal-fin origin on vertical between 12th and 15th vertebra. Two rays associated with first proximal radial of dorsal fin, and three associated with first proximal radial of anal fin. First dorsal-fin proximal radials slightly widened on proximal portion, first anal- fin proximal radials widened and dorso-posteriorly directed. Middle and distal radials of dorsal and anal fins ossified.

FIGURE 4. Fin support of Pterolebias longipinnis, UFRJ 6140, male, 48.3 mm SL: a, anterior portion of dorsal-fin support, left lateral view; b, anterior portion of anal-fin support, left lateral view; c, left pelvic girdle, dorsal view; d, left shoulder girdle, left lateral view. A1 = first anal-fin ray; CL = cleithrum; CO = coracoid; D1 = first dorsal-fin ray; DR = distal radial; MR = median radial; P3 = post-cleithrum 3; PB = pelvic bone; PLR = pelvic-fin rays; PR = proximal radials; PT = posttemporal; PTR = pectoral-fin rays; SC = supracleithrum; SP = scapula. Larger stippling indicates cartilage. Scale bar 1 mm.

Shoulder and pelvic girdles (Figs. 4c–d): Dorsal portion of cleithrum short, without posterior flange. Ventral process of posttemporal moderate to long. Posttemporal and supracleithrum co-ossified, limits almost inconspicuous. Prominent keel along posttemporal and supracleithrum. Ventral tip of coracoid dorsoposteriorly placed to ventral tip of cleithrum. Coracoid separated from scapula by narrow cartilage. Proximal radials of pectoral fin cubic, ventralmost radial with ventrally expanded posterior portion.

12 © 2005 Magnolia Press COSTA Third postcleithrum rod-shaped. Pelvic bones medially in contact, ischial process ZOOTAXA indistinct. 1067

Pterolebias longipinnis Garman, 1895 (Figs. 5–7)

Pterolebias longipinnis Garman, 1895: 142 (type locality: Santarém [Amazon River basin, Estado do Pará, Brazil]). Pterolebias bokermanni Travassos, 1955: 35 (type locality: Guajará-Mirim River [correctly city of Guajará-Mirim, Mamoré River floodplains, Madeira River drainage, Amazonian basin], Ter- ritório de Guaporé [now Estado de Rondônia], Brazil). Rivulichthys luelingi Meinken, 1969: 423 (type locality: forest pool in the region of Chapare River [Mamoré River drainage, a part of the Madeira River drainage, Amazonian basin] about 4 km from Todos Santos [Departamento de Cochabamba], Bolivia).

Material examined. Lower Amazonas River basin. Brazil: Pará: USNM 120429, lectotype, male, 44.7 mm SL (lectotype designated by Thomerson, 1984: 528). MZUSP 5048, 2; Cachoeira do Arari, Marajó Island; Excursão Departamento de Zoologia, 12 June 1966. UFRJ 3676, 2 (c&s); Marajó Island; no date on collectors. Madeira River drainage, Brazil: Rondônia: MNRJ 8672, holotype of P. bokermanni Travassos, male, 34.4 mm SL; Guajará-Mirim; H. Schultz, August 1953. MNRJ 8697, paratype, male, 39.7 mm SL; Forte Príncipe da Beira, Guaporé River floodplains; H. Schultz, October 1953. UFRJ 5882, 41; UFRJ 5883, 6 (c&s); temporary pool in Guajará- Mirim, Mamoré River floodplains, 20o4553.5”S 65o2021.7”W; W. J. E. M. Costa, S. Lima and A. Pinto, 7 June 2003. Brazil: Mato Grosso: UFRJ 3644, 7; UFRJ 6139, 2 (c&s); 2 km from Vila Bela, Guaporé River floodplains; W. J. E. M. Costa, M. I. Landim, C. Moreira and R. D'Arrigo, 28 April 1996. Bolivia: Beni: CBF 2041, 3; CBF 2047, 6 (2 c&s); Ballivian, Espíritu, Yacuma River drainage; J. Sarmiento & W. Hanagarth, 5–6 May 1987. CBF 2021, 3 (1 c&s); idem; J. Sarmiento & W. Hanagarth, 2–3 May 1987. CBF 2102, 1; idem; W. Hanagarth, 29 January 1983. CBF 1877, 6; CBF 1716, 4; idem; J. Sarmiento & W. Hanagarth, 23 April 1987. CBF 0032, 5; Cercado, Trinidad; G. Loubens et al., 27 June 1986. UFRJ 4385, 2; road Santa Cruz-Trinidad, Santa Cruz, San Pablo River floodplains; W. J. E. M. Costa, C. P. Bove and S. Barrera, 22 February 1997. MZUSP 27856, 2; San Juan Creek, Trinidad; ORSTOM-UTB, March 1983. Paraná–Paraguay River system. Argentina: Corrientes: MZUSP 10241, 10; Pirajuí Creek; E. Arriguti, 1973. Brazil: Mato Grosso: UFRJ 4901, 71; UFRJ 4896, 91; UFRJ 4905, 108; UFRJ 4889, 59; UFRJ 5583, 2; UFRJ 6140, 6 (c&s); temporary swamp near Porto Cercado; W. J. E. M. Costa, A. C. Bacellar, M. A. Barbosa and F. Autran, 9–10 June 1999. UFRJ 4911, 9; UFRJ 4894, 3; about 5 km N from Porto Cercado; W. J. E. M. Costa, A. C. Bacellar, M. A. Barbosa and F. Autran, 11 June 1999. UFRJ 6141, 2; idem; W. J. E. M. Costa, B. B. Costa, C. P. Bove, M. A. Barbosa & R. Cunha, 18 July 2003. UFRJ 4884,

PTEROLEBIAS © 2005 Magnolia Press 13 ZOOTAXA 17; swamp 5 km NW from Posto São Luís, RPPN Sesc-Pantanal; W. J. E. M. Costa, A. C. 1067 Bacellar, M. A. Barbosa and F. Autran, 12 June 1999. UFRJ 5455, 9; UFRJ 5456, 37; Porto Cercado; W. J. E. M. Costa, M. A. Barbosa, R. Cunha and S. Lima, 12 June 2002. UFRJ 3645, 4; swamp 5 km S from Santo Antônio do Leverger; W. J. E. M. Costa, M. I. Landim, C. Moreira and R. D'Arrigo, 26 April 1996. Brazil: Mato Grosso do Sul: MZUSP 49059, 3; Corumbá: Passo do Lontra, Miranda River floodplains; O. Froehlich, 28 January 1992. MZUSP 38397, 1; Cáceres; P. D. Cardoso, March 1987. UFRJ 5888, 7; Camburizal near Posto Espírito Santo, RPPN Sesc-Pantanal; W. J. E. M. Costa, B. B. Costa, C. P. Bove, M. A. Barbosa & R. Cunha, 15 July 2003.

FIGURE 5. Pterolebias longipinnis, UFRJ 6141, male, 44.6 mm SL (one day after collection); Brazil: Mato Grosso: Porto Cercado: rio Paraguay basin.

FIGURE 6. Pterolebias longipinnis, UFRJ 5882, male, 35.7 mm SL (one day after collection); Brazil: Rondônia: Guajará-Mirim: rio Mamoré floodplains.

FIGURE 7. Pterolebias longipinnis, UFRJ 5583, female, 33.5 mm SL (one day after collection); Brazil: Mato Grosso: Porto Cercado: rio Paraguay basin.

14 © 2005 Magnolia Press COSTA Diagnosis: Distinguished from P. phasianus by having a deeper body (body depth ZOOTAXA 25.0–28.8 % SL vs. 19.5–23.0 % SL in males, 25.3–27.0 % SL vs. 19.4–23.1 % SL in 1067 females; caudal peduncle depth 15.8–17.8 % SL vs. 11.8–14.8 % SL in males, 14.9–16.8 % SL vs. 11.9–13.7 % SL in females), three to seven rows of scales on anal-fin base in males (vs. a single row), one or two rows of scales on dorsal-fin base in males (vs. no scales), pelvic-fin bases medially united (vs. separated), more neuromasts in the infraorbital series (2–3 + 19–21 + 1, vs. 1–2 + 16–17 + 1), preopercular series (2 + 11–14, vs. 2 + 9–10), and mandibular series (6–7 + 2–4, vs. 5 + 2), flank light brown with oblique rows of green dots (vs. blue with dark brown stripes), absence of a subdistal zigzag bluish white line on anal fin in male (vs. presence). Pterolebias longipinnis usually has more dorsal and anal fin rays than P. phasianus (9–11 and 17–22, respectively, vs. 7–9 and 15–17). Description. Morphometric data appear in Table 2. Males larger than females, largest male 50.5 mm SL. Dorsal profile slightly convex from snout to end of dorsal-fin base, gently concave on caudal peduncle. Ventral profile slightly convex from lower jaw to end of anal-fin base, weakly concave along caudal peduncle. Body somewhat deep, subcylindrical anteriorly, deeper than wide, to compressed posteriorly. Greatest body depth at level of pelvic-fin base. Jaws short, snout slightly pointed.

TABLE 2. Morphometric data of Pterolebias longipinnis and P. phasianus.

P. longipinnis P. phasianus males females males females (n = 15) (n = 15) (n = 9) (n = 9) Standard length (mm) 33.8–47.6 28.7–47.1 31.4–41.8 25.7–32.3 Percents of standard length Body depth 25.0–28.8 25.3–27.0 19.5–23.0 19.4–23.1 Caudal peduncle depth 15.8–17.8 14.9–16.8 11.8–14.8 11.9–13.7 Predorsal length 76.8–79.8 79.5–82.6 78.6–82.5 80.4–83.3 Prepelvic length 46.1–51.8 48.5–54.5 47.8–51.0 50.3–52.9 Length of dorsal-fin base 10.2–13.1 8.7–12.0 7.9–9.9 6.7–9.1 Length of anal-fin base 26.7–31.2 22.5–28.9 23.1–24.6 19.7–23.1 Caudal-fin length 68.5–80.3 45.6–49.0 59.6–69.4 44.9–50.8 Pectoral-fin length 25.2–26.5 22.5–26.3 22.2–26.2 22.0–22.8 Pelvic-fin length 43.5–71.6 24.4–34.3 33.2–49.5 17.3–20.8 Head length 26.8–29.3 27.9–30.8 25.0–27.5 26.1–27.8 Percents of head length Head depth 70.9–78.9 66.4–73.4 60.7–66.0 61.4–65.9 Head width 66.1–71.2 67.8–72.4 63.6–70.0 68.4–75.7 Snout length 14.7–16.9 13.7–15.7 14.8–16.8 13.3–15.3 Lower jaw length 16.5–20.7 15.9–18.4 16.6–19.7 17.6–19.2 Eye diameter 34.5–40.2 33.9–42.3 35.7–41.0 37.3–41.0

PTEROLEBIAS © 2005 Magnolia Press 15 ZOOTAXA Tip of dorsal fin pointed with short filamentous rays in males, rounded to slightly 1067 pointed in females. Anal fin approximately rectangular in males, longer than deep, and long, reaching vertical through middle of caudal fin; tip of anal fin pointed, with filamentous rays. Anal fin rounded, short, and without filamentous rays in females. Caudal fin rounded to subtruncate, long in males, with posterior margin terminating in fringe-like posterior extensions; caudal fin short, elliptical, and without posterior extensions in females. Pectoral fin elliptical, posterior margin reaching vertical between pelvic-fin base and anus. Pelvic fin pointed, terminating in long tip, reaching between middle of anal-fin base and caudal-fin base in males, and between base of 4th and 13th anal-fin rays in females. Pelvic-fin bases medially united. Dorsal-fin origin on vertical between base of 12th and 14th anal-fin rays, and between neural spines of 18th and 22nd vertebrae. Anal-fin origin between pleural ribs of 12th and 15th vertebrae. Dorsal-fin rays 9–11; anal-fin rays 17–22; caudal-fin rays 24–28; pectoral-fin rays 15–16; pelvic-fin rays 8–9. Scales large, cycloid. Body and head entirely scaled, except anterior ventral surface of head. Body squamation slightly extending on caudal-fin base, with 3–5 transverse rows of scales onto caudal-fin base; 1–2 rows of scales on dorsal-fin base, 4–7 on anal-fin base. Frontal squamation E-patterned; E-scales not overlapping medially; scales arranged in irregular circular pattern around A-scale without exposed margins. Longitudinal series of scales 30–33; transverse series of scales 8; scale rows around caudal peduncle 16. Contact organs absent. Cephalic neuromasts: supraorbital 3 + 3, parietal 1, anterior rostral 1, posterior rostral 1, infraorbital 2–3 + 19–21 + 1, preorbital 3–5, otic 1, postotic 1–2, supratemporal 1, median opercular 1, ventral opercular 2–3, preopercular 2 + 11–14, mandibular 6–7 + 2–4, lateral mandibular 3. One neuromast on each scale of lateral series. Coloration: Males: Side of body light brown, with oblique rows of bright blue to green dots; often small bright orange spot on humeral region; sometimes black humeral blotch, more frequent in individuals from Mamoré River basin in Bolivia. Dorsum light brown, sometimes orangish brown in individuals from Mamoré River basin in Brazil; venter purplish white. Side of head light green with faint purplish gray spots. Jaws light gray. Infraorbital region and ventral surface of head light gray. Iris yellowish green, with dark brown bar crossing center of eye. Dorsal fin greenish yellow with small dark reddish brown spots, darker on basal portion of fin. Anal fin light greenish yellow, basal region light blue, with pale greenish brown spots, darker on posterior portion of fin, then alternating with bluish white spots; often distal portion of fin pink; about 60% of individuals from Mamoré, Guaporé and Paraguay river basins in Brazil with narrow distal black stripe and narrow light blue subdistal stripe, but about 40% of individuals without vestige of such stripes; sometimes orange subdistal stripe in individuals from Mamoré River basin in Brazil; often melanophores concentrated on distal portion of fin in individuals from Mamoré River basin in Bolivia. Caudal fin greenish yellow with small

16 © 2005 Magnolia Press COSTA pale greenish brown spots and light green dots; often dorsal and ventral margins and ZOOTAXA filaments dark orange. Pelvic fin greenish yellow, base light blue with dark reddish brown 1067 spots, filament light orange, sometimes tip white; sometimes dark orange pigmentation along fin. Pectoral fin hyaline with narrow gray bars. Females: Side of body gray with oblique rows of pale green dots. Dorsum gray, venter light gray. Side of head pale green. Jaws light gray. Infraorbital region and ventral surface of head light gray. Iris yellowish, with dark brown bar crossing center of eye. Unpaired fins hyaline with faint gray spots. Paired fins hyaline. Distribution: Lower Amazonian basin, Madeira River drainage, and Paraná–Paraguay River system, in Brazil, Bolivia, Paraguay and Argentina (Fig. 8). Habitat: Temporary lagoons in open sunny areas, sometimes at borders of forests.

FIGURE 8. Geographic distribution of Pterolebias longipinnis and P. phasianus. Triangles = P. longipinnis; stars = P. longipinnis and P. phasianus; inverted triangles = occurrence of P. longipin- nis recorded in literature; dot = occurrence of P. longipinnis and P. phasianus recorded in literature; 1 = type locality of P. longipinnis; 2 = type locality of P. bokermanni; 3 = type locality of Rivulich- thys luelingi; 4 = type locality of P. phasianus. Some symbols may represent more than one locality.

PTEROLEBIAS © 2005 Magnolia Press 17 ZOOTAXA Pterolebias phasianus Costa, 1988 1067 (Figs. 9–10)

Pterolebias phasianus Costa, 1988a: 658 (type locality: lagoon at Cáceres, Cuaiabá River [correctly Paraguay River] floodplains, Mato Grosso, Brazil).

Material examined. All from Paraguay River basin. Brazil: Mato Grosso: MZUSP 38109, holotype, male, 31.4 mm SL; MZUSP 38110, 1 paratype, female, 29.2 mm SL (c&s); MZUSP 38111, 1 paratype, female, 25.7 mm SL; lagoon in Cáceres, Paraguay River floodplains, Mato Grosso; P. D. Cardoso-Filho, March 1987. UFRJ 3649, 39; UFRJ 3673, 4 (c&s); Paraguay River floodplains, Cáceres; W. J. E. M. Costa, M. I. Landim, R. DArrigo & C. L. Moreira, 30 April 1996. UFRJ 318, 7; UFRJ 273, 1 (c&s); Cuiabá River basin, Mato Grosso; J. D. Soares, March 1990. UFRJ 4899, 8; UFRJ 4903, 2; swamp near Porto Cercado, Cuaibá River floodplains; W. J. E. M. Costa, F. Autran, A. C. Bacellar & A. Barbosa, 9 June 1999. UFRJ 5182, 2; same locality and collectors as UFRJ 4899, 11 June 1999. UFRJ 4914, 1; beach on Cuiabá River, RPPN SESC-Pantanal; idem, 10 June 1999. UFRJ 4885, 9; temporary pool at Posto São Luís, São Lourenço River drainage, RPPN SESC-Pantanal; idem, 12 June 1999. Mato Grosso do Sul: MZUSP 38112, 1; MZUSP 38113, 1; Ladário; E. K. Bastos, 10 March 1985. MZUSP 49027, 4; Passo do Lontra, Corumbá; O. Froehlich & E. A. Gonçalves, 23 May 1990. UFRJ 3674, 9; UFRJ 3675, 3 (c&s); floodplains of Miranda River, Miranda; W. J. E. M. Costa, M. I. Landim, R. DArrigo & C. L. Moreira, 21 April 1996. Diagnosis: Readily distinguished from P. longipinnis by having slender body (body depth 19.5–23.0 % SL vs. 25.0–28.8 % SL in males, 19.4–23.1 % SL vs. 25.3–27.0 % SL in females; caudal peduncle depth 11.8–14.8 % SL vs. 15.8–17.8 % SL in males, 11.9–13.7 % SL vs. 14.9–16.8 % SL in females), a single row of scales on the anal-fin base in males (vs. three to seven rows), no scales on the dorsal-fin base in males (vs. one or two rows of scales), pelvic-fin bases separated (vs. medially united), fewer neuromasts in the infraorbital series (1–2 + 16–17 + 1, vs. 2–3 + 19–21 + 1), preopercular series (2 + 9–10, vs. 2 + 11–14), and mandibular series (5 + 2, vs. 6–7 + 2–4), flank blue with dark brown stripes (vs. light brown with oblique rows of green dots), and presence of a subdistal zigzag bluish white line on anal fin in males (vs. absence). Pterolebias phasianus usually has fewer dorsal and anal fin rays than P. longipinnis (7–9 and 15–17, respectively, vs. 9–11 and 17–22). Description. Morphometric data appear in Table 2. Males larger than females, largest male 41.8 mm SL. Dorsal profile slightly convex from snout to end of dorsal-fin base, approximately straight on caudal peduncle. Ventral profile slightly convex from lower jaw to end of anal-fin base, nearly straight on caudal peduncle. Body slender, subcylindrical anteriorly, slightly deeper than wide, to compressed posteriorly. Greatest body depth at level of pelvic-fin base. Jaws short, snout slightly pointed.

18 © 2005 Magnolia Press COSTA ZOOTAXA 1067

FIGURE 9. Pterolebias phasianus, UFRJ 5182, male, 35.8 mm SL; Brazil: Mato Grosso: Porto Cercado: rio Paraguay basin.

FIGURE 10. Pterolebias phasianus, UFRJ 5182, female, 32.0 mm SL; Brazil: Mato Grosso: Porto Cercado. Photo by W. J. E. M. Costa.

Tip of dorsal fin pointed with short filamentous rays in males, rounded to slightly pointed in females. Anal fin approximately rectangular, longer than deep; tip pointed with filamentous rays, long, reaching vertical through middle of caudal fin in males, rounded and short in females. Caudal fin elliptical, long; filamentous rays on posterior edge of male caudal fin. Pectoral fin elliptical, posterior margin reaching vertical just posterior to pelvic-fin base. Pelvic fin long, its tip reaching between middle of anal-fin base and caudal-fin base in males, and between urogenital papilla and base of 2nd anal-fin ray in females. Dorsal-fin origin on vertical between base of 11th and 12th anal-fin ray, and between neural spines of 20th and 22nd vertebrae. Anal-fin origin between pleural ribs of 14th and 16th vertebrae. Dorsal-fin rays 7–9; anal-fin rays 15–17; caudal-fin rays 24–25; pectoral-fin rays 14–15; pelvic-fin rays 8. Scales large, cycloid. Body and head entirely scaled, except anterior ventral surface of head. Body squamation slightly extending on caudal-fin base, with 3–5 transverse rows of scales on caudal-fin base; row of scales along anal fin in males; no scales on dorsal fin.

PTEROLEBIAS © 2005 Magnolia Press 19 ZOOTAXA Frontal squamation E-patterned; E-scales not overlapping medially; scales arranged in 1067 circular pattern around central A-scale without exposed margins. Longitudinal series of scales 31–33, transverse series of scales 8, scale rows around caudal peduncle 16. Contact organs absent. Supraorbital neuromasts 3 + 3. Cephalic neuromasts: supraorbital 3 + 3, parietal 1–2, anterior rostral 1, posterior rostral 1, infraorbital 1 + 16–17 + 1, preorbital 2–3, otic 1, postotic 2, supratemporal 1, median opercular 1, ventral opercular 2, preopercular 2 + 9–10, mandibular 5 + 2, lateral mandibular 3. One neuromast on each scale of lateral series. Coloration: Males: Side of body iridescent blue with three reddish brown stripes along flank, more dorsal area lighter, often short brown stripe or row of spots between pelvic-fin base and posterior portion of anal-fin base; few small brown spots on anterior portion of flank between stripes; usually horizontal row with one to five bright orange rounded spots on humeral region, sometimes surrounded by black pigmentation. Dorsum pale brown, venter white. Opercular region golden with two or three dark brown to black oblique stripes, sometimes fragmented forming irregular blotches. Snout and jaws dark brown. Iris brown, with dark brown to black bar. Dorsal fin green with transverse rows of dark brownish red spots. Anal fin light blue on basal portion, yellow on median portion and black on distal portion; transverse row of reddish brown spots, sometimes coalesced forming stripe, between light blue and yellow zones; zigzag bluish white line on black distal zone. Caudal fin greenish blue with reddish brown spots; dorsal and ventral portions greenish yellow, edges brownish orange; sometimes narrow ventral black stripe. Pectoral fin hyaline with dark gray bars. Pelvic fin yellow with brown dots; filament yellow to light orange. Females: Side of body pale blue with dark brown stripes arranged in pattern similar to male pattern. Head color pattern as in males, but colors fainter. Unpaired fins hyaline with dark brown spots, sometimes spots coalesced forming stripe on median portion of anal fin; anal-fin base pale blue. Paired fins hyaline. Distribution: Middle Paraguay River basin, in Brazil and Bolivia (Fig. 8). Habitat notes: Temporary lagoons in the Pantanal. Often sympatric with P. longipinnis, in which cases P. phasianus is usually found in shaded areas and P. longipinnis is found in sunny areas.

Discussion

Taxonomy. Pterolebias possibly poses the greatest taxonomic problems within the Rivulidae (see History above). This is due to the wide geographic range of P. longipinnis, which is unique for an annual fish, and overlaps the type locality of two similar, controversial species, P. bokermanni and P. luelingi. Travassos (1955) distinguished P. bokermanni from P. longipinnis by the former having pelvic-fin bases medially united, dorsal and anal fins placed more posteriorly, a

20 © 2005 Magnolia Press COSTA slender body, pelvic fin longer, fewer scales in the transverse series, more scales in the ZOOTAXA longitudinal series, and greater development of cephalic “pores” (i.e. neuromasts). 1067 However, Travassos never examined any specimens of P. longipinnis from the type locality or from any other region, and based comparisons on the limited data provided in Garmans (1895) description. None of these features differs among populations formerly identified as P. longipinnis, P. bokermanni, and P. luelingi. The reasons that P. luelingi was first placed in Rivulichthys by Meinken (1969) and not in Pterolebias are unknown. The few characters presented in the original description of R. luelingi are congruent with those included in the original description of P. longipinnis. Thomerson (1984) considered R. luelingi to be a synonym of P. longipinnis, although he recorded eight pelvic-fin rays for the types of P. longipinnis, in contrast to seven rays recorded by Meinken for R. luelingi. However, Schindler (2004) used this putative difference in number of pelvic-fin rays to distinguish P. luelingi, occurring in the Mamoré River basin, from P. bokermanni, from the Guaporé and Paraguay river basins, and from P. longipinnis. In fact, the type locality of P. bokermanni is Guajará-Mirim, on floodplains of the Mamoré River, although it is also recorded from the Guaporé River floodplains (Travassos, 1955). All of the approximately 400 specimens of Pterolebias examined in the present study have eight pelvic-fin rays, except for a few specimens with nine rays. Schindler (2004) also noted a reduced squamation on the anal-fin base in males, including two or three rows of scales in P. longipinnis and P. luelingi, in contrast to four or more rows in P. bokermanni. However, all adult males of Pterolebias examined during the present study have four to seven rows, except P. phasianus, which has a single irregular row. Therefore, diagnostic features recorded by Schindler (2004) are not useful for distinguishing P. luelingi and P. bokermanni from P. longipinnis. Characters examined were not useful for recognizing P. bokermanni and P. luelingi as valid species, nor for distinguishing geographically distant populations previously identified as P. longipinnis from the Paraná–Paraguay system. Some distinctive color patterns, such as stripes on male anal fin, were noted in individuals from the Mamoré and Paraguay river basins in Brazil (see above description). However, these color features are not constant, are absent from a large percentage of individuals from those regions, and therefore are not useful as diagnostic characters in this particular instance. Phylogenetic relationships. Both monophyly of Pterolebias and the sister group relationship between Pterolebias and Gnatholebias are strongly supported by bootstrap values (Fig. 1). The clade Pterolebias was previously corroborated by morphological data (Costa, 1998a) and molecular studies (Murphy et al., 1999; Hrbek & Larson, 1999). However, the clade Pterolebias plus Gnatholebias, already reported in morphological analysis (Costa, 1998a), is in conflict with molecular phylogenetic studies, which strongly support the monophyly of a clade including Gnatholebias and other taxa endemic to northern South America (Murphy et al., 1999; Hrbek & Larson, 1999) (Fig. 11).

PTEROLEBIAS © 2005 Magnolia Press 21 ZOOTAXA 1067

FIGURE 11. Molecular hypotheses of relationships among species of Pterolebias, Gnatholebias and other rivulids: a, modified from Murphy et al. (1999); b, modified from Hrbek & Larson (1999). Numbers left of branches are bootstrap values.

Monophyly of Pterolebias is herein supported by four unambiguous synapomorphies: ventral process of angulo-articular narrow and pointed (16.2) (Figs. 3a, c), medial flap of second pharyngobranchial expanded (33.1) (Fig. 3g), small metallic orange spots on humeral region in males (82.1) (Figs. 5, 6, 9), and bars on pectoral fin in males (90.2) (more conspicuous in the example of Fig. 9). The Pterolebias clade is further corroborated by four synapomorphies homoplastically occurring in other rivulids: maxilla greatly twisted (13.2) (Figs. 3a–b), dorsal portion of metapterygoid strongly constricted (19.2) (Fig. 3a), posterior process of quadrate long (20.1) (Fig. 3a), and basihyal long (25.1) (Fig. 3a); and one reversal: pectoral fin rounded (62.0). Monophyly of the clade comprising Pterolebias and Gnatholebias is supported by three unambiguous synapomorphies: interarcual cartilage minute (31.1) (Figs. 3d, g), anterior proximal radials of anal fin curved and posteriorly directed (48.1) (Fig. 4b), and anal-fin base with scales (75.1). This assemblage is also supported by four synapomorphies homoplastically occurring in other rivulid lineages: external median teeth of premaxilla and dentary displaced laterally (17.1) (Figs. 3a–c), ischial process of pelvic girdle vestigial (55.1) (Fig. 4c), eight pelvic-fin rays (56.1), and pelvic fin long in males (64.1) (Figs. 5, 6, 9, 12). However, in a study of mitochondrial DNA by Hrbek & Larson (1999), Gnatholebias zonatus was hypothesized to be more closely related to Rachovia stellifer than to other rivulids, and Terranatos dolichopterus was hypothesized to be the

22 © 2005 Magnolia Press COSTA sister group of the clade including G. zonatus plus R. stellifer (Fig. 11b). A similar ZOOTAXA hypothesis was obtained from another mitochondrial DNA study (Murphy et al., 1999), in 1067 which G. zonatus was considered to be the sister group of T. dolichopterus, but R. stellifer was not included in the analysis (Fig. 11a). I have found no morphological support for an assemblage including Gnatholebias zonatus, Terratanos dolichopterus, and Rachovia stellifer, since all apomorphic conditions shared by these three genera are present in several other rivulid lineages. Both R. stellifer and T. dolichopterus have the pelvic fin slightly elongated in males, which could be considered homologous to the long pelvic fin of G. zonatus and G. hoignei. However, the pelvic fin is much longer in Gnatholebias and Pterolebias. In addition, in the latter two genera, the pelvic fin elongation is due to a long fourth ray (of eight rays) always occurring in the fin. In T. dolichopterus, however, the longer ray is the third (of eight), thus not considered a homologous condition; and in R. stellifer the longest ray is the fourth, but since there are only seven pelvic-fin rays in that species homology is doubtful. Gnatholebias zonatus and T. dolichopterus share a medial fusion on the pelvic fin (65.2), but this is not significant when compared to the many morphological synapomorphies shared by Gnatholebias and Pterolebias (see above). Gnatholebias zonatus and T. dolichopterus share reduction or loss of the ischial pelvic process (55.1), but this condition is also present in Pterolebias. Like Gnatholebias species, T. dolichopterus has filamentous rays on the unpaired fins (67.1, 68.1), but this feature is also present in Pterolebias and in some species of Rachovia and Austrofundulus (e. g., R. brevis (Regan), A. leohoignei Hrbek, Taphorn & Thomerson, A. leoni Hrbek, Taphorn & Thomerson), which would be more closely related to Terranatos according to the morphological studies. In addition, T. dolichopterus, R. maculipinnis, and A. transilis share four synapomorphies: dorsally directed process on the base of anterior epipleural ribs (40.1), dorsal profile of the anterior portion of body convex (59.1), supraorbital spot adjacent to eye (85.1), and dark pigmentation reduced on flank and fins in females (92.1). The first feature, already described and illustrated by Costa (1998a: fig. 28a), seems to be unique among aplocheiloid . Although morphological and molecular studies often produce slightly different estimates of phylogenetic relationships among species of the same group, Hillis & Wiens (2000) considered significant incongruence rare, pointing out some common causes, such as undersampling of characters or taxa, differences in phylogenetic methods and differences in rooting techniques. However, none of these causes seems to be applicable, since all studies are based on parsimony analysis, similar outgroup analysis methods, and considerable numbers of characters and representative taxa, with conflicting nodes having high bootstrap values. It still not possible to satisfactorily understand the incongruence found between morphological and molecular hypotheses of relationships among Pterolebias, Gnatholebias and other rivulids, making it difficult to establish which data set is

PTEROLEBIAS © 2005 Magnolia Press 23 ZOOTAXA misleading. It would be desirable to improve estimates of phylogenetic relationships 1067 through the addition of characters from unsampled structures in the morphological analysis or introduction of new genes in the molecular analysis, but this is beyond the scope of the present study. Classification. Based upon a morphological phylogenetic analysis of the Rivulidae, Costa (1998a) proposed a new classification, in which Pterolebias was divided into four genera (Pterolebias, Gnatholebias, Aphyolebias, and Micromoema). The groups recognized as genera were corroborated by subsequent molecular phylogenetic studies (Murphy et al., 1999; Hrbek & Larson, 1999), and the general classification has been employed both in scientific and aquaristic literature. However, Schindler (2004) considered Gnatholebias to be a junior synonym of Pterolebias, claiming that Pterolebias plus Gnatholebias form a monophyletic group, and that “... it would be better to adopt a more conservative nomenclature than to change the systematic categories after every new analysis ...” (Schindler, 2004: 74). This proposal is not followed here by the following reasons: 1) Pterolebias including species placed in Pterolebias plus Gnatholebias sensu Costa (1998a) is not a conservative nomenclature as proposed by Schindler (2004), since this generic composition would be new. Pterolebias was a more inclusive genus prior to publication of Costas (1998a) work, and included species today placed in Aphyolebias, Moema, and Micromoema, thus forming a paraphyletic assemblage, as estimated both by morphological and molecular studies (Costa, 1998a; Murphy et al., 1999; Hrbek & Larson, 1999). Therefore, Pterolebias including species placed in Gnatholebias but excluding species of Aphyolebias, Moema, and Micromoema should not be viewed as conservative, especially considering that Costas classification has been consistently employed during the last seven years. 2) Although the clade Pterolebias plus Gnatholebias is strongly supported by morphological characters (Costa, 1998a; the present study), it is still controversial, since it is not corroborated in two independent molecular phylogenetic analyses (Murphy et al., 1999; Hrbek & Larson, 1999) (Fig. 11). Thus, Pterolebias, as proposed by Schindler (2004), would be paraphyletic according to molecular studies. 3) Both Pterolebias and Gnatholebias sensu Costa (1998a) are monophyletic assemblages, well corroborated by morphological and molecular studies (Costa, 1998a; the present study). In addition, Pterolebias and Gnatholebias are readily distinguished even by external morphological characters, easily accessible both by museum systematists and amateur ichthyologists (e. g., short snout, rounded pectoral fin, and dark bars on the pectoral fin in Pterolebias, vs. long snout, pointed pectoral fin, and pectoral fin with black ventral margin in Gnatholebias) (Fig. 12). 4) These two monophyletic groups (i. e., Pterolebias and Gnatholebias) are potential indicators of historic biogeography, since, although not necessarily sister groups, they are closely related taxa with completely disjunct biogeographic distribution patterns.

24 © 2005 Magnolia Press COSTA ZOOTAXA 1067

FIGURE 12. Gnatholebias hoignei, UFRJ 6116, male, 52.4 mm SL (one day after collection); Venezuela: Portuguesa: Papelón: río Orinoco basin.

Acknowledgments

Thanks are due to B. B. Costa, C. P. Bove, G. C. Brasil, M. A. Barbosa, A. C. Bacellar, F. Autran, S. M. Q. Lima, R. D. Cunha, R. Paiva, A. Pinto, C. Moreira, M. I. Landim, R. DArrigo, S. Barrera, K. Marchetto, L. Martinez, L.F. Jara, and J. Coronel, for help in collecting trips; to D. Taphorn, G. Nunan, D. Moraes Jr., N. Menezes, H. Britski, O. Oyakawa, L. Parenti, S. Jewett, L. Palmer, R. Vari, J. Thomerson, M. Kottelat, J. Huber, G. Brasil, D. Nielsen, J. Carvalho, A. Cyrino, and J. Sarmiento for the loan, exchange, or donation of specimens, or support during visits to their institutions; and to L. Brandão and M.E. Abib for the logical support in the RPPN SESC-Pantanal. The manuscript benefited from the suggestions by M. Ghedotti, C. Gilbert, and two anonymous reviewers. Collecting trips were supported by SESC-Pantanal and Fundação o Boticário de Proteção à Natureza. This study was funded by CNPq-MCT and FAPERJ.

References

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26 © 2005 Magnolia Press COSTA Ramos, T.C. (1996) Tree Gardener (computer program), Version 2.3beta. Univ. São Paulo, São ZOOTAXA Paulo. 1067 Regan, C.T. (1912) A revision of the poeciliid fishes of the genera Rivulus, Pterolebias, and Cyn- olebias. Annals & Magazine of Natural History, series 8, 10, 494–508. Seegers, L. (1983) Pterolebias wischmanni nov. spec. aus dem Ucayali-Einzug in Peru (Pisces: Atheriniformes: Rivulinae). Deutsche Killifisch Gemeinschaft, 15, 67–74. Seegers, L. (1984) Ein neuer Rivuline aus Peru: Pterolebias rubrocaudatus (Pisces: Atherini- formes: Cyprinodontidae). Bonner Zoologische Beitrage, 35, 243–250. Seegers, L. (1987) Die Gattung Pterolebias Garman, 1895 mit der Beschreibung von Pterolebias staecki nov. spec. Die Aquarien- und Terrarienzeitschrft, 40, 199–204. Schindler, I. (2004) Pterolebias luelingi (Meinken, 1969), a valid species of annual killifish from Bolivia (Teleostei: Rivulidae). Zeitschrift für Fischkunde, 7, 71–75. Schindler, I. & Staeck, W. (1993a) Killifisch aus dem Fluβsystem des Rio Paraguay: 1. Pterolebias phasianus. Die Aquarien- und Terrarienzeitschrft, 46, 35–39. Schindler, I. & Staeck, W. (1993b) Killifisch aus dem Fluβsystem des Rio Paraguay: 2. Pterolebias longipinnis. Die Aquarien- und Terrarienzeitschrft, 46, 462–468. Swofford, D.L. (1998) PAUP*. Phylogenetic analysis using parsimony (*and other methods), Beta version 4.0b2. Sinauer, Sunderland, Massachusetts. Taylor, W.R. & Van Dyke, G.C. (1985) Revised procedures for staining and clearing small fishes and other vertebrates for bone and cartilage study. Cybium, 9, 107–109. Taphorn. D.C. & Thomerson, J.E. (1978) A revision of the South American cyprinodont fishes of the genera Rachovia and Austrofundulus, with the description of a new genus. Acta Biologica Venezolana, 9, 377–452. Terceira, A.C. (1973) Aquarium observation of Pterolebias NSC-1. Journal of the American Kil- lifish Association, 8, 22–24. Thomerson, J.E. (1974) Pterolebias hoignei, a new annual cyprinodont fish from Venezuela, with a redescription of Pterolebias zonatus. Copeia, 1974, 30–38. Thomerson, J.E. (1984) Rivulichthys luelingi, a junior synonym of Pterolebias longipinnis (Pisces: Rivulidae). Copeia, 1984, 528–529. Thomerson, J.E. & Taphorn, D.C. (1992) Two new annual killifishes from Amazonas Territory, Venezuela (Cyprinodontiformes: Rivulidae). Ichthyological Exploration of Freshwaters, 3, 377–384. Travassos, H. (1955) Notas Ictiológicas VIII: uma nova espécie do gênero Pterolebias Garman, 1895 (Actinopterygii, Cyprinodontiformes). Revista Brasileira de Biologia, 15, 33–39. Weitzman, S.H. & Wourms, J.P. (1967) South American cyprinodont fishes allied to Cynolebias with the description of a new species of Austrofundulus from Venezuela. Copeia, 1967, 89–100. Wiens, J.J. (ed.) (2000) Phylogenetic analysis of morphological data. Smithsonian Institution Press, Washington and London, 220 pp.

PTEROLEBIAS © 2005 Magnolia Press 27 ZOOTAXA Appendix 1 1067 The list below includes material examined of species used as terminal taxa in the present study, except those of the genus Pterolebias, which are listed in the “Taxonomic accounts” above. Data on material is organized in the following sequence: catalog number, number of specimens, locality. Abbreviations are: c&s, specimens cleared and stained for bone and cartilage, H, holotype, and P, paratype(s). Institutional acronyms are: MCNG, Museo de Ciencias Naturales de la UNELLEZ, Guanare; and, UFRJ, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro.

Rivulidae: Aphyolebias boticarioi: UFRJ 5986, H; UFRJ 5987, 11 P; UFRJ 5988, 5 P; Brazil: Estado do Acre: Porto Acre. Austrofundulus transilis: MCNG 7532, 9; Venezuela: Apure: Boca del Jobito. UFRJ 6120, 6; UFRJ 6121, 4 (c&s); Venezuela: Portuguesa: Papelón. Gnatholebias hoignei: MCNG 1116, 18; Venezuela: Portuguesa: La Trinidad. UFRJ 6116, 6; UFRJ 6117, 5 (c&s); Venezuela: Portuguesa: Papelón. Gnatholebias zonatus: MCNG 872, 6; Venezuela: Portuguesa: road Guanare-Barinas. MCNG 17808, 13; Venezuela: Guarico: 10 km N of El Sombrero. UFRJ 6123, 1; Venezuela: Portuguesa: Papelón. UFRJ 3166, 2 (c&s); aquarium material. Kryptolebias brasiliensis: UFRJ 3458, 32; UFRJ 3682, 1 (c&s); UFRJ 4603, 2 ex.; Brazil: Rio de Janeiro: Magé. Micromoema xiphophora: MCNG 26440, 20; Venezuela: Amazonas: Isla Ratón. UFRJ 3165, 1 (c&s); aquarium material. Moema apurinan: UFRJ 5980, H; UFRJ 5981, 9 P; UFRJ 5982, 7 P (c&s); Brazil: Estado do Acre: Porto Acre. Neofundulus paraguayensis: UFRJ 3647, 10; UFRJ 3648, 4 (c&s); Brazil: Mato Grosso do Sul, about 70 km NW from de Aquidauana. Pituna compacta: UFRJ 3563, 33; UFRJ 3564, 4 (c&s); Brazil: Tocantins: Barreira do Piqui. Rachovia maculipinnis: MCNG 35565, 2; Venezuela: Portuguesa: Sabaneta. UFRJ 6118, 7; UFRJ 6119, 4 (c&s); Venezuela: Portuguesa: Papelón. Rachovia stellifer: MCNG 25828, 9; Venezuela: Portuguesa: La Capilla. UFRJ 245, 5 (c&s); Venezuela: Cojedes: 1 km N of Caño Benito. Renova oscari: MCNG 35926, 2; Venezuela: Amazonas: Isla Ratón. UFRJ 4606, 3 (c&s): aquarium material. Rivulus janeiroensis: UFRJ 5333, 8; UFRJ 130, 2 P (c&s); UFRJ 5416, 7 (c&s); Brazil: Rio de Janeiro: Magé. Terranatos dolichopterus: UFRJ 6122, 2; Venezuela: Portuguesa: Guanarito. UFRJ 3910, 1; UFRJ 3911, 3 (c&s); Venezuela: Cojedes: 40 km S of El Pao. MCNG 27069, 29; Venezuela: Amazonas: Isla Ratón. Trigonectes rubromarginatus: UFRJ 3553, 13; UFRJ 3554, 3 (c&s); Brazil: Tocantins: Barreira do Piqui.

Nothobranchidae: Scriptaphyosemion guignardi: UFRJ 3883, 8; UFRJ 4110, 4 (c&s); Guinea: Dalaba.

Aplocheilidae: Aplocheilus panchax: UFRJ 3140, 2; UFRJ 3141, 2 (c&s); Indonesia: Sulawesi: Desa Radda.

28 © 2005 Magnolia Press COSTA Appendix 2 ZOOTAXA 1067 Characters (between brackets) and character states (between parentheses) used to erect the phylogenetic hypothesis among species of Pterolebias and other rivulids are listed below, with the respective reference to papers where the character is first described or discussed. Distribution of character states among terminal taxa is presented in the data matrix in Appendix 3.

Superficial dermal bones and neurocranium [1] Lacrymal (Parenti, 1981; Costa, 1998a, 1998b) (CI: 1.00; RI: 1.00): (0) flat, posterior rim wide; (1) slightly twisted, posterior rim reduced, bone formed mainly by canal; (2) very twisted and narrow, slender, canal vestigial. [2] Ventral portion of lachrymal (Costa, 1998a) (CI: 0.33; RI: 0.50): (0) short; (1) expanded. [3] Number of median vomerine teeth (Costa, 1998a) (CI: 0.33; RI: 0): (0) usually 1–4, sometimes 5–6; (1) 6–12; (?) teeth absent. [4] Anterior retrorse process of lateral ethmoid (Costa, 1990) (CI: 1.00; RI: 1.00): (0) short; (1) moderate to elongate. [5] Lateral process of sphenotic (CI: 0.50; RI: 0.50): (0) narrow; (1) wide. [6] Anterolateral process of parasphenoid (Costa, 1998a) (CI: 0.20; RI: 0.50): (0) short, free; (1) long, attached to pterosphenoid. [7] Posterior portion of parasphenoid (CI: 0.50; RI: 0.50): (0) wide; (1) narrow. [8] Lateral border of frontal (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) well-ossified, approximately straight; (1) poorly ossified, concave. Jaws, jaw suspensorium and opercular apparatus [9] General shape of the premaxilla and dentary (Costa, 1998a) (CI: 0.25; RI: 0.25): (0) elongate, snout profile sharply pointed; (1) short, snout profile blunt. [10] General shape of ascending process of premaxilla (modified from Parenti, 1981; Costa, 1998b) (CI: 1.00; RI: 1.00): (0) curved, posterior portion medially directed; (1) approximately straight. [11] Ascending process of premaxilla (CI: 0.50; RI: 0.50): (0) wide; (1) narrow. [12] Ventral process of maxilla (Parenti, 1981; Costa, 1998a) (CI: 1.00; RI: 1.00): (0) slightly curved, anterior margin rounded; (1) bent, anterior margin triangular. [13] Maxilla (modified from Parenti, 1981) (CI: 0.50; RI: 0.60): (0) approximately straight; (1) slightly twisted; (2) greatly twisted. [14] Rostral cartilage (Costa, 1998a) (CI: 0.50; RI: 0.66): (0) approximately rounded; state 1: longitudinal length longer than transversal length. [15] Coronoid process of dentary (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) broad; (1) narrow. [16] Ventral process of angulo-articular (CI: 0.75, RI: 0.75) (modified from Costa, 1990, 1998a): (0) large and broad; (1) large, somewhat narrowed; (2) moderate, narrow and

PTEROLEBIAS © 2005 Magnolia Press 29 ZOOTAXA pointed; (3) vestigial. 1067 [17] External medial teeth of premaxilla and dentary (Costa, 1998a) (CI: 0.50; RI: 0.75): (0) approximately directed as other teeth; (1) laterally displaced, strongly contrasting to other teeth. [18] Ventral portion of palatine (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) long, overlapping dorsal portion of quadrate; state 1: short, not or slightly contacting quadrate. [19] Metapterygoid (modified from Costa, 1998a) (CI: 0.50; RI: 0.75): (0) about rectangular, dorsal and ventral portions wide and approximately equal in width; (1) dorsal portion slightly constricted; (2) about triangular, dorsal portion strongly constricted. [20] Posterior process of quadrate (Costa, 1998a) (CI: 0.50; RI: 0.66): (0) short, about 50% of quadrate length; (1) long, about 70% of quadrate length. [21] Preopercle (Costa, 1990) (CI: 1.00; RI: 1.00): (0) robust, L-shaped, with a well- developed anteromedian rim; (1) thin, C-shaped, with a reduced anteromedian rim. [22] Dorsal arm of preopercle (Costa, 1990) (CI: 1.00; RI: 1.00): (0) broad; (1) narrow and pointed. Hyoid and branchial arches [23] Anterior process of urohyal (Costa, 1998a) (CI: 0.50; RI: 0): (0) short; (1) elongate. [24] Dorsal process of urohyal (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) short; (1) elongate. [25] Basihyal (Costa, 1998a) (CI: 0.50; RI: 0.50): (0) shorter than space occupied by basibranchials; (1) longer than space occupied by basibranchials. [26] Basihyal ossification (Costa, 1998b) (CI: 1.00; RI: 1.00): (0) more than half length cartilaginous; (1) more than half length ossified. [27] Interhyal (Parenti, 1981) (CI: 1.00; RI: 1.00): (0) ossified; (1) cartilaginous. [28] Interhyal (CI: 1.00; RI: 1.00): (0) large; (1) minute. [29] Subdistal process of second epibranchial (Costa, 2004) (CI: 1.00; RI: 1.00): (0) present; (1) absent. [30] Uncinate process of third epibranchial (modified from Costa, 1998a, b) (CI: 1.00; RI: 1.00): (0) long; (1) moderate; (2) short. [31] Interarcual cartilage (modified from Parenti, 1981) (CI: 1.00; RI: 1.00): (0) not reduced; (1) reduced (CI: 1.00; RI: 1.00). [32] Number and arrangement of second pharyngobranchial teeth (Costa, 2004) (CI: 0.66; RI: 0.50): (0) numerous teeth arranged in two rows; (1) few teeth arranged in single row; (2) teeth absent. [33] Medial flap of second pharyngobranchial (CI: 1.00; RI: 1.00): (0) short; (1) expanded. [34] Teeth on third pharyngobranchial (modified from Costa, 1990) (CI: 1.00; RI: 1.00): (0) base of teeth not widened, all conical; (1) base of teeth of the anterior portion of the dentigerous plate widened, some teeth sub-molariform. [35] Proximal edge of first hypobranchial (Costa, 1998a) (CI: 0.33; RI: 0.33): (0) plain,

30 © 2005 Magnolia Press COSTA terminating in single cartilage united to second basibranchial; (1) bifid, terminating in ZOOTAXA cartilage united to second basibranchial and another smaller cartilage united to first 1067 basibranchial. [36] Distal edge of first hypobranchial (Costa, 2004) (CI: 1.00; RI: 1.00): (0) articular face restricted to cartilaginous head of first ceratobranchial; (1) articular face anteriorly expanded. [37] Fifth ceratobranchial (CI: 1.00; RI: 1.00): (0) narrow; (1) broad. [38] Orientation of anterior tip of fifth ceratobranchial (CI: 0.50; RI: 0.50): (0) anterior; (1) anterolateral. Vertebrae and caudal skeleton [39] Pointed, anteriorly directed process on first vertebra (modified from Costa, 1990) (CI: 1.00; RI: 1.00): (0) absent; (1) present. [40] Dorsally directed process on the base of anterior epipleural ribs (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) absent; (1) present. [41] Hypurals (modified from Costa, 1998a) (CI: 0.60; RI: 0.75): (0) two dorsal plates and one ventral plate separated by gap; (1) two plates separated by wide gap; (2) two plates in close proximity, sometimes ankylosed; (3) single plate. [42] Proximal end of parahypural (Costa, 1998b) (CI: 1.00; RI: 1.00): (0) robust with paired dorsal processes overlapping preural centrum; (1) shortened and laminar, without dorsal paired process, not contacting preural centrum. [43] Hemal spine of preural centrum two (Costa, 1998b) (CI: 0.50; RI: 0): (0) distinctively wider than hemal spines anterior to it; (1) slightly wider or equal in width to hemal spines anterior to it. [44] Number of vertebrae (modified from Costa, 1990) (CI: 0.33; RI: 0.55): (0) 29–32; (1): 33–35; (2) 36–38. [45] Number of caudal-fin rays (modified from Costa, 1990) (CI: 0.50; RI: 0): (0) 26–31; (1) 32–36; (2) 24–25 [not ordered]. Dorsal and anal-fin squeleton [46] First dorsal-fin ray (modified from Parenti, 1981) (CI: 1.00; RI: 1.00): (0) single long first ray connected to two proximal radials; (1) long fin ray connected to two proximal radials, preceded by one or two short fin rays. [47] Anterior proximal radials of dorsal and anal fins (CI: 0.50; RI: 0.75): (0) slender; (1): wide. [48] Orientation of anterior proximal radials of anal fin (modified from Costa, 1998a) (CI: 1.00; RI: 1.00): (0) anteriorly or dorsally directed; (1) posteriorly directed. Shoulder and pelvic girdle [49] Pectoral-fin insertion (Costa, 1998b) (CI: 1.00; RI: 1.00): (0) lateral; (1) ventrolateral. [50] Supracleithrum and posttemporal (modified from Parenti, 1981; Costa, 1998b) (CI: 0.66; RI: 0.50): (0): separated; (1) co-ossified, limits almost inconspicuous; (2) fused to form a single structure.

PTEROLEBIAS © 2005 Magnolia Press 31 ZOOTAXA [51] Keel along supracleithrum-posttemporal (CI: 1.00; RI: 1.00): (0) absent; (1) present. 1067 [52] Posterior flange of cleithrum (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) present; (1) absent. [53] First postcleithrum (Parenti, 1981) (CI: 1.00; RI: 1.00): (0) present; (1) absent. [54] Fourth pectoral radial (Costa, 1998a) (CI: 0.25; RI: 0.40): (0) not expanded; (1) ventrally expanded. [55] Ischial process of pelvic girdle (CI: 0.33; RI: 0.60): (0) prominent; (1) vestigial. [56] Number of pelvic-fin rays (Costa, 1990) (CI: 0.50; RI: 0.71): (0) six; (1) seven; (2) eight. External morphology of body and head [57] Mouth (Costa, 1998b) (CI: 1.00; RI: 1.00): (0) terminal; (1) superior. [58] Orbital rim (modified from Parenti, 1981) (CI: 1.00; RI: 1.00): (0) free dorsally, attached ventrally; (1) completely attached. [59] Fatty predorsal ridge in older males (modified from Taphorn & Thomerson, 1978; Parenti, 1981) (CI: 1.00; RI: 1.00): (0) absent; (1) present. [60] Branchiostegal and opercular membranes (Parenti, 1981) (CI: 1.00; RI: 1.00): (0) not united; (1) united. [61] Skin folder on corner of preopercular region (CI: 1.00; RI: 1.00): (0) present; (1) absent. External morphology of fins [62] Pectoral fin (Costa, 1990) (CI: 0.50; RI: 0.80): (0) rounded; (1) pointed. [63] Pectoral-fin length (modified from Parenti, 1981) (CI: 0.33; RI: 0.60): (0) 18.6–23.8 % SL; (1) 24.1–31.1 % SL. [64] Extent of pelvic fin in males (Costa, 1998a) (CI: 0.50; RI: 0.75): (0) short, its tip not surpassing anterior portion of anal fin; (1) long, its tip reaching the central or the posterior portion of the anal fin. [65] Pelvic-fin (CI: 0.66; RI: 0.50): (0) bases separated or in contact; (1) bases united; (2) pelvic fins united along proximal portion of medial margin. [66] Dorsal and anal fins (modified from Costa, 1998a) (CI: 0.40; RI: 0.75): (0) short, tip rounded; (1) somewhat elongated, tip pointed; (2) long, tip sharply pointed. [67] Filaments on tip of dorsal and anal fins in males (Costa, 1998a) (CI: 0.33; RI: 0.66): (0) absent; (1) present. [68] Filaments on posterior border of caudal fin in males (Costa, 1998a) (CI: 0.33; RI: 0.75): (0) absent; (1) present. [69] Caudal-fin length in males (Costa, 1990) (CI: 0.50; RI: 0.83): (0) 32.5-41.5; (1) 42.0- 49.0; (2) 52.5-81.0. [70] Dorsal and ventral extensions on caudal fin in males (Costa, 1998a) (CI: 0.33; RI: 0): (0) absent; (1) present. Squamation [71] General arrangement of frontal scales (Hoedeman, 1958) (CI: 0.66; RI: 0.50): (0)

32 © 2005 Magnolia Press COSTA transverse; (1) circular; (2) irregular [not ordered]. ZOOTAXA [72] Predominant frontal squamation-pattern (CI: 0.50; RI: 0.40): (0) G; (1) E; (2) D; (3) F 1067 [not ordered]. [73] Arrangement of E-scales (Costa, 1990) (CI: 0.50; RI: 0.66): (0) overlapped; (1) not overlapped. [74] Caudal-fin squamation in older males (Costa, 1990, 1998a) (CI: 0.28; RI: 0.16): (0) approximately on anterior 10–30 % of fin; (1) approximately on 40 % of fin; (2) approximately on 50–80 % of fin. [75] Anal-fin base squamation in males (CI: 1.00; RI: 1.00): (0) no scales on anal-fin base; (1) 1–7 rows of scales on anal-fin base. [76] Dorsal-fin base squamation in males (CI: 1.00; RI: 1.00): (0) no scales on anal-fin base; (1) 1–2 rows of scales on anal-fin base. Laterosensory system [77] Cephalic canals (modified from Parenti, 1981) (CI: 1.00; RI: 1.00): (0) anterior and posterior infraorbital, preopercular and posterior mandibular closed, supraorbital, rostral, anterior mandibular open with skin trenches around neuromasts; (1) anterior and posterior infraorbital, and preopercular closed, supraorbital, rostral, anterior and posterior mandibular open with skin trenches around neuromasts; (2) anterior and posterior infraorbital, dorsal preopercular, supraorbital, rostral, anterior and posterior mandibular open with skin trenches around neuromasts; (3) no vestige of canals, all neuromasts completely exposed. [78] Number of anterior supraorbital neuromasts (Costa, 1990) (CI: 0.42; RI: 0.42): (0) 3; (1) 4; (2) 5; (3) 6–7. Contact organs [79] Contact on flank scales in males (CI: 0.50; RI: 0.50): (0) absent; (1) present. [80] Pectoral-fin contact organs in males (CI: 1.00; RI: 1.00): (0) absent; (1) present. Male color patterns [81] General color pattern on flank (CI: 0.33; RI: 0.50): (0) vertical; (1) oblique; (2) longitudinal [not ordered] [82] Orange pigmentation pattern on humeral region (modified from Costa, 1998a) (CI: 1.00; RI: 1.00): (0) not distinctively colored; (1) small metallic orange spots. [83] Dark pigmentation pattern on iris (Parenti, 1981) (CI: 1.00; RI: 1.00): (0) no distinctive mark; (1) bar crossing eye. [84] Pigmentation pattern on suborbital region (Costa, 1998a) (CI: 0.66; RI: 0.83): (0) no distinctive mark; (1) gray to black suborbital bar; (2) red suborbital spot [not ordered]. [85] Dark pigmentation pattern on supraorbital region (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) no distinctive mark; (1) spot adjacent to eye. [86] Pigmentation pattern on post-orbital and preopercular region (CI: 0.60; RI: 0): (0) no distinctive pattern; (1) two oblique bars; (2) post-orbital vertical bar; (3) two oblique stripes [not ordered].

PTEROLEBIAS © 2005 Magnolia Press 33 ZOOTAXA [87] Yellow to orange pigmentation pattern on ventral portion of caudal fin (modified from 1067 Costa, 1998a) (CI: 0.50; RI: 0.80). (0) not distinctive concentrated; (1) concentrated to form stripe; (?) variable. [88] Red pigmentation pattern on subventral portion of caudal fin (CI: 1.00; RI: 1.00): (0) not distinctive concentrated; (1) concentrated to form stripe. [89] Dark pigmentation on dorsal portion of caudal fin (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) not distinctive concentrated; (1) concentrated to form dark brown stripe; (?) variable. [90] Melanophore pattern on pectoral fin (Costa, 1998a) (CI: 0.40; RI: 0.40): (0) no distinctive marks; (1) spots; (2) bars [unordered]. [91] Melanophore pattern on ventral margin of pectoral fin (Costa, 1998a) (CI: 1.00; RI: 1.00): (0) not distinctive concentrated; (1) concentrated to form stripe. Female color patterns [92] Dark pigmentation pattern on flank and fins (Costa, 1998a) (CI: 0.50; RI: 0.66): (0) forming bars, stripes or spots, according to the pattern occurring in males; (1) flank almost plain, pigmentation reduced to minute dots, not presenting the general pattern as in males. [93] Dark pigmentation pattern on upper portion of caudal-fin base (Costa, 1990) (CI: 0.33; RI: 0): (0) no distinctive mark; (1) one black spot.

34 © 2005 Magnolia Press COSTA Appendix 3 ZOOTAXA 1067 Matrix of 93 morphological characters for 19 aplocheiloid species. Characters and states are according to Appendix II. 0 = plesiomorphic state; 1–4 = apomorphic states; ? = not pertinent or unknown state.

1–10 11–20 21–30 31–40 41–50

Aplocheilus panchax 0000000000 0000000000 0000000000 0000000000 0000000002

Scriptaphyosemion guignardi 1000000010 1000000000 0000000000 0200000000 1110110012

Kryptolebias brasiliensis 2001000110 1111100100 1000010001 0100100000 1100010010

Rivulus janeiroensis 2001000111 1121110100 1000011112 0100110010 1111010011

Pterolebias longipinnis 20-1010111 1121121121 1001111112 1110111010 1110011111

Pterolebias phasianus 2011010111 1121121121 1001111112 1110110010 1110211111

Gnatholebias zonatus 2001001101 0110111110 1001011112 1200-10010 3112011111

Gnatholebias hoignei 2001001101 0110111110 1001011112 1200010010 3112011111

Rachovia stellifer 2101010111 1111110110 1011011112 0100110010 2111010011

Rachovia maculipinnis 2101010111 1111110110 1011011112 0100110011 2110010011

Austrofundulus transilis 2101000111 1111110110 1001011112 0100010011 2110010011

Terranatos dolichopterus 2001000111 1111110110 1001011112 0100110011 2110011011

Pituna compacta 2101100111 1111130110 1101011112 0100110010 2110010011

Aphyolebias boticarioi 2101010111 1111110120 1001011112 0100110110 2111010011

Moema apurinan 20-1011101 1111110120 1001011112 0100110110 1111010011

Micromoema xiphophora 2001000111 1121111111 1001111112 0100110010 1110010011

Renova oscari 2011010111 1121110111 1001011112 0100110110 1110110011

Neofundulus paraguayensis 2001110111 1111110100 1001011112 0100110010 1111010011

Trigonectes rubromarginatus 2011110101 1111110100 1001011112 0101110010 1112110011 ...... continued on the next page

PTEROLEBIAS © 2005 Magnolia Press 35 ZOOTAXA Appendix 3 (continued). 1067

51–60 61–70 71–80 81–90 91–93 Aplocheilus panchax 0000000000 0000000000 0000000000 0000000000 000 Scriptaphyosemion guignardi 0000001100 1000000000 0000001000 1000010000 000 Kryptolebias brasiliensis 0010011101 1000000000 1100002000 0001000000 000 Rivulus janeiroensis 1011101101 1000000000 1110002000 ?000000000 001 Pterolebias longipinnis 1111121101 1011121120 1110112000 1110000002 000 Pterolebias phasianus 1111121101 1011021120 1110102000 2110030002 000 Gnatholebias zonatus 1111121101 1111221120 1310102000 0010000010 100 Gnatholebias hoignei 1111121101 1111221120 1310102000 0010000010 100 Rachovia stellifer 1110011101 1000010000 1112002000 1011000000 001 Rachovia maculipinnis 1111021111 1010010110 1312002001 101112-000 010 Austrofundulus transilis 1111011111 1010000000 2112002200 1011100000 010 Terranatos dolichopterus 1110121111 1010221111 0100002300 1011100000 010 Pituna compacta 1111011101 1010010000 1310003310 1011000001 000 Aphyolebias boticarioi 1111011101 1110021111 1110002010 1010001101 010 Moema apurinan 1111011101 1110021120 1110002010 2010011100 000 Micromoema xiphophora 1111011101 1110020011 1110002000 2012011100 000 Renova oscari 1111011101 1110020110 1310002000 2012001100 001 Neofundulus paraguayensis 1110011101 1010010000 1111002100 2010001001 000 Trigonectes rubromarginatus 1111011101 1001020000 1211002100 2010001000 000

36 © 2005 Magnolia Press COSTA