ZooKeys 954: 109—| 56 (2020) A peer-reviewed open-access journal doi: 10.3897/zookeys.954.50667 RESEARCH ARTICLE #Zookey S https:/ / ZOO keys. pensoft.net Launched to accelerate biodiversity research Systematics of Pholidobolus lizards (Squamata, Gymnophthalmidae) from southern Ecuador, with descriptions of four new species Vanessa Parra', Pedro M. Sales Nunes”, Omar Torres-Carvajal! | Museo de Zoologia, Escuela de Ciencias Biolégicas, Pontificia Universidad Catélica del Ecuador, Avenida 12 de Octubre 1076 y Roca, Quito, Ecuador 2. Departamento de Zoologia, Centro de Biociéncias, Universidade Federal de Pernambuco, Avenida Professor Moraes Rego, s/n. Cidade Universitdria CEP 50670-901, Recife, PE, Brazil Corresponding author: Omar Torres-Carvajal (omartorcar@gmail.com) Academic editor: Anthony Herrel | Received 30 January 2020 | Accepted 28 May 2020 | Published 29 July 2020 http://zoobank.org/A2A9BE21-F571-42ED-979D-FAD7D8151721 Citation: Parra V, Nunes PMS, Torres-Carvajal O (2020) Systematics of Pholidobolus lizards (Squamata, Gymnophthalmidae) from southern Ecuador, with descriptions of four new species. ZooKeys 954: 109-156. https:// doi.org/10.3897/zookeys.954.50667 Abstract Four new species of Pholidobolus lizards are described from poorly explored areas in the Andes of southern Ecuador based on morphological and genetic evidence. Among other morphological characters, Pho- lidobolus samek sp. nov. and P condor sp. nov. differ from their congeners in having green dorsolateral stripes on head. Males of P condor sp. nov. differ from those of P samek sp. nov. in having reddish flanks and venter. P dolichoderes sp. nov. is distinguished by having a long neck, with more scales between orbit and tympanum, whereas P fascinatus sp. nov. is distinguished by lacking enlarged medial scales on collar and a conspicuous vertebral stripe. In addition, the phylogenetic position of the new species is inferred using DNA sequences of mitochondrial and nuclear genes. The phylogeny supports strongly monophyly of each of the new species and renders P macbrydei paraphyletic and split into six subclades. Available data suggest that the new species have restricted distribution ranges (< 100 km? each), and it is proposed that their classification be as Data Deficient or Critically Endangered species. The results reveal unexpected levels of diversity within Pholidobolus in the Andes of southern Ecuador and highlight the importance of improving scientific collections and conservation efforts in this area. Keywords Andes, Cordillera del Condor, diversity, phylogeny, taxonomy Copyright Vanessa Parra et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 110 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Introduction The uplift of the Andes mountains was one of the most influential geological events for the evolution and diversification of the South American biota during the Cenozoic. For example, it created many habitats and microclimates that became important centers of biodiversity and endemism (Pérez-Escobar et al. 2017). Therefore, the evolution of diverse Andean taxa is a complex research topic that has attracted the attention of many scientists (Castoe et al. 2004, Torres-Carvajal et al. 2015, Betancourt et al. 2018, Moravec et al. 2018, Lehr et al. 2019). With more than 250 species, Gymnophthalmidae is one of the most diverse lizard clades in the Neotropics. The uplift of the Andes had a strong influ- ence on the radiation of gymnophthalmid lizards, resulting in high levels of diversity and endemism along the Tropical Andes (Torres-Carvajal et al. 2016; Moravec et al. 2018). Pholidobolus lizards are among the most prominent gymnophthalmids in the northern Andes. They are small (SVL < 60 mm), terrestrial, oviparous, and restricted to the Andes of Colombia, Ecuador, and northern Peru at elevations between 1800 and 4100 m (Hurtado-Gémez et al. 2018; Torres-Carvajal et al. 2014; Venegas et al. 2016). Pholidobolus is currently known to include ten species: P affinis, P anomalus, P dicrus, P. hillisi, P macbrydei, P montium, P paramuno, P. prefrontalis, P ulisesi, and P vertebralis, of which three were described in recent years. Remarkably, P anomalus is the only species in the genus that occurs in southern Peru (Cusco), but its generic identity remains questionable (Torres-Carvajal and Mafla-Endara 2013). The study of Pholidobolus and other gymnophthalmid taxa has been often ham- pered by the paucity of specimens in collections. For example, the recent description of P paramuno reveals the importance of increased sampling effort in the Paramo eco- system in the northern Andes of Colombia. Similarly, recent collections in poorly ex- plored areas of the southern Andes of Ecuador yielded new specimens of Pholidobolus lizards, which we were unable to assign to any of the currently recognized species. Based on these specimens, here we combine evidence from morphology and DNA sequences to describe four new species of Pholidobolus and infer their phylogenetic affinities. Materials and methods Genetic data Total genomic DNA was digested and extracted from liver or muscle tissue using a guanidinium isothiocyanate extraction protocol. Tissue samples were first mixed with Proteinase K and a lysis buffer and digested overnight prior to extraction. DNA sam- ples were quantified using a Nanodrop ND-1000 (NanoDrop Technologies, Inc.), re- suspended and diluted to 25 ng/ul in ddH2O prior to amplification. Using primers and amplification protocols from the literature (Pellegrino et al. 2001; Torres-Carvajal and Mafla-Endara 2013), we obtained 1,493 aligned nucleotides (nt) encompassing three mitochondrial genes, 12S (339 nt), 16S (533 nt), and ND4 (621 nt) from 16 individuals of the four new species herein described, as well as 21 individuals of Systematics of Pholidobolus lizards 111 Pholidobolus macbrydei. In addition, we obtained 411 nucleotides of the Dynein Axone- mal Heavy Chain 3 (DNAH3) nuclear gene from 65 individuals of Anadia rhombifera, Macropholidus annectens, M. huancabambae, M. labiopunctatus, M. ruthveni, Pholidobolus affinis, P dicrus, P. hillisi, P macbrydei, P montium, P. prefrontalis, P ulisesi, P vertebralis, and the four new species. DNAH3 was amplified using the primers DNAH3_fl (GG- TAAAATGATAGAAGAYTACTG) and DNAH3_1r6 (CTKGAGTTRGAHACAAT- KATGCCAT). The amplification protocol consisted of 1 cycle of initial denaturation for 5 min at 95 °C, 40 cycles of denaturation for 35s at 94 °C, annealing for 1 min at 72 °C, and extension for 1 min at 72 °C, as well as a final extension for 10 min at 72 °C (Townsend et al. 2008). Positive PCR products were visualized in agarose electrophoretic gels and treated with ExoSAP-IT to remove unincorporated primers and dNTPs. Cycle sequencing reactions were carried out by Macrogen Inc. GenBank accession numbers of sequences generated in this study are shown in Table 1. After incorporating GenBank se- quences, our data matrix for phylogenetic analyses contained 74 taxa and 1904 characters. Phylogenetic analyses Data were assembled and aligned in Geneious v5.4.6. (Kearse et al. 2012) under default settings for MAFFT Multiple Alignment (Katoh and Toh 2010). ND4 and DNAHS3 sequences were translated into amino acids for confirmation of alignment. The best-fit nucleotide substitution models and partitioning scheme were chosen si- multaneously using PartitionFinder v2.1.1 (Lanfear et al. 2012) under the Bayesian Information Criterion (BIC). Genes were combined into a single dataset with four partitions: (i) 1“ codon position of ND4 and 12S [GTR + I + G]; (ii) 2" codon posi- tion of ND4, 1* codon and 2" codon positions of DNAH3 [HKY + I + G]; (iii) 3" codon position of ND4 [GTR + G]; (iv) 16S and 3 codon position of DNAH3 [SYM + 1+ G]. Both maximum likelihood (ML) and Bayesian inference (BI) methods were used to obtain the optimal tree topology of the combined, partitioned dataset using the programs RAxML v.8.2.12 (Stamatakis 2014) and MrBayes v3.2.6 (Ronquist et al. 2012), respectively. The ML analysis was performed under the GTRGAMMA model for all partitions. Nodal support (BS) was assessed with the rapid bootstrapping algo- rithm under the MRE-based Boot-stopping criterion (252 replicates). For BI analysis, all parameters were unlinked between partitions (except topology and branch lengths), and rate variation (prset ratepr = variable) was invoked. Four independent runs, each with four MCMC chains, were set for ten million generations, sampling every 10,000 generations. All analyses were performed using the CIPRES platform (Miller et al. 2010). Results were analyzed in Tracer 1.6 (Rambaut and Drummond 2007) to assess convergence and effective sample sizes (ESS) for all parameters, based on which the first 10% of trees were removed from each run. The remaining trees were used to calcu- late posterior probabilities (PP) for each bipartition in a Maximum Clade Credibility Tree. The phylogenetic trees were visualized and edited using Fig Iree v1.4.2 (Rambaut 2014). In order to address interspecific genetic differentiation, uncorrected genetic distances were calculated in MEGA 7 (Kumar et al. 2016) after removing ambiguous positions for each sequence pair (pairwise deletion option). 112 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Table |. Vouchers, locality data, and GenBank accession numbers of taxa included in this study. Se- quences added in this study are in bold. Taxon Voucher Locality GenBank number GenSeq as DNAH3_| Nomenclature Anadia QCAZ 11862 | QCAZ 11862; KU902135 | KU902216 | KU902291 |MN849427| — genseq-4 rhombifera Ecuador: Cotopaxi: San Francisco de Las Pampas Macropholidus | QCAZ 11120 | Ecuador: Loja: 15 km| KC894341 | KC894355 | KC894369 |MN849430| — genseq-4 annectens E Loja QCAZ 11121 | Ecuador: Loja: 15 km} KC894342 | KC894356 | KC894370 | MN849431| — genseq-4 E Loja Macropholidus | CORBIDI Peru: Piura: KC894343 | KC894357 | KC894371 | MN849428) — genseq-4 huancabambae 10492 Huancabamba: Las Pozas CORBIDI Peru: Piura: KC894344 | KC894358 | KC894372 - genseq-4 10493 Huancabamba: Las Pozas CORBIDI Peru: Piura: KC894345 | KC894359 | KC894373 |MN849429) — genseq-4 10496 Huancabamba: Las Pozas Macropholidus | CORBIDI | Peru: Piura: Ayabaca | KP874774 MN849432| — genseq-4 labiopunctatus 12932 Macropholidus | CORBIDI | Peru: Lambayeque: | KC894354 | C894368 | C894382 |MN849433| — genseq-4 ruthveni 4281 El Totora Pholidobolus QCAZ 9641 | Ecuador: Cotopaxi: | KC894348 | C894362 | C894376 |MN849435| — genseq-4 affinis San Miguel de Salcedo, Cutuchi River QCAZ 9900 Ecuador: KC894349 - genseq-4 Chimborazo: Colta Pholidobolus QCAZ 16788 | Ecuador: Morona- | MN724005 MN849464| — genseq-2 condor sp. nov. Santiago: el Quimi QCAZ 16789 | Ecuador: Morona- | MN724006 MN849465| —_genseq-2 Santiago: el Quimi QCAZ 16790 | Ecuador: Morona- | MN724007 MN849466| _—_ genseq-2 Santiago: el Quimi QCAZ 15844 | Ecuador: Morona- | MN723996 MN849434| _— genseq-1 Santiago: el Quimi Pholidobolus QCAZ 5304 | Ecuador: Morona- | KP874776 MN849436| _—_ genseq-4 dicrus Santiago: Guarumales QCAZ 6936 Ecuador: - KP874829 | KP874939 |MN849437| — genseq-4 Tungurahua: Rio Blanco Pholidobolus QCAZ 16349 | Ecuador: Cafar: Ofa | MN724000 | MN720234 | MN717129 | MN849459| _ genseq-2 dolichoderes sp. | QCAZ, 16350 | Ecuador: Cafar: Ofia| MN724001 MN849460| _genseq-2 nov. QCAZ 16351 | Ecuador: Cafar: Offa | MN724002 | MN720236 | MN717131|MN849461| _genseq-2 QCAZ 16352 | Ecuador: Cafar: Offa | MN724003 | MN720237 | MN717132 | MN849462| _genseq-2 QCAZ 16353 | Ecuador: Cafiar: Offa | MN724004 | MN720238 | MN717133 | MN849463 senseq-1 Pholidobolus QCAZ 15118 | Ecuador: El Oro: =| MN724017|MN720251 | MN717146|MN849476| —_ genseq-2 Jfascinatus sp. Chillacocha nov. QCAZ 15120] Ecuador: El Oro: =| MN724018 | MN720252 | MN717147 | MN849477| _ genseq-1 Chillacocha QCAZ 15122 | Ecuador: El Oro: | MN724019 | MN720253 = MN849478| _ genseq-2 Chillacocha QCAZ 15170 | Ecuador: El Oro: =| MN724020 | MN720254 | MN717148 | MN849479| —genseq-2 Chillacocha Systematics of Pholidobolus lizards 113 Taxon Voucher Locality GenBank number GenSeq 12S 16S ND4 DNAH3_| Nomenclature Pholidobolus QCAZ 4998 | Ecuador: Zamora- | KP090167 | KP090170 | KP090173 |MN849438| — genseq-4 hillisi Chinchipe: near San Francisco Research Station QCAZ 4999 | Ecuador: Zamora- | KP090169 | KP090172 | KP090175 |MN849439| — genseq-4 Chinchipe: near San Francisco Research Station QCAZ 5000 | Ecuador: Zamora- | KP090168 | KP090171 | KP090174 |MN849440/ — genseq-4 Chinchipe: near San Francisco Research Station “Pholidobolus KU 218406 Ecuador: Azuay: | AY507848 | AY507867 | AY507886 - genseq-4 macbrydei” Cuenca QCAZ 9914 | Ecuador: Azuay: | KC894352 | KC894366 | KC894380 |MN849441| — genseq-4 Guablid QCAZ 9932 | Ecuador: Azuay: 20 | KC894353 | KC894367 | KC894381 |MN849442| — genseq-4 km on road Cuenca- El Cajas QCAZ 9947 | Ecuadro: Cafiar: | MN724012 | MN720246|MN717141|MN849474| — genseq-4 Cafar QCAZ 10051 | Ecuador: Cafar: | MN724014|MN720248 | MN717143 | MN849472| — genseq-4 Rio Guallicanga, quebrada Juncal QCAZ 10052 | Ecuador: Cafar: |MN724015 |MN720249 | MN717144|MN849473| — genseq-4 Rio Guallicanga, quebrada Juncal QCAZ 10050 | Ecuador: Cafar: | MN724013 |MN720247 | MN717142|MN849471| — genseq-4 A 1000 m de la Panamericana Juncal QCAZ 15811 | Ecuador: Cafiar: | MN724021 | MN720255 | MN717149 | MN849480| —_ genseq-4 Mazar QCAZ 15812 | Ecuador: Cafiar: | MN724022 |MN720256 | MN717150|MN849481| — genseq-4 Mazar QCAZ 15813 | Ecuador: Cafar: | MN724023 | MN720257 | MN717151|MN849482| —_ genseq-4 Mazar QCAZ 15814 | Ecuador: Cafiar: | MN724024|MN720258 | MN717152 - genseq-4 Mazar QCAZ 15815 | Ecuador: Cafiar: | MN724025 | MN520259 | MN717153 E genseq-4 Mazar QCAZ 15816 | Ecuador: Cafar: | MN724026|MN720260 | MN717154| MN849483| — genseq-4 Mazar QCAZ 15817 | Ecuador: Cafar: | MN724027|MN720261 | MN717155 | MN849484| —_genseq-4 Mazar QCAZ 15818 | Ecuador: Cafar: | MN724028 | MN720262 | MN717156| MN849485| _—_genseq-4 Mazar QCAZ 15819 | Ecuador: Cafar: | MN724029|MN720263 | MN717157 | MN849486| —_ genseq-4 Mazar QCAZ 15820 | Ecuador: Cafar: | MN724030|MN720264 | MN717158 | MN849487| _—_ genseq-4 Mazar QCAZ 15823 | Ecuador: Cafiar: | MN724031|MN720265 | MN717159 | MN849488| _—_ genseq-4 Mazar QCAZ 15824 | Ecuador: Cafar: | MN724032 | MN720266 | MN717160| MN849489| _—_genseq-4 Mazar QCAZ 6945 Ecuador: Loja: | MN724008 | MN720242 | MN717137 | MN849467| —_ genseq-4 Jimbura QCAZ 6946 Ecuador: Loja: genseq-4 Jimbura MN724009 | MN720243 | MN717138 | MN849468 114 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Taxon Voucher Locality GenBank number GenSeq 12S DNAH3__| Nomenclature “Pholidobolus | QCAZ 10054 Ecuadro: Loja‘ | MN724016|MN720250 | MN717145 | MN849475| —_genseq-4 macbrydei” Colambo Yacuri Forest QCAZ 7894 | Ecuador: El Oro: | MN724011 MN849470| _—_genseq-4 Guanazan QCAZ 7891 | Ecuador: ElOro: | MN724010 MN849469| _—_genseq-4 Guanazan Pholidobolus QCAZ 4051 | Ecuador: Pichincha: | KC894346 MN849443| —__genseq-4 montium Quito QCAZ 9044 | Ecuador: Pichincha: | KC894347 MN849444| — genseq-4 Tababela Pholidobolus MHUAR _ | Colombia: Antoquia | MK215018 - genseq-4 paramuno 12451 MHUAR _ | Colombia: Antoquia | MK215019 - genseq-4 12480 MHUAR _ | Colombia: Antoquia | MK215020 - genseq-4 12481 Pholidobolus QCAZ 9908 Ecuador: KC894350 - genseq-4 prefrontalis Chimborazo: Alausi QCAZ 9951 Ecuador: KC894351 | KC894365 | KC894379 | MN849448| — genseq-4 Chimborazo: Tixan Pholidobolus QCAZ 14954 | Ecuador: Zamora | MN723997 |MN720231 | MN717126|MN849445| —_ genseq-2 samek sp. nov. Chinchipe: Cerro Plateado QCAZ 14955 | Ecuador: Zamora | MN723998 | MN720332 | MN717127 | MN849446| —_ genseq-1 Chinchipe: Cerro Plateado QCAZ 14956 | Ecuador: Zamora | MN723999 |MN720233 | MN717128 | MN849447| —_ genseq-2 Chinchipe: Cerro Plateado Pholidobolus CORBIDI Peru: Cajamarca: | KP874787 | KP874839 | KP874948 |MN849449| — genseq-4 ulisesi 12735 Jaen: Huamantanga Forest CORBIDI Peru: Cajamarca: | KP874788 | KP874840 | KP874949 = genseq-4 12702 Jaen: Huamantanga Forest CORBIDI Pert: Chota: La | KP874786 MN849450| —genseq-4 1679 Granja Pholidobolus QCAZ 10667 | Ecuador: Pichincha: | KP874784 | KP874836 | KP874946 |MN849455| __ genseq-4 vertebralis Santa Lucia de Nanegal QCAZ 10750 | Ecuador: Pichincha: | KP874785 | KP874837 | KP874947 |MN849458| _ genseq-4 Santa Lucia de Nanegal QCAZ 5057 | Ecuador: Carchi: | KP874778 | KP874830 | KP874940 |MN849451| — genseq-4 Chilma Bajo QCAZ 8687 | Ecuador: Carchi: | KP874779 | KP874831 | KP874941 |MN849452| — genseq-4 Chilma Bajo QCAZ 8688 | Ecuador: Carchi: | KP874780 | KP874832 | KP874942 |MN849453| _— genseq-4 Chilma Bajo QCAZ 8689 | Ecuador: Carchi: | KP874781 | KP874833 | KP874943 |MN849454| — genseq-4 Chilma Bajo QCAZ 8717 | Ecuador: Carchi: | KP874782 | KP874834 | KP874944 |MN849456/ _— genseq-4 next to Chilma Bajo QCAZ 8724 | Ecuador: Carchi: | KP874783 | KP874835 | KP874945 |MN849457| — genseq-4 next to Chilma Bajo Systematics of Pholidobolus lizards 115 Specimens and morphological data We examined 98 specimens of Pholidobolus macbrydei (Appendix I) and 41 of the new species described herein (see corresponding type series). All specimens are deposited in the herpetological collection at Museo de Zoologia, Pontificia Universidad Catdlica del Ecuador, Quito (QCAZ). The following measurements were taken with a digital caliper (to the nearest 0.1 mm): AGD .axilla-groin distance; ShL © shank length; HL head length; SVL and snout-vent length. HW __ head width; Tail length (TL) was measured with a ruler. Sex was determined by dissection or by noting the presence of everted hemipenes. We followed the terminology of Monta- nucci (1973) and Kizirian (1996) for morphological characters. Because the new species are similar in morphology to Pholidobolus macbrydei, we assessed the degree of differentiation among them with a Principal Components Anal- ysis (PCA) in R (R Core Team 2018). The PCA was based on 16 quantitative mor- phological characters: (1) number of supraoculars (NSO), (2) number of scales along margin of upper jaw (SUJ), (3) number of scales along margin of lower jaw (SLJ), (4) number of gular and jaw scales (SGJ), (5) number of ventrals (SGV), (6) number of dorsals (DEL), (7) number of temporals (NTS), (8) number of scales around body (SAB), (9) number of scales around tail (SAT), (10) number of supradigital scales of third finger (SF3), (11) number of supradigital scales of fifth finger (SF5), (12) number of supradigital scales of third toe (ST3), (13) number of supradigital scales of fourth toe (ST4), (14) number of supradigital scales of fifth toe (ST5), (15) lower eyelid scales (LES), and (16) collar scales (i.e., posterior transverse row of gulars; SGC) (Peters 1964, Montanucci 1973). Hemipenes were prepared following the procedures described by Manzani and Abe (1988), as modified by Pesantes (1994) and Zaher (1999). Organs were everted after immersion in a potassium hydroxide solution, the retractor muscles were manually separated, and the everted organs filled with blue-stained petroleum jelly. Hemipenes were then immersed in an alcoholic solution of Alizarin Red for 24 hours in order to stain eventual calcified structures (e.g., spines or spicules), in an adaptation proposed by Nunes et al. (2012) on the procedures described by Uzzell (1973) and Harvey and Embert (2008). The terminology of hemipenial structures follows previous literature (Dowling and Savage 1960; Hurtado-Gomez et al. 2018; Nunes et al. 2012; Sanchez- Pacheco et al. 2017; Savage 1997; Venegas et al. 2016). Systematics The taxonomic conclusions of this study are based on the observation of morpho- logical features and color pattern, as well as inferred phylogenetic relationships. We 116 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) consider this information as species delimitation criteria following a general lineage or unified species concept (de Queiroz 1998; 2007). The new species share with all known species of Pholidobolus the presence of a ventrolateral fold between fore and hind limbs and the absence of a single transparent palpebral disc (Montanucci 1973). Results Phylogenetic relationships and genetic distances Tree topologies under ML and BI approaches were generally similar; here we describe the maximum clade credibility tree (Fig. 1). Our hypothesis supports the monophyly of Pholidobolus (BS= 60, PP = 0.99) and is congruent with previous molecular phylogenies in that P lisesi and P hillisi form a clade (BS = 62, PP = 0.92) sister to all other congeners (Torres-Carvajal et al. 2015, 2016; Hurtado-Gémez et al. 2018). Following branching order, the strongly supported species pair P affinis, P montium is sister to all remaining species, which form a clade where (P prefrontalis (P paramuno (P. dicrus, P. vertebralis))) is sister to a subclade containing the new species described in this paper and a paraphyletic P macbrydei. Hereafter we refer to the latter subclade as the “P macbrydei” species complex. The “P macbrydei” species complex (BS = 81, PP = 1) is divided into two allopatric and strongly supported clades (Fig. 1) that include four new species described below and a paraphyletic “P macbrydei” divided in six subclades (Clades A’-F). A southeastern clade (BS = 99, PP = 1) contains P condor sp. nov. as sister to (P samek sp. nov., “P. macbrydei” Clade A [Loja province]). The ML tree recovered P condor as sister to “P. macbrydei” Clade A with low support (BS = 58). A northwestern clade (BS = 85, PP = 0.96) is composed of “P macbryder” Clade B from Canar province as sister to a clade that includes all remaining samples, in which P fascinatus sp. nov. is nested along with “P macbrydei” Clades C, D, and E (Azuay and Canar provinces) in a strongly supported subclade (BS = 71, PP = 1) sister to the maximally supported (P dolichoderes, “P macbrydei” Clade F [El Oro prov- ince]). All new species are strongly supported as monophyletic (BS > 98, PP = 1). Uncorrected p-genetic distances for 16S, 12S, and ND4 are presented in Tables 2, 3, and 4, respectively. Distance values among all recognized species of Pholidobolus, the four new species described in this paper, and the six “P macbrydei” clades range between 1 (e.g., P condor sp. nov. vs. P samek sp. nov., Clade C vs. Clade D)—10% (e.g., P paramuno vs. P dicrus) for 12S (average = 5% + 0.01 SD); 1 (P dolichoderes sp. nov. vs. Clade F)—6% (e.g., P dicrus vs. P ulisesi) for 16S (average = 4% + 0.01 SD); and 4 (P dolichoderes sp. nov. vs. Clade F)—19% (e.g., P dicrus vs. P vertebralis) for ND4 (average = 14% + 0.03 SD). Maximum distance values within the “P macbrydei” complex are 5% (Clade A vs. Clade F) for 12S, 4% (P condor sp. nov. vs. Clade F) for 16S, and 14% (Clade A vs. Clade F) for ND4. The genetic distances for the nuclear gene NDH3 are generally low (0-3%, average = 1% + 0.01 SD). Systematics of Pholidobolus lizards Pholidobolus ulisesi CORBIDI12737 Pholidobolus ulisesi CORBIDI12735 ioe 100 Pholidobolus ulisesi CORBIDI1679 62 Pholidobolus hillisi QCAZ4999 1 Pholidobolus hillisi QCAZ5000 100 Pholidobolus hillisi QCAZ4998 1 Pholidobolus affinis QCAZ9900 1 100 Pholidobolus affinis QCAZ9641 Pholidobolus montium QCAZ4051 Pholidobolus montium KU196355 100 Pholidobolus montium QCAZ9044 1 Pholidobolus prefrontalis QCAZ9908 100 Pholidobolus prefrontalis QCAZ9951 Pholidobolus vertebralis QCAZ10667 Pholidobolus vertebralis QCAZ10750 1 Pholidobolus vertebralis QCAZ8689 100 Pholidobolus vertebralis QCAZ8687 Pholidobolus vertebralis QCAZ8724 0.99} Pholidobolus vertebralis QOAZ5057 5 Pholidobolus vertebralis QOCAZ8688 Pholidobolus vertebralis QCAZ8717 1 B 1 Pholidobolus dicrus QCAZ6936 68 100 Pholidobolus dicrus QCAZ5304 Pholidobolus paramuno MHUAR12480 Pholidobolus paramuno MHUAR12481 Pholidobolus paramuno MHUAR12451 1, “Pholidobolus macbrydei” QCAZ7891 El Oro | Clade F , foo! “Pholidobolus macbrydei” QCAZ7894 El Oro ve Pholidobolus dolichoderes sp. n. QCAZ16351 Azuay Pholidobolus dolichoderes sp. n. QCAZ16349 Azuay Pholidobolus dolichoderes sp. n. QCAZ16353 Azuay Pholidobolus dolichoderes sp. n. QCAZ16352 Azuay Pholidobolus dolichoderes sp. n. QCAZ16350 Azuay Pholidobolus fascinatus sp. n. QOAZ15120 El Oro TAG? 0.52 1 100 100 0.96} 71 1 100 1 100 99 1 100 0.03 Pholidobolus fascinatus sp. n. QOAZ15118 El Oro Pholidobolus fascinatus sp. n. QCAZ15170 El Oro Pholidobolus fascinatus sp. n. QOAZ15122 El Oro “Pholidobolus macbrydei” QCAZ15824 Cafar “Pholidobolus macbrydei” QCAZ15820 Cafar “Pholidobolus macbrydei” QCAZ15819 Cafar “Pholidobolus macbrydei” QCAZ10050 Cafiar “Pholidobolus macbrydei” QCAZ15816 Cafar “Pholidobolus macbrydei” QCAZ15812 Cafar “Pholidobolus macbrydei” QCAZ15813 Cafar “Pholidobolus macbrydei” QCAZ15815 Cafar “Pholidobolus macbrydei” QCAZ15814 Cafar “Pholidobolus macbrydei” QCAZ15818 Cafar “Pholidobolus macbrydei” QCAZ15817 Cafar “Pholidobolus macbrydei” QCAZ15823 Cafiar “Pholidobolus macbrydei” QCAZ15811 Cafar “Pholidobolus macbrydei” QCAZ9914 Azuay I Clade D Clade E “Pholidobolus macbrydei” QCAZ9932 Azuay | “Pholidobolus macbrydei” KU218406 Azuay Clade C Pholidobolus samek sp. n. QCAZ14954 Zamora Chinchipe Pholidobolus samek sp. n. QCAZ14956 Zamora Chinchipe Pholidobolus samek sp. n. QCAZ14955 Zamora Chinchipe Pholidobolus condor sp. n. QCAZ16790 Morona Santiago Pholidobolus condor sp. n. QCAZ15844 Morona Santiago Pholidobolus condor sp. n. QCAZ16788 Morona Santiago Pholidobolus condor sp. n. QCAZ16789 Morona Santiago “Pholidobolus macbrydei” QCAZ10051 Cafar “Pholidobolus macbrydei” QCAZ10052 Cafar Clade B “Pholidobolus macbrydei” QCAZ9947 Cafiar “Pholidobolus macbrydei” QCAZ6945 Loja “Pholidobolus macbrydei” QCAZ6946 Loja Clade A “Pholidobolus macbrydei” QCAZ10054 Loja Figure |. Phylogeny of Pholidobolus. Maximum clade credibility tree derived from a partitioned analysis of 1904 bp of mitochondrial and nuclear DNA. Bayesian posterior probabilities are shown above branches and bootstrap values (RAxML analysis) below branches; values < 0.5 and 50, respectively, are not shown. For clarity, outgroup taxa and values on short branches are not shown. Species outside the “P macbrydei” complex are in grey; new species described in this paper are in color matching the distribution records of the map in Figure 7. The species name followed by voucher number and province (“P macbrydei” complex only) are provided for each terminal. Photographs from top to bottom: P dolichoderes sp. nov. holotype, P fascinatus sp. nov. holotype, “P macbrydei” (Clade B) QCAZ 15824, P samek sp. nov. holotype, P condor sp. nov. holotype. Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) 118 900 y00 O00 00 Sso0 so0 sd00 OO S00 O00 OO FOO S00 O00 YOO FOO S0°0 we Pag arta SHIOGUP TOU 8I soo 6©6Ss00 6800 S00 2S 9070S S00 SO'N-—s SDD—s*sO9070—s-«d 900s SD'N—s SD'D-—s SDN SO'D—s« SOD SO pepe eG CE Sad. 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Oo “sro. SL:0° ZrO. 40 S0= STO: S10 SEER OG CR OG CY ero, LPO PO. SLO SPrO- ZrO SO. IQ £10 suf smoqopyogd 11 600 O10 600 ¢€10 10 800 00 cro cro d Pr. OL 90°0" “900 TPO Tl0- -90°0. OTTO: O10, OO dP. 6 90°70: FTO” EPO. 40°C, LOF0. SEO: “LTO Pro: WO er: S00 <0O00. =EEO “UL SD ePeO” 2 EDO OFO 600. 1.0 were extracted from the PCA (Table 5). These components accounted for 50.7% of the total variation. The highest loadings cor- responded to supratympanic temporals (NTS) and number of scales along margin of upper jaw (SUJ) for PC I, and number of scales around the tail (SAT) and number of scales around the body (SAB) for PC II (Table 5). In general, there is wide overlap in morphological space among species of the “P. macbrydei” complex (Fig. 2). Comparative hemipenial morphology Hemipenes of holotypes of the four new species described herein are approximately 4—5 mm and 5-7 subcaudal scales long. The organs are fully everted in specimens of P fascinatus, P condor, and P samek and partially everted in P dolichoderes; the hemipenes of the holotype of P fascinatus and P condor are fully expanded, whereas the organs of P dolichoderes and P samek are partially expanded (Fig. 3). All hemi- penes have two small lobes detached from the hemipenial body when the organ is fully everted. The hemipenis of P condor presents a distinctive capitular groove originating at the median hemipenial body and extending toward the lobes. The lobes of P fascinatus, P condor, and P samek present folds on their tips, which are not visible in P dolichoderes due to the partial eversion. The hemipenial body is 122 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) i=) ® P. condor sp. n. “ P. dolichoderes sp. n. ™ P. fascinatus sp. n. “P. macbrydei” P. samek sp. n. Component 2 (9. 99%) -4. 5.0 Component 1 (40.69%) Figure 2. Principal components analysis of 16 morphological variables and 140 specimens of the “P/o- lidobolus macbrydei” species complex. See Table 5 for character loadings on each component. cylindrical in P dolichoderes and P condor, whereas in P samek and P fascinatus the body is conical, with the basis distinctly thinner than the rest of the body. The sulcus spermaticus is broad in P fascinatus, P dolichoderes, and P samek, narrower in P condor; in P fascinatus and P condor, the sulcus spermaticus is deeper than in P. dolichoderes and P. samek. The sulcus originates medially at the base of the organ and extends in a straight line throughout the body towards the lobes in all species. However, unlike P dolichoderes and P condor, the sulcus originates between thick lips in P samek and P fascinatus. In all species, the sulcus spermaticus bifurcates at the lobular crotch, with each branch extending along the medial face of each lobe. The sides and borders of the sulcate and asulcate faces are ornamented with a series of roughly equidistant and chevron-shaped flounces, with the chevron vertices aligned medially on each side and directed proximally. All flounces bear calcified comb-like series of spicules, distinctively stained in red with Alizarin. The number of flounces extending along the hemipenial body varies slightly among species: 21 in P condor and P. samek and 22 in P dolichoderes and P fascinatus. The base of the asulcate face bears three medial flounces in P condor, P dolichoderes, and P samek, and four in P fascinatus. All species have a conspicuous unevenness forming a bulge along the margins of the asulcate face. Systematics of Pholidobolus lizards 123 Figure 3. Comparative hemipenial morphology of Pholidobolus. Sulcate (left), lateral (center) and asul- cate (right) views of: A Pholidobolus samek sp. nov. (QCAZ 14955) B Pholidobolus condor sp. nov. (QCAZ 15844) C Pholidobolus dolichoderes sp. nov. (QCAZ 16353) D Pholidobolus fascinatus sp. nov. (QCAZ 15120). Scale bar: 1 mm. 124 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Systematic accounts Pholidobolus samek sp. nov. http://zoobank.org/43 1 C8AD2-3 164-405 1-B7DC-1459C4949F51 Figures 4—6 Proposed standard English name: Green-striped cuilanes Proposed standard Spanish name: Cuilanes de franjas verdes Holotype. QCAZ 14955 (Figs 4, 5), adult male, Ecuador, Provincia Zamora-Chinchi- pe, Cerro Plateado Biological Reserve, Cerro Plateado plateau, 4.6159S, 78.7870W, WGS84, 2844 m, 23 September 2016, collected by Diego Almeida, Eloy Nusirquia, Fernando Ayala, Javier Pinto, Alex Achig and Malki Bustos. Paratypes (6). Ecuapor: Provincia Zamora-Chinchipe: QCAZ 14954 (adult fe- male), same data as holotype; QCAZ 14956 (adult female), Cerro Plateado Biological Reserve, 4.6050S, 78.8167W, WGS84, 2320 m, 28 September 2016; QCAZ 14969- 70, 14976-77(hatchlings) Cerro Plateado Biological Reserve, 4.6179S, 78.7838W, WGS84, 2873 m, 24 September 2016, same collectors as holotype. Diagnosis. Pholidobolus samek is unique among its congeners, except P condor sp. nov., in having green dorsolateral stripes on the head. However, adult males of P. samek differ from those of P condor sp. nov. in having brighter dorsolateral head stripes and lacking a reddish venter. In addition, P affinis, P prefrontalis, P macbrydei, P dolichoderes sp. nov., and P. montium differ from P samek (character states of P samek in parentheses) in having a loreal scale frequently in contact with the supralabials (lo- real scale not in contact with supralabials), and dorsal scales finely wrinkled (slightly keeled). Pholidobolus ulisesi and P hillisi differ from P samek in having a diagonal white bar along the rictal region (white rictal bar absent). Pholidobolus samek can be distin- guished from P dicrus by lacking a bifurcating vertebral stripe at midbody. Pholidobolus affinis, P prefrontalis, P. dicrus, P hillisi, and P. vertebralis further differ from P samek in having well defined prefrontal scales (if present, prefrontal scales poorly differenti- ated). Additionally, P samek has fewer dorsal scales (27-29) than P affinis (45-55), P montium (35-50), P prefrontalis (37-46), P macbrydei (31-43), P fascinatus sp. nov. (32-37), and P dolichoderes sp. nov. (35-40). Pholidobolus samek can be further distinguished from P fascinatus by having widened medial scales on collar, and from P. dolichoderes sp. nov. by having fewer temporals (4—5 and 7-9, respectively), fewer ventrals (19—21 and 25-27), and fewer gulars (15-18 and 22-23). Characterization. (1) Two (rarely three) supraoculars, anteriormost slightly larger than posterior one; (2) prefrontals present or absent; (3) femoral pores absent in both sexes; (4) four to five opaque lower eyelid scales; (5) scales on dorsal surface of neck striated, becoming slightly keeled from forelimbs to tail; (6) two or three rows of lateral granules at midbody; (7) 27—29 dorsal scales between occipital and posterior margin of hindlimb; (8) lateral body fold present; (9) keeled ventrolateral scales on each side absent; (10) dorsum grayish brown with a distinct golden gray middorsal stripe, slen- der at midbody, becoming pale gray towards tail; (11) labial stripe white or orange; Systematics of Pholidobolus lizards 125 (12) flanks of body dark brown; (13) conical hemipenial body, with sulcus spermaticus originating between thick lips. Description of holotype. Adult male (QCAZ 14955) (Figs 4, 5); SVL 46.7 mm; TL 80.9 mm; dorsal and lateral head scales juxtaposed, finely wrinkled; rostral hexago- nal, 2.06 times as wide as high; frontonasal irregularly quadrangular, wider than long, laterally in contact with nasal, loreal and first superciliary, slightly bigger than frontal; prefrontal scales absent; frontal longer than wide, in contact with one supraocular on the left side, and two on the right side; frontoparietals pentagonal, longer than wide, slightly wider posteriorly, each in contact laterally with supraocular IJ; interparietal roughly heptagonal; parietals slightly bigger than interparietal, hexagonal, and posi- tioned anterolaterally to interparietal, each in contact anteriorly with supraocular II (and supraocular III on right side) and dorsalmost postocular; postparietals three, me- dial scale smaller than laterals; seven supralabials, fourth one longest and below center of eye; six infralabials, fourth one shortest and below center of eye; temporals enlarged, irregularly hexagonal, juxtaposed, smooth; two large supratemporal scales, smooth; nasal slightly divided, irregularly pentagonal, longer than high, in contact with rostral anteriorly, first and second supralabials ventrally, frontonasal dorsally, loreal postero- dorsally and frenocular posteroventrally; nostril on ventral aspect of nasal, directed lateroposteriorly; loreal rectangular, wider dorsally; frenocular higher than long, in contact with nasal, separating loreal from supralabials; two supraoculars on left side, three on right side (posteriormost much smaller), with the first one being the largest; four elongate superciliaries, first one enlarged, in contact with loreal; palpebral disc di- vided into four enlarged, pigmented scales; suboculars three (on the left side the medial subocular is fragmented), elongated and homogeneous in size; two postoculars, the dorsalmost wider than the other; ear opening vertically oval, without denticulate mar- gins; tympanum recessed into a shallow auditory meatus; mental semicircular, wider than long; postmental pentagonal, slightly wider than long, followed posteriorly by three pairs of genials, the anterior two in contact medially and the posterior one sepa- rated by postgenials; all genials in contact with infralabials; gulars imbricate, smooth, posteriorly widened in two longitudinal rows; posterior row of gulars (collar) with six scales, the medial two widened. Nuchal scales similar in size to dorsals, except for the anteriormost that are wid- ened; scales on sides of neck small and granular; dorsal scales hexagonal, elongate, imbricate, arranged in transverse rows; scales on dorsal surface of neck striated, be- coming progressively keeled from forelimbs to tail; number of dorsal scales between occipital and posterior margin of hindlimbs 27; dorsal scale rows in a transverse line at midbody 26; one longitudinal row of smooth, enlarged ventrolateral scales on each side; dorsals separated from ventrals by two rows of small scales at level of 13° row of ventrals; lateral body fold between fore and hindlimbs present; ventrals smooth, wider than long, arranged in 20 transverse rows between collar fold and preanals; six ventral scales in a transverse row at midbody; subcaudals smooth; axillary region with granular scales; scales on dorsal surface of forelimb striated, imbricate; scales on ventral surface of forelimb granular; two thick, smooth thenar scales; supradigitals (left/right) 126 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Figure 4. Holotype of Pholidobolus samek sp. nov. (QCAZ 14955) in dorsal (A), ventral (B), and lateral (C) views. Male, SVL = 46.7 mm. (A, B): preserved specimen; (C): live specimen. Photographs by Darwin Nufiez and Valeria Chasiluisa. Systematics of Pholidobolus lizards 127 Figure 5. Head of holotype of Pholidobolus samek sp. nov. (QCAZ 14955) in lateral (A), dorsal (B), and ventral (C) views. Photographs by Valeria Chasiluisa. Scale bar: 5 mm. 128 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) 3/3 on finger I, 6/7 on II, 8/8 on HI, 9/9 on IV, 6/6 on V; supradigitals 3/4 on toe I, 6/6 on IL, 10/9 on III, 11/12 on IV, 7/7 on V; subdigital lamellae of fingers I and II single, paired on HI (except the four distalmost), paired at base on IV, on finger V all single; subdigital lamellae 5/5 on finger I, 11/12 on H, 15/16 on HI, 17/16 on IV, 9/10 on V; subdigital lamellae on toes I and II single, on toe III, IV and V all paired, except for the three distalmost subdigitals; subdigital lamellae 6/6 on toe I, 11/10 on II, 16/15 on IL, 21/21 on IV, 14/14 on V; groin region with small, imbricate scales; scales on dorsal surface of hindlimbs smooth and imbricate; scales on ventral surface of hindlimbs smooth; scales on posterior surface of hindlimbs granular; femoral pores absent; preanal pores absent; cloacal plate paired, bordered by four scales anteriorly, of which the two medialmost are enlarged. Additional measurements (mm) and proportions of the holotype: HL 11.4; HW 7-43 Shlt.7:0; AGD 23:9; TL/ISVIE1 5; AL SVL 0:2; SEENW/SV.L-0:2; Shle/SVL.085 AGD/SVL 0.5. Color of holotype in life. Dorsal background from head to base of tail grayish brown, with a golden light brown vertebral stripe extending from occiput to tail; bright green dorsolateral stripes on head; cream white longitudinal stripe extending from first supralabial to shoulder; sides of neck, flanks and limbs dark brown; reddish brown narrow stripe extending from tympanum to arm insertion; ventrolateral region of body grayish brown; throat cream; chest, belly and base of tail cream orange (Figs 4C, 6B). Color of holotype in preservative. Dorsal background uniformly grayish brown, with a golden-gray vertebral stripe extending from occiput to tail; vertebral stripe wider anteriorly, becoming slightly slender at most posterior part of body; dorsal and lateral surfaces of head brown (rostral, frontonasal, frontal, frontoparietals, and supraoculars); bluish white longitudinal stripe extending from first supralabial to shoulder and fad- ing on flanks; ventrolateral aspect of neck dark brown with a dorsolateral light brown stripe extending posteriorly along flanks to hindlimbs; forelimbs with scattered ocelli (black with white center); flanks grayish brown with two dorsolateral stripes on each side, the dorsal one dark brown and the most ventral one brown diffuse with dark brown spots; tail brown dorsally; ventral surface of head gray, chest and venter dark gray, ventral surface of tail slightly gray, with scattered dark brown marks. Variations. Measurements and scale counts of Pholidobolus samek are presented in Table 6. Supralabials 8/7 (left/right) and temporals five in specimen QCAZ 14956; small and separated prefrontals on both sides in QCAZ 14954 and one prefrontal on right side in QCAZ 14956; little intrusive scales between parietal and postparietal in QCAZ 14954; frontal hexagonal in QCAZ 14956; roughly decagonal interparietal in QCAZ 14954. Usually two scales on posterior cloacal plate, four in QCAZ 14954 and 14956. Male is larger (SVL 46.7 mm, NV = 1) than females (maximum SVL 45.4 mm, WN = 2). Hatchlings (QCAZ 14969, 14970, 14976) with eight or seven (QCAZ 14976) posterior gular (collar) scales. Unlike the male holotype, females have an orange-brown longitudi- nal stripe extending from third supralabial to shoulder and fading on the flanks (Fig. 6). Distribution and natural history. Pholidobolus samek inhabits cloud forests in Cordillera del Céndor, southeastern Ecuador at elevations between 2324-2844 m (Fig. 7). The new species is known only from Zamora-Chinchipe province, on the Systematics of Pholidobolus lizards 129 Figure 6. Pholidobolus samek sp. nov. in life. A Adult female, paratype (QCAZ 14954) B adult male, holotype (QCAZ 14955). sandstone plateaus of Cerro Plateado Biological Reserve. The ground at the type local- ity is covered with mosses, roots, and bromeliads. Such ground cover is locally known as bamba. All specimens were found active at 11h30—17h00 under stones or terrestrial bromeliads (Fig. 8). Four eggs, collected under flat stones on 24-09-2016, were incu- bated in sphagnum and perlite in captivity for two months approximately. They were 130 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Table 6. Summary of morphological characters and measurements (mm) of Pholidobolus samek sp. nov., P condor sp. nov., P. dolichoderes sp. nov., and P fascinatus sp. nov. Range (first line) and mean + standard deviation (second line) are presented. Character P. samek sp. nov. N = 7| P. condor sp. nov. (adults = 3) N = 4 (adults = 1) 7-10 (9.14 + 1.07) Scales along margin of upper jaw 8-9 (8.75 + 0.5) P. dolichoderes sp. nov. N = 5 (adults = 3) 9-11 (10.2 + 0.84) P. fascinatus sp. nov. N = 27 (adults = 4) 7-10 (8.36 + 0.91) Scales along margin of lower jaw Gulars Ventrals in transverse row at midbody 8-9 (8.25 + 0.5) 15-18 (16.71 + 1.11) | 14-16 (15 + 0.82) 19-21 (20 + 0.82) 5-10 (7.14 + 2.67) 18-20 (19 = 1.15) 10-11 (10.2 + 0.45) 22-23 (22.8 + 0.48) 25-27 (25.8 + 0.84) 4-10 (7.4 + 1.58) 14-17 (15.72 + 0.89) 21-25 (22.96 + 1.21) Dorsals from occiput to base of tail Temporals 27-29 (27.71 + 0.76) 4-5 (4.14 + 0.38) 26-30 (27.75 + 1.71) 4—5 (4.25 + 0.5) 35-40 (36.8 + 2.05) 7-9 (8 + 0.70) 32-37 (34.64 + 1.19) 3-5 (3.44 + 0.65) Scales around midbody Scales around tail 14-16 (15 + 0.81) 14—20 (17.86 + 2193) 5-30 (oF AES) 27-30 (28 + 1.41) 31-33 (32.2 + 0.84) 18-19 (18.6 + 0.55) 28-34 (30.96 + 1.79) 18-22 (20.32 + 1.18) Lower eyelid scales 4—5 (4.14 + 0.38) SI 4-6 (4.8 + 0.84) 4—6 (5.04 + 0.61) Gular (collar) 6-8 (7.14 + 0.9) 6-9 (7.75 + 1.26) 6-8 (6.4 + 0.89) 9-12 (10.28 + 0.73) scales Head length in 9.9-11.4 (10.76 + 11 9.7-10.6 (10.05 + 0.46) | 8.9-12.3 (10.22 + adults 0.77) 1.80) Head width in 6.5-7.4 (6.93 + 0.48) 6.6 6.2-6.3 (6.26 0.05) |6.6—9.2 (7.58 + 1.45) adults SVL in adults 41.6-49.3 (45.89 + 41.1-50.6 (45.75 + | 42.6-52.5 (47.3 + 3.89) 4.74) 4.98) 14.0-14.1 mm long, 8.0-8.5 mm wide, and weighted 0.4 g on average. Hatchlings (QCAZ 14969-70, 14976—77) weighted 0.3 g and were 24.7 mm in SVL on average. Conservation status. Pholidobolus samek is only known from Cordillera del Cén- dor. The population size for this species is unknown, but our sampling suggests low abundances. Because of the small known distribution, as well as habitat destruction through mining activities nearby (Van Teijlingen 2016), we suggest assigning P samek to the Critically Endangered category under criteria Bla, b(iii); C1; D, according to IUCN (2012) guidelines. Etymology. The specific epithet samek means green in the Shuar language, in al- lusion to the green dorsolateral head stripes distinguishing the new species from other congeners. The type locality of Pholidobolus samek lies within territory of Shuar indig- enous people, who inhabit the Amazonian rainforest in Ecuador and Peru. Remarks. Pholidobolus samek sp. nov. is very similar morphologically and genet- ically to P condor sp. nov. These species can be easily distinguished from each other by coloration in adult males, although we recognize that our sample size is small (N = 7 and 4, respectively) and includes only one adult male per species. However, Systematics of Pholidobolus lizards 13 ECUADO ~ Morona Pacific a if ere Santiago Ocean bf Elevation in m 0-1000 | 1001—2000 2001-3000 >3000 Chinchip Figure 7. Distribution of samples of the “Pholidobolus macbrydei” species complex included in phylo- genetic analyses. Circles correspond to four new species described in this paper: P samek sp. nov. (red), P condor sp. nov. (blue), P dolichoderes sp. nov. (brown), and P fascinatus sp. nov. (green). Triangles are “Pholidobolus macbrydei” clades as illustrated in the phylogenetic tree (Fig. 1): A (green) B (white) C (red) D (blue) E (black) F (turquoise). Orange diamond corresponds to type locality of P macbrydei. This map was created in QGIS v3.10. further evidence supports recognition of P samek and P condor as different species. First, they are reciprocally monophyletic and they are not sister taxa, with P samek being sister to “P macbrydei” Clade A (Fig. 1), which is very different in color pat- terns from either P samek or P condor (V. Parra and O. Torres-Carvajal, personal ob- servation). Second, unlike the 12S gene (the less variable gene in this study), genetic distances between P samek and P condor for 16S and ND4 are not the lowest (Tables 2 and 4, respectively) within Pholidobolus. For example, the 16S distance between P. samek and P. condor (3%) is the same as the distance between the well-recognized species P paramuno and P affinis. In addition, genetic exchange among P samek, P. condor and Clade A is very unlikely as they are isolated from each other on moun- taintops above 2000 m (Fig. 7). Pholidobolus condor sp. nov. http://zoobank.org/BB38EC4E-634D-47 28-BA30-412913E7D0E0 Figures 9, 10 Proposed standard English name: Condor cuilanes Proposed standard Spanish name: Cuilanes del Condor 132 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Figure 8. Habitat of Pholidobolus samek sp. nov. A Vegetation around type locality, Cerro Plateado Bio- logical Reserve, Ecuador B habitat where holotype was found. Photographs by Alvaro Pérez. Systematics of Pholidobolus lizards 1:33 Holotype. QCAZ 15844 (Figs 9, 10), adult male, Ecuador, Provincia Morona San- tiago, buffer zone of El Quimi Biological Reserve, plateau on the eastern side of El Quimi river valley, 3.51892S, 78.3690W, WGS84, 2209 m, 11 July 2017, collected by Diego Almeida, Darwin Nufiez, Eloy Nusirquia, Alex Achig and Ricardo Gavilanes. Paratypes (3). Ecuapor: Provincia Morona Santiago: QCAZ 16790 (hatchling), El Quimi Biological Reserve, base camp towards old heliport (high zone), 3.51894S, 78.36897W, WGS84, 2226 m, 17 April 2018; QCAZ 16788-89 (hatchlings), El Quimi Biological Reserve, near base camp, 3.5182S, 78.3913W, WGS84, 1994 m, 12 April 2018, collected by Diego Almeida, Darwin Nunez, Eloy Nusirquia, Alex Achig and Maria del Mar Moretta. Diagnosis. Pholidobolus condor is unique among its congeners, except P samek sp. nov., in having green dorsolateral stripes on the head. However, adult males of P condor differ from those of P samek sp. nov. in having lighter dorsolateral head stripes, and reddish flanks and venter. In addition, P ulisesi, P dicrus, P hillisi, and P. vertebralis dif- fer from P condor (character states of P condor in parentheses) in having a conspicuous light vertebral stripe (light vertebral stripe absent). Pholidobolus affinis, P prefrontalis, P dicrus, P. hillisi, and P. vertebralis further differ from P condor in having prefrontal scales (prefrontal scales absent). Additionally, P condor sp. nov. has fewer dorsal scales (26-30) than P affinis (45-55), P montium (35-50), P prefrontalis (37-46), R. mac- brydei (31-43), and P dolichoderes sp. nov. (35-40). Pholidobolus condor can be further distinguished from P fascinatus sp. nov. by having widened medial scales on collar, and from P dolichoderes sp. nov. by having fewer temporals (7—9 and 4—5, respectively), fewer ventrals (18-20 and 25-27), and fewer gulars (14-16 and 22-23). Characterization. (1) Two (rarely three) supraoculars, anteriormost larger than posterior one; (2) prefrontals absent; (3) femoral pores absent; (4) four opaque lower eyelid scales; (5) scales on dorsal surface of neck striated or smooth, progressively stri- ated from forelimbs to tail; (6) two rows of lateral granules at midbody; (7) 27-31 dorsal scales between occipital and posterior margin of hindlimb; (8) lateral body fold present; (9) keeled ventrolateral scales on each side absent; (10) dorsum dark brown with a narrow, pale brown stripe; (11) labial stripe white; (12) flanks of body dark brown or gray; (13) hemipenial body cylindrical with distinctive capitular groove. Description of holotype. Adult male (QCAZ 15844) (Figs 9, 10); SVL 42.7 mm; TL 74.8 mm; dorsal and lateral head scales juxtaposed, finely wrinkled; rostral hexago- nal, 1.67 times as wide as high; frontonasal quadrangular, slightly bigger than frontal, laterally in contact with nasal, loreal and first superciliary; prefrontal scales absent; frontal pentagonal, longer than wide, wider anteriorly, in contact with first superciliary and su- praocular; frontoparietals hexagonal, longer than wide, slightly wider in the middle, each in contact laterally with supraocular I; interparietal octagonal, with a short medial suture posteriorly, lateral borders nearly parallel to each other; parietals larger than interparietal, hexagonal and positioned anterolaterally to interparietal, each in contact laterally with supraocular II and dorsalmost postocular; postparietals three, medial scale smaller than lateral ones; eight supralabials, fourth one longest and below center of eye; six infralabials, fourth one below center of eye; temporals enlarged, irregularly hexagonal, smooth; two 134 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) : ee Figure 9. Holotype of Pholidobolus condor sp. nov. (QCAZ 15844) in dorsal (A), ventral (B), and lateral (C) views. Male, SVL = 42.7 mm. Preserved specimen (A); live specimen (B, C). Photographs by Malki Bustos. Systematics of Pholidobolus lizards 135 Figure 10. Head of holotype of Pholidobolus condor sp. nov. (QCAZ 15844) in lateral (A), dorsal (B), and ventral (C) views. Photographs by Valeria Chasiluisa. Scale bar: 5 mm. 136 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) large and smooth supratemporals; nasal shield slightly divided above nostril, irregularly pentagonal, longer than high, in contact with rostral anteriorly, first and second suprala- bials ventrally, frontonasal dorsally, loreal posterodorsally and frenocular posteroventral- ly; nostril on ventral aspect of nasal, directed laterally; loreal quadrangular, slightly wider dorsally, not in contact with supralabials; frenocular higher than long, in contact with na- sal; nasal separating loreal from supralabials; two supraoculars, anteriormost one the wid- est; four elongate superciliaries, anteriormost enlarged, in contact with loreal; palpebral disc divided into five pigmented scales; four suboculars, anteriormost three elongated and homogeneous in size, posteriormost widest; two postoculars, the dorsalmost wider than the other; ear opening vertically oval, without denticulate margins; tympanum re- cessed into a shallow auditory meatus; mental wider than long; postmental pentagonal, slightly wider than long, followed posteriorly by three pairs of genials, the anterior two pairs in contact medially and the third pair separated by postgenials; all genials in contact with infralabials; gulars imbricate, smooth, widened in two longitudinal rows; posterior row of gulars (collar) with nine scales, the medial three slightly widened. Nuchal scales slightly smaller than dorsals, except for the anteriormost that are widened; scales on sides of neck small and granular; dorsal scales elongate, imbricate, arranged in transverse rows; scales on dorsal surface of neck striated, becoming progres- sively keeled from forelimbs to tail; dorsal scales between occipital and posterior mar- gin of hindlimbs 27; dorsal scale rows in a transverse line at midbody 27; one longitu- dinal row of smooth, enlarged ventrolateral scales on each side; dorsals separated from ventrals by two rows of small scales at the level of 13" row of ventrals; lateral body fold between fore and hindlimbs present; ventrals smooth, wider than long, arranged in 20 transverse rows between collar fold and preanals; six ventral scales in a transverse row at midbody; subcaudals smooth; axillary region with granular scales; scales on dorsal surface of forelimb striated, imbricate; scales on ventral surface of forelimb granular; two thick, smooth thenar scales; supradigitals (left/right) 3/3 on finger I, 6/6 on UH, 8/8 on HI, 9/9 on IV, 6/6 on V; supradigitals 3/3 on toe I, 6/6 on II, 9/9 on IL, 12/12 on IV, 7/7 on V; subdigital lamellae of finger I, H, HI, and V single, on finger IV few scales in the middle paired; subdigital lamellae 6/6 on finger I, 11/11 on HJ, 15/15 on III, 17/16 on IV, 10/10 on V; subdigital lamellae on toes I and II single, on toes HI, IV and V paired, except for two or three distalmost subdigitals; subdigital lamellae 7/6 on toe I, 12/12 on H, 15/16 on HI, 22/22 on IV, 12/12 on V; groin region with small, juxtaposed scales; scales on dorsal surface of hindlimbs striated and imbricate; scales on ventral surface of hindlimbs smooth; scales on posterior surface of hindlimbs granular; femoral pores absent; preanal pores absent; cloacal plate paired, bordered by four scales anteriorly, of which the two medialmost are enlarged. Additional measurements (mm) and proportions of the holotype: HL 11.0; HW 6.6; ShL 5.8; AGD 20.4; TL/SVL 1.7; HL/SVL 0.3; HW/SVL 0.2; ShL/SVL 0.1; AGD/SVL 0.5. Color of holotype in life. Dorsal background from head to base of tail dark brown, with a golden brown vertebral stripe extending from occiput to tail; greenish cream dorsolateral stripes on head, becoming light brown on posterior part of body; white longitudinal stripe extending from first supralabial to shoulder; sides of neck, flanks Systematics of Pholidobolus lizards 137 and limbs dark brown; chocolate brown narrow stripe extending from tympanum to arm insertion; ventrolateral region of body grayish brown; throat reddish cream; chest, belly, base of tail and lateral region of tail bright orange, with brown marks in some scales; ventral surface of hind limbs with orange diffuse marks (Fig. 9B,C). Color of holotype in preservative. Dorsal background uniformly dark brown with a grayish brown middorsal stripe extending from occiput onto tail; dorsolateral stripe distinct, pale gray, extending from snout to near base of tail; head brown dorsally (rostral, frontonasal, frontal, frontoparietals and supraoculars) and dark brown lateral- ly; white longitudinal stripe extending from first supralabial to forelimb; lateral aspect of neck dark brown with a dorsolateral light brown stripe extending posteriorly along flanks to hindlimbs; flanks grayish brown; tail dark brown dorsally and bronze later- ally; ventral surface of head gray, with dirty cream genials and scattered black marks; chest, belly and ventral surface of tail light gray with light red spots; ventral surface of limbs dark gray (Fig. 9A). Variations. Measurements and scale counts of Pholidobolus condor are presented in Table 6. Supraoculars three on left side in specimen QCAZ 16789; supralabials six in QCAZ 16789 and 16790, and seven in QCAZ 16788; two quadrangular frontonasals in QCAZ 16788; transverse rows of ventral scales between collar fold and preanals 18 in QCAZ 16788 and 19 in QCAZ 16790. Hatchlings with eight (QCAZ 16788-89) or six (QCAZ 16790) posterior gular (collar) scales. Unlike the adult male, hatchlings lack reddish color on tail. Distribution and natural history. Pholidobolus condor occurs in Cordillera del Condor in southeastern Ecuador at elevations between 1994—2226 m. The new species is known from El Quimi Biological Reserve in Morona Santiago province (Fig. 7). The holotype was found active at 21h14 at the base of a bromeliad on a sandstone plateau of shrub vegetation (Fig. 11). Several eggs were found within a bromeliad, suggesting that females of P condor lay their eggs in communal nests. Four eggs that were found on the ground at the base of bromeliads and under a trunk were incubated in sphagnum and perlite in captiv- ity for approximately three months. On average, hatchlings weighted 0.4 g and were 23.7 mmunSV1-. Conservation status. Pholidobolus condor is only known from Cordillera del Cén- dor in southeastern Ecuador. This area is currently threatened by mining activities (Ron et al. 2018; Valencia et al. 2017; Van Teijlingen 2016). Habitat destruction and fragmentation is evident at a distance of ~11 km from the collection sites (Mazabanda et al. 2018). Because of the small known distribution and habitat disturbance, we sug- gest assigning P condor to the Critically Endangered category under criteria Bla, b(iii); C1; D, according to IUCN (2012) guidelines. Etymology. ‘The specific epithet condor refers to Cordillera del Condor, where the new species was discovered. The Cordillera del Condor is an eastern outlier of the main Andean chain, where a significant number of species have been discovered in the last decade (Brito et al. 2017; Huamantupa-Chuquimaco and Neill 2018; Ron et al. 2018; Torres-Carvajal et al. 2009; Valencia et al. 2017). Remarks. See remarks on Pholidobolus samek sp. nov. above. 138 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Figure | 1. Habitat of Pholidobolus condor sp. nov. at El Quimi Biological Reserve, Ecuador. Photographs by Alvaro Pérez. Pholidobolus dolichoderes sp. nov. http://zoobank.org/95D82201-D761-40F4-8395-4AAC38F24563 Figures 12-14 Proposed standard English name: Long-necked cuilanes Proposed standard Spanish name: Cuilanes de cuello largo Holotype. QCAZ 16353 (Figs 12, 13), adult male, Ecuador, Provincia Azuay, San Felipe de Offa, 3.4292S, 79.2364W, WGS84, 2672 m, 16 March 2018, collected by Diego Almeida, Darwin Nunez, Eloy Nusirquia, Alex Achig and Katherine Nicolalde. Paratypes (4). Ecuapor: Provincia Azuay: QCAZ 16349, 16352 (adult females), San Felipe de Ona, Susudel-Poetate road, 3.4322S, 79.2369W, WGS84, 2506 m, 16 March 2018; QCAZ 16350—51 (juveniles), San Felipe de Ofa, 3.4275S, 79.2339W, WGS84, 2675 m, 16 March 2018, same collectors as holotype. Diagnosis. Pholidobolus dolichoderes is unique among its congeners in having a long neck with granular scales between the posterior corner of the orbit and the ante- rior edge of the tympanum, as well as an inconspicuous ventrolateral fold between fore and hindlimbs. In addition, P ulisesi, P dicrus, P hillisi, and P. vertebralis differ from P dolichoderes in having a conspicuous light vertebral stripe. The new species further dif- fers from P affinis in lacking ocelli on flanks, and from P condor sp. nov., P macbrydei, and P montium in having prefrontal scales. Pholidobolus dolichoderes has more dorsals Systematics of Pholidobolus lizards 139 Figure 12. Holotype of Pholidobolus dolichoderes sp. nov. (QCAZ 16353) in life in dorsal (A), ventral (B), and lateral (C) views. Male, SVL = 41.1 mm. Photographs by Gustavo Pazmifo. (35-40) and ventrals (25-27) than P samek sp. nov. (27-29 and 19-21, respectively) and P condor sp. nov. (26-30 and 18-20), and, unlike P fascinatus sp. nov., it has wid- ened medial scales on collar. In addition, P dolichoderes has more temporals (7-9) and gulars (22—23) than P samek sp. nov. (4—5 and 15-18, respectively), P condor sp. nov. (4-5 and 14-16), and P fascinatus sp. nov. (3-5 and 14-17). 140 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Figure 13. Head of holotype of Pholidobolus dolichoderes sp. nov. (QCAZ 16353) in lateral (A), dorsal (B), and ventral (C) views. Photographs by Valeria Chasiluisa. Scale bar: 5 mm. Systematics of Pholidobolus lizards 141 Characterization. (1) Three supraoculars, anteriormost larger than posterior ones; (2) prefrontals present; (3) femoral pores present in both sexes; (4) four to six opaque lower eyelid scales; (5) scales on dorsal surface of neck smooth, becoming slightly keeled from forelimbs to tail; (6) two or three rows of lateral granules at midbody; (7) 35-20 dorsal scales between occipital and posterior margin of hindlimb; (8) lateral body fold present but inconspicuous; (9) keeled ventrolateral scales on each side ab- sent; (10) dorsum dark brown with a diffuse pale brown vertebral stripe that becomes grayish brown towards tail; (11) labial stripe white; (12) flanks of body gray brown; (13) white stripe along forelimb present; (14) hemipenial body cylindrical, with sulcus spermaticus originating between thick lips. Description of holotype. Adult male (QCAZ 16353) (Figs 12, 13); SVL 41.1 mm; TL 96.3 mm; dorsal and lateral head scales imbricated, smooth; rostral hexagonal, 1.75 times as wide as high; frontonasal heptagonal, slightly wider than long, laterally in con- tact with nasal, similar in size to frontal; prefrontals present, in wide contact medially, and in contact with loreal and first superciliary laterally; frontal hexagonal, longer than wide, wider anteriorly, in contact with first and second supraoculars; frontoparietals hexagonal, longer than wide, slightly wider posteriorly, each in contact with second and third supraoculars, parietals and interparietal; interparietal heptagonal, lateral bor- ders nearly parallel to each other; parietals wider than interparietal, heptagonal, and positioned anterolaterally to interparietal, each in contact with third supraocular and dorsalmost postocular; postparietals three, medial scale smaller than lateral ones; seven supralabials, fourth one the longest and below center of eye; five infralabials, fourth one below center of eye; temporals small, irregularly, smooth; supratemporal scales not well differentiated, smooth; nasal shield divided above the nostril, longer than high, in con- tact with rostral anteriorly, first and second supralabials ventrally, frontonasal dorsally, loreal posteriorly; loreal pentagonal, slightly wider dorsally, in contact with second and third supralabials; frenocular longer than high, in contact with loreal; three supraocu- lars, with the first one being the widest; four elongate superciliaries, anteriormost one enlarged, in contact with loreal; palpebral disc oval, pigmented, divided into four scales; four suboculars, two elongated and similar in size, the anteriormost and posteriormost larger than the others; three postoculars, dorsalmost wider than the others; ear opening vertically oval, without denticulate margins; tympanum recessed into a shallow audi- tory meatus; mental wider than long; postmental pentagonal, slightly wider than long, followed posteriorly by three pairs of genials, the anterior two pairs in contact medially and the third pair separated by postgenials; all genials in contact with infralabials; gulars imbricate, smooth, widened in two longitudinal rows; gular fold complete, posterior row of gulars (collar) with six scales, the medial two distinctly widened. Nuchal scales slightly smaller than dorsals, except for the anteriormost that are widened; scales on sides of neck small and granular; dorsal scales elongate, juxtaposed, arranged in transverse rows; scales on dorsal surface of neck striated, becoming slightly keeled from forelimbs to tail; dorsal scales between occipital and posterior margin of hindlimbs 35; dorsal scale rows in a transverse line at midbody 32; one longitudinal row of smooth, enlarged ventrolateral scales on each side; dorsals separated from ven- 142 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) trals by three rows of granular scales at level of 13" row of ventrals; lateral body fold between fore and hindlimbs poorly defined; ventrals smooth, arranged in 26 transverse rows between collar fold and preanals; six ventral scales in a transverse row at midbody; subcaudals smooth; axillary region with granular scales; scales on dorsal surface of forelimb smooth, imbricate; scales on ventral surface of forelimb granular; two thick, smooth thenar scales; supradigitals (left/right) 3/0 on finger I, 7/7 on II, 9/8 on IU, 10/10 on IV, 5/5 on V; supradigitals 4/4 on toe I, 7/7 on II, 11/11 on III, 12/11 on IV, 9/8 on V; subdigital lamellae of fingers I and II mostly single, IM and IV paired proxi- mally, on finger V all single; subdigital lamellae 5 on left finger I (right finger missing), 10/10 on H, 14/14 on HI, 14/14 on IV, 9/9 on V; subdigital lamellae on toe I single, on toe II paired at the middle, on toe II and IV paired along proximal half, and on toe V paired proximally; subdigital lamellae 5/5 on toe I, 10/10 on II, 14/14 on III, 18/19 on IV, 11/11 on V; groin region with small, imbricate scales; scales on dorsal surface of hindlimbs striated and imbricate; scales on ventral surface of hindlimbs smooth; scales on posterior surface of hindlimbs granular; femoral pores present, three on left leg and five on right leg; preanal pores absent; cloacal plate paired, bordered by four scales anteriorly, of which the two medialmost are enlarged. Additional measurements (mm) and proportions of the holotype: HL 9.8; HW 6.2; ShL 5.4; AGD 20.7; TL/SVL 2.4; HL/SVL 0.2; HW/SVL 0.1; ShL/SVL 0.1; AGD/SVL 0.5. Color of holotype in life. Dorsal background of head dark brown; diffuse pale brown vertebral stripe that becomes grayish brown towards tail; creamy white dor- solateral stripes on head extending posteriorly and fading away at midbody; white longitudinal stripe extending from first supralabial to shoulder; sides of neck brown; flanks grayish brown with diffuse dark brown marks; limbs brown; ventrolateral region of body grayish brown; throat and chest cream; belly grayish cream; base of tail gray with dark little spots (Figs 12, 14B). Color of holotype in preservative. Dorsal background uniformly brown with a diffuse light brown vertebral stripe extending from occiput onto tail, but fading at posterior end of body; dorsal and ventral surface of head brown; flanks light brown, with scattered dark brown spots; head and neck with two distinct white longitudinal stripes, the ventral one extending from first supralabial to forelimb, and the dorsal one from canthus rostralis to scapular region, posterior to which if fades into a light brown stripe; lateral aspect of neck dark brown; tail grayish brown; gular, chest and venter regions pale gray; ventral surface of tail and limbs gray. Variations. Measurements and scutellation data of Pholidobolus dolichoderes are presented in Table 6. Superciliaries 4/5 (left/right) in specimen QCAZ 16350; palpe- bral disc divided into 5/6 scales in QCAZ 16352 and 3/5 in QCAZ 16351; frontona- sal pentagonal in QCAZ 16349-52; prefrontals pentagonal in QCAZ 16349, 16350 and 16352; two rows of lateral granules at midbody in QCAZ 16439, 16350 and 16351. Usually six gular (collar) scales, eight in QCAZ 16349. Male is smaller (SVL 41.1 mm, N = 1) than females (maximum SVL 48.1 mm, N = 2). Systematics of Pholidobolus lizards 143 ‘= 2S SP OF Sete aVaewe |! Figure | 4. Close-up of head and neck of Pholidobolus dolichoderes sp. nov. in life. QCAZ 16349 (A adult female); QCAZ 16353 (B male holotype). Photographs by Gustavo Pazmifo. Adult females differ from holotype in having a grayish brown vertebral stripe, fading away posteriorly, and grayish brown flanks (Fig. 14). Juvenile QCAZ 16350 differs from holotype in having grayish brown flanks, without scattered dark brown spots; juvenile QCAZ 16351 is unique in having white spots on flanks and over forelimbs. Distribution and natural history. Pholidobolus dolichoderes is known to occur be- tween 2506-2675 m in San Felipe de Ofa, southwestern Azuay province (Fig. 7). This area is composed of many different landscapes including small valleys, desert areas and wet paramo. Most specimens were found active at day (10h26—15h30), mostly on the ground or near spiny ground bromeliads known as achupallas (Puya sp.). Conservation status. Pholidobolus dolichoderes is only known from unprotected localities around Ona. The population size of this species is unknown, but our sam- pling suggests low abundances. Because of the small known distribution and lack of additional data, we suggest assigning P dolichoderes to the Data Deficient category ac- cording to IUCN (2012) guidelines. Etymology. The specific epithet dolichoderes derives from the Greek words do- likhés, meaning long, and deré, meaning neck, in allusion to the distinctively long neck of this species. 144 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Pholidobolus fascinatus sp. nov. http://zoobank.org/C5EC3F40-41 DF-4A7D-A3AA-C2C8AGEF81C1 Figures 15—17 Proposed standard English name: Haunted cuilanes Proposed standard Spanish name: Cuilanes encantados Holotype. QCAZ 15120 (Figs 15, 16), adult male, Ecuador, Provincia El Oro, Lake Chillacocha, 3.4984S, 79.6188W, WGS84, 3382 m, 20 November 2016, collected by Diego Almeida, Darwin Nufez, Eloy Nusirquia, Santiago Guaman and Guadalupe Calle. Paratypes (26). Ecuapor: Provincia El Oro: QCAZ 15122 (adult male), QCAZ 15121 (adult female), QCAZ 15169-73, 15177-78, 15180, 15193, 15221, 15243-44, 15396-15405 (juveniles), same data as holotype; QCAZ 15118 (adult fe- male), Lake Chillacocha, 3.4986S, 79.6187W, WGS84, 3348 m, 17 November 2016, same collectors as holotype. Diagnosis. Pholidobolus fascinatus is unique among its congeners in lacking wid- ened medial scales on collar (posterior row of gulars). In addition, P fascinatus differs from P affinis, P prefrontalis, P macbrydei, P dolichoderes sp. nov., and P montium in having a loreal scale frequently in contact with the supralabials (loreal scale, if present, not in contact with supralabials in the other species). Pholidobolus ulisesi, P dicrus, P. hillisi, P paramuno, and P. vertebralis differ from P fascinatus in having a conspicuous light vertebral stripe. Pholidobolus samek sp. nov. and P condor sp. nov. differ from P fascinatus in having bright green dorsolateral stripes on the head. In addition, P fasci- natus has more dorsals (32-37) and ventrals (21-25) than P samek sp. nov. (27-29 and 19-21, respectively) and P condor sp. nov. (26-30 and 18-20); and it has fewer tempo- rals (3—5) and gulars (14-17) than P dolichoderes sp. nov. (7-9 and 22-23, respectively). Characterization. (1) Two (rarely three) supraoculars, anteriormost larger than posterior one; (2) prefrontals present or absent; (3) femoral pores absent in both sexes; (4) four to six opaque lower eyelid scales; (5) scales on dorsal surface of neck smooth, becoming striated from forelimbs to tail; (6) one row of lateral granules at midbody; (7) 32-37 dorsal scales between occipital and posterior margin of hindlimb; (8) lateral body fold present; (9) dorsum brown with a diffused chocolate brown middorsal stripe that fades away towards tail; (11) labial stripe white or cream; (12) flanks of body brown; (13) conical hemipenial body, with sulcus spermaticus originating between distinctly thick lips; (14) 22 flounces extending along hemipenial body. Description of holotype. Adult male (QCAZ 15120) (Figs 15, 16); SVL 52.5 mm; TL 37.6 mm; dorsal and lateral head scales juxtaposed, finely wrinkled; rostral hex- agonal, 2.27 times as wide as high; frontonasal hexagonal, wider than long, in contact with nasal laterally, slightly larger than frontal; prefrontal scales irregularly pentagonal; frontal heptagonal, longer than wide, slightly wider anteriorly, in contact with pre- frontals and frontonasal anteriorly, two supraoculars laterally, and frontoparietals pos- teriorly; frontoparietals pentagonal, longer than wide, slightly wider posteriorly, each in contact laterally with supraocular H; interparietal heptagonal, lateral borders nearly Systematics of Pholidobolus lizards 145 parallel to each other; parietals hexagonal, each in contact laterally with supraocular II and dorsalmost postocular; postparietals four, with medial scales less than half the size of lateral ones; eight supralabials, fourth one the longest and below center of eye; eight infralabials, third and fourth one below center of eye; temporals enlarged, irregularly hexagonal, juxtaposed, smooth; two large, smooth supratemporal scales; nasal divided, irregularly pentagonal, longer than high, in contact with rostral anteriorly, first and second supralabials ventrally, frontonasal dorsally, loreal posterodorsally and frenocular posteroventrally; nostril in center of nasal, directed lateroposteriorly; loreal rectangular, wider ventrally; frenocular longer than high, higher anteriorly, in contact with nasal, separating loreal from supralabials; two supraoculars, homogeneous in size; four super- ciliaries, anteriormost enlarged and in contact with loreal; palpebral disc divided into five pigmented scales; suboculars elongated, four on right side and three on left side; two postoculars, dorsalmost wider than the other; ear opening vertically oval, with- out denticulate margins; tympanum recessed into a shallow auditory meatus; mental semicircular, longer than wide; postmental pentagonal, slightly longer than wide, fol- lowed posteriorly by three pairs of genials, the anterior two in contact medially and the posterior one separated by postgenials; all genials in contact with infralabials; gulars imbricate, smooth, widened in two longitudinal rows; posterior row of gulars (collar) with 11 scales that are similar in size. Nuchal scales similar in size to dorsals, except for the anteriormost that are wid- ened; scales on sides of neck small and slightly granular; dorsal scales hexagonal, elon- gate, imbricate, arranged in transverse rows; scales on dorsal surface of neck smooth, becoming progressively striated from forelimbs to tail; dorsal scales between occipital and posterior margin of hindlimbs 33; dorsal scale rows in a transverse line at midbody 25; dorsals separated from ventrals by one row of small scales at level of 13° row of ventrals; lateral body fold between fore and hindlimbs present; ventrals smooth, wider than long, arranged in 25 transverse rows between collar fold and preanals; six ventral scales in a transverse row at midbody; subcaudals smooth; axillary region with granular scales; scales on dorsal surface of forelimb striated, imbricate; scales on ventral surface of forelimb granular; two thick, smooth thenar scales; supradigitals (left/right) 3/3 on finger I, 7/6 on IL, 8/8 on III, 10/10 on IV, 5/5 on V; supradigitals 3/3 on toe I, 6/6 on II, 8/9 on III, 11/11 on IV, 8/8 on V; subdigital lamellae of finger I single, on finger II all paired, except by the three distalmost, on finger II (proximal half) paired, on finger IV slightly paired at the middle, on finger V all single in right finger and three paired in left finger; subdigital lamellae 5/5 on finger I, 9/9 on II, 13/13 on III, 14/15 on IV, 9/9 on V; subdigital lamellae on toes I and II paired proximally and single distally, on toes III, IV and V paired, except for the three to five distalmost subdigitals; subdigi- tal lamellae 5/5 on toe I, 10/10 on II, 14/13 on II, 18/18 on IV, 11/12 on V; groin region with small, imbricate scales; scales on dorsal surface of hindlimbs smooth and imbricate; scales on ventral surface of hindlimbs smooth; scales on posterior surface of hindlimbs granular; femoral pores absent; preanal pores absent; cloacal plate paired, bordered anteriorly by two enlarged scales. 146 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Figure 15. Holotype of Pholidobolus fascinatus sp. nov. (QCAZ 15120) in life in dorsal (A), ventral (B), and lateral (C) views. Male, SVL = 52.5 mm. Photographs by Diego Quirola. Additional measurements (mm) and proportions of the holotype: HL 12.3; HW 9.2; ShL 6.7; AGD 26.5; TL/SVL 0.7; HL/SVL 0.2; HW/SVL 0.2; ShL/SVL 0.1; AGD/SVL 0.5. Color in life of the holotype. Dorsal background from head to base of tail brown, with a diffuse chocolate-brown middorsal stripe that fades away towards tail; light brown dorsolateral stripes on head extending posteriorly and fading away at midbody; white longitudinal stripe extending from third supralabial to shoulder; sides of neck, flanks, and limbs brown; reddish brown narrow stripe extending from tympanum to arm inser- tion; ventrolateral region of body grayish brown; throat and chest gray; belly background gray with conspicuous orange marks; tail orange anteriorly and laterally (Figs 15, 17A). Systematics of Pholidobolus lizards 147 Figure | 6. Head of holotype of Pholidobolus fascinatus sp. nov. (QCAZ 15120) in lateral (A), dorsal (B), and ventral (C) views. Photographs by Valeria Chasiluisa. Scale bar: 5 mm. 148 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Color in preservative of the holotype. Dorsal background uniformly brown with a cream brown vertebral stripe extending from head onto tail; vertebral stripe slender anteriorly, becoming slightly wider posteriorly; head light brown with black dots dor- sally (rostral, frontonasal, frontal, frontoparietals and supraoculars) and brown later- ally; cream longitudinal stripe extending from third supralabial to shoulder; ventrolat- eral aspect of neck brown; forelimbs with scattered black dots; flanks brown; tail brown dorsally; ventral surface of head light gray, chest and venter dark gray, ventral surface of tail slightly brown, with scattered dark brown marks. Variations. Measurements and scale counts of Pholidobolus fascinatus are presented in Table 6. Supralabials 9/9 (left/right) in specimens QCAZ 15118 and 15122, and supraoculars 3/3 in QCAZ 15118; loreal scale absent in QCAZ 15118; prefrontals ab- sent in QCAZ 15122 and 15173; little intrusive scales between postparietal and fron- toparietals in QCAZ 15118, 15121 and 15122; frontonasal quadrangular in QCAZ 15122; frontal nonagonal and pentagonal in QCAZ 15118 and 15173, respectively; interparietal hexagonal in QCAZ 15122; parietal pentagonal in QCAZ 15170. Four posterior cloacal scales in QCAZ 15118. Males are slightly smaller (SVL 47.6 mm, NV = 2) than female (maximum SVL 48.2 mm, N = 2). Adult male QCAZ 15122 differs from holotype in having sides of tail and chest dark brown without gray spots. Adult female QCAZ 15118 differs from holotype in having a light gray chest, a dark gray ventral surface of tail, dark brown sides of tail, and in lacking orange or red brown color on sides of neck (Fig. 17). Distribution and natural history. Pholidobolus fascinatus inhabits wet paramo in the western slopes of the Andes of southern Ecuador (Fig. 7). The new species is known only from El Oro province, at 3348-3382 m. All specimens were found active at 14h00—17h00 mostly under stones. We found 41 eggs (17 as fragmented eggshells) in a communal nest next to male QCAZ 15120. We incubated the 24 unhatched eggs in soil and perlite in captivity. They were 11.9-13.2 mm long, 5.5—9.2 mm wide, and weighted 0.5 g on average. Hatchlings (V = 20) weighted 0.4 g and were 26.2 mm in SVL on average. Conservation status. P/holidobolus fascinatus is only known from localities around Lake Chillacocha. The population size for this species is unknown, but our sampling suggests average abundances. Because of the small known distribution and lack of ad- ditional data, we suggest assigning P fascinatus to the Data Deficient category, accord- ing to IUCN (2012) guidelines. Etymology. The species epithet fascinatus is a Latin word meaning enchanted, in allusion to Lake Chillacocha, also known as the Enchanted Lake. According to local belief, this lake is enchanted and has healing powers. Discussion The systematics of Pholidobolus and its sister taxon Macropholidus have been contro- versial partly because morphological evidence has been misinterpreted. Nonetheless, the recent use of molecular phylogenies has reshaped the systematics and taxonomy Systematics of Pholidobolus lizards 149 Figure 17. Color variation in live specimens of Pholidobolus fascinatus sp. nov. A male holotype (QCAZ 15120, SVL = 52.5 mm) B male paratype (QCAZ 15122, SVL = 42.6 mm) C female paratype (QCAZ 15118, SVL = 46.7 mm). of this clade (Torres-Carvajal and Mafla-Endara 2013; Torres-Carvajal et al. 2015). In addition, recent collections in poorly explored areas along the Andes of Colombia, Ecuador, and Peru have led to the discovery and description of new species (Hurtado- Goémez et al. 2018; Torres-Carvajal et al. 2014; Venegas et al. 2016). In this paper we use morphological and molecular evidence to describe four new species of Pholidobo- lus, all except P dolichoderes sp. nov. from remote highlands, based mostly on recent collections in southern Ecuador. Unexpectedly, allocating the new species described herein within the phylogenetic tree of Pholidobolus rendered P macbrydei paraphyletic, suggesting that populations currently assigned to this taxon represent multiple spe- cies, some of which (e.g., Clades A and F) match the evolutionary significant units identified by Mafla-Endara (2011). Nonetheless, we refrain from describing any of these putative species (Clades A—F) in this paper as we believe that further sampling and analysis are necessary. According to our PCA results, three of the four new species are morphologically different from other “Pholidobolus macbrydei” (Fig. 2). Compo- nents I and II in the PCA, however, explain less than 50% of the variation within the “P macbrydei” clade (Table 5). Thus, it is necessary to study additional morphologi- cal characters and increase sample size to better elucidate morphological differences. 150 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Mafla-Endara (2011) also suggested hybridization between P macbrydei from Caniar province and P prefrontalis based on both the relatively great variation in morphology within the Cafar populations, and their morphological similarity to P prefrontalis. Nevertheless, our nuclear phylogenetic tree does not suggest hybridization between P macbrydei and P prefrontalis (Appendix I). Current evidence prevents us from assigning the name P macbrydei to any of the recovered clades. However, we suspect that P macbrydei belongs or is more closely related to Clades C, D, and E for two reasons (Fig. 1). First, adult males in these clades match closely the description of P macbrydei (Montanucci 1973). Second, Clades C, D, and E lie nearby the type locality of P macbrydei. It is noteworthy that Clade B also lies near the type locality of P macbrydei (Fig. 7), although males in Clade B lack the red lateral stripes characteristic of P macbrydei. DNA samples from the type locality should help clarify the taxonomy of this group. The Cordillera del Condor is a sub-Andean mountain chain geologically similar to the Tepuis of the Guiana region. It is composed of marine and continental sediments (Neill 2005). This area is presently threatened by mining activities, despite discovery of a significant number of new species in the last ten years (Brito et al. 2017; Huamantupa- Chuquimaco and Neill 2018; Mashburn et al. 2020; Ron et al. 2018; Torres-Carvajal et al. 2009; Valencia et al. 2017) suggesting that Cordillera del Condor is a diversity hotspot. Our discovery of P samek and P condor further supports this idea. ‘Therefore, we strongly advise authorities to improve conservation efforts for Cordillera del Condor. The discovery of four new species and a paraphyletic P macbrydei reveals high levels of unexpected diversity within Pholidobolus from southern Ecuador. This study supports the idea that Andean herpetofauna in this region is more diverse in species numbers than previously thought (Sanchez-Pacheco et al. 2012), especially for poorly explored areas like Cordillera del Condor. Collections in this area are usually scarce due to complex logistics. However, we recommend more intensive sampling efforts. Future studies should include larger samples and other types of evidence (e.g., genomic data, environmental variables) that might prove useful for species delimitation within Pholidobolus and other vertebrate taxa. Acknowledgements This research was funded by the Secretaria de Educacion Superior, Ciencia, Tecnologia e Innovacién del Ecuador SENESCYT (‘Iniciativa Arca de Noé, PIs Omar Torres- Carvajal and Santiago Ron), and the Pontificia Universidad Catdlica del Ecuador. P.M.S.N. is grateful to the Conselho Nacional de Desenvolvimento Cientifico e Tec- noldgico (CNPq) for financial support (Fellowship #313622/2018-3). We thank the QCAZ field and laboratory teams. Fernando Ayala, Marcel Caminer, Maria José Nav- arrete, Emilio Oviedo, Valeria Chasiluisa, Santiago Ron, and Andrés Merino provided valuable advice and help during the research. 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ECUADOR: Provincia Azuay: Cuenca-Azogues, 2.895222S, 78.95822W, 2486 m, QCAZ 6985; Cuenca- Chaucha, 2.861209S, 79.37869W, 2943 m, QCAZ 9668; Cuenca-Cochapamba, 2.797120S, 79.41562W, 3548 m, QCAZ 10133-10135; Cuenca-El Cajas, 2.777448, 79.17001W, 3508 m, QCAZ 9932, 9936; 2.74105S, 79.23479W, 4092 m, QCAZ 8010; 2.776299S, 79.23743W, 4068 m, QCAZ 8011; 3.04155S, 79.21567W, 3766 m, QCAZ 8897, 8899-8903, 8906; Cuenca-Mazan Forest, 2.875225, 79.12923W, 3189 m, QCAZ 8008, 8013; Cutchil, 3.133999S, 78.81300W, 2900 m, QCAZ 823; Guablid, 2.77488S, 78.69758W, 2453 m, QCAZ 9915, 9919-9920; Gualaceo, 2.909767S, 78.73436W, 2625 m, QCAZ 10875; Gualaceo-Limén; 2.948S, 78.71200W, 3110 m, QCAZ 819-20, 822; 2.964S, 78.70199W, 3140 m, QCAZ 825; Patacocha hill, 3.121109S, 79.065W, 3340 m, QCAZ 6144; Pucara, 3.213678, 79.46739W, QCAZ 11038; Quinoas river, 3.087267S, 79.27762W, 3200 m, QCAZ 1564, 1566; Sigsig, 2.99900S, 78.80700W, 2890 m, QCAZ 1537; 3.129500S, 78.80400W, 2969 m, QCAZ 5605-5606, 5608; Sigsig-Gualaqui- za, 3.106875S, 78.79558W, 2935 m, QCAZ 8646-8647; Tarqui, 3.015880S, 79.04447W/, 2680 m, QCAZ 8512. Provincia Canar: Cajas National Park, 2.70654S, 79.22765W, 3651 m, QCAZ 8946; Cahar, 2.560760S, 78.93077W, QCAZ 9947; Guallicanga river, 2.473189S, 78.97289W, 3048 m, QCAZ 10051-53; Gualaceo, 2.882159S, 78.77536W, 2298 m, QCAZ 9606; Juncal, 2.432109S, 78.90223W, Systematics of Pholidobolus lizards 155 3960 m, QCAZ 10048; 2.473189S, 78.97289W, 3048 m, QCAZ 10050; Mazar, 2.54508S, 78.70078W, 2839 m, QCAZ 15811—13; 2.54649S, 78.69826W, 2924 m, QCAZ 15814-16; 2.57138S, 78.746W, 3442 m, QCAZ 15817-15823; 2.57088, 78.74586W, 3451 m, QCAZ 15824; 2.545804S, 78.69611W, 2842 m, QCAZ 10970. Provincia Chimborazo: Frutatian lake, 2.215848, 78.50136W, 3700 m, QCAZ 9217-9218; Magdalena lake, 2.187416S, 78.50686W, 3556 m, QCAZ 9214; Ozogoche, 2.368733S, 78.68871W, 4040 m, QCAZ 6006; Riobamba-Melan, 1.875020S, 78.54773W, 3564 m, QCAZ 9626-9628; Riobamba-Timbo, 1.929219S, 78.53718W, 3408 m, QCAZ 9616-9620; Shulata, 2.339309S, 78.84322W, 3228 m, QCAZ 5597-5598. Provincia El Oro: Guanazan, 3.440034S, 79.48695W, 2638 m, QCAZ 7894. Provincia Loja: Fierro Hurco, 3.710421S, 79.30498W, 3439 m, QCAZ 6949-6950; Jimbura-Jimbura lake, 4.708868S, 79.44657W, 3036 m, QCAZ 6947-6948; Jimbura- path to Jimbura lake, 4.709469S, 79.43558W, 3348 m, QCAZ 10054-10055, 10057-10062; Jimbura-Lagunillos, 4.628244S, 79.46353W, 3450 m, QCAZ 6146-6147; 4.817000S, 79.36199W, 3600 m, QCAZ 3785; San Lucas, 3.731853S, 79.26059W, 2470 m, QCAZ 2861; Saraguro, 3.62025S, 79.23581W, 3100 m, QCAZ 3606; 3.679457S, 79.23769W, 3190 m, QCAZ 3673-3674; Tar- qui, QCAZ 5545. Provincia Morona Santiago: Sangay National Park, 1.960939S, 78.43198W, 3345 m, QCAZ 9612. Provincia Tungurahua: Patate-El Corral, 1.2725S, 78.46805W, 3468 m, QCAZ 9995-9996. Provincia Zamora Chinchipe: Podocarpus National Park, 4.484149S, 79.14875W, 1800 m, QCAZ 3743. 156 Vanessa Parra et al. / ZooKeys 954: 109-156 (2020) Appendix Il P. vertebralis QOAZ10750 P. vertebralis QCAZ8724 P. vertebralis QCAZ8717 P. vertebralis QCAZ10667 P. vertebralis QCAZ8689 P. vertebralis QCAZ8688 P. vertebralis QCAZ8687 P. vertebralis QCAZ5057 63 P. montium QCAZ4051 0.99 P. montium QCAZ9044 83 P. dierus QCAZ5304 o.o7 + P. dicrus QCAZ6936 38 P. affinis QCAZ9641 58 P. macbrydei QCAZ10051 aa 28 0.64 P. macbrydei QCAZ10052 0.52 * P. macbrydei QCAZ9947 0.95 P. prefrontalis QCAZ9951 y P. condor sp. n. QCAZ15844 62 P. hillisi QCAZ4998 P. hillisi QCAZ4999 51 * P. hillisi QC AZ5000 P, samek sp. n. QCAZ14954 0.51 0.66 P. samek sp. n. QCAZ14955 51 P. samek sp. n. QCAZ14956 P. macbrydei QCAZ6945 0.60 P. macbrydei QCAZ6946 44 P. macbrydei QCAZ10054 P. macbrydei QCAZ9914 0.93 P. condor sp. n. QCAZ16788 P, condor sp. n. QCAZ16789 P. condor sp. n. QCAZ16790 P. macbrydei QCAZ9932 54 P. fascinatus sp. n. QCAZ15118 P. fascinatus sp. n. QGAZ15120 0.84 P, fascinatus sp. n. QGAZ15122 P. fascinatus sp. n. QCAZ15170 31 P. dolichoderes sp. n. QCAZ16349 P. dolichoderes sp. n. QCAZ16350 0.58 53 —— P. dolichoderes sp. n. QGAZ16351 | 9.99 P. dolichoderes sp. n. QCAZ16352 P. dolichoderes sp. n. QCAZ16353 68 — P. macbrydei QCAZ7891 0.97 P. macbrydei QCAZ7894 — P maebrydei QCAZ10050 Juncal 61 P. macbrydei QCAZ15819 Mazar | 0.88 P. macbrydei QCAZ15820 Mazar 0.68 P, macbrydei QCAZ15823 Mazar P,. maebrydei QCAZ15811 Mazar P. macbrydei QCAZ15812 Mazar P. maebrydei QCAZ15813 * } P. macbrydei QCAZ15816 Mazar P. macbrydei QCAZ15817 P,. macbrydei QCAZ15818 P, macbrydei QCAZ15824 M. ruthveni CORBIDI4281 M. annectens QGAZ11120 M. annectens QCAZ11121 0.82 M. labiopunctatus CORBIDI12932 k P. ulisesi CORBIDI12735 * P. ulisesi CORBIDI1679 74 M. huancabambae CORBIDI10492 0.85 { M. huancabambae CORBIDI10496 0.0020 Phylogeny of Pholidobolus. ML phylogram derived from the analysis of 411 bp of nuclear DNA. Bootstrap values are shown above branches and Bayesian posterior probabilities below branches (< 50% not shown). Asterisks indicate maximum values. The outgroup taxon (Anadia rhombifera) is not shown. Species names followed by voucher numbers are shown.