Print
Systematics of Pholidobolus lizards (Squamata, Gymnophthalmidae) from southern Ecuador, with descriptions of four new species
expand article infoVanessa Parra, Pedro M. Sales Nunes§, Omar Torres-Carvajal
‡ Pontificia Universidad Católica del Ecuador, Quito, Ecuador
§ Universidade Federal de Pernambuco, Recife, Brazil
Open Access

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, Pholidobolus 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 km2 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 Cóndor, diversity, phylogeny, taxonomy

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 influence 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-Gómez 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 hampered 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 ecosystem in the northern Andes of Colombia. Similarly, recent collections in poorly explored 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 samples were quantified using a Nanodrop ND-1000 (NanoDrop Technologies, Inc.), re-suspended and diluted to 25 ng/µl 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 Pholidobolus macbrydei. In addition, we obtained 411 nucleotides of the Dynein Axonemal 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_f1 (GGTAAAATGATAGAAGAYTACTG) and DNAH3_r6 (CTKGAGTTRGAHACAATKATGCCAT). 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 sequences, our data matrix for phylogenetic analyses contained 74 taxa and 1904 characters.

Table 1.

Vouchers, locality data, and GenBank accession numbers of taxa included in this study. Sequences added in this study are in bold.

Taxon Voucher Locality GenBank number GenSeq Nomenclature
12S 16S ND4 DNAH3
Anadia rhombifera QCAZ 11862 QCAZ 11862; Ecuador: Cotopaxi: San Francisco de Las Pampas KU902135 KU902216 KU902291 MN849427 genseq-4
Macropholidus annectens QCAZ 11120 Ecuador: Loja: 15 km E Loja KC894341 KC894355 KC894369 MN849430 genseq-4
QCAZ 11121 Ecuador: Loja: 15 km E Loja KC894342 KC894356 KC894370 MN849431 genseq-4
Macropholidus huancabambae CORBIDI 10492 Peru: Piura: Huancabamba: Las Pozas KC894343 KC894357 KC894371 MN849428 genseq-4
CORBIDI 10493 Peru: Piura: Huancabamba: Las Pozas KC894344 KC894358 KC894372 genseq-4
CORBIDI 10496 Peru: Piura: Huancabamba: Las Pozas KC894345 KC894359 KC894373 MN849429 genseq-4
Macropholidus labiopunctatus CORBIDI 12932 Peru: Piura: Ayabaca KP874774 KP874826 KP874936 MN849432 genseq-4
Macropholidus ruthveni CORBIDI 4281 Peru: Lambayeque: El Totora KC894354 C894368 C894382 MN849433 genseq-4
Pholidobolus affinis QCAZ 9641 Ecuador: Cotopaxi: San Miguel de Salcedo, Cutuchi River KC894348 C894362 C894376 MN849435 genseq-4
QCAZ 9900 Ecuador: Chimborazo: Colta KC894349 KC894363 KC894377 genseq-4
Pholidobolus condor sp. nov. QCAZ 16788 Ecuador: Morona-Santiago: el Quimi MN724005 MN720239 MN717135 MN849464 genseq-2
QCAZ 16789 Ecuador: Morona-Santiago: el Quimi MN724006 MN720240 MN717134 MN849465 genseq-2
QCAZ 16790 Ecuador: Morona-Santiago: el Quimi MN724007 MN720241 MN717136 MN849466 genseq-2
QCAZ 15844 Ecuador: Morona-Santiago: el Quimi MN723996 MN720230 MN717125 MN849434 genseq-1
Pholidobolus dicrus QCAZ 5304 Ecuador: Morona-Santiago: Guarumales KP874776 KP874828 KP874938 MN849436 genseq-4
QCAZ 6936 Ecuador: Tungurahua: Río Blanco KP874829 KP874939 MN849437 genseq-4
Pholidobolus dolichoderes sp. nov. QCAZ 16349 Ecuador: Cañar: Oña MN724000 MN720234 MN717129 MN849459 genseq-2
QCAZ 16350 Ecuador: Cañar: Oña MN724001 MN720235 MN717130 MN849460 genseq-2
QCAZ 16351 Ecuador: Cañar: Oña MN724002 MN720236 MN717131 MN849461 genseq-2
QCAZ 16352 Ecuador: Cañar: Oña MN724003 MN720237 MN717132 MN849462 genseq-2
QCAZ 16353 Ecuador: Cañar: Oña MN724004 MN720238 MN717133 MN849463 genseq-1
Pholidobolus fascinatus sp. nov. QCAZ 15118 Ecuador: El Oro: Chillacocha MN724017 MN720251 MN717146 MN849476 genseq-2
QCAZ 15120 Ecuador: El Oro: Chillacocha MN724018 MN720252 MN717147 MN849477 genseq-1
QCAZ 15122 Ecuador: El Oro: Chillacocha MN724019 MN720253 MN849478 genseq-2
QCAZ 15170 Ecuador: El Oro: Chillacocha MN724020 MN720254 MN717148 MN849479 genseq-2
Pholidobolus hillisi QCAZ 4998 Ecuador: Zamora-Chinchipe: near San Francisco Research Station KP090167 KP090170 KP090173 MN849438 genseq-4
QCAZ 4999 Ecuador: Zamora-Chinchipe: near San Francisco Research Station KP090169 KP090172 KP090175 MN849439 genseq-4
QCAZ 5000 Ecuador: Zamora-Chinchipe: near San Francisco Research Station KP090168 KP090171 KP090174 MN849440 genseq-4
Pholidobolus macbrydei KU 218406 Ecuador: Azuay: Cuenca AY507848 AY507867 AY507886 genseq-4
QCAZ 9914 Ecuador: Azuay: Guablid KC894352 KC894366 KC894380 MN849441 genseq-4
QCAZ 9932 Ecuador: Azuay: 20 km on road Cuenca-El Cajas KC894353 KC894367 KC894381 MN849442 genseq-4
QCAZ 9947 Ecuadro: Cañar: Cañar MN724012 MN720246 MN717141 MN849474 genseq-4
QCAZ 10051 Ecuador: Cañar: Río Guallicanga, quebrada Juncal MN724014 MN720248 MN717143 MN849472 genseq-4
QCAZ 10052 Ecuador: Cañar: Río Guallicanga, quebrada Juncal MN724015 MN720249 MN717144 MN849473 genseq-4
QCAZ 10050 Ecuador: Cañar: A 1000 m de la Panamericana Juncal MN724013 MN720247 MN717142 MN849471 genseq-4
QCAZ 15811 Ecuador: Cañar: Mazar MN724021 MN720255 MN717149 MN849480 genseq-4
QCAZ 15812 Ecuador: Cañar: Mazar MN724022 MN720256 MN717150 MN849481 genseq-4
QCAZ 15813 Ecuador: Cañar: Mazar MN724023 MN720257 MN717151 MN849482 genseq-4
QCAZ 15814 Ecuador: Cañar: Mazar MN724024 MN720258 MN717152 genseq-4
QCAZ 15815 Ecuador: Cañar: Mazar MN724025 MN520259 MN717153 genseq-4
QCAZ 15816 Ecuador: Cañar: Mazar MN724026 MN720260 MN717154 MN849483 genseq-4
QCAZ 15817 Ecuador: Cañar: Mazar MN724027 MN720261 MN717155 MN849484 genseq-4
QCAZ 15818 Ecuador: Cañar: Mazar MN724028 MN720262 MN717156 MN849485 genseq-4
QCAZ 15819 Ecuador: Cañar: Mazar MN724029 MN720263 MN717157 MN849486 genseq-4
QCAZ 15820 Ecuador: Cañar: Mazar MN724030 MN720264 MN717158 MN849487 genseq-4
QCAZ 15823 Ecuador: Cañar: Mazar MN724031 MN720265 MN717159 MN849488 genseq-4
QCAZ 15824 Ecuador: Cañar: Mazar MN724032 MN720266 MN717160 MN849489 genseq-4
QCAZ 6945 Ecuador: Loja: Jimbura MN724008 MN720242 MN717137 MN849467 genseq-4
QCAZ 6946 Ecuador: Loja: Jimbura MN724009 MN720243 MN717138 MN849468 genseq-4
Pholidobolus macbrydei QCAZ 10054 Ecuadro: Loja: Colambo Yacuri Forest MN724016 MN720250 MN717145 MN849475 genseq-4
QCAZ 7894 Ecuador: El Oro: Guanazán MN724011 MN720245 MN717140 MN849470 genseq-4
QCAZ 7891 Ecuador: El Oro: Guanazán MN724010 MN720244 MN717139 MN849469 genseq-4
Pholidobolus montium QCAZ 4051 Ecuador: Pichincha: Quito KC894346 KC894360 KC894374 MN849443 genseq-4
QCAZ 9044 Ecuador: Pichincha: Tababela KC894347 KC894361 KC894375 MN849444 genseq-4
Pholidobolus paramuno MHUAR 12451 Colombia: Antoquia MK215018 MK215032 MK215046 genseq-4
MHUAR 12480 Colombia: Antoquia MK215019 MK215033 MK215047 genseq-4
MHUAR 12481 Colombia: Antoquia MK215020 MK215034 MK215048 genseq-4
Pholidobolus prefrontalis QCAZ 9908 Ecuador: Chimborazo: Alausí KC894350 KC894364 KC894378 genseq-4
QCAZ 9951 Ecuador: Chimborazo: Tixán KC894351 KC894365 KC894379 MN849448 genseq-4
Pholidobolus samek sp. nov. QCAZ 14954 Ecuador: Zamora Chinchipe: Cerro Plateado MN723997 MN720231 MN717126 MN849445 genseq-2
QCAZ 14955 Ecuador: Zamora Chinchipe: Cerro Plateado MN723998 MN720332 MN717127 MN849446 genseq-1
QCAZ 14956 Ecuador: Zamora Chinchipe: Cerro Plateado MN723999 MN720233 MN717128 MN849447 genseq-2
Pholidobolus ulisesi CORBIDI 12735 Peru: Cajamarca: Jaen: Huamantanga Forest KP874787 KP874839 KP874948 MN849449 genseq-4
CORBIDI 12737 Peru: Cajamarca: Jaen: Huamantanga Forest KP874788 KP874840 KP874949 genseq-4
CORBIDI 1679 Perú: Chota: La Granja KP874786 KP874838 KP874947 MN849450 genseq-4
Pholidobolus vertebralis QCAZ 10667 Ecuador: Pichincha: Santa Lucía de Nanegal KP874784 KP874836 KP874946 MN849455 genseq-4
QCAZ 10750 Ecuador: Pichincha: Santa Lucía de Nanegal KP874785 KP874837 KP874947 MN849458 genseq-4
QCAZ 5057 Ecuador: Carchi: Chilma Bajo KP874778 KP874830 KP874940 MN849451 genseq-4
QCAZ 8687 Ecuador: Carchi: Chilma Bajo KP874779 KP874831 KP874941 MN849452 genseq-4
QCAZ 8688 Ecuador: Carchi: Chilma Bajo KP874780 KP874832 KP874942 MN849453 genseq-4
QCAZ 8689 Ecuador: Carchi: Chilma Bajo KP874781 KP874833 KP874943 MN849454 genseq-4
QCAZ 8717 Ecuador: Carchi: next to Chilma Bajo KP874782 KP874834 KP874944 MN849456 genseq-4
QCAZ 8724 Ecuador: Carchi: next to Chilma Bajo KP874783 KP874835 KP874945 MN849457 genseq-4

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 DNAH3 sequences were translated into amino acids for confirmation of alignment. The best-fit nucleotide substitution models and partitioning scheme were chosen simultaneously 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) 1st codon position of ND4 and 12S [GTR + I + G]; (ii) 2nd codon position of ND4, 1st codon and 2nd codon positions of DNAH3 [HKY + I + G]; (iii) 3rd codon position of ND4 [GTR + G]; (iv) 16S and 3rd codon position of DNAH3 [SYM + I + 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 algorithm 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 calculate posterior probabilities (PP) for each bipartition in a Maximum Clade Credibility Tree. The phylogenetic trees were visualized and edited using FigTree 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).

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 Zoología, Pontificia Universidad Católica del Ecuador, Quito (QCAZ). The following measurements were taken with a digital caliper (to the nearest 0.1 mm):

AGD axilla-groin distance;

HL head length;

HW head width;

ShL shank length;

SVL and snout-vent length.

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 Montanucci (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 Analysis (PCA) in R (R Core Team 2018). The PCA was based on 16 quantitative morphological 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-Gómez et al. 2018; Nunes et al. 2012; Sánchez-Pacheco et al. 2017; Savage 1997; Venegas et al. 2016).

Systematics

The taxonomic conclusions of this study are based on the observation of morphological features and color pattern, as well as inferred phylogenetic relationships. We 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. ulisesi and P. hillisi form a clade (BS = 62, PP = 0.92) sister to all other congeners (Torres-Carvajal et al. 2015, 2016; Hurtado-Gómez 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.

Figure 1. 

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.

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. macbrydei” Clade B from Cañar 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 Cañar provinces) in a strongly supported subclade (BS = 71, PP = 1) sister to the maximally supported (P. dolichoderes, “P. macbrydei” Clade F [El Oro province]). 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).

Table 2.

Pairwise genetic distances (uncorrected p) of 16S DNA sequences among species and clades of Pholidobolus included in this study. This analysis involved 66 nucleotide sequences and 533 positions.

Taxon 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
1 Pholidobolus condor sp. nov.
2 Pholidobolus samek sp. nov. 0.03
3 Pholidobolus dolichoderes sp. nov. 0.04 0.03
4 Pholidobolus fascinatus sp. nov. 0.03 0.03 0.03
5 Clade A 0.03 0.02 0.04 0.03
6 Clade B 0.03 0.03 0.03 0.03 0.02
7 Clade C 0.02 0.02 0.03 0.02 0.03 0.03
8 Clade D 0.03 0.02 0.03 0.03 0.03 0.03 0.02
9 Clade E 0.03 0.03 0.04 0.02 0.04 0.03 0.02 0.03
10 Clade F 0.04 0.03 0.01 0.04 0.04 0.02 0.03 0.03 0.04
11 Pholidobolus affinis 0.04 0.03 0.03 0.03 0.04 0.03 0.03 0.02 0.03 0.03
12 Pholidobolus dicrus 0.04 0.04 0.04 0.04 0.04 0.04 0.04 0.03 0.05 0.04 0.05
13 Pholidobolus hillisi 0.05 0.05 0.05 0.05 0.04 0.04 0.04 0.03 0.05 0.04 0.04 0.05
14 Pholidobolus montium 0.03 0.02 0.03 0.03 0.02 0.02 0.02 0.02 0.04 0.03 0.03 0.04 0.04
15 Pholidobolus paramuno 0.04 0.04 0.04 0.03 0.04 0.03 0.03 0.03 0.04 0.04 0.03 0.04 0.04 0.03
17 Pholidobolus prefrontalis 0.03 0.03 0.03 0.03 0.03 0.02 0.03 0.03 0.03 0.03 0.03 0.04 0.04 0.03 0.03
17 Pholidobolus ulisesi 0.05 0.05 0.05 0.05 0.05 0.05 0.06 0.06 0.05 0.05 0.05 0.06 0.04 0.05 0.05 0.05
18 Pholidobolus vertebralis 0.05 0.04 0.04 0.04 0.05 0.04 0.04 0.04 0.05 0.04 0.05 0.05 0.05 0.04 0.04 0.04 0.06
Table 3.

Pairwise genetic distances (uncorrected p) of 12S DNA sequences among species and clades of Pholidobolus included in this study. This analysis involved 65 nucleotide sequences and 339 positions.

Taxon 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
1 Pholidobolus condor sp. nov.
2 Pholidobolus samek sp. nov. 0.01
3 Pholidobolus dolichoderes sp. nov. 0.04 0.04
4 Pholidobolus fascinatus sp. nov. 0.03 0.03 0.03
5 Clade A 0.03 0.03 0.05 0.04
6 Clade B 0.03 0.03 0.04 0.03 0.03
7 Clade C 0.02 0.02 0.02 0.02 0.03 0.03
8 Clade D 0.02 0.02 0.03 0.02 0.04 0.03 0.01
9 Clade E 0.02 0.02 0.02 0.02 0.03 0.03 0.01 0.01
10 Clade F 0.04 0.04 0.01 0.04 0.05 0.04 0.03 0.03 0.03
11 Pholidobolus affinis 0.05 0.04 0.06 0.05 0.05 0.05 0.05 0.05 0.05 0.06
12 Pholidobolus dicrus 0.07 0.07 0.08 0.07 0.07 0.07 0.07 0.08 0.07 0.08 0.08
13 Pholidobolus hillisi 0.05 0.04 0.05 0.05 0.06 0.05 0.04 0.04 0.05 0.05 0.06 0.08
14 Pholidobolus montium 0.03 0.04 0.05 0.05 0.06 0.04 0.04 0.04 0.04 0.05 0.02 0.07 0.05
15 Pholidobolus paramuno 0.06 0.06 0.07 0.03 0.07 0.07 0.06 0.06 0.06 0.07 0.07 0.10 0.07 0.06
16 Pholidobolus prefrontalis 0.02 0.02 0.04 0.04 0.03 0.03 0.02 0.03 0.03 0.05 0.03 0.05 0.04 0.02 0.06
17 Pholidobolus ulisesi 0.04 0.04 0.04 0.04 0.04 0.04 0.03 0.04 0.03 0.04 0.04 0.07 0.04 0.04 0.06 0.03
18 Pholidobolus vertebralis 0.08 0.08 0.09 0.08 0.08 0.07 0.08 0.08 0.08 0.09 0.08 0.06 0.10 0.08 0.10 0.06 0.08
Table 4.

Pairwise genetic distances (uncorrected p) of ND4 DNA sequences among species and clades of Pholidobolus included in this study. This analysis involved 64 nucleotide sequences and 621 positions.

Taxon 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
1 Pholidobolus condor sp. nov.
2 Pholidobolus samek sp. nov. 0.08
3 Pholidobolus dolichoderes sp. nov. 0.13 0.12
4 Pholidobolus fascinatus sp. nov. 0.09 0.10 0.09
5 Clade A 0.09 0.09 0.13 0.11
6 Clade B 0.12 0.12 0.12 0.10 0.13
7 Clade C 0.10 0.11 0.10 0.05 0.12 0.11
8 Clade D 0.11 0.11 0.10 0.05 0.11 0.11 0.06
9 Clade E 0.10 0.10 0.10 0.06 0.11 0.11 0.06 0.06
10 Clade F 0.12 0.12 0.04 0.08 0.14 0.13 0.09 0.10 0.09
11 Pholidobolus affinis 0.13 0.14 0.15 0.12 0.13 0.15 0.12 0.11 0.12 0.15
12 Pholidobolus dicrus 0.15 0.15 0.15 0.15 0.17 0.15 0.15 0.16 0.16 0.15 0.16
13 Pholidobolus hillisi 0.17 0.17 0.17 0.14 0.16 0.18 0.14 0.16 0.14 0.16 0.17 0.19
14 Pholidobolus montium 0.13 0.14 0.15 0.12 0.12 0.17 0.13 0.13 0.13 0.14 0.11 0.17 0.17
15 Pholidobolus paramuno 0.14 0.14 0.16 0.14 0.15 0.16 0.14 0.13 0.13 0.16 0.14 0.17 0.17 0.15
16 Pholidobolus prefrontalis 0.12 0.12 0.14 0.11 0.12 0.14 0.13 0.13 0.12 0.14 0.13 0.16 0.17 0.13 0.13
17 Pholidobolus ulisesi 0.15 0.15 0.15 0.16 0.15 0.16 0.14 0.15 0.14 0.15 0.15 0.17 0.16 0.16 0.17 0.16
18 Pholidobolus vertebralis 0.17 0.17 0.17 0.17 0.17 0.17 0.15 0.16 0.15 0.17 0.16 0.19 0.18 0.18 0.16 0.17 0.17

Morphological comparisons among species

Two components with eigenvalues > 1.0 were extracted from the PCA (Table 5). These components accounted for 50.7% of the total variation. The highest loadings corresponded 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).

Table 5.

Character loadings, eigenvalues, and percentage of variance explained by Principal Components (PC) I and II. The analysis was based on 16 morphological characters of specimens of “Pholidobolus macbrydei”, Pholidobolus samek sp. nov., Pholidobolus condor sp. nov., Pholidobolus dolichoderes sp. nov. and P. fascinatus sp. nov. Highest loadings are in bold.

Variable PCA
PC I PC II
NSO 0.13 -0.11
SUJ 0.32 -0.15
SLJ 0.30 -0.10
SGJ 0.29 -0.07
SGV 0.31 0.25
DEL 0.30 0.33
NTS 0.32 -0.20
SAB 0.26 0.39
SAT 0.18 0.55
SF3 0.18 -0.11
SF5 0.22 -0.31
ST3 0.24 0.04
ST4 0.20 -0.12
ST5 0.29 -0.02
LES -0.05 0.20
SGC -0.22 0.35
Eigenvalue 6.51 1.60
% 40.69 9.99
Figure 2. 

Principal components analysis of 16 morphological variables and 140 specimens of the “Pholidobolus macbrydei” species complex. See Table 5 for character loadings on each component.

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 hemipenes 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 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.

Figure 3. 

Comparative hemipenial morphology of Pholidobolus. Sulcate (left), lateral (center) and asulcate (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.

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.

Systematic accounts

Pholidobolus samek sp. nov.

Figures 4, 5, 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-Chinchipe, 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.

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 Nuñez and Valeria Chasiluisa.

Paratypes (6)

Ecuador: Provincia Zamora-Chinchipe: QCAZ 14954 (adult female), 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.

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.

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 (loreal 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 distinguished 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 differentiated). 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, slender at midbody, becoming pale gray towards tail; (11) labial stripe white or orange; (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 hexagonal, 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 II; interparietal roughly heptagonal; parietals slightly bigger than interparietal, hexagonal, and positioned anterolaterally to interparietal, each in contact anteriorly with supraocular II (and supraocular III on right side) and dorsalmost postocular; postparietals three, medial 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 posterodorsally 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 divided 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 margins; 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 separated 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 widened; scales on sides of neck small and granular; dorsal scales hexagonal, elongate, imbricate, arranged in transverse rows; scales on dorsal surface of neck striated, becoming 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 13th 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, 6/7 on II, 8/8 on III, 9/9 on IV, 6/6 on V; supradigitals 3/4 on toe I, 6/6 on II, 10/9 on III, 11/12 on IV, 7/7 on V; subdigital lamellae of fingers I and II single, paired on III (except the four distalmost), paired at base on IV, on finger V all single; subdigital lamellae 5/5 on finger I, 11/12 on II, 15/16 on III, 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 III, 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 7.4; ShL 7.0; AGD 23.9; TL/SVL 1.5; HL/SVL 0.2; 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 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).

Figure 6. 

Pholidobolus samek sp. nov. in life. A Adult female, paratype (QCAZ 14954) B adult male, holotype (QCAZ 14955).

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 fading 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, N = 1) than females (maximum SVL 45.4 mm, N = 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 longitudinal 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 Cóndor, southeastern Ecuador at elevations between 2324−2844 m (Fig. 7). The new species is known only from Zamora-Chinchipe province, on the sandstone plateaus of Cerro Plateado Biological Reserve. The ground at the type locality 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 incubated in sphagnum and perlite in captivity for two months approximately. They were 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.

Figure 7. 

Distribution of samples of the “Pholidobolus macbrydei” species complex included in phylogenetic 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.

Conservation status

Pholidobolus samek is only known from Cordillera del Cóndor. 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 B1a, b(iii); C1; D, according to IUCN (2012) guidelines.

Figure 8. 

Habitat of Pholidobolus samek sp. nov. A Vegetation around type locality, Cerro Plateado Biological Reserve, Ecuador B habitat where holotype was found. Photographs by Álvaro Pérez.

Etymology

The specific epithet samek means green in the Shuar language, in allusion to the green dorsolateral head stripes distinguishing the new species from other congeners. The type locality of Pholidobolus samek lies within territory of Shuar indigenous people, who inhabit the Amazonian rainforest in Ecuador and Peru.

Remarks

Pholidobolus samek sp. nov. is very similar morphologically and genetically 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, 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 patterns from either P. samek or P. condor (V. Parra and O. Torres-Carvajal, personal observation). 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 mountaintops above 2000 m (Fig. 7).

Pholidobolus condor sp. nov.

Figures 9, 10 Proposed standard English name: Condor cuilanes Proposed standard Spanish name: Cuilanes del Cóndor

Holotype

QCAZ 15844 (Figs 9, 10), adult male, Ecuador, Provincia Morona Santiago, 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 Núñez, Eloy Nusirquia, Alex Achig and Ricardo Gavilanes.

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.

Paratypes (3)

Ecuador: 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 Núñez, Eloy Nusirquia, Alex Achig and María del Mar Moretta.

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.

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 differ 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), P. macbrydei (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 striated 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 hexagonal, 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 supraocular; frontoparietals hexagonal, longer than wide, slightly wider in the middle, each in contact laterally with supraocular II; 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 large and smooth supratemporals; nasal shield slightly divided above nostril, irregularly pentagonal, longer than high, in contact with rostral anteriorly, first and second supralabials ventrally, frontonasal dorsally, loreal posterodorsally and frenocular posteroventrally; 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 nasal; nasal separating loreal from supralabials; two supraoculars, anteriormost one the widest; 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 recessed 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 progressively keeled from forelimbs to tail; dorsal scales between occipital and posterior margin of hindlimbs 27; dorsal scale rows in a transverse line at midbody 27; one longitudinal row of smooth, enlarged ventrolateral scales on each side; dorsals separated from ventrals by two rows of small scales at the level of 13th 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 II, 8/8 on III, 9/9 on IV, 6/6 on V; supradigitals 3/3 on toe I, 6/6 on II, 9/9 on III, 12/12 on IV, 7/7 on V; subdigital lamellae of finger I, II, III, and V single, on finger IV few scales in the middle paired; subdigital lamellae 6/6 on finger I, 11/11 on II, 15/15 on III, 17/16 on IV, 10/10 on V; subdigital lamellae on toes I and II single, on toes III, IV and V paired, except for two or three distalmost subdigitals; subdigital lamellae 7/6 on toe I, 12/12 on II, 15/16 on III, 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 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 laterally; 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 laterally; 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.

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 (adults = 3) P. condor sp. nov. N = 4 (adults = 1) P. dolichoderes sp. nov. N = 5 (adults = 3) P. fascinatus sp. nov. N = 27 (adults = 4)
Scales along margin of upper jaw 7–10 (9.14 ± 1.07) 8–9 (8.75 ± 0.5) 9–11 (10.2 ± 0.84) 7–10 (8.36 ± 0.91)
Scales along margin of lower jaw 8–9 (8.25 ± 0.5) 5–10 (7.14 ± 2.67) 10–11 (10.2 ± 0.45) 4–10 (7.4 ± 1.58)
Gulars 15–18 (16.71 ± 1.11) 14–16 (15 ± 0.82) 22–23 (22.8 ± 0.48) 14–17 (15.72 ± 0.89)
Ventrals in transverse row at midbody 19–21 (20 ± 0.82) 18–20 (19 ± 1.15) 25–27 (25.8 ± 0.84) 21–25 (22.96 ± 1.21)
Dorsals from occiput to base of tail 27–29 (27.71 ± 0.76) 26–30 (27.75 ± 1.71) 35–40 (36.8 ± 2.05) 32–37 (34.64 ± 1.19)
Temporals 4–5 (4.14 ± 0.38) 4–5 (4.25 ± 0.5) 7–9 (8 ± 0.70) 3–5 (3.44 ± 0.65)
Scales around midbody 25–32 (27.71 ± 2.75) 27–30 (28 ± 1.41) 31–33 (32.2 ± 0.84) 28–34 (30.96 ± 1.79)
Scales around tail 14–16 (15 ± 0.81) 14–20 (17.86 ± 2.73) 18–19 (18.6 ± 0.55) 18–22 (20.32 ± 1.18)
Lower eyelid scales 4–5 (4.14 ± 0.38) 5 4–6 (4.8 ± 0.84) 4–6 (5.04 ± 0.61)
Gular (collar) scales 6–8 (7.14 ± 0.9) 6–9 (7.75 ± 1.26) 6–8 (6.4 ± 0.89) 9–12 (10.28 ± 0.73)
Head length in adults 9.9–11.4 (10.76 ± 0.77) 11 9.7–10.6 (10.05 ± 0.46) 8.9–12.3 (10.22 ± 1.80)
Head width in adults 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)
SVL in adults 41.6–49.3 (45.89 ± 3.89) 42.7 41.1–50.6 (45.75 ± 4.74) 42.6–52.5 (47.3 ± 4.98)

Distribution and natural history

Pholidobolus condor occurs in Cordillera del Cóndor 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).

Figure 11. 

Habitat of Pholidobolus condor sp. nov. at El Quimi Biological Reserve, Ecuador. Photographs by Álvaro Pérez.

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 captivity for approximately three months. On average, hatchlings weighted 0.4 g and were 23.7 mm in SVL.

Conservation status

Pholidobolus condor is only known from Cordillera del Cóndor 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 suggest assigning P. condor to the Critically Endangered category under criteria B1a, b(iii); C1; D, according to IUCN (2012) guidelines.

Etymology

The specific epithet condor refers to Cordillera del Cóndor, where the new species was discovered. The Cordillera del Cóndor 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.

Pholidobolus dolichoderes sp. nov.

Figures 12, 13, 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 Oña, 3.4292S, 79.2364W, WGS84, 2672 m, 16 March 2018, collected by Diego Almeida, Darwin Núñez, Eloy Nusirquia, Alex Achig and Katherine Nicolalde.

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 Pazmiño.

Paratypes (4)

Ecuador: Provincia Azuay: QCAZ 16349, 16352 (adult females), San Felipe de Oña, Susudel-Poetate road, 3.4322S, 79.2369W, WGS84, 2506 m, 16 March 2018; QCAZ 16350–51 (juveniles), San Felipe de Oña, 3.4275S, 79.2339W, WGS84, 2675 m, 16 March 2018, same collectors as holotype.

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.

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 anterior 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 differs 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 (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 widened 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).

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 absent; (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 contact 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 borders 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 contact 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 supraoculars, 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 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; 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 ventrals by three rows of granular scales at level of 13th 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 III, 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, III and IV paired proximally, on finger V all single; subdigital lamellae 5 on left finger I (right finger missing), 10/10 on II, 14/14 on III, 14/14 on IV, 9/9 on V; subdigital lamellae on toe I single, on toe II paired at the middle, on toe III 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 dorsolateral 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).

Figure 14. 

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 Pazmiño.

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; palpebral disc divided into 5/6 scales in QCAZ 16352 and 3/5 in QCAZ 16351; frontonasal 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).

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 between 2506−2675 m in San Felipe de Oña, 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 Oña. The population size of this species is unknown, but our sampling suggests low abundances. Because of the small known distribution and lack of additional data, we suggest assigning P. dolichoderes to the Data Deficient category according to IUCN (2012) guidelines.

Etymology

The specific epithet dolichoderes derives from the Greek words dolikhós, meaning long, and derē, meaning neck, in allusion to the distinctively long neck of this species.

Pholidobolus fascinatus sp. nov.

Figures 15, 16, 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 Núñez, Eloy Nusirquia, Santiago Guamán and Guadalupe Calle.

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.

Paratypes (26)

Ecuador: 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 female), Lake Chillacocha, 3.4986S, 79.6187W, WGS84, 3348 m, 17 November 2016, same collectors as holotype.

Figure 16. 

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.

Diagnosis

Pholidobolus fascinatus is unique among its congeners in lacking widened 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. fascinatus 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 temporals (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 hexagonal, 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 prefrontals and frontonasal anteriorly, two supraoculars laterally, and frontoparietals posteriorly; frontoparietals pentagonal, longer than wide, slightly wider posteriorly, each in contact laterally with supraocular II; interparietal heptagonal, lateral borders nearly 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 superciliaries, 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, without denticulate margins; tympanum recessed into a shallow auditory meatus; mental semicircular, longer than wide; postmental pentagonal, slightly longer than wide, followed 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 widened; scales on sides of neck small and slightly granular; dorsal scales hexagonal, elongate, 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 13th 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 II, 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 III (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; subdigital lamellae 5/5 on toe I, 10/10 on II, 14/13 on III, 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.

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 insertion; 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).

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).

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 dorsally (rostral, frontonasal, frontal, frontoparietals and supraoculars) and brown laterally; cream longitudinal stripe extending from third supralabial to shoulder; ventrolateral 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 absent in QCAZ 15122 and 15173; little intrusive scales between postparietal and frontoparietals 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, N = 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 (N = 20) weighted 0.4 g and were 26.2 mm in SVL on average.

Conservation status

Pholidobolus 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 additional data, we suggest assigning P. fascinatus to the Data Deficient category, according 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 controversial partly because morphological evidence has been misinterpreted. Nonetheless, the recent use of molecular phylogenies has reshaped the systematics and taxonomy 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-Gómez 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 Pholidobolus, 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 species, 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). Components 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 morphological characters and increase sample size to better elucidate morphological differences. Mafla-Endara (2011) also suggested hybridization between P. macbrydei from Cañar province and P. prefrontalis based on both the relatively great variation in morphology within the Cañar populations, and their morphological similarity to P. prefrontalis. Nevertheless, our nuclear phylogenetic tree does not suggest hybridization between P. macbrydei and P. prefrontalis (Appendix II).

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 Cóndor 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 Cóndor 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 Cóndor.

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 (Sánchez-Pacheco et al. 2012), especially for poorly explored areas like Cordillera del Cóndor. 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 Secretaría de Educación Superior, Ciencia, Tecnología e Innovación del Ecuador SENESCYT (‘Iniciativa Arca de Noé’, PIs Omar Torres-Carvajal and Santiago Ron), and the Pontificia Universidad Católica del Ecuador. P.M.S.N. is grateful to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for financial support (Fellowship #313622/2018-3). We thank the QCAZ field and laboratory teams. Fernando Ayala, Marcel Caminer, María José Navarrete, Emilio Oviedo, Valeria Chasiluisa, Santiago Ron, and Andrés Merino provided valuable advice and help during the research. We are especially thankful to Tiffany Doan and Santiago Sánchez-Pacheco for their critical reviews and helpful comments. Photographs were taken from Bioweb (www.bioweb.bio) unless otherwise noted.

References

  • Betancourt R, Reyes-Puig C, Lobos SE, Yánez-Muñoz MH, Torres-Carvajal O (2018) Sistemática de los saurios Anadia Gray, 1845 (Squamata: Gymnophthalmidae) de Ecuador: límite de especies, distribución geográfica y descripción de una especie nueva. Neotropical Biodiversity 4: 83–102. https://doi.org/10.1080/23766808.2018.1487694
  • Brito JM, Tinoco N, Chávez D, Moreno-Cárdenas P, Batallas D, Ojala-Barbour R (2017) New species of arboreal rat of the genus Rhipidomys (Cricetidae, Sigmodontinae) from Sangay National Park, Ecuador. Neotropical Biodiversity 3:1, 65–79. https://doi.org/10.1080/23766808.2017.1292755
  • Castoe TA, Doan TM, Parkinson CL (2004) Data partitions and complex models in Bayesian analysis: The phylogeny of gymnophthalmid lizards. Systematic Biology 53: 448–469. https://doi.org/10.1080/10635150490445797
  • de Queiroz K (1998) The general lineage concept of species, species criteria, and the process of speciation. In: Howard DJ, Berlocher SH (Eds) Endless Forms: Species and Speciation. Oxford University Press, Oxford, 57–75.
  • Dowling HG, Savage JM (1960) A guide to the snake hemipenis: a survey of basic structure and systematic characteristics. Zoologica 45: 17–28.
  • Harvey MB, Embert D (2008) Review of Bolivian Dipsas (Serpentes: Colubridae), with comments on other South American species. Herpetological Monographs 22: 54–105. https://doi.org/10.1655/07-023.1
  • Hurtado-Gómez J, Arredondo J, Nunes PMS, Daza JM (2018) A New Species of Pholidobolus (Squamata: Gymnophthalmidae) from the Paramo Ecosystem in the Northern Andes of Colombia. South American Journal of Herpetology 13(3): 271–286. https://doi.org/10.2994/SAJH-D-15-00014.1
  • IUCN (2012) IUCN Red List Categories and Criteria: Version 3.1. Second edition. IUCN, Gland, Switzerland and Cambridge, UK, iv + 32 pp.
  • Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C (2012) Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28: 1647–1649. https://doi.org/10.1093/bioinformatics/bts199
  • Kizirian DA (1996) A review of Ecuadorian Proctoporus (Squamata: Gymnophthalmidae) with descriptions of nine new species. Herpetological Monographs 10: 85–155. https://doi.org/10.2307/1466981
  • Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular biology and evolution 33(7): 1870–1874. https://doi.org/10.1093/molbev/msw054
  • Lanfear R, Calcott B, Ho SYW, Guindon S (2012) Partition-Finder: Combined selection of partitioning schemes and substitution models for phylogenetic analyses. Molecular Biology and Evolution 29: 1695–1701. https://doi.org/10.1093/molbev/mss020
  • Lehr E, Moravec J, Šmíd J, Lundberg M, Köhler G, Catenazzi A (2019) A new genus and species of arboreal lizard (Gymnophthalmidae: Cercosaurinae) from the eastern andes of Peru. Salamandra 55: 1–13.
  • Mafla-Endara P (2011) Filogeografía de las lagartijas andinas del género Pholidobolus (Squamata: Gymnophthalmidae) en Ecuador. Disertación previa a la obtención del título de Licenciada en Ciencias Biológicas. Pontificia Universidad Católica del Ecuador.
  • Manzani PR, Abe AS (1988) Sobre dois novos métodos de preparo do hemipênis de serpentes. Memorias do Instituto Butantan 50: 15–20.
  • Mashburn B, Pérez Á, Persson C, Zapata N, Cevallos D, Muchhala N (2020) Burmeistera quimiensis (Lobelioideae, Campanulaceae): A new species from the Cordillera del Cóndor range in southeast Ecuador. Phytotaxa 433: 67–74. https://doi.org/10.11646/phytotaxa.433.1
  • Mazabanda C, Kemper R, Thieme A, Hettler B, Finer M (2018) Impacts of Mining Project “Mirador” in the Ecuadorian Amazon. Monitoring Andean Amazon Project (MAAP). https://maaproject.org/mirador-ecuador
  • Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Proceedings of the Gateway Computing Environments Workshop (GCE), 14 Nov. 2010, New Orleans, LA, 1–8. https://doi.org/10.1145/2016741.2016785
  • Montanucci RR (1973) Systematics and evolution of the Andean lizard genus Pholidobolus (Sauria: Teiidae). Miscellaneous Publication. Museum of Natural History, University of Kansas 59: 1–52.
  • Moravec J, Šmíd J, Štundl J, Lehr E (2018) Systematics of Neotropical microteiid lizards (Gymnophthalmidae, Cercosaurinae), with the description of a new genus and species from the Andean montane forests. ZooKeys 774: 105–139. https://doi.org/10.3897/zookeys.774.25332
  • Neill DA (2005) Cordillera del Cóndor: Botanical treasures between the Andes and the Amazon. Plant Talk 41: 17–21.
  • Nunes PMS, Fouquet A, Curcio FF, Kok PJR, Rodrigues MT (2012) Cryptic species in Iphisa elegans Gray, 1851 (Squamata: Gymnophthalmidae) revealed by hemipenial morphology and molecular data. Zoological Journal of the Linnean Society 166: 361–376. https://doi.org/10.1111/j.1096-3642.2012.00846.x
  • Pellegrino KCM, Rodrigues MT, Yonenaga-Yassuda Y, Sites JW (2001) A molecular perspective on the evolution of microteiid lizards (Squamata, Gymnophthalmidae), and a new classification for the family. Biological Journal of the Linnean Society 74: 315–338. https://doi.org/10.1006/bijl.2001.0580
  • Pérez-Escobar O, Gottschling M, Chomicki G, Condamine F, Klitgård B, Pansarin E, Gerlach G (2017) Andean Mountain Building Did Not Preclude Dispersal of Lowland Epiphytic Orchids in the Neotropics. Scientific Reports 7: 4919. https://doi.org/10.1038/s41598-017-04261-z
  • Peters AJ (1964) Dictionary of Herpetology. Hafner Publishing Company, New York, 392 pp.
  • R Core Team (2018) R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna. https://www.R-project.org
  • Ron SR, Caminer MA, Varela-Jaramillo A, Almeida-Reinoso D (2018) A new treefrog from Cordillera del Cóndor with comments on the biogeographic affinity between Cordillera del Cóndor and the Guianan Tepuis (Anura, Hylidae, Hyloscirtus). ZooKeys 809: 97–124. https://doi.org/10.3897/zookeys.809.25207
  • Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, H√∂hna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: 539–542. https://doi.org/10.1093/sysbio/sys029
  • Sánchez-Pacheco SJ, Aguirre-Peñafiel V, Torres-Carvajal O (2012) Lizards of the genus Riama (Squamata: Gymnophthalmidae): the diversity in southern Ecuador revisited. South American Journal of Herpetology 7: 259–275. https://doi.org/10.2994/057.007.0308
  • Sánchez-Pacheco SJ, Torres-Carvajal O, Aguirre-Peñafiel V, Nunes PMS, Verrastro L, Rivas GA, Rodrigues MT, Grant T, Murphy RW (2017) Phylogeny of Riama (Squamata: Gymnophthalmidae), impact of phenotypic evidence on molecular datasets, and the origin of the Sierra Nevada de Santa Marta endemic fauna. Cladistics 34: 260–291. https://doi.org/10.1111/cla.12203
  • Savage JM (1997) On terminology for the description of the hemipenis of squamate reptiles. Herpetological Journal 7: 23–25.
  • Torres-Carvajal O, de Queiroz K, Etheridge R (2009) A new species of iguanid lizard (Hoplocercinae, Enyalioides) from southern Ecuador with a key to eastern Ecuadorian Enyalioides. Zookeys 27: 59–71. https://doi.org/10.3897/zookeys.27.273
  • Torres-Carvajal O, Lobos SE, Venegas PJ (2015) Phylogeny of Neotropical Cercosaura (Squamata: Gymnophthalmidae) lizards. Molecular Phylogenetics and Evolution 93: 281–288. https://doi.org/10.1016/j.ympev.2015.07.025
  • Torres-Carvajal O, Lobos SE, Venegas PJ, Chávez G, Aguirre-Peñafiel V, Zurita D, Echevarría LY (2016) Phylogeny and biogeography of the most diverse clade of South American gymnophthalmid lizards (Squamata, Gymnophthalmidae, Cercosaurinae). Molecular Phylogenetics and Evolution 99: 63–75. https://doi.org/10.1016/j.ympev.2016.03.006
  • Torres-Carvajal O, Mafla-Endara P (2013) Evolutionary history of Andean Pholidobolus and Macropholidus (Squamata: Gymnophthalmidae) lizards. Molecular Phylogenetics and Evolution 68: 212–217. https://doi.org/10.1016/j.ympev.2013.03.013
  • Torres-Carvajal O, Venegas PJ, Lobos SE, Mafla-Endara P, Nunes PMS (2014) A new species of Pholidobolus (Squamata: Gymnophthalmidae) from the Andes of southern Ecuador. Amphibian & Reptile Conservation 8: 76–88.
  • Townsend TM, Alegre RE, Kelley ST, Wiens JJ, Reeder TW (2008) Rapid development of multiple nuclear loci for phylogenetic analysis using genomic resources: An example from squamate reptiles. Molecular Phylogenetics and Evolution 47: 129–142. https://doi.org/10.1016/j.ympev.2008.01.008
  • Uzzell T (1973) A revision of the genus Prionodactylus with a new genus for P. leucostictus and notes on the genus Euspondylus (Sauria, Teiidae). Postilla 159: 1–67. https://doi.org/10.5962/bhl.part.11535
  • Valencia JH, Dueñas MR, Székely P, Batallas D, Pulluquitín F, Ron SR (2017) A new species of direct-developing frog of the genus Pristimantis (Anura: Terrarana: Craugastoridae) from Cordillera del Cóndor, Ecuador, with comments on threats to the anuran fauna in the region. Zootaxa 4353: 447–466. https://doi.org/10.11646/zootaxa.4353.3.3
  • Van Teijlingen K (2016) The ´will to improve´ at the mining frontier: Neo-extractivism, development and governmentality in the Ecuadorian Amazon. Extractive Industries and Society 3: 902–911. https://doi.org/10.1016/j.exis.2016.10.009
  • Venegas PJ, Echeverría LY, Lobos SE, Nunes PMS, Torres-Carvajal O (2016) A new species of Andean microteiid lizard (Gymnophthalmidae: Cercosaurinae: Pholidobolus) from Peru, with comments on P. vertebralis. Amphibian & Reptile Conservation 10(1): 21–33.
  • Zaher H (1999) Hemipenial morphology of the South American xenodontine snakes, with a proposal for a monophyletic Xenodontinae and a reappraisal of colubroid hemipenes. Bulletin of the American Museum of Natural History 240: 1–168. http://hdl.handle.net/2246/1646

Appendix I

Additional specimens examined of Pholidobolus macbrydei. 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, 2.77744S, 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, 2.87522S, 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-Limón; 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, 3.21367S, 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-Gualaquiza, 3.106875S, 78.79558W, 2935 m, QCAZ 8646–8647; Tarqui, 3.015880S, 79.04447W, 2680 m, QCAZ 8512. Provincia Cañar: Cajas National Park, 2.70654S, 79.22765W, 3651 m, QCAZ 8946; Cañar, 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, 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; 2.5708S, 78.74586W, 3451 m, QCAZ 15824; 2.545804S, 78.69611W, 2842 m, QCAZ 10970. Provincia Chimborazo: Frutatián lake, 2.21584S, 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-Melán, 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: Guanazán, 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; Tarqui, 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.

Appendix II

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.