A molecular phylogeny of Equatorial African Lacertidae, with the description of a new genus and species from eastern Democratic Republic of the Congo

Abstract

Currently, four species of the lacertid lizard genus Adolfus are known from Central and East Africa. We sequenced up to 2,825 bp of two mitochondrial (16S and cyt b) and two nuclear (c-mos and RAG1) genes from 41 samples of Adolfus (representing every species), two species each of Gastropholis and Holaspis, and in separate analyses combined this data with GenBank sequences of all other Eremiadini genera and four Lacertini outgroups. Data from DNA sequences were analyzed with maximum parsimony (PAUP), maximum-likelihood (RAxML) and Bayesian inference (MrBayes) criteria. Results demonstrated that Adolfus is not monophyletic: A. africanus (type species), A. alleni and A. jacksoni are sister taxa, whereas A. vauereselli and a new species from the Itombwe Plateau of Democratic Republic of the Congo are in a separate lineage. Holaspis and Gastropholis were recovered in separate clades. Based on this molecular data, relatively substantial sequence divergence and multiple morphological differences, we describe a new genus of lacertid for the lineage including A. vauereselli and the new Itombwe species. The recognition of this new, endemic genus underscores the conservation importance of the Albertine Rift, especially the Itombwe Plateau, a unique region that is severely threatened by unchecked deforestation, mining and poaching.

INTRODUCTION

Meadow and forest lizards of the lacertid genus Adolfus are currently known from Central and East Africa, including A. africanus (mid- to low elevation forests from Cameroon to Kenya), A. alleni (montane moorlands of Kenya and Uganda), and A. jacksoni and A. vauereselli, which are both known from mid- to high elevation forests in countries surrounding the Albertine Rift (Spawls et al., 2002; Köhler et al., 2003). Adolfus are medium-sized (total size to 25.6 cm), relatively slim lizards, and tend to be good climbers on standing and fallen timber, rocky walls, holes and crevices (A. africanus is also known to climb twiggy and herbaceous plants), but tend to hunt on the ground (Arnold, 1989a, 1998; Spawls et al., 2002). Recent work on this genus has included aspects of reproduction (A. jacksoni, Goldberg, 2009), endoparasites (A. jacksoni, Goldberg & Bursey, 2009), geographic distribution (A. africanus, Köhler et al., 2003), and morphology and color pattern (A. jacksoni, Poblete, 2002).

The taxonomic status and affinities of the currently recognized species of Adolfus have changed considerably over time. The genus Adolfus was first proposed by Sternfeld (1912) for the taxon A. fridericianus, which was presumably in honor of Adolphus Frederick, Duke of Mecklenburg, who led the German East Africa Expedition in 1907–08 when the specimens were collected (Frederick, 1910). In his opus on the Family Lacertidae, Boulenger (1920) considered Adolfus fridericianus to be a synonym of Algiroides africanus (= Algyroides africanus), a species he described in 1906, and recognized Algiroides alleni, Lacerta jacksonii (a species he described in 1899), and L. vauereselli. Based on morphological characters, Arnold (1973) resurrected the genus Adolfus for A. africanus, A. alleni and A. vauereselli, and noted a close relationship between this genus and Bedriagaia, Gastropholis and Lacerta jacksoni. In morphology-based parsimony and compatibility analyses, Arnold (1989a) transferred Lacerta jacksoni to the genus Adolfus, synonymized Bedriagaia with Gastropholis, recognized a clade called the “Equatorial African group” including Adolfus, Gastropholis and Holaspis (a well-supported clade recovered in a later morphology-based phylogeny by Harris, Arnold & Thomas, 1998), and discussed the problematic relationship of Holaspis to the paraphyletic genus Adolfus; if the latter two genera were to be joined, Holaspis would have priority. Arnold (1989a,b) admitted that Adolfus was poorly defined, and considered A. jacksoni to be the most plesiomorphic member of the Equatorial African clade. In a more extensive morphological analysis of the entire Family Lacertidae, Arnold (1989b) grouped the Equatorial African Clade with Lacerta jayakari (now Omanosaura jayakari), Lacerta australis (now Australolacerta australis) and several other genera (e.g., Tropidosaura, Poromera, Nucras) in an “Ethiopian and advanced Saharo-Eurasian forms” (ESE) group, which was later included in an “Armatured Clade” (Afrotropical species plus Eremias, Acanthodactylus, Mesalina and Ophisops) in recognition of the members’ unique supporting structure of the male hemipenis (Arnold, 1986a, 1998; Harris et al., 1998). Mayer & Benyr (1994) used an albumin-based analysis of most lacertid genera to imply paraphyly of the ESE group, with some of the Saharo-Eurasian genera grouping with European lacertids. Based on a combination of morphology and mtDNA data that contradicted several findings of Mayer & Benyr (1994), Harris et al. (1998) assigned the subfamily Eremiainae (Szczerbak, 1975) to the Armatured Clade.

More recent analyses of lacertids with mitochondrial data have done little to clarify the position of Adolfus in relation to other members of the Equatorial African clade, or the ESE group as a whole. Although Fu (1998) recovered a monophyletic “African clade” in a mitochondrial phylogeny of lacertids, no members of the Equatorial African clade were included. Harris (1999) combined the mitochondrial data of Fu (1998) and Harris et al. (1998) with some new data to produce a phylogeny of Lacertidae, but support for the ESE clade (still recognized as Eremiainae) was weak; two samples of Adolfus (A. africanus and A. jacksoni) were not supported as sister taxa. Fu (2000) published another phylogeny of Lacertidae with six mitochondrial genes (4.7 kb of DNA data), with most trees supporting the monophyly of the ESE clade, but with the exception of three closely related genera (Nucras, Latastia and Heliobolus), relationships among ESE genera were unclear, and the monophyly of two samples of Adolfus (A. jacksoni and A. vauereselli) was again not supported.

Mayer & Pavlicev (2007) published the first lacertid phylogeny based on nuclear data (c-mos and RAG1), and recovered two clades within a well-supported ESE (Eremiainae) group: clade B₁, mainly from sub-Saharan Africa, including Poromera, Nucras, Latastia, Philochortus, Pseuderemias, Heliobolus, Tropidosaura, Pedioplanis, Ichnotropis and Meroles; and clade B₂, mainly from the Saharo-Eurasian region, including Ophisops, Omanosaura, Acanthodactylus, Eremias, Mesalina, Adolfus and Holaspis, with the latter two Central African genera as well-supported sister taxa. Arnold, Arribas & Carranza (2007) re-analyzed the datasets of Harris et al. (1998) and Fu (2000), and published yet another lacertid phylogeny based on two mitochondrial genes (12S and cyt b). Although their main focus was not on the ESE group, they redefined the Eremiainae as the tribe Eremiadini, and placed the North African monotypic genus Atlantolacerta as the most basal member of the Eremiadini. Pavlicev & Mayer (2009) criticized the dataset of the latter study as “relatively short mitochondrial sequences when all taxa are considered,” rejected the tribe Eremiadini (instead recognizing it as subfamily Eremiadinae), but confirmed the placement of Atlantolacerta as the most basal member of the group. Hipsley et al. (2009) used mitochondrial and nuclear data from several previous studies to confirm the main findings of Mayer and Pavlicev (2007), but continued to recognize the tribe Eremiadini (sensu Arnold et al., 2007) and revised the date of its origin to the mid- to late Eocene, when the group could have invaded northwestern Africa via small island chains.

Three of the four species of Adolfus can be found in eastern Democratic Republic of the Congo (DRC), which harbors a panoply of habitats ranging from lowland rainforest to alpine grassland (Bastin et al., 2004; Vande weghe, 2004). Based on fieldwork in the poorly known Itombwe Plateau (eastern DRC) by EG, CK and MMA, we collected several specimens of an Adolfus that does not fit the description of any currently recognized species. To clarify the position of the Itombwe population to other Adolfus, we sequenced multiple genes from several members of the Equatorial African group of lacertids (Adolfus, Gastropholis and Holaspis), and discovered that the Itombwe population is a new species belonging to a lineage that deserves recognition as a distinct genus. We follow the General Lineage Species Concept (de Queiroz, 1998, 1999), an extension of the Evolutionary Species Concept (Wiley, 1981), which provides a consistent philosophical framework for taxonomic decisions, and rejects the premise of subspecies as natural groups. Our species recognition criteria (de Queiroz, 2007; Wiens and Penkrot, 2002) correspond in part to traditional morphological species, which are diagnosed by unique morphological characters, size and color pattern. We utilize a molecular estimate of phylogenetic relationships that is based on multiple, unlinked markers from multiple individuals within species to guide species delimitation and diagnosis, and identify relevant comparisons for species diagnoses (Barraclough & Davies, 2005; Brown et al., 2009).

MATERIALS AND METHODS

DNA EXTRACTION, PCR AMPLIFICATION AND SEQUENCING

Two mitochondrial (16S and cyt b) and two nuclear (c-mos and RAG1) genes were sequenced from all genera in the Equatorial African Group, including 41 samples of all species of Adolfus, Holaspis guentheri, H. laevis, Gastropholis prasina, G. vittatus, and five outgroup taxa, including: Acanthodactylus erythrurus (clade B₂ of Mayer and Pavlicev, 2007), the basal-most member of Eremiadini (Atlantolacerta andreanskyi, Arnold et al., 2007), and three Lacertini genera (Iberolacerta cyreni, Podarcis muralis, Timon tangitanus). Some samples (e.g., Adolfus alleni) did not amplify for all genes; all sequences were deposited in GenBank (Appendix I). Genomic DNA was isolated from alcohol-preserved liver or muscle tissue samples with the Qiagen DNeasy tissue kit (Qiagen Inc., Valencia, CA, USA). We used 25 µl PCR reactions with gene-specific primers (Table 1) with an initial denaturation step of 95°C for 2 min, followed by denaturation at 95°C for 35s, annealing at 50°C for 35s, and extension at 72°C for 95s with 4s added to the extension per cycle for 32 (mitochondrial genes) or 34 (nuclear genes) cycles. Amplicons were visualized on a 1.5% agarose gel stained with SYBR Safe DNA gel stain (Invitrogen Corporation, Carlsbad, CA, USA), and target products were purified with AMPure magnetic bead solution (Agencourt Bioscience, Beverly, MA, USA) and sequenced with BigDye® Terminator Cycle Sequencing Kits (Applied Biosystems, Foster City, CA, USA). Sequencing reactions were purified with CleanSeq magnetic bead solution (Agencourt Bioscience) and sequenced with an ABI 3130xl automated sequencer at the DNA Core Facility at the University of Texas at El Paso (UTEP). Forward and reverse sequence contigs for each sample were assembled and edited using SeqMan (DNAStar, Maison, WI, USA) to ensure accuracy. Four samples of Adolfus showed evidence of pseudogenes (i.e., six codon insertion relative to all other lacertids with a reading frame shift) for c-mos, including A. jacksoni (CAS 201598), A. vauereselli (UTEP 20294, 20296), and the new species (UTEP 20263); Pavlicev and Mayer (2006) also reported c-mos pseudogenes in three species of Lacerta. Our pseudogene sequences were excluded from the data set of this study.

Table 1.

Primer sequences used in this study.

Name	Source	Sequence	Gene
16SA-L	Palumbi et al. (1991)	5’-CGCCTGTTTATCAAAAACAT-3’	16S
16SB-H	Palumbi et al. (1991)	5’-CCGGTCTGAACTCAGATCACGT-3’	16S
CytbF700	Bauer et al. (2007)	5'-CTTCCAACACCAYCAAACATCTCAGCATGATGAAA-3'	cyt b
CytbR700	Bauer et al. (2007)	5'-ACTGTAGCCCCTCAGAATGATATTTGTCCTCA-3'	cyt b
Hcmos3	Mayer and Pavlicev (2007)	5’-GGTGATGGCAAATGAGTAGAT-3’	c-mos
L-1zmos	Mayer and Pavlicev (2007)	5’-CTAGCTTGGTGTTCTATAGACTGG-3’	c-mos
Hcmos1	Mayer and Pavlicev (2007)	5’-GCAAATGAGTAGATGTCTGCC-3’	c-mos
R13	Groth and Barrowclough (1999)	5'- TCTGAATGGAAATTCAAGCTGTT-3'	RAG1
R18	Groth and Barrowclough (1999)	5'-GATGCTGCCTCGGTCGGCCACCTTT-3'	RAG1
RAG1f700	Bauer et al. (2007)	5'-GGAGACATGGACACAATCCATCCTAC-3'	RAG1
RAG1r700	Bauer et al.(2007)	5'-TTTGTACTGAGATGGATCTTTTTGCA-3'	RAG1
RAG-R1	Mayer and Pavlicev (2007)	5’-AAAATCTGCCTTCCTGTTATTG-3’	RAG1
RAG-fo	Mayer and Pavlicev (2007)	5’-GAAAAGGGCTACATCCTGG-3’	RAG1
RAG-re	Mayer and Pavlicev (2007)	5’-CCAGTTATTGCTTTTACAGTTC-3’	RAG1

SEQUENCE ALIGNMENT AND PHYLOGENETIC ANALYSES

An initial alignment of each gene was produced in MEGALIGN (DNA Star) with the Clustal W algorithm, and manual adjustments were made in MacClade 4.08 (Maddison & Maddison, 2005). Protein-coding genes were translated to amino acids with MacClade to confirm conservation of the amino acid reading frame, ensure alignment and check for premature stop codons. No ambiguously aligned regions were observed, and as a result, no data were excluded from phylogenetic analyses. Phylogenetic relationships among the samples were assessed with maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) optimality criteria in the programs PAUP* 4.0b10 (Swofford, 2002), RAxML (Stamatakis, 2006) and MrBayes 3.1 (Ronquist & Huelsenbeck, 2003), respectively. For MP analyses, the heuristic search algorithm was used with 100 random-addition replicates, accelerated character transformation and tree bisection-reconnection branch swapping, zero-length branches collapsed to polytomies, and gaps treated as missing data; we used non-parametric bootstraps (1,000 pseudoreplicates) to assess node support in resulting topologies from these parsimony searches (Felsenstein, 1985). The Akaike Information Criterion (Posada & Buckley, 2004) in jModelTest (Posada, 2008) was used to find the model of evolution that best fit the data for subsequent BI analyses. RAxML analyses were executed with partitioned datasets (one for 16S, and one for each codon position of all other protein-coding genes), and 100 replicate ML inferences were peformed for each analysis. Each analysis was initiated with a random starting tree, included the GTRGAMMA option (−m) and employed the rapid hill-climbing algorithm (− x) (Stamatakis et al., 2007). Clade support was assessed with 1,000 bootstrap replicates, with the rapid-hill climbing algorithm (Stamatakis, Hoover & Rougemont, 2008). Phylogenetic trees were visualized with FigTree (http://tree.bio.ed.ac.uk/software/figtree/).

Partitioned Bayesian analyses were conducted with default priors. Analyses were initiated with random starting trees and run for 10,000,000 generations; Markov chains were sampled every 1000 generations. Convergence was checked by importing the trace files (p files) from the MrBayes output to the computer program Tracer v1.3 (http://tree.bio.ed.ac.uk/software/tracer/), which plots the likelihood values against generation number. Once the graphical plot leveled off, convergence had been met; we conservatively discarded 25% of trees as “burn in.” Four separate analyses with two independent chains were executed to check for convergence of log-likelihoods in stationarity (Huelsenbeck & Ronquist, 2001; Leaché & Reeder, 2002). To test the monophyly of polyphyletic lineages recovered in our phylogenetic analyses of the four-gene dataset, we used the Shimodaira-Hasegawa (SH) and approximately unbiased (AU) tests as implemented in CONSEL V0.1i (Shimodaira & Hasegawa, 2001; Shimodaira, 2002). We tested the hypothesis of zero-length branches for polyphyletic lineages of the Equatorial African lacertids by comparing the likelihood of the optimal ML tree from the four-gene dataset to the likelihood of the optimal tree with one branch collapsed with the “describe trees” function in PAUP* (sensu Poe & Chubb, 2004), and a Bonferroni-corrected p value of 0.025.

Combining data from multiple mitochondrial genes is appropriate because the entire animal mitochondrial genome is inherited as a single unit, and different mitochondrial genes are not independent estimates of organismal phylogeny (Moore, 1995; Page, 2000). We combined mitochondrial and nuclear gene datasets if there was no strong bootstrap support for conflicting nodes (exceeding 70% for MP analyses [Hillis & Bull, 1993] and 95% for ML and BI analyses [Leaché & Reeder, 2002; Wilcox et al., 2002]) when these datasets were analyzed independently. After preliminary analyses confirmed there was no conflict between mitochondrial and nuclear gene datasets (data not shown), we conducted two analyses: (1) c-mos and a 1,012-bp fragment of RAG1 (primers from Mayer and Pavlicev, 2007) for samples from this study and previously sequenced lacertids from GenBank (Appendix 1) with Gallotia as the outgroup; and (2) both mitochondrial (16S and cyt b) genes, c-mos and a 1,394-bp fragment of RAG1 (primers from Groth & Barrowclough [1999] and Bauer et al., [2007]) for every sample from this study (hereafter referred to as the four-gene dataset) with three Lacertini outgroups.

MORPHOLOGY

Specimens examined for this study (Appendix 2) were preserved in 10% buffered formalin in the field, and transferred to 70% ethanol at the conclusion of each expedition. Tissues were harvested before formalin fixation from the liver or hind limb muscle of lizards, and preserved in 95% ethanol. Institutional abbreviations are listed at http://www.asih.org/codons.pdf. The senior author recorded morphometric data from these preserved specimens with vernier calipers to the nearest 0.1 mm under a stereomicroscope. Color descriptions are based on preserved specimens, field notes, and color digital images in life. Sex was determined by direct examination of gonads, or from the presence of everted hemipenes as noted in field notes. X-rays for descriptions of the post-cranial skeleton were taken with a Kodak Image Station In-Vivo FX (Carestream Health, Inc., Rochester, NY, USA) under the following conditions: f-stop: 8.0; FOV: 198 mm; focal plane: 0; exposure time: 288 sec; Kilovolt Potential Energy: 35; filter: 600WB.

Meristic and mensural characters were chosen from lacertid studies by Arnold (1989b) and Lue and Lin (2008). Measurements were taken on the right side of the lizard and include: snout–vent length (SVL, from tip of snout to anterior margin of vent); tail length (TL, from posterior margin of vent to tail tip, measured only from specimens with complete and original tails); head length (HL, from tip of snout to anterior margin of ear opening); maximum head width (HW, measured at the broadest point); head height (HH, measured at the jaw rictus); skull length (SKL, from tip of snout to posterior margin of occipital); snout–eye length (SEL, from tip of snout to anterior margin of eye); mouth length (ML); snout–arm length (SAL, from tip of snout to anterior margin of forelimb); axilla–groin distance (AGD, from posterior edge of forelimb insertion to anterior edge of hind limb insertion); humerus length (HML); radius–ulna length (RUL); femur length (FL); tibia–fibula length (TFL); and longest toe length (LTL, length of fourth toe on hind limb).

Meristic data were taken from the right side of each lizard, except for femoral pore counts if field/museum tags were tied to the right leg. Definition of scales follow those of Arnold (1989b) and Arnold et al. (2007), and include: chin shields (CS); femoral pores (FP); supralabials (SL); infralabials (IL); supraoculars (SO): supraciliaries (SC); supraciliary granules (SG); supratemporals (ST); anterior dorsal scale rows (ADS, counted transversely at posterior insertion of forelimbs); posterior dorsal scale rows (PDS, counted transversely at anterior insertion of hind limbs); dorsal scale rows at midbody (DSR, counted transversely at midpoint between fore- and hind limbs); dorsal scale numbers (DSN, counted longitudinally from posterior margin of occipital to posterior margin of hind limbs); ventral rows (VR, counted transversely at midbody); ventral scale numbers (VN, counted longitudinally from posterior margin of collars to anteior margin of preanal scales, took average from the middle two rows); caudal scales (CDS, counted around the tail at the position of the 11^th and 15^th scale to avoid the difference between males and females); and subdigital lamellae on fingers (SDF1 to SDF5) and toes (SDT1 to SDT5).

RESULTS

MOLECULAR PHYLOGENETICS

Relationships among members of the Equatorial African Group of lacertid lizards are shown in Figures 1–2; MP, ML and BI analyses produced nearly identical topologies for each dataset, with only minor differences in bootstrap support for each analysis. For the four-gene dataset (Fig. 2), we noted a six-codon deletion in the RAG1 gene (between positions 134–151) in multiple samples of Adolfus africanus and A. jacksoni.

Figure 1.

Maximum likelihood phylogeny (RAxML tree) of lacertid lizards in the Equatorial African Group, based on the combined nuclear c-mos/RAG1 dataset from this study and Genbank samples from Mayer and Pavlicev (2007). Bootstrap and posterior probability values for each well-supported node are listed in the order: maximum parsimony/maximum likelihood/Bayesian inference.

Figure 2.

Maximum likelihood phylogeny (RAxML tree) of the Equatorial African clade of lizards based on the combined 16S, cyt b, c-mos and RAG1 genes. Bootstrap and posterior probability values for each well-supported node are listed in the order: maximum parsimony/maximum likelihood/Bayesian inference.

The following models of nucleotide substitution were selected by jModeltest for BI analyses: 16S (GTR + I + G); cyt b 1^st codon (TIM2ef + I); cyt b 2^nd codon (GTR + I); cyt b 3^rd codon (GTR + I + G); c-mos 1^st codon (HKY + G); c-mos 2^nd codon (TIM3 + G); c-mos 3^rd codon (TrN + G); RAG1 1^st codon (TrN + I); RAG1 2^nd codon (TPM1uf + G); RAG1 3^rd codon (TPM3uf + I + G). The MP analysis of the c-mos/RAG1 dataset (Fig. 1), included 1605 base pairs (933 constant, 429 parsimony-informative) and resulted in 28,908 most parsimonious trees (length = 1511, CI = 0.574, RI = 0.768); the ML analysis likelihood score was –11052.633819. The MP analysis of the four-gene dataset (Fig. 2), included 2,825 base pairs (2,185 constant, 444 parsimony-informative) and resulted in 5,368 most parsimonious trees (length = 1588, CI = 0.520, RI = 0.777); the ML analysis likelihood score was –11185.625563.

The c-mos/RAG1 tree (Fig. 1) showed strong support for a monophyletic Eremiadini, and a well-supported clade of Ethiopian lacertids (corresponding to clade B₁ of Mayer and Pavlicev, 2007). The remaining Eremiadini lineages were recovered with the following well-supported clades: Eremias (two species), Acanthodactylus (three species), Adolfus vauereselli + A. sp. nov. (Itombwe Plateau), and Adolfus africanus + A. jacksoni. The four-gene dataset (Fig. 2) also shows well-supported clades for Adolfus vauereselli + A. sp. nov. (Itombwe Plateau), and Adolfus africanus + A. alleni + A. jacksoni, with both of these lineages included in a clade with Acanthodactylus, Gastropholis and Holaspis, and a well-supported sister relationship of all of these taxa to Atlantolacerta, again confirmed as the most basal member of Eremiadini.

Among genera of previously recognized lacertids, uncorrected p sequence divergence for the c-mos/RAG1 dataset (Table 2) ranged from 2.4% (Ichnotropis vs. Meroles) to 8.5% (Heliobolus vs. Ophisops). Among previously recognized genera of the Equatorial African Group, uncorrected p sequence divergence for the c-mos/RAG1 dataset ranged from 2.2–3.7% (Adolfus sensu strico vs. Gastropholis) to 3.5–4.1% (Adolfus sensu stricto vs. Holaspis); divergences between the two well-supported lineages of Adolfus (A. africanus + A. alleni + A. jacksoni vs. A. vauereselli + A. sp. nov. [Itombwe Plateau]) ranged from 2.7–3.4% (Table 2). Uncorrected p sequence divergence for the c-mos/RAG1 dataset ranged from 0.0–0.1% within populations of Adolfus vauereselli and A. sp. (Itombwe Plateau), but ranged from 1.3–2.2% between these well-supported taxa; equivalent 16S mitochondrial data ranged from 0.0–1.5% within populations of each taxon to 5.9–6.3% between these taxa (data not shown). Among the two disjunct, montane populations of A. alleni, cyt b divergence (the only gene that amplified for both samples) was 10.9% (data not shown). Hypothesis tests that constrained the monophyly of Adolfus were not significantly different from our preferred tree (AU: p = 0.381; SH: p = 0.382). Tests for zero-length branches for the lineage containing Holaspis + Adolfus vauereselli + A. sp. nov. (p = 0.263) and the lineage containing Gastropholis + A. africanus + A. alleni + A. jacksoni (p = 0.139) were not significantly different from zero.

Table 2.

Uncorrected p sequence divergence (c-mos/RAG1 dataset) for selected samples of Adolfus and other Eremiadini genera included in this study. When more than one sample was sequenced for a given species, specific locality information is provided for the sample included in this table.

	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21	22	23
1 Acanthodactylus erythrurus	—
2 Adolfus africanus (Hombo, DRC)	0.04 219	—
3 Adolfus jacksoni (Rukiva, Rwanda)	0.04 085	0.02 244	—
4 Adolfus jacksoni (Arusha, Tanzania)	0.03 840	.019 26	0.00 835	—
5 Adolfus vauereselli(Kahuzi Biega NP, DRC)	0.04 440	0.02 735	0.03 468	0.03 390	—
6 Adolfus sp. nov. (HOLOTYPE)	0.03 952	0.03 264	0.03 085	0.03 144	0.01 610	—
7 Atlantolacerta andreanskyi	0.03 963	0.03 011	0.02 969	0.02 696	0.03 358	0.03 337	—
8 Eremias arguta	0.04 087	0.03 452	0.03 015	0.02 817	0.03 466	0.03 271	0.03 095	—
9 Gastropholis vittatus	0.04 130	0.03 077	0.02 514	0.02 183	0.03 854	0.03 323	0.03 082	0.03 206	—
10 Heliobolus lugubris	0.07 169	0.06 906	0.06 469	0.06 277	0.07 254	0.06 668	0.06 189	0.06 529	0.06 534	—
11 Holaspis laevis (Usambaras, Tanzania)	0.05 157	0.04 085	0.03 455	0.03 519	0.04 305	0.03 964	0.03 788	0.03 955	0.03 833	0.07 220	—
12 Ichnotropis squamulosa	0.05 354	0.05 375	0.05 100	0.04 813	0.05 742	0.05 487	0.04 363	0.05 160	0.05 039	0.07 489	0.05 350	—
13 Latastia longicaudata	0.06 032	0.06 008	0.05 526	0.05 187	0.06 104	0.05 789	0.05 115	0.05 587	0.05 340	0.05 022	0.06 403	0.06 421	—
14 Meroles suborbitalis	0.05 024	0.04 919	0.04 710	0.04 420	0.05 211	0.04 968	0.04 229	0.04 771	0.04 838	0.07 282	0.05 148	0.02 392	0.05 964	—
15 Mesalina guttulata	0.05 473	0.05 182	0.05 150	0.04 872	0.05 979	0.05 602	0.05 244	0.05 336	0.05 216	0.08 035	0.05 964	0.06 860	0.07 282	0.06 591	—
16 Nucras lalandii	0.05 089	0.05 177	0.04 775	0.04 485	0.05 343	0.04 974	0.04 357	0.04 648	0.04 776	0.05 967	0.05 465	0.05 478	0.04 396	0.05 276	0.06 721	—
17 Omanosaura jayakari	0.03 586	0.03 258	0.03 078	0.03 008	0.03 209	0.03 021	0.03 158	0.03 076	0.03 205	0.06 842	0.03 829	0.04 784	0.05 399	0.04 457	0.05 524	0.04 648	—
18 Ophisops elegans	0.05 285	0.05 491	0.05 214	0.05 054	0.05 140	0.04 847	0.05 303	0.04 896	0.05 342	0.08 475	0.05 712	0.06 608	0.07 156	0.06 089	0.06 905	0.06 281	0.04 708	—
19 Pedioplanis undata	0.05 859	0.05 364	0.04 838	0.04 734	0.05 393	0.05 226	0.04 677	0.04 896	0.04 967	0.07 156	0.05 650	0.05 478	0.05 964	0.05 210	0.07 282	0.05 340	0.04 457	0.06 717	—
20 Philochortus spinalis	0.06 041	0.05 757	0.05 785	0.05 515	0.06 237	0.05 757	0.05 246	0.05 781	0.05 472	0.05 089	0.06 660	0.06 234	0.03 958	0.05 906	0.07 164	0.05 153	0.05 654	0.07 664	0.05 904	—
21 Poromera fordii	0.04 717	0.04 981	0.04 900	0.04 734	0.04 941	0.04 908	0.04 292	0.04 583	0.04 836	0.06 340	0.05 399	0.05 602	0.05 399	0.05 461	0.06 591	0.04 646	0.04 143	0.06 152	0.05 461	0.05 529	—
22 Pseuderemias smithi	0.06 599	0.06 073	0.05 402	0.05 318	0.06 040	0.05 600	0.05 304	0.05 713	0.05 278	0.04 772	0.06 530	0.06 734	0.03 893	0.06 404	0.07 534	0.04 837	0.05 462	0.07 785	0.05 713	0.04 147	0.05 839	—
23 Tropidosaura gularis	0.04 273	0.04 338	0.04 083	0.03 838	0.04 438	0.04 153	0.03 532	0.03 955	0.03 959	0.06 215	0.04 457	0.03 966	0.04 959	0.03 766	0.05 838	0.04 020	0.03 515	0.05 524	0.03 766	0.05 090	0.03 892	0.05 086	—

TAXONOMIC IMPLICATIONS

Our molecular datasets indicate Adolfus is polyphyletic (with weak support) with regard to Acanthodactylus, Gastropholis and Holaspis (Figs. 1–2), there is a six-codon deletion in the RAG1 gene for the lineage including A. alleni, A. africanus and A. jacksoni, and c-mos/RAG1 uncorrected p sequence divergence between the two well-supported Adolfus lineages is equal to or exceeds divergences noted for previously recognized lacertid genera (Table 2; Mayer & Pavlicev, 2007). Although our hypothesis tests that constrained the monophyly of Adolfus were not significant, these results are not surprising given the zero-length branches separating the lineages of Equatorial African lacertids. Because there are numerous mensural, meristic and qualitative differences between the well-established genera of Equatorial African lacertids (Table 3; Arnold, 1989a), and considerable taxonomic instability would be created by grouping this diverse assemblage of lizards into one genus, we recognize each well-supported lineage of Adolfus as a distinct genus. Accounts for both genera are provided below, and follow the format of Arnold et al. (2007).

Table 3.

Comparison of selected mensural, meristic and qualitative diagnostic characters for genera in the Equatorial African group of lacertid lizards. SVL/TL given as percentage data. Data are from this study, Arnold (1989b), Kroniger & in den Bosch (2001), Schmidt (1919) and Spawls et al. (2002). + = present, − = absent. — = data not available.

Character	Adolfus	Congolacerta gen. nov.	Gastropholis	Holaspis
Adult SVL (mm)	55–84	50–58	80–110	38–52
SVL/TL	49–60	44–52	42–45	71–93
Ventral Scale Count (transversely)	6	6	10–14	6
Femoral pores	11–19	11–16	13–15	18–24
Frontoparietal scales	+	+	+	−
Vertebral series of enlarged scales	−	−	−	+
Tail strongly depressed and fringed laterally	−	−	−	+
Tail prehensile	−	−	+	−
Ventrals keeled	−	−	+	−
Dorso-anterior border of quadrate bone	rounded	rounded	angular	rounded
Size of long free ribs immediately posterior to thoracic ribs	moderately elongated	very elongated	moderately elongated	very elongated
Posterior border of medial loop of clavicle	present and slender	present and thickened	present and slender	absent
Intramuscular portion of hemipenial armature	not deeply cleft	deeply cleft anteriorly	not deeply cleft	deeply cleft anteriorly
Shape of hemipenial clavulae	simple	complexly lobed	simple	simple
Female genital sinus	unlobed	bilobed	unlobed	unlobed
Habitat	forest and clearings, grassland	forest clearings, grassland	forest canopy	forest
Clutch size	3–5	—	5	2
Ventral coloration	yellow, orange, green or blue	yellow or unpigmented	yellow-green	orange to orange-gray

Our data also suggest species diversity within Adolfus sensu stricto is currently underestimated. The sequence divergence (cyt b) between the samples of A. alleni from the Aberdares and Mt. Kenya suggest these populations are not conspecific, and Arnold (1989a: table 2) provided mensural and meristic data that showed marked differences between populations from Mt. Kenya, Mt. Elgon, and the Aberdares. Loveridge (1957) did not recognize any of these populations as taxonomically distinct, but additional sampling is needed before taxonomic recognition of these populations would be warranted. Further study is also needed on the Arusha, Tanzania population of A. jacksoni, which has a color pattern that is noticeably different from populations in the Albertine Rift (see also Poblete, 2002; Spawls et al., 2002).

Adolfus

Sternfeld, 1912

Type species

Adolfus africanus (Boulenger, 1906) [=Adolfus fridericianus Sternfeld, 1912].

Synonymy

• -Algiroides Duméril & Bibron, 1839 (part); Boulenger, 1906. Proceedings of the Zoological Society of London 1906:570 [Algiroides africanus]; Barbour, 1914. Proceedings of the New England Zoological Club, Boston 4:97 [Algiroides alleni].
• -Lacerta Linnaeus, 1758 (part); Boulenger, 1899. Proceedings of the Zoological Society of London 1899:96 [Lacerta jacksoni]; Lönnberg in Sjöstedt, 1907. Wissenschaftliche Ergebnisse der Swedischen Zoologischen Expedition nach dem Kilimandjaro, dem Meru und den umgebenden Massai-steppen Deutsch-Ostafrikas 4:5 [Lacerta jacksoni kibonotensis]; Boulenger, 1920. Monograph of the Lacertidae. Vol. 1:295. [Lacerta jacksonii].

Content

Adolfus africanus (Boulenger, 1906); Adolfus alleni (Barbour, 1914); Adolfus jacksoni (Boulenger, 1899).

Distribution

Western Cameroon east to southern Sudan, Uganda, Kenya, and Tanzania, and south to northwestern Zambia (Köhler et al., 2003), with isolated montane populations in the Aberdare Mountains, Mt. Kenya and Mt. Elgon (Spawls et al., 2002).

Diagnosis

Several mensural, meristic and qualitative characters that diagnose Adolfus are shown in Tables 3–5, including: relatively large SVL (55–84 mm); dorso-anterior border of quadrate bone rounded; size of long free ribs immediately posterior to thoracic ribs moderately enlarged; posterior border of medial loop of clavicle present and slender; small post-femoral mite pockets absent (except in A. jacksoni); intramuscular portion of hemipenial armature not deeply cleft; shape of hemipenial clavulae simple; female genital sinus unlobed; habitat in forest, forest clearings and grasslands; clutch size 3–5; and ventral coloration yellow, blue, orange or green.

Table 5.

Meristic characters of adult species in the genera Adolfus and Congolacerta gen. nov. Data are averages ± one standard deviation, with ranges in parentheses. Abbreviations are explained in the Materials and Methods; m = male, f = female, n = unknown gender. Data for A. alleni are taken from single individuals from Mt. Elgon (Uganda, 1n) and the Aberdare Mountains (Kenya, 1m), which are likely not conspecific.

Characters	A. africanus (4 m, 3 f)	A. alleni (1n, 1m)	A. jacksoni (10m, 6f)	C. asukului (3 m, 1f)	C. vauereselli (6 m, 5 f)
CS	6	6, 5	6	6	6
FP	15.29 ± 1.11 (14–17)	11, 11	17.27 ± 1.28 (15–19)	13.25 ± 2.22 (11–16)	10.0 ± 1.00 (8–11)
SL	7.14 ± 0.38 (7–8)	6, 5	6.13 ± 0.34 (6–7)	7	6.36 ± 0.51 (6–7)
IL	6	4, 5	6	6	5.91 ± 0.30 (5–6)
SO	4	3, 4	4.06 ± 0.25 (4–5)	3.25 ± 0.50 (3–4)	4.09 ± 0.54 (3–5)
SC	6	5, 3	5.00 ± 0.37 (4–6)	4.75 ± 0.50 (4–5)	5.55 ± 0.82 (4–7)
SG	6.43 ± 0.79 (6–8)	0, 0	3.53 ± 1.06 (2–5)	3.75 ± 0.50 (3–4)	6.36 ± 1.21 (4–8)
ST	4.86 ± 0.90 (4–6)	2, 3	4.81 ± 0.91 (3–6)	4.50 ± 1.00 (3–5)	3.20 ± 0.63 (2–4)
ADS	49.29 ± 10.03 (36–60)	31, 35	61.06 ± 6.61 (51–74)	63.00 ± 4.36 (60–68)	63.36 ± 7.55 (47–73)
PDS	25.43 ± 2.82 (20–28)	19, 22	40.06 ± 2.08 (37–44)	34.25 ± 2.50 (31–37)	39.82 ± 4.14 (32–44)
DSR	24.14 ± 1.22 (23–26)	19, 22	40.25 ± 2.38 (35–44)	31.25 ± 2.22 (28–33)	39.64 ± 5.41 (31–48)
DSN	48.00 ± 3.46 (42–53)	48, 46	95.38 ± 4.51 (90–105)	81.00 ± 5.42 (73–85)	73.18 ± 9.39 (54–84)
VR	6	6, 6	6	6	6
VN	23.14 ± 0.90 (22–24)	26, 25	27.34 ± 2.37 (24.5–31.5)	25.88 ± 1.65 (24–28)	22.55 ± 0.96 (21.5–24.0)
CDS- 11th scale	15.43 ± 0.98 (14–16)	21, 21	24.31 ± 1.25 (22–27)	23.25 ± 1.71 (21–25)	19.67 ± 2.50 (16–24)
CDS- 15th scale	15.14 ± 1.07 (14–16)	21, 21	24.00 ± 1.27 (21–26)	23.50 ± 1.73 (21–25)	18.13 ± 1.81 (16–21)
SDF1	8.14 ± 1.07 (7–10)	7, 6	8.25 ± 0.58 (7–9)	6.50 ± 0.58 (6–7)	7.64 ± 0.67 (7–9)
SDF2	13.17 ± 0.98 (12–14)	10, 9	13.31 ± 0.87 (12–15)	10.75 ± 0.50 (10–11)	11.55 ± 0.82 (11–13)
SDF3	16.71 ± 1.38 (15–18)	14, 12	18.00 ± 1.16 (16–20)	14.33 ± 1.16 (13–15)	15.00 ± 1.55 (13–17)
SDF4	16.71 ± 0.76 (16–18)	12, 12	19.44 ± 1.37 (17–22)	15.50 ± 0.58 (15–16)	16.91 ± 0.94 (16–19)
SDF5	11.57 ± 0.54 (11–12)	8, 9	12.38 ± 1.03 (11–14)	10.00 ± 1.41 (9–12)	10.46 ± 0.69 (9–11)
SDT1	8.29 ± 0.76 (7–9)	8, 6	8.69 ± 1.08 (6–10)	7	8.09 ± 0.54 (7–9)
SDT2	12.67 ± 1.03 (11–14)	11, 10	13.44 ± 0.89 (11–15)	10.75 ± 0.50 (10–11)	11.46 ± 1.13 (10–13)
SDT3	16.00 ± 0.58 (15–17)	15, −	18.79 ± 1.37 (17–21)	15.25 ± 0.50 (15–16)	15.27 ± 1.49 (12–17)
SDT4	19.00 ± 0.58 (18–20)	19, 18	23.64 ± 1.69 (21–27)	19.25 ± 0.50 (19–20)	20.18 ± 1.99 (17–22)
SDT5	13.71 ± 0.76 (13–15)	12, 11	16.13 ± 0.92 (15–17)	12.50 ± 0.58 (12–13)	13.73 ± 1.27 (12–16)

Description

Size and proportions

Relatively large member of the Equatorial African group of lizards (55–84 mm SVL), with no sexual dimorphism and a long tail (SVL/TL = 49–60%; Tables 4–5) that is cylindrical without lateral fringes.

Table 4.

Measurements (in mm) of adult species in the genera Adolfus and Congolacerta gen. nov. Data are averages ± one standard deviation, with ranges in parentheses. Abbreviations and measurements are explained in the Materials and Methods. SVL/TL given as percentage data; m = adult male, f = adult female, n = unknown gender. Data for A. alleni are taken from single individuals from Mt. Elgon (Uganda, 1n) and the Aberdare Mountains (Kenya, 1m), which are likely not conspecific.

Characters	A. africanus (4 m, 3 f)	A. alleni (1n, 1m)	A. jacksoni (10 m, 6f)	C. asukului (3 m, 1f)	C. vauereselli (6 m, 5 f)
SVL (m)	56.95 ± 1.13 (55.7–58.4)	48.0, 46.0	70.46 ± 6.42 (64.0–84.3)	55.40 ± 2.52 (53.7–58.3)	53.42 ± 2.15 (50.0–55.5)
SVL (f)	60.90 ± 0.95 (59.9–61.8)	—	70.28 ± 5.46 (62.3–76.6)	51.9	55.46 ± 3.89 (50.0–60.4)
TL (m)	104.6, 113.6	—	101.3, 112.6	111.4	114.74 ± 3.89 (50.0–60.4)
TL (f)	103.0	—	—	—	112.10 ± 4.10 (109.2–115.0)
SVL/TL	54.93 ± 5.55 (49.0–60.0)	—	56.8, 63.3	52.3	46.95 ± 2.66 (44.4–51.7)
HL	14.36 ± 0.61 (13.4–15.1)	10.0, 10.3	16.33 ± 2.07 (12.4–20.2)	11.35 ± 2.53 (8.4–14.0)	12.61 ± 0.82 (11.1–13.8)
HW	8.74 ± 0.44 (8.2–9.5)	6.4, 6.4	10.63 ± 1.61 (7.8–14.2)	8.18 ± 1.43 (6.5–10.0)	8.12 ± 1.41 (6.8–11.9)
HH	6.21 ± 0.20 (6.0–6.6)	5.3, 5.6	7.19 ± 1.24 (5.6–9.4)	8.18 ± 3.12 (4.8–11.3)	5.57 ± 0.45 (5.0–6.5)
SKL	14.06 ± 0.57 (13.1–14.7)	9.7, 11.0	16.75 ± 2.32 (12.5–20.5)	12.58 ± 1.48 (10.4–13.7)	12.57 ± 0.77 (11.2–13.8)
SEL	6.31 ± 0.43 (5.9–7.0)	3.7, 4.4	6.50 ± 0.83 (5.3–7.7)	4.90 ± 0.62 (4.0–5.4)	5.51 ± 0.47 (4.7–6.1)
ML	11.14 ± 0.48 (10.7–11.8)	8.4, 8.9	12.10 ± 1.45 (9.3–14.1)	10.33 ± 1.46 (8.2–11.5)	10.11 ± 0.74 (8.7–11.0)
SAL	20.76 ± 1.62 (19.0–23.7)	17.9, 17.8	25.74 ± 3.69 (19.5–32.0)	19.47 ± 2.63 (16.2–22.1)	19.98 ± 1.90 (16.4–22.5)
AGD	26.39 ± 2.64 (23.0–30.4)	23.3, 22.7	32.10 ± 4.22 (25.9–40.2)	27.83 ± 0.95 (26.8–28.9)	24.91 ± 2.38 (21.8–29.5)
HML	7.47 ± 0.85 (6.6–8.9)	4.8, 4.5	7.53 ± 1.11 (5.5–9.2)	5.48 ± 0.76 (4.4–6.1)	7.00 ± 1.31 (6.0–10.4)
RUL	7.84 ± 0.22 (7.5–8.1)	4.9, 4.6	7.73 ± 0.97 (6.0–9.1)	5.73 ± 1.17 (4.2–6.9)	7.99 ± 1.35 (6.2–10.7)
FL	10.06 ± 0.51 (9.3–11.0)	5.9, 6.4	10.11 ± 0.96 (8.6–11.9)	6.93 ± 1.10 (5.3–7.7)	8.69 ± 1.10 (7.2–10.6)
TFL	10.20 ± 0.51 (9.3–10.9)	6.0, 6.2	10.01 ± 1.27 (7.6–11.7)	7.10 ± 1.29 (5.2–8.1)	9.06 ± 0.88 (8.2–10.3)
LTL	10.40 ± 0.84 (9.3–11.7)	6.4, 6.5	10.51 ± 1.10 (8.6–12.3)	7.78 ± 1.09 (6.3–8.9)	8.76 ± 0.99 (6.8–9.8)

Skull

Premaxilla without anterior boss; postfrontal and postorbital bones fused; shape of squamosal bone slender; squamosal and parietal not in contact; dorso-anterior border of quadrate bone rounded; temporal osteoderms absent (except in A. alleni, which is variable); maxilla not extending to coronoid notch; and 14 scleral ossicles in each eye (Arnold, 1989a).

Post-cranial skeleton

Average number of presacral vertebrae in males 26–27 (except A. africanus, which is 25 or less); 7–9 long free dorsal ribs immediately posterior to thoracic ribs (except A. africanus, which is 6–7); moderately elongated long free dorsal ribs immediately posterior to thoracic ribs; posterior border of medial loop of clavicle present and slender; and transverse process of anterior autotomic caudal vertebrae directed roughly laterally (Arnold, 1989a).

Scaling

Contact between postnasal and supranasal scales below level of nostril absent; two loreal scales on each side (except A. alleni, which has one); supraciliary granules present (except A. alleni); lower eyelid opaque and covered with relatively small scales; parietal scales without lateral corner erosion; temporal scaling relatively fine (except A. alleni, which is very coarse, with 13 or fewer scales on each side, excluding the supratemporals and tympanic); keeling on temporal scales absent (A. alleni), present (A. africanus) or variable (A. jacksoni); keeling on collar scales absent (except A. africanus, which is variable); granules beneath collar scattered or absent (except A. jacksoni, which has many); dorsal scales more or less uniform in size (except A. africanus, which has flank scales that are distinctly smaller than the mid-dorsals); micro-ornamentation of dorsal scales smooth (except for A. africanus, which has pustullate scales with minute tubercles); flank scales in close contact; six or eight longitudinal rows of ventral body scales (except A. africanus, which has four complete rows and an outer row on each side that is strongly reduced anteriorly); keeling on ventrals absent (except A. africanus, which has keeling on the outer longitudinal row); preanal scale entire and without keeling; no keeling on scales beneath limbs; row of femoral pores long, extending almost to knee (except A. africanus, which has a shortened row of femoral pores, well separated from the knee); scales bearing femoral pores not or only slightly projecting, close together in males; hind toes without fringes; no pad of spinous scales on dorsum of tail base (Arnold, 1989a). In contrast to the latter author, we observed gular folds (as indicated by a heavy crease between the ear openings on the throat of adult animals) in A. jacksoni; the character was noted as absent in A. alleni and A. jacksoni, and variable in A. africanus by Arnold (1989a).

Coloring

Adolfus africanus: the entire head is metallic copper bronze with a continuous mid-dorsal band of the same color and width of the head continuing to the end of the tail. Within the mid-dorsal band are numerous randomly distributed black spots, usually beginning near the origin of the forelimbs and extending slightly beyond the base of the tail. A longitudinal series of white round spots border the mid-dorsal metallic band laterally; these coalesce into thin narrow stripes on the tail. The lateral sides of the body have dark brown bands originating on the side of the head and extending posteriorly onto the tail; some specimens have additional, diffuse rounded white spots aligned along the lower edge of the dark lateral band. Venter immaculate lime green. Adolfus alleni: ground color brown or olive, with a broad or fine dark vertebral stripe. Two black-edged, lime-green or red-brown dorsolateral stripes extend from the posterior edge of the parietals to about the hind limb insertions, and may continue as brown lines onto the tail. The lateral sides of the body are rufous or light brown; the belly varies from orange or orange-pink to blue. Adolfus jacksoni: brown to olive on the dorsum of the head, with a continuous mid-dorsal band of the same color (occasionally light green) and width of the head continuing to the end of the tail. Within the band are randomly scattered black spots or oblique black dashes. The lateral sides of the body are much darker than the dorsum, usually brown but sometimes black, and usually contain several series of white or blue, black-edged ocelli, the uppermost and most lateral of which are usually arranged in longitudinal rows and may comprise scattered blue and black scales. The venter is sometimes spotted but more frequently immaculate, and varies from yellow to dull blue (Spawls et al., 2002), or bright orange in breeding males from Tanzania (WRB, pers. obs.). Poblette (2002) described a Kenyan specimen with an “army green” dorsum with black, irregular medial dots and flanks with black lateral stripes that were spotted with a luminescent cyan color.

Distinctive internal features

Tongue surface mainly squamate; tongue color in alcohol dark; a continuous ulnar nerve present but connected to the brachial trunk by a bridge in the lower arm (except A. africanus, which has a variable ulnar nerve pattern); exit of oviducts into genital sinus dorsal; female genital sinus unlobed (Arnold, 1989a).

Hemipenis

Size relatively large; intramuscular portion of hemipenial armature not deeply cleft; medial side of hemipenial armature not reduced; size of hemipenial clavulae large; shape of hemipenial clavulae simple (Arnold, 1989a).

Ecology

Adolfus africanus is known from primary Guineo-Congolean forest (580–2200 m) and has been observed basking in dappled sunlight on fallen tree limbs, trunks and exposed roots within a few meters of ground clearings in forest (only a few were observed on tree trunks above 3 m from the ground), suggesting this species is primarily an inhabitant of undergrowth (Spawls et al., 2002; Köhler et al., 2003). It has been collected in highly disturbed forest in northeastern DRC (EG, CK and MMA pers. obs.) and Kenya (Köhler et al., 2003). Adolfus alleni is known from alpine moorland, heather and Hagenia-Hypericum zones from 2,700–4,500 m, and is more terrestrial than other members of the genus, living in tussock grass and open patches in between (Spawls et al., 2002). Adolfus jacksoni is known from clearings, forest edges, gallery forest, and disturbed habitats, even occurring in the middle of the city of Bukavu (DRC) on slopes that have been cleared of forest for centuries (EG, CK and MMA, pers. obs., Schaller, 1964), and in suburban gardens in Arusha, Tanzania (WRB, pers. obs). The species has been recorded from 450–3,000 m (Spawls et al., 2002).

Reproduction

No reproductive data are available for Adolfus africanus or A. alleni, but A. jacksoni has been observed nesting communally in crevices on exposed vertical road cut walls, and lays clutches of 3–5 eggs (Spawls et al., 2002). Goldberg (2009) confirmed the range of clutch size for A. jacksoni as 3–5 eggs (mean = 4.1 ± 0.90 standard deviation), noted reproductively active males and females at opposite ends of the year (February–March and September), and documented evidence of multiple clutches in females.

Remarks

Several morphological features (e.g., osteology, hemipenis) are shared with Gastropholis, but not other Equatorial African genera (Table 3), lending support for the weakly supported placement of Gastropholis as sister to Adolfus in our phylogenetic analyses (Figs. 1–2).

Congolacerta

Greenbaum, Villanueva, Kusamba, Aristote & Branch gen. nov.

Type species

Lacerta vauereselli Tornier, 1902.

Etymology

A feminine name derived from Democratic Republic of the Congo, where the genus occurs along most of the eastern montane border (Albertine Rift), and lacerta, a lizard.

Synonymy

• -Lacerta Linnaeus, 1758 (part); Tornier, 1902. Zoologische Anzeiger 25:701. [Lacerta vauereselli].
• -Algiroides Duméril & Bibron, 1839 (part); Peracca, 1917. Atti della Reale Accademia delle Scienze di Torino 52:351 [Algiroides boulengeri].
• -Adolfus Sternfeld, 1912 (part); Arnold, 1973. Bulletin of the British Museum (Natural History), Zoology 25:357 [Adolfus vauereselli].

Content

Congolacerta asukului sp. nov. (described below); C. vauereselli (Tornier, 1902).

Distribution

Occurs from the Blue Mountains (west of Lake Albert in DRC) along the Albertine Rift and its foothills through Uganda, Rwanda and Tanzania as far south as the Kabobo Plateau at the border of South Kivu and Katanga Provinces, DRC (Spawls et al., 2002; Appendix 2).

Diagnosis

Several mensural, meristic and qualitative characters that diagnose Congolacerta are shown in Tables 3–5, including: modest SVL (50–58 mm); dorso-anterior border of quadrate bone rounded; size of long free ribs immediately posterior to thoracic ribs very elongated; posterior border of medial loop of clavicle present and thickened; small to very small postfemoral mite pockets present (Arnold, 1986b); intramuscular portion of hemipenial armature deeply cleft anteriorly; shape of hemipenial clavulae complexly lobed; female genital sinus bilobed; habitat forest clearings and grasslands; and ventral coloration usually unpigmented (C. vauereselli) or yellow with black or brown blotches (C. asukului).

Description

Size and proportions

Relatively modest-sized member of the Equatorial African group of lizards (50–58 mm SVL), with no sexual dimorphism and a modest-sized tail (SVL/TL = 44–52%; Tables 4–5) that is cylindrical without lateral fringes.

Skull

Congolacerta vauereselli premaxilla without anterior boss; postfrontal and postorbital bones fused; shape of squamosal bone slender; squamosal and parietal not in contact; dorso-anterior border of quadrate bone rounded; temporal osteoderms absent; maxilla not extending to coronoid notch; and 14 scleral ossicles in each eye (Arnold, 1989a).

Post-cranial skeleton

Average number of presacral vertebrae in males 25 or less (both species); Congolacerta vauereselli has 6–7 long free dorsal ribs immediately posterior to thoracic ribs; very elongated long free dorsal ribs immediately posterior to thoracic ribs, about twice the length of other free dorsal ribs; posterior border of medial loop of clavicle present and thickened; and transverse process of anterior autotomic caudal vertebrae directed roughly laterally (Arnold, 1989a).

Scaling

Contact between postnasal and supranasal scales below level of nostril absent; two loreal scales on each side; supraciliary granules present; lower eyelid opaque and covered with relatively small scales; parietal scales without lateral corner erosion; temporal scaling relatively fine; keeling on temporal scales variable, but usually absent; keeling on collar scales absent; granules beneath collar scattered or absent; dorsal scales somewhat enlarged; micro-ornamentation of dorsal scales smooth; flank scales in close contact; four complete rows of ventral body scales and an outer row on each side that is strongly reduced anteriorly; keeling on ventrals absent; preanal scale entire and without keeling; no keeling on scales beneath limbs; row of femoral pores long, extending almost to knee (C. asukului) or shortened row of femoral pores, well separated from the knee (C. vauereselli); scales bearing femoral pores not or only slightly projecting, close together in males; hind toes without fringes; no pad of spinous scales on dorsum of tail base (Arnold, 1989a). In contrast to the latter author, we did not observe a gular fold on any specimens of C. vauereselli, and only faint indications of a gular fold on three adult specimens of C. asukului.

Coloring

Congolacerta vauereselli: the dorsum of the head is light yellow to copper bronze with a continuous mid-dorsal band of the same color and width of the head continuing to the end of the tail. Within the mid-dorsal band are small dark brown to black spots, sometimes forming a vertebral stripe. The lateral sides of the body are reddish brown, edged in black above, with one or two series of white, black-edged ocellar spots. A cream or white streak extends from the cheek to the side of the neck and passes through the ear opening. Venter usually immaculate and unpigmented. Coloring of C. asukului is generally similar to that of C. vauereselli (one major exception is yellow ventral pigmentation with black or brown blotches), and details are given in the species description below.

Distinctive internal features

Congolacerta vauereselli tongue surface mainly squamate; tongue color in alcohol dark; a “Varanidae” ulnar nerve pattern with no continuous independent ulnar nerve and all fibers to lower limb passing through the branchial trunk; exit of oviducts into genital sinus dorsal; female genital sinus bilobed (Arnold, 1989a).

Hemipenis

Congolacerta vauereselli size relatively large; intramuscular portion of hemipenial armature very deeply cleft anteriorly; medial side of hemipenial armature not reduced; size of hemipenial clavulae large; shape of hemipenial clavulae complexly lobed (Arnold, 1989a).

Ecology

Congolacerta vauereselli is found in clearings and openings within Guineo-Congolian forests from 1,000–2,675 m. Little is known of its natural history, but based on observations made in Bwindi National Park (Uganda), Spawls et al. (2002) suggested it is likely similar to Adolfus africanus. Congolacerta asukului is known from high elevations (> 2,650 m) grasslands of the Itombwe Plateau, and has been found in small burrows among tussocks of grass.

Reproduction

No reproductive data are available for either species of Congolacerta.

Remarks

Several mensural, meristic, qualitative and molecular divergence characters distinguish the Itombwe population of Congolacerta from its congener C. vauereselli. The Itombwe population is described as a new species below.

Congolacerta asukului

Greenbaum, Villanueva, Kusamba, Aristote & Branch sp. nov.

Asukulu’s grass lizard

Holotype

UTEP 20263 (field no. EBG 2025, Figs. 3 A,B, 4), an adult male, from footpath south of Rurambo village, Itombwe Plateau, South Kivu Province (SKP), DRC (02.99437°S, 28.87620°E, 2,876 m; see Figs. 6–7). Collected ca. 08:00 hrs on 23 May 2009 by MMA, EG, CK, Wandege Mastaki Monigan, Maurice Luhumio and Asukulu M’Mema.

Figure 3.

Photographs of Congolacerta in life. Dorsal (A) and ventral (B) view of C. asukului holotype UTEP 20263 (adult male, 58.3 mm SVL), dorsal view (C) of C. asukului paratype UTEP 20265 (adult male, 53.7 mm SVL), dorsal view (D) of C. asukului paratype UTEP 20267 (subadult male, 42.7 mm SVL), and dorsal (E) and ventral (F) view of C. vauereselli UTEP 20289 (adult male, 54.4 mm SVL).

Figure 4.

Photographs of the holotype of Congolacerta asukului (UTEP 20263, adult male, 58.3 mm SVL) after preservation. Dorsal (A) and ventral (B) view of whole specimen, lateral (C), dorsal (D) and ventral (E) view of head, and ventral view of cloacal region (F) illustrating femoral pores.

Figure 6.

Map of the Itombwe Plateau, showing collection localities for Congolacerta asukului sp. nov. (open squares). The type locality is indicated by a star symbol.

Figure 7.

Photograph of the type locality of Congolacerta asukului sp. nov., showing grassland habitat with rocky outcrops.

Paratopotype

UTEP 20264 (field no. EBG 2028), a subadult male, with same date, locality, collectors, and circumstances of capture as holotype.

Other paratypes

UTEP 20265 (field no. EBG 2082, Fig. 3 C), an adult male, collected by MMA, EG and CK 25 May 2009 at Komesha village, Itombwe Plateau, SKP, DRC (03.0870°S, 28.8101°E, 2,891 m); UTEP 20266 (field no. EBG 2114), an adult female, collected by MMA, EG and CK 26 May 2009 at Mugegema village, Itombwe Plateau, SKP, DRC (03.06940°S, 28.76813°E, 2,765 m); UTEP 20267–68 (field nos. EBG 1715–16), one adult male and one subadult male, collected by EG, WMM, MMA, CK, ML, and AM 30 June 2008 at Ruhuha, Itombwe Plateau, SKP, DRC (03.37871°S, 29.01293°E, 2,886 m).

Diagnosis

Congolacerta asukului can be distinguished from all other species in the Equatorial African group of lacertids by the following combination of characters: (1) medium body size (SVL 53.7–58.3 for adult males; 51.9 in one adult female); (2) dorsum brown, rusty brown or tan with several dark brown to black blotches forming a vertebral line from occipital to first quarter of tail, and a dark brown line with cream or grayish white blotches extending from lateral side of rostral through eye and flanks to lateral side of tail; (3) moderate numbers of femoral pores (11–16); (4) low numbers of supraciliary granules (3–4); (5) moderate numbers of dorsal scale rows at midbody (28–33); (6) moderate numbers of dorsal scales in a longitudinal row from occipital to posterior insertion of hind limb (73–85); (7) high numbers of ventral scales from collar to preanal (24–28); (8) high numbers of caudal scale rows at 15^th scale (21–25); (9) smooth dorsal scales; and (10) yellow ventral coloration with black or brown blotches.

Differential diagnosis from similar species

Because the genera Adolfus and Congolacerta have similar external morphology, the new species is diagnosed from all species in each genus. Congolacerta asukului differs from its partially sympatric and phenotypically similar congener C. vauereselli (Fig. 3 E,F) by a higher SVL/TL ratio of 52.3 (vs. 44.4–51.7), a smaller HML (4.4–6.1 vs. 6.0–10.4), a smaller TFL (5.2–8.1 vs. 8.2–10.3), a higher number of femoral pores (11–16 vs. 8–11), a smaller number of supraciliary granules (3–4 vs. 4–8; Fig. 5), a smaller number of dorsal scale rows at midbody (28–33 vs. 31–48), a higher number of VN (24–28 vs. 21.5–24), a higher number of caudal scales at the 15^th scale row (21–25 vs. 16–21), a smaller number of subdigital lamellae on digits 1 (6–7 vs. 7–9), 2 (10–11 vs. 11–13), and 4 (15–16 vs. 16–19), dorsal scale keeling (smooth vs. keeled), ventral pigmentation (yellow with black or brown blotches vs. usually unpigmented), and habitat (montane grassland vs. forest clearings and openings). Algiroides boulengeri, Peracca, 1917, described from Fort Portal, Uganda (east of Ruwenzori Mountains) was synonymized with C. vauereselli by Loveridge (1957:229), with which it shares keeled dorsal scales and a strip of metallic bronze in the middle 7–8 longitudinal scale rows (Peracca, 1917), and is clearly not conspecific with Congolacerta asukului. Most examined specimens of C. vauereselli have unpigmented venters, but UTEP 20295 (adult male) from the Kabobo Plateau (most basal population of this species in all analyses, Figs. 1–2) has a yellow venter.

Figure 5.

Dorsal views of the heads of Congolacerta asukului sp. nov. (holotype, UTEP 20263) and C. vauereselli (UTEP 20291). Supraciliary granules are shaded in gray. Scale bars = 1 mm.

The new species differs from Adolfus africanus in having a smaller SEL (4.0–5.4 vs. 5.9–7.0), a smaller HML (4.4–6.1 vs. 6.6–8.9), a smaller RUL (4.2–6.9 vs. 7.5–8.1), a smaller FL (5.3–7.7 vs. 9.3–11.0), a smaller TFL (5.2–8.1 vs. 9.3–10.9), a smaller LTL (6.3–8.9 vs. 9.3–11.7), a smaller number of supraciliaries (4–5 vs. 6), a smaller number of supraciliary granules (3–4 vs. 6–8), a higher number of dorsal scale rows at midbody (28–33 vs. 23–26), a higher number of DSN (73–85 vs. 42–53), a higher number of VN (24–28 vs. 22–24), a higher number of caudal scales at the 11^th and 15^th scale rows (21–25 vs. 14–16), fewer numbers of subdigital lamellae for each digit (Table 5), dorsal scale keeling (smooth vs. strongly keeled), ventral coloration (yellow with black or brown blotches vs. green), and habitat (montane grassland vs. forest clearings and openings).

The new species differs from our two examined specimens of Adolfus alleni in having two loreals (vs. one), supraciliary granules (present vs. absent), dorsal scale shape (rhombic vs. lanceolate), dorsal scale keeling (smooth vs. strongly keeled), granules beneath the collar (present vs. absent), and ventral coloration (yellow with black or brown blotches vs. orange, orange-pink or blue). The new species also has larger numbers of anterior, posterior and mid-dorsal scale rows, femoral pores, and subdigital lamellae under Toe IV (Table 5). Arnold (1989a: table 2) noted several interesting mensural, meristic and qualitative differences among three allopatric populations of A. alleni, which suggests that our two samples from Mt. Elgon (Uganda) and the Aberdare Mountains (Kenya) are likely not conspecific (Appendix 2).

The new species differs from Adolfus jacksoni in having a smaller SVL (53.7–58.3 vs. 64.0–84.3), smaller SEL (4.0–5.4 vs. 5.3–7.7), smaller FL (5.3–7.7 vs. 8.6–11.9), a smaller number of femoral pores (11–16 vs. 15–19), a smaller number of PDS (31–37 vs. 37–44), a smaller number of DSR (28–33 vs. 35–44), a smaller number of DSN (73–85 vs. 90–105), a smaller number of subdigital lamellae of several fingers and toes (Table 5), flank color pattern (spots vs. ocelli), and habitat (montane grassland vs. forest clearings and openings). It is not known to tolerate anthropogenically disturbed habitats.

Description of holotype

Measurements of the holotype are provided in Table 6. Rostral separated from frontonasal by supranasals; nostril surrounded by supranasal, postnasal and first supralabial; supralabials seven (fourth largest) and infralabials six on each side; supraoculars three on each side, the posteriormost ones much smaller than others; supraciliaries five on each side, first supraciliary largest and continuing to dorsum of head to contact first supraocular (likely fused to former first supraocular), relative lengths 1 > 2 > 5 > 4 > 3; second supraciliary in contact with first supraocular, posterior three supraciliaries separated from posterior supraoculars by two (left) or four (right) supraciliary granules; postnasal one, followed by two loreals at each side, anterior loreal smaller than posterior one; two prefrontals separated by medial contact of frontal and frontonasal; frontal hexagonal, contacting supranasals, postnasals, anteior loreals, prefrontals and frontal; frontoparietals two and connected; parietals two, separated by two interparietals (anterior interparietal largest) and occipital; supratemporals five on each side, the first one largest; temporal scales non-imbricate, much larger than scales posterior to ear opening; six pairs of chin shields, anteriormost three pairs in contact medially; faint indication of gular fold; collar with seven plates, granules present beneath collar; dorsal scales on body enlarged, imbricate, smooth and rhombic, extending anteriorly beyond forelimbs on to neck, slightly larger than lateral scales at midbody, much larger than lateral scales near limb insertions; 61 anterior dorsal scale rows, 37 posterior dorsal scale rows, 32 scale rows at midbody; 85 scales counted longitudinally from occipital to the posterior margin of hind limb on middle-left and middle-right rows, respectively; lateral body scales at midbody smooth and rhombic, arranged in disorder; lateral body scales at limb insertions small, smooth and granular, arranged in disorder; small post-femoral mite pockets present (containing larvae of trombiculid mites); ventral scales rectangular, smooth, in six longitudinal rows at midbody, median and outer longitudinal rows smaller than others, outermost rows incomplete and smooth; 26 (left) and 25 (right) scales counted longitudinally from collars to preanal on middlemost two rows; preanal single, ovoid, enlarged and smooth; 14 femoral pores on each side; scales on anterior and dorsal surfaces of forelimbs enlarged, smooth and imbricate; scales on posterior and ventral surfaces of forelimbs mostly small and granular, with two rows of enlarged, smooth and imbricate scales; two rows of enlarged, smooth and imbricate scales on antero-ventral side of hind limbs, the other areas with small, smooth granular scales; relative lengths of appressed fingers IV > III > II = V > I; subdigital lamellae seven (left) – seven (right), 11–11, 14–15, 16–16, 10–9 on fingers I, II, III, IV and V, respectively; relative lengths of appressed toes IV > III > V > II > I; subdigital lamellae eight (left) – seven (right), 11–11, 15–16, 19–19, 13–13 on toes I, II, III, IV and V, respectively; tail long (191% of SVL) and complete, covered with strongly keeled scales on lateral and dorsal sides, in 33 rows at base, decreased to 24 rows at 15^th scale.

Table 6.

Continuous (mm) and meristic variation among six specimens of Congolacerta asukului sp. nov Abbreviations and measurements are explained in the Materials and Methods; m = adult male, f = adult female, s = subadult male, SD = standard deviation. Tail length (TL) data is not provided for specimens with broken or regenerated tails.

Character	EBG 2025, m	EBG 1716, m	EBG 2082, m	EBG 2114, f	EBG 1715, s	EBG 2028, s	Average	SD
SVL	58.3	54.2	53.7	51.9	42.7	47.2	51.33	5.56
TL	111.4	—	—	—	—	—	—	—
HL	14.0	12.8	8.4	10.2	9.9	11.3	11.10	2.04
HW	10.0	8.0	8.2	6.5	6.3	6.9	7.65	1.39
SAL	22.1	21.0	18.6	16.2	15.6	16.8	18.38	2.67
AGD	28.3	27.3	26.8	28.9	20.4	23.6	25.88	3.26
HML	6.1	5.5	5.9	4.4	4.9	4.5	5.22	0.72
RUL	6.9	5.5	6.3	4.2	4.1	5.1	5.35	1.12
FL	7.3	7.4	7.7	5.3	5.6	5.6	6.48	1.09
TFL	8.1	7.6	7.5	5.2	6.6	6.8	6.97	1.03
FP	14	16	12	11	15	13	13.50	1.87
SO	3	4	3	3	4	3	3.33	0.52
SC	5	5	5	4	6	4	4.83	0.75
SG	4	4	3	4	1	3	3.17	1.17
ST	5	5	5	3	4	5	4.50	0.84
DSR	32	32	28	33	32	32	31.50	1.76
DSN	85	83	73	83	79	82	80.83	4.31
VN	25.5	26	24	28	23	24.5	25.17	1.75
SDF4	16	15	15	14	15	16	15.17	0.75
SDT4	19	19	20	19	19	20	19.33	0.52

Coloration in life

From photographs of holotype before preservation (Fig. 3A,B). Dorsal ground color brown to rusty brown with several dark brown to black blotches on the head scales, mid-dorsal area (forming a vertebral line from occipital to first ¼ of tail), limbs and tail. A narrow line of cream to white spots forms a border between the dorsum and flanks from the parietals to the insertion of the hind limb. A dark brown line with cream blotches extends from the lateral side of the rostral through the eye and flanks to the lateral side of the tail. Lateral side of snout and neck (below dark brown line) cream with dark brown blotches. Chin shields white, remainder of venter light yellow anteriorly, becoming increasingly darker yellow posteriorly, with dark brown to black blotches on the lateral sides of the venter and limbs. Coloration in preservative (70% ethanol) similar to coloration in life.

Variation

Variation of mensural and meristic data in the paratypes of Congolacerta asukului are shown in Table 6. The holotype is the only specimen with separated prefrontals; the prefrontals are in broad contact in UTEP 20267–68 and 20266, and the prefrontals are in narrow contact in EBG 2028 and 2082. Three specimens (UTEP 20265–67) have only one interparietal; the interparietal in UTEP 20266 is unusually elongate, and is about 2.5 times longer than wide. Coloration in life of UTEP 20265 (Fig. 3C) differed from the holotype in having a tan dorsal ground color, and grayish white flanks below the dark brown lateral stripe. Coloration in life of UTEP 20267 (Fig. 3D) differed from the holotype in having rusty brown head plates, and an olive brown ground color on the remainder of the dorsum.

Ecology and natural history

Congolacerta asukului is a diurnal species that occurs in high-elevation (> 2,650 m) grasslands (often near rocky outcrops) of the Itombwe Plateau (Figs. 6–7) in the Albertine Rift Montane Forest ecoregion as defined by Burgess et al. (2004). At least two individuals were observed basking on rocks, and one individual was captured after it retreated to a small burrow among tussocks of grass. Laurent (1964) described this habitat as high-elevation meadows that are common between swamps, rivers and subalpine scrub forests, which are dominated by tree heathers and other Ericacea (Doumenge, 1998).

Etymology

The new species is named in honor of Itombwe native Asukulu M’Mema, an aspiring zoologist and conservationist who was our colleague and guide through the Itombwe Plateau in 2008 and 2009, when all the known specimens of C. asukului were collected. Asukulu was killed by militia during his investigation into the killing of a gorilla on August 22, 2009 in Mulombozi village (on the route between Miki and Mwenga) in the western slopes of the Itombwe Plateau. For his service to conservation in Africa, he was posthumously honored with the Medail de Bravoure from the Alexander Abraham Foundation in 2010.

Conservation

Congolacerta asukului is currently known from four localities, which have a total extent of occurrence of about 550 square kilometers. Because of mining activity, widespread cattle/goat grazing (EG, CK and MMA, pers. obs.) and dry-season burning of grasslands for agriculture (CK, pers. obs.), the area, extent and quality of the grassland habitat is declining. Applying the IUCN Red List criteria to C. asukului results in a classification of endangered, EN B1ab(iii).

DISCUSSION

Using the Bayesian phylogenetic analysis program TreeTime, Hipsley et al. (2009) estimated that the common ancestor of Eremiadini lacertids dispersed to northwestern Africa via small island chains from Europe in the mid- to late Eocene, substantially earlier than the previous estimates of ca. 14 Myr by Pavlicev & Mayer (2009). Separation of the “Saharo-Eurasian” and “Ethiopian” lineages (equivalent to clades B₁ and B₂ of Mayer & Pavlicev, 2007) occurred shortly thereafter. Given the pattern we recovered for the Equatorial African group of lacertids near the base of the Eremiadini clade, it is likely that the genera Adolfus, Congolacerta, Gastropholis and Holaspis diverged from each other shortly after the group’s arrival in Central Africa, either in the late Eocene or the Miocene. The Miocene is a relatively dry period marked by expansion of grasslands (Lovett et al., 2005), and is synchronous with orogeny of the Albertine Rift, which began forming via doming in the early Miocene (Vande weghe, 2004; Partridge, 2010).

Because Congolacerta asukului is known from a small area (550 km²) at the highest elevations of the Itombwe Plateau, potential negative effects from global warming are of paramount concern. In general, the African continent warmed by 0.5° C since 1900, but global climate models for future temperature and precipitation trends are problematic (Hulme et al., 2005). Hernes et al. (1995) and Ringius et al. (1996) constructed climate change scenarios that predicted equatorial African countries would warm by 1.4° C by the 2050’s. Hulme et al. (2001) also constructed climate change scenarios that predicted a 0.2–0.5° C/decade increase in temperature, with the least amount of warming in equatorial latitudes and coastal environments. These authors examined the inter-model range of their global climate models to assess their levels of agreement, and found the smallest range over northern Africa and the equator, suggesting warming estimates for equatorial Africa are relatively robust. However, estimates of the magnitude and direction of future rainfall changes in Africa are not precise, because models have not accounted for the roles of land-cover change, and dust and biomass aerosols in inducing regional climate changes (Hulme et al., 2005). This is especially relevant to Itombwe, where human activity during Africa’s World War (Prunier, 2008) destroyed large areas of natural habitat on the plateau (EG, CK and MMA pers. obs.).

Alward, Detling & Milchunas (1999) noted that average annual minimum temperatures (T_MIN) have increased at twice the rate of average annual maximum temperatures at a global scale. These authors analyzed a 23-year dataset from a montane grassland long-term ecological research site in Colorado (USA), and demonstrated that increased T_MIN was correlated with decreased net primary production by the dominant C₄ grass, rendering the habitat more susceptible to invasion by exotic species and less tolerant of drought and grazing. If similar effects are occurring at Itombwe, it is likely that the high-elevation grassland habitat will be devastated, possibly leading to the extinction of Congolacerta asukului. Moreover, in a study that focused on the potential effects of global climate change on Mexican lizards, Sinervo et al. (2010) noted extinction risk was significantly related to low latitudinal and altitudinal range limits, where thermal physiology and/or ecological interactions limit species, especially viviparous species in cool, montane habitats. Although C. asukului is likely oviparous, it is restricted to a limited latitudinal and elevation range in a cool, montane habitat, rendering it especially vulnerable to extinction risk from global warming.

Recognition of the genus Congolacerta adds another distinct evolutionary lineage to a long list of taxa that are endemic to the Albertine Rift mountains in eastern DRC, Uganda, Rwanda, Burundi and Tanzania (Plumptre et al., 2007). Overall, the Albertine Rift contains more than half of continental Africa’s bird species (Omari et al., 1999), and nearly 40% of its mammal species (Plumptre et al., 2007), including endangered mountain gorillas (Gorilla beringei; Schaller, 1964; Eckhart and Lanjouw, 2008). Many of these high-elevation forests are threatened by deforestation, and on a global scale, montane forests make up 12% of existing tropical forests worldwide, but are being cleared at twice the rate of the global average (Sodhi, Brook & Bradshaw, 2007).

Among other Albertine Rift sites, Itombwe (under consideration for a national park) has the most threatened species of amphibians (Laurent, 1964, 1983; Evans et al., 2008; Stuart et al., 2008; Roelke et al., in press), and the second-highest number of endemic amphibians (most not recorded for 60 years), rendering it among the most important sites for amphibian conservation in continental Africa (Burgess et al., 2004). Two amphibian genera, Laurentophryne, Tihen, 1960 and Chrysobatrachus, Laurent, 1951, are endemic to Itombwe, and the latter genus is only found above 2,400 m, with a peak in activity during the dry season (Laurent, 1951, 1964). Congolacerta asukului underscores the high levels of reptile endemism of the plateau—Itombwe also contains large numbers of endemic and threatened plants, butterflies, birds, and mammals (Louette, 1990; Omari et al., 1999; Plumptre et al., 2003). This diversity is remarkable given the superficial herpetological exploration of the plateau; Laurent (1954) mentioned only two reptiles from the upper plateau of Itombwe (2,500–3,000 m), including taxa in the lizard genera Trioceros (Chamaeleonidae) and Leptosiaphos (Scincidae).

In October, 2006, parts of the Itombwe Plateau were recognized as the Réserve Naturelle d’Itombwe (RNI), by decree of the Ministry of Environment, Conservation of Nature and Tourism (CK, pers. comm.). An administration with a managing warden was established for RNI in 2008, but armed militias have impeded conservation efforts (EG, CK and MMA, pers. obs., and see C. asukului etymology above). The reserve faces severe anthropogenic pressure from a growing human population (Barnes & Lahm, 1997), and park rangers are not yet established to ameliorate poaching and deforestation (elephants were likely extirpated recently), which is currently not monitored. Based on preliminary vegetation surveys, Doumenge (1998) remarked that Itombwe is, “undoubtedly the second most important, if not the most important, location of highland forests in continental Africa.” Because multiple, additional new species of amphibians and reptiles await description (EG, unpubl. data), it is likely that Itombwe’s importance as a center of endemism and conservation concern will increase as biological exploration continues.

APPENDIX 1

List of specimens, geographical origin, voucher number, and GenBank accession numbers for mitochondrial and nuclear gene sequences used in this study. DRC = Democratic Republic of the Congo; CER = Corey E. Roelke field series; IBE-S and Sa field numbers were kindly provided by Salvador Carranza; CRT field number was kindly provided by Zoltan Nagy and Jos Kielgast; NM Nairobi and field numbers without letters, William R. Branch tissue collection.

Species	Geographic origin	Museum/field voucher number	16S	cyt b	c-mos	RAG1
Acanthodactylus boskianus	Egypt (animal trade)	—	—	—	EF632251	EF632206
Acanthodactylus erythrurus	Morocco: Foret de Cedres (Azrou)	IBES 2917	HQ605790	HQ605832	HQ605874	HQ616540
Acanthodactylus scutellatus	Egypt: Abu Simbel	—	—	—	EF632252	EF632207
Adolfus africanus	DRC: South Kivu Province, Hombo	UTEP 20269	HQ605799	HQ605846	HQ605887	HQ616552
Adolfus africanus	DRC: South Kivu Province, Mashaba	UTEP 20271	HQ605828	HQ605870	HQ605911	HQ616576
Adolfus alleni	Kenya: Aberdares	NM Nairobi	—	HQ605841	HQ605882	—
Adolfus alleni	Kenya: Mt. Kenya	ZFMK 82078	HQ605779	HQ605840	—	—
Adolfus jacksoni	DRC: South Kivu Province, Lwiro	UTEP 20276	HQ605792	HQ605842	HQ605883	HQ616548
Adolfus jacksoni	DRC: South Kivu Province, Bitale	UTEP 20279	HQ605793	HQ605843	HQ605884	HQ616549
Adolfus jacksoni	Uganda: Kabale District, Bwindi Impenetrable National Park	CAS 201598	HQ605794	HQ605844	HQ605885	HQ616550
Adolfus jacksoni	Rwanda: North Province, Ruhengeri	CER 227	HQ605826	HQ605868	HQ605909	HQ616574
Adolfus jacksoni	Rwanda: North Province, Ruhengeri	UTA R55679	HQ605827	HQ605869	HQ605910	HQ616575
Adolfus jacksoni	Rwanda: North Province, Buhanga Forest	UTA R56311	HQ605825	HQ605867	HQ605908	HQ616573
Adolfus jacksoni	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20272	HQ605813	HQ605858	HQ605899	HQ616564
Adolfus jacksoni	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20273	HQ605814	HQ605859	HQ605900	HQ616565
Adolfus jacksoni	DRC: Orientale Province, Aboro	UTEP 20283	HQ605806	HQ605852	HQ605893	HQ616558
Adolfus jacksoni	DRC: South Kivu Province, Itombwe Plateau	UTEP 20280	HQ605809	HQ605854	HQ605895	HQ616560
Adolfus jacksoni	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20274	HQ605816	HQ605861	HQ605902	HQ616567
Adolfus jacksoni	DRC: Orientale Province, Aboro	UTEP 20285	HQ605823	HQ605866	HQ605907	HQ616572
Adolfus jacksoni	DRC: Orientale Province, Aboro	UTEP 20282	HQ605811	HQ605856	HQ605897	HQ616562
Adolfus jacksoni	DRC: South Kivu Province, Lwiro	UTEP 20275	HQ605815	HQ605860	HQ605901	HQ616566
Adolfus jacksoni	DRC: South Kivu Province, Lwiro	UTEP 20277	HQ605817	HQ605862	HQ605903	HQ616568
Adolfus jacksoni	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20278	HQ605818	HQ605863	HQ605904	HQ616569
Adolfus jacksoni	DRC: South Kivu Province, Bukavu	UTEP 20281	HQ605805	HQ605851	HQ605892	HQ616557
Adolfus jacksoni	Rwanda: Rukiva	—	—	—	EF632253	EF632208
Adolfus jacksoni	Arusha, Tanzania	761	HQ605785	HQ605839	HQ605881	HQ616547
Algyroides fitzingeri	Italy: Nuoro: Sardinia, Lula, direction to Conca e Crapa, monte Turuddo	Sa 35	HQ605789	—	—	—
Algyroides moreoticus	Greece: Korinthia, Feneos	—	—	—	EF632254	EF632209
Algyroides nigropunctatus	Greece: Preveza, Parga	—	—	—	EF632255	EF632210
Anatolacerta danfordi	Turkey: Icel, Camliyayla	—	—	—	DQ461743	EF632224
Apathya cappadocica	Turkey: Kayseri, Mt. Ercyas	—	—	—	EF632268	EF632223
Archaeolacerta bedriagae	France: Corsica	—	—	—	EF632256	EF632211
Atlantolacerta andreanskyi	Morocco: Oukaimeden	IBES 1699	HQ605787	HQ605830	HQ605872	HQ616538
Congolacerta asukului	DRC: South Kivu Province, Itombwe Plateau	UTEP 20268	HQ605801	—	HQ616585	HQ616577
Congolacerta asukului	DRC: South Kivu Province, Itombwe Plateau	UTEP 20265	HQ605804	HQ605850	HQ605891	HQ616556
Congolacerta asukului	DRC: South Kivu Province, Itombwe Plateau	UTEP 20264	HQ605803	HQ605849	HQ605890	HQ616555
Congolacerta asukului	DRC: South Kivu Province, Itombwe Plateau	UTEP 20263	HQ605802	HQ605848	HQ605889	HQ616554
Congolacerta asukului	DRC: South Kivu Province, Itombwe Plateau	UTEP 20267	HQ605800	HQ605847	HQ605888	HQ616553
Congolacerta asukului	DRC: South Kivu Province, Itombwe Plateau	UTEP 20266	HQ605808	HQ605853	HQ605894	HQ616559
Congolacerta vauereselli	DRC: South Kivu/Katanga Province border, Kabobo Plateau	UTEP 20295	HQ605810	HQ605855	HQ605896	HQ616561
Congolacerta vauereselli	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20289	HQ605795	—	HQ616586	HQ616578
Congolacerta vauereselli	DRC: South Kivu Province, Itombwe Plateau	UTEP 20293	HQ605798	HQ605845	HQ605886	HQ616551
Congolacerta vauereselli	DRC: Uganda: Kabale District, Bwindi Impenetrable National Park	CAS 201613	HQ605797	—	HQ616587	HQ616579
Congolacerta vauereselli	Rwanda: North Province, Bisate	UTA R55658	HQ605824	—	HQ616592	HQ616584
Congolacerta vauereselli	DRC: South Kivu Province, Itombwe Plateau	UTEP 20292	HQ605796	—	HQ616588	HQ616580
Congolacerta vauereselli	DRC: Orientale Province, Aboro	UTEP 20297	HQ605822	HQ605865	HQ605906	HQ616571
Congolacerta vauereselli	DRC: Orientale Province, Aboro	UTEP 20296	HQ605812	HQ605857	HQ605898	HQ616563
Congolacerta vauereselli	DRC: South Kivu Province, Itombwe Plateau	UTEP 20294	HQ605807	—	HQ616589	HQ616581
Congolacerta vauereselli	DRC: Orientale Province, Aboro	UTEP 20298	HQ605821	HQ605864	HQ605905	HQ616570
Congolacerta vauereselli	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20288	HQ605820	—	HQ616591	HQ616583
Congolacerta vauereselli	DRC: South Kivu Province, Kahuzi-Biega National Park	UTEP 20287	HQ605819	—	HQ616590	HQ616582
Dalmatolacerta oxycephala	Croatia: Hvar Island	—	—	—	EF632271	EF632228
Darevskia valentini	Armenia: Rasdan	—	—	—	EF632257	EF632212
Dinarolacerta mosorensis	Montenegro: Durmitor Mts.	—	—	—	EF632270	EF632227
Eremias arguta	Ukraine	—	—	—	EF632258	EF632213
Eremias pleskei	Armenia: Ararat region	—	—	—	EF632259	EF632214
Gallotia galloti	Spain: Tenerife Island	—	—	—	EF632260	EF632215
Gastropholis prasina	Tanzania: East Usambaras	760	HQ605781	HQ605835	HQ605877	HQ616543
Gastropholis vittatus	Tanzania: Amani, East Usambaras (captive specimen)	691	HQ605780	HQ605834	HQ605876	HQ616542
Heliobolus lugubris	Namibia: Haruchas	—	—	—	EF632261	EF632216
Heliobolus spekii	Tanzania: Lake Natron	—	—	—	EF632262	EF632217
Hellenolacerta graeca	Greece: Lakonia, Monemvasia	—	—	—	EF632269	EF632225
Holaspis guentheri	DRC: Orientale Province: Yaekela near Yangambi, Congo River	CRT 3660	HQ605784	HQ605838	HQ605880	HQ616546
Holaspis laevis	Tanzania: E. Usambaras	763	HQ605783	HQ605837	HQ605879	HQ616545
Holaspis laevis	Tanzania: Handeni	764	HQ605782	HQ605836	HQ605878	HQ616544
Holaspis laevis	Tanzania: Usambara Mts.	—	—	—	EF632263	EF632218
Iberolacerta cyreni	Spain: Avila: Track to Laguna Grande de Gredos	IBES 1010	HQ605788	HQ605831	HQ605873	HQ616539
Iberolacerta horvathi	Austria: Carinthia, Rattendorf	—	—	—	EF632264	EF632219
Iberolacerta monticola	Portugal: Sierra Estrela	—	—	—	EF632265	EF632220
Ichnotropis squamulosa	Mozambique (animal trade)	—	—	—	EF632266	EF632221
Lacerta agilis	Austria: Lower Austria, Weitra	—	—	—	EF632267	EF632222
Latastia longicaudata	Eritrea: Nakfa	—	—	—	EF632272	EF632229
Meroles suborbitalis	Namibia: Rosh Pinah	—	—	—	EF632273	EF632230
Mesalina guttulata	Tunisia: Tamerza	—	—	—	EF632274	EF632231
Mesalina rubropunctata	Egypt: Hurghada	—	—	—	EF632275	EF632232
Nucras lalandii	South Africa: Stellenbosch	—	—	—	EF632276	EF632233
Omanosaura jayakari	United Arab Emirates: Fujayrah	—	—	—	EF632277	EF632234
Ophisops elegans	Greece: Evros, Gianuli	—	—	—	EF632278	EF632235
Parvilacerta parva	Turkey: Malatya	—	—	—	EF632279	EF632236
Pedioplanis undata	Namibia: Nauchas	—	—	—	EF632280	EF632237
Philochortus spinalis	Eritrea: Ghinda	—	—	—	EF632281	EF632238
Phoenicolacerta laevis	Cyprus: Pafos	—	—	—	DQ461715	EF632226
Podarcis muralis	France: Languedoc-Roussillon: Lac du Salut de Vesoles	IBES 1181	HQ605791	HQ605833	HQ605875	HQ616541
Podarcis muralis	Austria: Lower Austria, Gumpoldskirchen	—	—	—	EF632282	EF632239
Poromera fordii	Cameroon: Mt. Nlonako	—	—	—	EF632283	EF632240
Psammodromus algirus	Spain: Lerida	—	—	—	EF632284	EF632241
Psammodromus hispanicus	Spain: Barcelona	—	—	—	EF632285	EF632242
Pseuderemias smithi	Kenya: Lake Turkana	—	—	—	EF632286	EF632243
Takydromus amurensis	Russia: Amur Region	—	—	—	EF632287	EF632244
Takydromus sexlineatus	Indonesia (animal trade)	—	—	—	EF632288	EF632245
Teira dugesii	Portugal: Madeira Island	—	—	—	EF632289	EF632246
Timon lepidus	Spain: Alicante	—	—	—	EF632290	EF632247
Timon tangitanus	Morocco: Foret de Cedres (Azrou)	IBES 2892	HQ605786	HQ605829	HQ605871	HQ616537
Tropidosaura gularis	South Africa: SW-Cape	—	—	—	EF632291	EF632248
Zootoca vivipara	Austria: Lower Austria, Schneeberg	—	—	—	EF632292	EF632249

APPENDIX 2

SPECIMENS EXAMINED

Adolfus africanus

CAS 54812 (male), Democratic Republic of the Congo (DRC), Medje; CAS 176853 (female), Uganda, Rukungiri District, Impenetrable (Bwindi) Forest Reserve, Buhoma Rd, 2 km S of forest reserve boundary, 1.0098°S, 29.6207E, 1,585m; CAS 176858 (female), Uganda, Rukungiri District, Impenetrable (Bwindi) Forest Reserve, Ngoto Swamp, 0.8855°S, 29.7295°E, 1,280 m; CAS 204389 (male), Uganda, Toro District, Kibale National Park, Makerere University Biological Field Station; UTEP 20269 (male), UTEP 20270 (female), DRC, South Kivu Province, Hombo, 1.8446°S, 28.4435°E, 800 m; UTEP 20271 (male), DRC, South Kivu Province, Mashaba village near Irangi, 1.8745°S, 28.4518°E, 800 m.

Adolfus alleni

CAS 162680, Uganda, Mt. Elgon, Arugot, 2,896 m; USNM 49411 (male), Kenya, Aberdare Range summit, 0.3167°S, 36.6167°E.

Adolfus jacksoni

CAS 122729 (male), Kenya, Western Province, Kakamega District, Kakamega Forest, near Kaimosi; CAS 152783 (male), Kenya, Nyanza Province, Kisumu District, Chemelil; CAS 201598 (female), Uganda, Kabale District, Bwindi Impenetrable National Park, Institute for Tropical Forest Conservation (Ruhizha), 1.0466°S, 29.7746°E, 2,362 m; CAS 201610 (female), Uganda, Kabale District, Bwindi Impenetrable National Park, Kabale-Kayonza rd, 1.0436°S, 29.7682°E, 2,347 m; CAS 204386 (male), Uganda, Masaka District, Lake Nabagabo; UTEP 20272 (male), UTEP 20273 (male), DRC, South Kivu Province, Kahuzi-Biega National Park, Tshivanga, 2.3128°S, 28.7552°E, 2,200 m; UTEP 20274 (subadult), DRC, South Kivu Province, Mbayo, 2.2663°S, 28.7838°E, 1,943 m; UTEP 20275 (male), South Kivu Province, Lwiro, 2.2457°S, 28.8126°E, 1,678 m; UTEP 20276 (subadult), UTEP 20277 (male), DRC, South Kivu Province, Lwiro, 2.2409°S, 28.8496°E, 1,530 m; UTEP 20278 (female), DRC, South Kivu Province, Kahuzi-Biega National Park, Mugaba, 2.2675°S, 28.6621°E, 2,264 m; UTEP 20279 (subadult), DRC, South Kivu Province, Bitale, 2.2078°S, 28.6296°E, 1,770 m; UTEP 20280 (female), DRC, South Kivu Proince, Itombwe Plateau, Kizuka, 03.0066°S, 28.7501°E, 2,450 m; UTEP 20281 (subadult), DRC, South Kivu Province, Bukavu, 02.5066°S, 28.8558°E, 1,529 m; UTEP 20282 (female), UTEP 20283 (male), UTEP 20284 (female), DRC, Orientale Province, Aboro, 2.0164°N, 30.8336°E, 2,088 m; UTEP 20285 (male), UTEP 20286 (male), DRC, Orientale Province, summit of Mt. Aboro, 2.0056°N, 30.5190°E, 2,445 m.

Congolacerta vauereselli

CAS 201613 (subadult), CAS 201614 (female), Uganda, Kabale District, Bwindi Impenetrable National Park, Mubwindi Swamp, NW end, S trail, 1.0710°S, 29.7536°E; CAS 204387 (female), Uganda, Toro District, Kibale National Park, Makerere University Biological Field Station, Kanyawara, Lower Camp; UTEP 20287 (subadult), DRC, South Kivu Province, Kahuzi-Biega National Park, Bwindi, 2.2769°S, 28.6613°E, 2,333 m; UTEP 20288 (male), DRC, South Kivu Province, Kahuzi-Biega National Park, Chinya, 2.2739°S, 28.6600°E, 2,297 m; UTEP 20289 (male), UTEP 20290 (female), UTEP 20291 (male), DRC, South Kivu Province, Kahuzi-Biega National Park, Mugaba, 2.2675°S, 28.6621°E, 2,264 m; UTEP 20292 (subadult), DRC, South Kivu Province, Itombwe Plateau, Bishaka, 03.3410°S, 28.7944°E, 2,208 m; UTEP 20293 (subadult), DRC, South Kivu Province, Itombwe Plateau, ca. Miki, 3.3746°S, 28.6426°E, 1,799 m; UTEP 20294 (female), DRC, South Kivu Province, Itombwe Plateau, Mugegema, 03.0618°S, 28.7786°E, 2,675 m; UTEP 20295 (male), DRC, South Kivu/Katanga Province border, Kabobo Plateau near Kilwemapante, 5.0538°S, 28.9917°E, 1,993 m; UTEP 20296 (female), UTEP 20297 (male), UTEP 20298 (female), UTEP 20299 (male), DRC, Orientale Province, Aboro, 2.0164°N, 30.8336°E, 2,088 m.