Liver samples from nine specimens from Phnom Banan, 16 specimens from Phnom Kamping Poi, five specimens from Phnom Khpoh, and 11 specimens from Phnom Sampeu were stored in 95% ethanol. Genomic DNA was isolated using the Qiagen DNeasyTM tissue kit (Valencia, CA, USA). NADH dehydrogenase subunit 2 gene (ND2) and downstream tRNA-Trp, tRNA-Ala, and tRNA-Asn was chosen for phylogeneic analyses with 41 specimens newly sequenced for this work. ND2 was amplified using a double-stranded Polymerase Chain Reaction (PCR) under the following conditions: 2.5 μl genomic DNA (~10–30 ng), 2.5 μl light strand primer (5 μM), 2.5 μl heavy strand primer (5 μM), 1.0 μl dinucleotide pairs (1.0 μM), 2.0 μl 5× buffer (2.0 μM), 1.0 MgCl 10× buffer (1.0 μM), 0.18 μl Taq polymerase (5u/μl), and 9.8s μl ultrapure H2O at n + 1. PCR reactions were executed on a BIO RAD T-100 Thermal Cycler under the following conditions: initial denaturation at 95 °C for 2 min, followed by a second denaturation at 95 °C for 35 s, annealing at 54 °C for 35 s, followed by a cycle extension at 72 °C for 1:35 min repeated for 34 cycles, followed by a final extension cycle run at 68 °C for 7 min. All PCR products were visualized on a 1.0% agarose electrophoresis gel. Successfully targeted PCR products were outsourced to GENEWIZ® for PCR purification, cycle sequencing, and sequencing. Primers used for amplification and sequencing are presented in Murdoch et al. (2019: table 2). Sequences were analyzed from both the 3’ and the 5’ ends separately to confirm congruence between the reads. Both the forward and the reverse sequences were uploaded and edited in GeneiousTM v. 11.1.5 (Drummond et al. 2011) and edited therein. The protein-coding region of the ND2 sequence was aligned by eye. MacClade v. 4.08 (Maddison and Maddison 2015) was used to calculate the correct amino acid reading frame and to confirm the lack of premature stop codons. GenBank accession numbers for all specimens are listed in Fig. 2.

Figure 2. 

A Maximum Likelihood consensus tree of the Cyrtodactylus intermedius group B enlarged section of the Maximum Likelihood consensus tree highlighting the Battambang clade C MFA plots and percent character contribution bar graphs based on the nearly total evidence data set of the Battambang clade.

Maximum likelihood (ML) and Bayesian inference (BI) were used to estimate the phylogenetic relationships of the aligned sequences. The ML phylogeny was estimated using the IQ-TREE webserver (Nguyen et al. 2015; Trifinopoulos et al. 2016) preceded by the selection of substitution models using the Bayesian Information Criterion (BIC) in ModelFinder (Kalyaanamoorthy et al. 2017), which supported HKY+F+G4 as the best fit model of evolution for codon positions 1 and 2, and TPM3u+F+G4 for position 3. Ten-thousand bootstrap pseudoreplicates via the ultrafast bootstrap (UFB; Hoang et al. 2018) approximation algorithm were employed. Nodes having UFB values of 95 and above were considered highly supported (Minh et al. 2013) and nodes with 90–94 UFB values were considered well-supported. The Bayesian inference (BI) analysis was carried out in MrBayes 3.2.3. (Ronquist et al. 2012) on XSEDE using the CIPRES Science Gateway (Cyberinfrastructure for Phylogenetic Research; Miller et al. 2010) employing models of evolution most similar to those above and default priors. Two independent Markov chain Monte Carlo (MCMC) simulations were performed each with four chains, three hot and one cold. We ran the MCMC simulation for 100 million generations, sampled every 10,000 generations, and discarded the first 10% of each run as burn-in. Convergence and stationarity of all parameters from both runs were checked in Tracer v. 1.6 (Rambaut and Drummond 2013) to ensure effective sample sizes (ESS) were above 200. Post-burn-in sampled trees from both runs were combined using the sumt function and a 50% majority rule consensus tree was constructed. Nodes with Bayesian posterior probabilities (BPP) of 0.95 and above were considered highly supported (Huelsenbeck et al. 2001; Wilcox et al. 2002) and nodes BPPs of 0.90–0.94 were considered well-supported. Retaining only ingroup taxa, uncorrected pairwise sequence divergences were calculated in MEGA X (Tamura et al. 2021) using the pairwise deletion option.

The general lineage concept (GLC: de Queiroz 2007) adopted herein proposes that a species constitutes a population of organisms evolving independently from other such populations owing to a general lack of gene flow. By “independently,” it is meant here that new mutations arising in one species cannot readily spread into another species (Barraclough et al. 2003; de Queiroz 2007). Under the GLC, molecular phylogenies were used to recover monophyletic mitochondrial lineages of individual(s) (i.e., populations) in order to develop initial species-level hypotheses – the grouping stage of Hillis (2019). Univariate, multivariate, and discrete color pattern and morphological data were analyzed to search for statistically significant differences in morphology and color patterns that were consistent with the previous species-level hypotheses designations – the construction of boundaries representing the hypothesis-testing step of Hillis (2019). This way, the inherent errors of simultaneously delimiting (phylogeny) and diagnosing (taxonomy) species are avoided (Frost and Hillis 1990; Frost and Kluge 1994; Hillis 2019).

All statistical analyses were conducted using R Core Team (2020). A Levene’s test for meristic and size-corrected morphometric characters was conducted to test for equal variances across all groups. Since all characters had equal variances (F ≥ 0.05), they were analyzed by an analysis of variance (ANOVA) and a TukeyHSD post hoc test.

A multiple factor analysis (MFA) using the R package FactorMineR (Husson et al. 2017) and visualized using the Factoextra package (Kassambara and Mundt 2017) was employed for individuals from the four new populations to compare their differences or similarities in morphospace. The MFA was conducted using a concatenated data set comprised of 12 meristic (SL, IL, PVT, LRT, VS, E4TL U4TL, T4TL, TFS, PS, PPS, and PCT), 14 size-corrected morphometric (SVL, FL, TBL, AG, HL, HW, HD, ED, EE, SN, EN, IO, EL, and IN) and four categorical (TT-wht, FS-sz, DP-fad, and BB-wd) characters for a nearly total evidence morphological data set (Suppl. material 1). Not all categorical characters could be used due to their invariability among the four populations. To remove potential effects of allometry in the morphometric characters (see Chan and Grismer 2022), size was corrected using the following equation: X_adj = log(X)-β[log(SVL)-log(SVL_mean)], where X_adj = adjusted value; X = measured value; β = unstandardized regression coefficient for each population; and SVL_mean = overall average SVL of all populations (Thorpe 1975, 1983; Turan 1999; Lleonart et al. 2000). The morphometrics of each population were size-corrected separately, then concatenated so as not to conflate intra- with interspecific variation (Reist 1986). All data were scaled to their standard deviation to insure they were analyzed based on correlation and not covariance.

MFA is a global, unsupervised, multivariate analysis that incorporates qualitative and quantitative data (Pagès 2015) simultaneously, making it possible to analyze different data types in a nearly total morphological evidence environment. For multispatial diagnostics, MFA is superior to principle component analysis (PCA) in that the data are standardized so the analysis is not biased by a single data type (e.g., morphometric). In an MFA, each individual is described by a different set of variables (i.e., characters) which are structured into different data groups in a global data frame, in this case, quantitative data (i.e., meristic and size-corrected morphometrics) and categorical data (i.e., color pattern and morphological characters). In the first phase of the analysis, separate multivariate analyses are carried out for each set of variables—principal component analyses (PCA) for each quantitative data set and a multiple correspondence analysis (MCA) for the categorical data. The data sets are then normalized separately by dividing all their elements by the square root of their first eigenvalue. For the second phase of the analysis, the normalized data sets are concatenated into a single data frame for a final global PCA. Standardizing the data in this manner prevents one data type from overleveraging another in the second phase. This way, the contributions of each data type to the overall variation in the data set are standardized (Pagès 2015; Kassambara and Mundt 2017).

In order to compare body shapes among the four populations, a PCA and a discriminant analysis of principal components (DAPC) of the 14 size-corrected morphometric characters were employed (Suppl. material 1). PCA is a dimension reducing unsupervised analysis (i.e., all individuals are treated independently) that recovers morphospatial relationships among the sampled individuals (i.e., data points) and how well they form clusters that may or may not align with the putative species boundaries delimited by phylogenetic analyses and diagnosed by the univariate analyses. On the other hand, DAPC from the adegent package 2.1.5 in R (Jombart 2021) is a supervised analysis (i.e., groups are specified a priori in the analysis) that relies on scaled data calculated from a PCA as a prior step to ensure that variables analyzed are not correlated and number fewer than the sample size. Dimension reduction of the DAPC prior to plotting, was accomplished using PCAtest (Camargo 2022) to recover the principle component (PC) axes which contributed significantly to the overall signal within the PCA. The function runs 1,000 random permutations and bootstrap replicates of the empirical data. Based on the bootstrap resampling and permutation, 95% confidence intervals around mean values are calculated. Significant p-values imply there is non-random correlational structure in the overall dataset and that the PCA is biologically meaningful. Statistically significant PC axes reflect non-random correlations amongst the variables that have a larger contribution beyond random noise. PCA and DAPC analyses were also performed on the meristic data set (Suppl. material 1) as well in order to determine which data type, morphometric or meristic, contributed most to the morphospatial distribution among the four new populations and why from an adaptive standpoint.

A PERMANOVA analysis from the vegan package 2.5-3 in R (Oksanen et al. 2020) was used to determine if the centroid locations and group clusters of each species/population from the MFA were statistically different from one another (Skalski et al. 2018) based on the load scores of dimensions 1–5. Using load scores as opposed to raw data, which are normally used, allows for the incorporation of the categorical characters which cannot be run untransformed in a PERMANOVA. All load scores for the PCAs, however, were used. The analysis calculates a Euclidean (dis)similarity matrix using 50,000 permutations. A pairwise post hoc test calculates the differences between the populations, generating a Bonferroni-adjusted p-value and a pseudo-F ratio (F statistic). A p < 0.05 is considered significant and larger F statistics indicate more pronounced group separation. A rejection of the null hypothesis (i.e., centroid positions and the spread of the data points [i.e., clusters] are no different from random) signifies a statistically significant difference between species/populations. This test results in a statistically defensible method of concluding which species/populations plots are actually different from one another and removes the ad hoc “eyeballing it” as is normally done.

Both the ML and BI analyses indicate that the four new populations form a strongly supported monophyletic group (UFB 97–100/BPP 1.00), referred to here as the Battambang clade, deeply nested within the intermedius group (Fig. 2A, B). The analyses also strongly support (100/1.00) the Battambang clade’s sister lineage relationship to Cyrtodactylus thylacodactylus. Additionally, individuals from all four respective populations are strongly recovered as monophyletic (100/1.00; Fig. 2B). Within the Battambang clade, the Phnom (P.) Banan population forms the well-supported sister population (100/1.00) to the remaining three populations collectively. Both analyses recover the P. Kamping Poi population as being the well-supported potential sister population of the P. Sampeu-P. Khpoh lineage (Fig. 2B). The sister population relationship between the P. Sampeu and P. Khpoh populations is poorly supported in the ML analysis and not supported in the BI analysis (80/0.00). Furthermore, the branch length subtending that relationship is so short that these populations, with the P. Kamping Poi population, could be considered as polytomous (Fig. 2A). The Battambang clade has an uncorrected pairwise sequence divergence from the remaining species of the intermedius group ranging from 5.0% between the P. Khoh population and C. cardamomensis to 24.5% between the P. Sampeu population and C. disjunctus (Suppl. material 2). The pairwise sequence divergence among the individuals of the Battambang clade ranges from 1.1–2.2% (Suppl. material 2).

The PCA of the morphometric data recovered all populations plotting separately in morphospace along the ordination of the first two PCs except for slight overlap between the P. Kamping Poi and P. Banan populations (Fig. 3A). However, the DAPC showed significant overlap in the 66% ellipsoids among all populations (Fig. 3B). Despite their morphospatial separation in the PCA, the PERMANOVA mirrored the DAPC and found significant differences only between the P. Kamping Poi and P. Sampeu populations and between the P. Banan and P. Sampeu populations (Fig. 3C). PC1 accounted for 23.5% of the variation and loaded most heavily for head length (HL) and snout length (SN) and PC2 accounted for 16.6% of the variation and loaded most heavily for eye to ear distance (EE) – all characters relating to dimensions of the head (Table 1).

Conversely, the PCA of the meristic data recovered nearly all populations plotting completely separately in morphospace along the ordination of the first two PCs which was mirrored in the DAPC (Fig. 4A, B). Corroborating these differences, the PERMANOVA analysis recovered all populations as being significantly different from one another except for P. Sampeu and P. Khpoh. Most were significantly different at the more discerning p-adjusted value (Fig. 4C). PC1 accounted for 30.7% of the variation and loaded most heavily for the total number of 4^th toe lamellae (T4TL), number of expanded 4^th toe lamellae (E4TL), and the number of paravertebral tubercles (PVT). PC2 accounted for 21.4% of the variation and loaded most heavily for the total number of enlarged precloacal scales (PS), number of longitudinal rows of tubercles (LRT), number of unexpanded 4^th toe lamellae (U4TL), and number of ventral scales (VS) (Table 2).

Figure 4. 

A, B PCA and DAPC plots, respectively, of the meristic data of the Battambang clade C summary statistics from the PERMANOVA analysis. Shaded cells denote populations whose morphospatial positions are statistically different.

The MFA results of the near total evidence morphological data set was very similar to the PCA and DAPC of the meristic data in that all populations plotted separately along the first two dimensions (Fig. 2C). It would be tempting to say that this was a result of the meristic data, however those data contributed to only ~37% of the 24.1% of the variation in dimension1 (Dim 1) (Fig. 2C). The categorical data contributed to the majority of variation in Dim 2 and Dim 3 and the morphometric data contributed the majority of variation in Dim 4. The PERMANOVA analysis also recovered all populations as significantly different from one another at the unadjusted p-value, except for except for the P. Sampeu and P. Khpoh populations, and most were significantly different at the p-adjusted value (Fig. 4C).

Results of the ANOVA analysis recovered significant differences in both meristic and morphometric characters among the four populations in a varying number of population comparisons across a varying number of characters (Table 3).

All the newly discovered populations are monophyletic and none are phylogenetically nested within any other species of the intermedius group nor are they sister to any of the group’s nominal species (Fig. 2A). With the exception of the P. Sampeu and P. Khpoh populations, they all bear statistically significant morphospatial differences from one another in the MFA and the PCAs (Figs 2–4) and have significantly different mean values from one another in varying numbers of morphometric and meristic characters (Table 3) as well as differing categorically (Suppl. material 3). The polytomous relationship among the P. Kamping Poi, P. Sampeu, and P. Khpoh populations and the absence of characters separating the latter two that differ only by a pairwise sequence divergence of only 1.1%, indicates they all could be considered the same species despite their allopatry and with no chance of dispersal (see discussion). The short branch lengths subtending the inter-nodes of the other populations in the Battambang clade may indicate the same. Therefore, at this point, all populations are considered conspecific and described as a single species. The type series is designated from the P. Kamping Poi population due to its larger sample size, while the remaining three populations are treated as geographic variants.

 Cyrtodactylus kampingpoiensis sp. nov.

https://zoobank.org/F5827770-1118-4C6D-8930-F3DC4E8FDD42

Common English name: Kamping Poi Bent-toed Gecko

Common Khmer name: តុកកែភ្នំកំពីងពួយ

Figs 5, 6, 7

Type material.

Holotype • Adult male (LSUHC 15206) collected from Phnom Kamping Poi, Banan District, Battambang Province, Cambodia at 13°5.795'N, 102°55.798'E, at 114 m and nearby areas on 22 March 2024 by Pablo Sinovas, Seiha Hun, Phyroum Chourn, Matthew L. Murdoch, L. Lee Grismer, Evan S. H. Quah, Sothearen Thi, Christian Ching, and Anthony Cobos. Paratypes • Two adult males (LSUHC 15203 and 15211) and five adult females (LSUHC 15205, 15207, 15209–10, and 15212) bear the same collection data as the holotype. The type series was collected from 1530–2200 hrs.

Figure 5. 

Adult male holotype of Cyrtodactylus kampingpoiensis sp. nov. LSUHC 15206 A dorsal view B ventral view of gular region, throat, forelimbs, and feet C ventral view D ventral view of tail, pelvic region, hind limbs, and feet.

Additional specimens examined.

• Six hatchlings (LSUHC 15196– 200, and 151202) bear the same collection data as the type series. The specimens were too small to recover reliable morphometric and meristic data but were included in the phylogenetic analyses.

Figure 6. 

Cyrtodactylus kampingpoiensis sp. nov. A adult male holotype LSUHC 15206 B gravid adult female LSUHC 15207 C adult female LSUHC 15205 D juvenile LSUHC 15176 E juvenile male LSUHC 15203.

Diagnosis.

Cyrtodactylus kampingpoiensis sp. nov. can be separated from all other species of the intermedius group by the combination of having a maximum SVL of 79.6 mm (female); 9–11 supralabials; nine or 10 infralabials; 30–37 paravertebral tubercles; 19–21 rows of longitudinally arranged tubercles; 38–46 longitudinal rows of ventrals; 5–7 expanded subdigital lamellae on the fourth toe; 11–13 unmodified subdigital lamellae on the fourth toe; 18–20 total subdigital lamellae on the fourth toe; 26–34 total number of enlarged femorals; no femoral pores; 5–9 enlarged precloacals; 7–9 precloacal pores in males (n = 3); three or four rows of large post-precloacal scales; 0–3 postcloacal tubercles; four dark body bands; 11–13 dark caudal bands (n = 7); 10–12 light caudal bands (n = 7); body tubercles not greatly reduced and moderately keeled; caudal tubercles extend beyond base of tail; subcaudals transversely expanded but not extending up onto side of tail; enlarged femorals and enlarged precloacals not continuous; enlarged proximal femorals equal (rarely subequal, one of seven) in size to distal femorals; no interdigital pockets; dorsal pattern not faded; no distinct dark markings on the top of head; lightened centers in dark body bands variable; no dark markings in light interspaces between body bands; dark body bands equal in width to light interspaces; light interspaces not reduced to a narrow thin white band; dark body bands bordered by prominent white tubercles; dark caudal bands slightly wider than light caudal bands; light caudal bands bearing dark markings in adults; posterior margin of nuchal loop not smoothly rounded (Table 4; Fig. 7; Suppl. materials 3, 4).

Table 4.

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Data for the type series of Cyrtodactylus kampingpoiensis sp. nov. from Phnom Kamping Poi, Battambang Province, Cambodia.

Specimen	LSUHC 15206 holotype	LSUHC 15210 paratype	LSUHC 15205 paratype	LSUHC 15209 paratype	LSUHC 15207 paratype	LSUHC 15211 paratype	LSUHC 15203 paratype	LSUHC 15212 paratype
sex	m	f	f	f	f	m	m	f
Meristic characters
supralabials (SL)	9	11	10	11	11	10	10	11
infralabials (IL)	9	9	10	9	9	10	9	10
paravertebral tubercles (PVT)	32	37	34	34	35	31	30	34
longitudinal rows of tubercles (LRT)	20	21	20	19	20	20	20	21
longitudinal rows of ventral scales (VS)	43	41	42	41	38	42	46	44
expanded subdigital lamellae on 4th toe (T4LE)	5	7	6	7	7	7	7	7
unmodified subdigital lamellae on 4th toe (T4LU)	13	13	12	11	12	12	11	12
total subdigital lamellae on 4th toe (T4TL)	18	20	18	19	19	19	18	19
enlarged femoral scales (R,L)	16+18	15+14	17+13	14+15	13+15	13+15	13+13	13+14
total enlarged femoral scales (TFS)	34	29	30	29	28	28	26	27
femoral pores	0	0	0	0	0	0	0	0
enlarged precolacal scales (PS)	9	5	6	7	6	9	8	9
precloacal pores (PP)	3L, 4R, gap 2	0	0	0	0	9	8	0
post-precloacal scale rows (PPS)	3	3	3	4	3	3	3	3
postcloacal tubercles (PCT)	3	0	2 small	3 small	3 small	3	3	3
Categorical characters
subcaudals expanded	yes	yes	yes	yes	yes	yes	yes	yes
subccaudals extend up onto lateral side of tail	no	no	no	no	no	no	no	no
body dark bands (BB)	4	4	4	4	4	4	4 broken	4
dark caudal bands (DCB)	/	11	12	12	13	11	11	12
light caudal bands (LCB)	/	10	11	11	12	10	10	11
body tubercles greatly reduced (Tub-red)	no	no	no	no	no	no	no	no
body tubercles moderately keeled (Tub-kld)	yes	yes	yes	yes	yes	yes	yes	yes
caudal tubercles extend beyond base of tail (CT-ext)	yes	yes	yes	yes	yes	yes	yes	yes
subcaudals expanded (SubC-exp)	yes	yes	yes	yes	yes	yes	yes	yes
subccaudals extend up onto lateral side of tail (SubC-lat)	no	no	no	no	no	no	no	no
enlarged femoral and precloacal scales continuous (FS-PS)	no	no	no	no	no	no	no	no
enlarged proximal femoral < 1/2 size of distal femorals (FS-sz)	equal	equal	equal	equal	equal	equal	equal	subequal
pocketing between digits of hind feet (Dig-pok)	no	no	no	no	no	no	no	no
dorsal pattern faded (CP_fd)	no	no	no	no	no	no	no	no
distinct dark pigmented blotches on top of head present (HD-mrk)	no	no	no	no	no	no	no	no
dark body bands with lightened centers (BB-cntr)	yes	weak	weak	weak	weak	weak	no	no
dark body markings in light interspaces (BB-intr)	no	no	no	no	no	no	no	no
dark dorsal bands thin or ~ same width as light interspaces (BB-wd)	equal	equal	equal	equal	equal	equal	equal	equal
light interspaces reduced to a narrow thin white band	no	no	no	no	no	no	no	no
dark dorsal bands bordered by prominently colored white tubercles (Wht-tub)	yes	yes	yes	yes	yes	yes	yes	yes
dark caudal bands wider than light caudal bands (DCB-wd)	yes	yes	yes	yes	yes	yes	yes	yes
light caudal bands bearing dark markings in adults (WCB-mrk)	yes	yes	yes	yes	yes	yes	yes	yes
Morphometric characters (mm)
SVL	78.9	71.1	71.4	79.6	77.9	71.0	53.0	79.0
TaiL	82.0	90.0	92.0	103.0	105.0	95.0	68.0	no tail
TaiW	7.3	6.6	5.8	7.4	6.8	5.8	4.4	6.4
FL	12.2	11.9	11.2	12.4	12.8	10.7	8.2	12.6
TBL	15	14.1	13.9	15.7	14.6	13.8	9.6	14.8
AG	35.6	34.5	32.8	36.6	34.9	32.6	23.4	36.0
HL	22.5	20.3	20.6	22.8	22.2	20.1	16.4	22.6
HW	15.0	13.9	13.4	15.6	15.3	14.3	10.3	16.3
HD	8.7	9.0	8.4	10.2	8.4	7.8	6.4	9.9
ED	5.1	4.5	4.6	4.9	4.9	4.8	3.7	5.2
EE	6.8	5.8	5.5	6.1	6.2	5.8	4.6	6.3
SN	8.5	7.9	8.0	8.8	8.6	8.0	6.3	9.0
EN	6.2	5.6	5.9	6.3	6.4	4.8	4.4	6.3
IO	5.1	4.8	4.4	5.1	5.4	4.2	3.9	5.2
EL	1.9	2.1	2.1	2.2	2.3	1.7	1.6	1.9
IN	2.2	2.5	2.3	2.4	2.3	1.9	1.7	2.3

Figure 7. 

Type series of Cyrtodactylus kampingpoiensis sp. nov.

Description of holotype.

(Fig. 5; Table 4). Adult male SVL 78.9 mm; head moderate in length (HL/SVL 0.29), width (HW/HL 0.67), somewhat flattened (HD/HL 0.39), distinct from neck, triangular in dorsal profile; lores weakly concave anteriorly, weakly inflated posteriorly; prefrontal concave; canthus rostralis rounded; snout elongate (SN/HL 0.38), slightly convex, rounded in dorsal profile; eye large (ED/HL 0.23); ear opening elliptical, obliquely oriented, moderate in size; eye to ear distance slightly greater than diameter of eye; rostral rectangular, partially divided, bordered posteriorly by large left and right supranasals and a small internasal, bordered laterally by first supralabials; external nares bordered anteriorly by rostral, dorsally by large supranasal, posteriorly by two moderately sized postnasals, bordered ventrally by first supralabial; nine (R,L) rectangular supralabials tapering abruptly to below posterior margin of eye, first–sixth supralabials largest; nine (R, L) infralabials tapering smoothly to slightly past the termination of enlarged supralabials to corner of mouth; scales of rostrum and lores flat, much larger than granular scales on top of head and occiput; scales of occiput intermixed with small, rounded, tubercles; superciliaries elongate, largest dorsally; mental triangular, bordered laterally by first infralabials and posteriorly by large left and right trapezoidal postmentals contacting medially for ~ 40% of their length posterior to mental; one row of enlarged, sublabials extending posteriorly to seventh infralabials (R, L); gular and throat scales small, granular, grading posteriorly into slightly larger, flatter, smooth, imbricate, pectoral and ventral scales.

Body relatively long (AG/SVL 0.45) with well-defined ventrolateral folds; dorsal scales small, granular, interspersed with moderately sized, keeled, semi-regularly arranged tubercles extending from occiput to beyond base of tail; ~ 20 longitudinal rows of tubercles at midbody; ~ 32 paravertebral tubercles; 43 flat, imbricate, ventral scales much larger than dorsal scales; seven enlarged pore-bearing precloacal scales, 4R, 3L separated by two poreless scales; no deep precloacal groove or depression; and three rows of large post-precloacal scales on midline.

Forelimbs moderate in length and stature (FL/SVL 0.15); granular scales of forelimbs larger than those on body, small rounded tubercles on dorsal surface of forearms; palmar scales rounded, juxtaposed; digits well-developed, inflected at basal interphalangeal joints, slightly narrower distal to inflections; subdigital lamellae transversely expanded, those proximal to joint inflections much wider than the nearly unmodified lamellae distal to inflections; claws well-developed, sheathed by a dorsal and ventral scale; hind limbs robust, wider and longer than forelimbs (TBL/SVL 0.19), covered dorsally by granular scales interspersed with moderately sized tubercles, larger and flatter scales anteriorly; ventral scales of thighs flat, imbricate, larger than dorsals; subtibial scales, flat, imbricate, slightly smaller than those of thigh; one row of 16(R)18(L) slightly enlarged femoral scales terminating distally before knee, not continuous with enlarged precloacal scales, and poreless; proximal femorals nearly same size as distal femorals, all femorals forming an abrupt union with smaller, granular, posteroventral scales of thigh; plantar scales flat, juxtaposed; digits well-developed, inflected at basal interphalangeal joints; claws well-developed, sheathed by a dorsal and ventral scale at base; and five (R,L) wide subdigital lamellae on fourth toe proximal to joint inflection, 13 (R,L) narrower lamellae distal to joint inflection, 18 total subdigital lamellae.

Tail long (TL/SVL 1.04), partially regenerated, original portion 9.6 mm, regenerated portion 72.4 mm, 7.3 mm wide at base, tapering to a point; dorsal caudal scales of original portion of tail small, generally square, juxtaposed; median row of subcaudals significantly larger than dorsal caudals, transversely expanded, not extending dorsally onto lateral side of tail; body tubercles extending slightly beyond base of tail; hemipenal swellings at base of tail, three large postcloacal tubercles on both sides; and postcloacal scales flat, imbricate.

Color and pattern.

(Figs 5–7). Ground color of top of head, limbs, and dorsum straw to light-brown; top of head immaculate; prominent dark-brown nuchal loop edged in light-yellow bearing a somewhat flat posterior border extends between postorbital regions across the nape; no dark-brown band on nape; four immaculate dark-brown, slightly wavy-edged, dorsal body bands edged with bright-white tubercles have slightly lightened centers and are equal in width to straw-colored interspaces, extend from shoulders to groin, terminating ventrally slightly above the ventrolateral folds; first dorsal band extends anteriorly across shoulders; light-colored dorsal interspaces immaculate; forelimbs and hind limbs generally immaculate; one dark-brown caudal band on original portion of tail; regenerated portion of tail straw-colored overlain with faint, dark, irregularly shaped markings; iris gold with thin black reticulations; venter beige with faint, dark shadowing on lateral edges of belly, limbs, and throat; and subcaudal region essentially unicolor light-brown.

Etymology.

The species name kampingpoiensis is in reference to the type locality at Phnom Kamping Poi, Banan District, Battambang Province, Cambodia (Fig. 1).

Distribution.

The type series of Cyrtodactylus kampingpoiensis sp. nov. is known only from the type locality at Phnom Kamping Poi, Banan District, Battambang Province, Cambodia (Fig. 1).

Variation.

(Figs 6, 7). The paratypes remarkedly approach the holotype in general coloration and pattern. The most notable variation pertains to the dorsal banding of the juvenile LSUHC 15203 whose third and fourth bands are broken on the midline and offset (Fig. 6). The overall pattern of LSUHC 15212 is far less bold than that of the holotype and the medial portion of its first band is posteriorly protracted. The juveniles have darker bands lacking lightened centers and are edged with yellow tubercles and there are no dark markings in the caudal interspaces. This is all accentuated in the hatchlings whose coloration is often slightly faded in the posterior ~ 25% of the tail. The precloacal pore series in the holotype is separated medially by two poreless scales whereas the pore-bearing precloacal scales on the paratypes are continuous. Differences in meristics, morphometrics, and categorical characters are detailed in Table 4 and Suppl. material 3.

Geographic variation.

(Tables 3, 5). Potential diagnostic differences in maximum SVL, meristics, and categorical characters between Cyrtodactylus kampingpoiensis sp. nov. and all other species in the intermedius group – except for species in the Battambang clade (used here in reference to all four populations) – are detailed in Suppl. material 4. Cyrtodactylus kampingpoiensis sp. nov. has an uncorrected pairwise sequence divergence from the intermedius group species ranging from 3.5–23.6% (Suppl. material 2). Cyrtodactylus kampingpoiensis sp. nov. bears a 1.1–2.2% uncorrected pairwise sequence divergence from the other three populations. In meristics, it differs further from the P. Banan population in having a statistically significant fewer number of enlarged precloacals (PS; 5–9 vs 9–12), unmodified subdigital lamellae (U4TL; 11–13 vs 13–16) and total number of subdigital lamellae (T4TL; 18–20 vs 20–23) (Table 5). It differs further in morphometrics in having a statistically significant longer ear-eye distances (EE), longer snout (SN), and a shorter foreleg (TBL). The PERMANOVA analyses indicate that Cyrtodactylus kampingpoiensis sp. nov. differs significantly in morphospace from the P. Banan population in the MFA (Figs 2B, 4C) and the meristic PCA (Fig. 4C). Categorically, C. kampingpoiensis sp. nov. differs from the P. Banan population in that its slightly enlarged femorals and enlarged precloacals are usually discontinuous versus being continuous (four of five specimens; Suppl. materials 3, 4); the dark body bands are equal in width to the light interspaces as opposed to being wider; and hatchings and small juveniles have only a slightly faded tail tip as opposed to a nearly immaculate white tail tip (Fig. 6D vs Fig. 9E).

Table 5.

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Summary statistics for C. kampingpoiensis sp. nov., the Phnom Banan population, the Phnom Sampeu population, and the Phnom Khpoh population.

Phnom Banan
meristics	SL	IL	PVT	LRT	VS	T4LE	T4LU	T4TL	TFS	PS	PPS	PCT
mean	10.2	9.8	35.4	19.2	41.6	7.4	14.4	21.8	24.8	10.2	2.8	3.2
sd	0.84	0.84	2.30	0.84	2.41	0.55	1.14	1.30	4.09	1.10	0.45	0.45
min	9	9	33	18	39	7	13	20	19	9	2	3
max	11	11	38	20	45	8	16	23	30	12	3	4
N	5	5	5	5	5	5	5	5	5	5	5	5
morphometrics	SVL	FL	TBL	AG	HL	HW	HD	ED	EE	SN	EN	IO	EL	IN
mean	1.8477	1.0691	1.1673	1.5267	1.3165	1.1360	0.9285	0.6818	0.7279	0.9067	0.7826	0.6307	0.2856	0.3507
sd	0.05514	0.02585	0.01116	0.03584	0.00607	0.01535	0.01345	0.00718	0.03343	0.00724	0.00917	0.04445	0.05414	0.01534
min	1.7536	1.0324	1.1505	1.4994	1.3092	1.1126	0.9133	0.6724	0.6718	0.8985	0.7768	0.5906	0.1998	0.3344
max	1.8971	1.0999	1.1793	1.5822	1.3255	1.1484	0.9488	0.6897	0.7579	0.9184	0.7986	0.6915	0.3422	0.3713
N	5	5	5	5	5	5	5	5	5	5	5	5	5	5
Cyrtodactylus kampingpoiensis sp. nov
meristics	SL	IL	PVT	LRT	VS	T4LE	T4LU	T4TL	TFS	PS	PPS	PCT
mean	10.38	9.38	33.38	20.13	42.13	6.63	12.00	18.75	28.88	7.38	3.13	2.51
±sd	0.7440	0.5175	2.2638	0.6409	2.3566	0.7440	0.7559	0.7071	2.4165	1.5980	0.3536	1.0357
min	9	9	30	19	38	5	11	18	26	5	3	1
max	11	10	37	21	46	7	13	20	34	9	4	3
N	8	8	8	8	8	8	8	8	8	8	8	8
morphometrics	SVL	FL	TBL	AG	HL	HW	HD	ED	EE	SN	EN	IO	EL	IN
mean	1.8585	1.0603	1.1437	1.5222	1.3214	1.1539	0.9337	0.6737	0.7701	0.9109	0.7581	0.6777	0.2946	0.3418
±sd	0.05861	0.01463	0.01153	0.01162	0.00527	0.01282	0.03000	0.01147	0.01681	0.00693	0.02826	0.02548	0.04026	0.03511
min	1.7243	1.0405	1.1302	1.5113	1.3117	1.1355	0.8960	0.6601	0.7469	0.9004	0.6909	0.6311	0.2371	0.2866
max	1.9009	1.0860	1.1603	1.5483	1.3263	1.1750	0.9714	0.6893	0.8038	0.9247	0.7787	0.7102	0.3428	0.4054
N	8	8	8	8	8	8	8	8	8	8	8	8	8	8
Phnom Khpoh
meristics	SL	IL	PVT	LRT	VS	T4LE	T4LU	T4TL	TFS	PS	PPS	PCT
mean	10.7	9.3	28.7	17.0	40.0	6.7	13.0	19.7	26.0	9.3	3.0	2.3
sd	0.58	0.58	1.15	1.00	2.65	0.58	0.00	0.58	0.00	0.58	0.00	1.15
min	10	9	28	16	37	6	13	19	26	9	3	1
max	11	10	30	18	42	7	13	20	26	10	3	3
N	3	3	3	3	3	3	3	3	3	3	3	3
morphometrics	SVL	FL	TBL	AG	HL	HW	HD	ED	EE	SN	EN	IO	EL	IN
mean	1.8421	1.0567	1.1363	1.5025	1.3072	1.1454	0.9266	0.6878	0.7404	0.8945	0.7504	0.6900	0.2695	0.3688
sd	0.09860	0.00346	0.00574	0.00557	0.00859	0.00040	0.02209	0.01171	0.01367	0.00371	0.00774	0.03742	0.06337	0.02250
min	1.72835	1.05317	1.13071	1.49710	1.29886	1.14503	0.90506	0.67638	0.72637	0.89092	0.74284	0.65175	0.20462	0.34579
max	1.90309	1.06008	1.14219	1.50824	1.31603	1.14582	0.94920	0.69977	0.75369	0.89833	0.75830	0.72652	0.33124	0.39074
N	3	3	3	3	3	3	3	3	3	3	3	3	3	3
Phnom Sampeu
meristics	SL	IL	PVT	LRT	VS	T4LE	T4LU	T4TL	TFS	PS	PPS	PCT
mean	10.7	10.0	29.9	16.0	37.6	6.1	12.7	18.9	26.0	8.0	2.6	1.4
sd	0.76	0.82	1.07	1.00	2.30	0.38	0.76	0.90	2.08	0.00	0.53	0.53
min	10	9	28	15	34	6	12	18	24	8	2	1
max	12	11	31	17	41	7	14	20	29	8	3	2
N	7	7	7	7	7	7	7	7	7	7	7	7
morphometrics	SVL	FL	TBL	AG	HL	HW	HD	ED	EE	SN	EN	IO	EL	IN
mean	1.8406	1.0502	1.1300	1.5054	1.2993	1.1328	0.9133	0.6655	0.7311	0.8915	0.7646	0.6706	0.2311	0.3475
sd	0.04343	0.02355	0.01813	0.00650	0.00566	0.00929	0.02377	0.01475	0.02156	0.00573	0.00415	0.02465	0.05812	0.03146
min	1.7513	1.0148	1.1137	1.4952	1.2942	1.1145	0.8739	0.6350	0.6936	0.8835	0.7577	0.6399	0.1326	0.2967
max	1.8887	1.0827	1.1648	1.5150	1.3108	1.1420	0.9501	0.6772	0.7608	0.9022	0.7698	0.7045	0.2956	0.3900
N	7	7	7	7	7	7	7	7	7	7	7	7	7	7

Table 6.

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Data for the referred series of the Phnom Banan population from Phnom Banan, Battambang Province, Cambodia.

Specimen	LSUHC 15174	LSUHC 15173	LSUHC 15170	LSUHC 15169	LSUHC 15171
sex	m	m	f	m	m
Meristic characters
supralabials (SL)	10	9	11	10	11
infralabials (IL)	10	9	10	11	9
paravertebral tubercles (PVT)	37	33	36	38	33
longitudinal rows of tubercles (LRT)	20	19	20	19	18
ventral scales (VS)	39	43	40	45	41
expanded subdigital lamellae on 4th toe (E4TL)	7	8	7	8	7
unmodified subdigital lamellae on 4th toe (U4TL)	16	14	14	15	13
total subdigital lamellae on 4th toe (T4TL)	23	22	21	23	20
enlarged femoral scales (FS) (R,L)	15+14	13+13	10+9	13+14	15+15
total of enlarged femoral scales (TFS)	29	24	19	27	30
femoral pores (FP)	0	0	0	0	0
enlarged precolacal scales (PS)	10	10	12	10	9
precloacal pores (PP)	10	10 1 scale gap	12 dimples	10	9
post-precloacal scale rows (PPS)	3	2	3	3	3
postcloacal tubercles (PCT)	4	3	3	3	3
dark body bands (BB)	4	4	4	4	4
dark caudal bands (DCB)	/	10	/	11	12
light caudal bands (LCB)	/	/	/	10	11
Categorical characters
body tubercles greatly reduced (Tub-red)	no	no	no	no	no
body tubercles moderately keeled (Tub-kld)	yes	yes	yes	yes	yes
caudal tubercles extend beyond base of tail (CT-ext)	yes	yes	yes	yes	yes
subcaudals expanded (SubC-exp)	yes	yes	yes	yes	yes
subccaudals extend up onto lateral side of tail (SubC-lat)	no	no	no	no	no
enlarged femoral and precloacal scales continuous (FS-PS)	yes	yes	no	yes	yes
enlarged proximal femoral < 1/2 size of distal femorals or equal in size (FS-sz)	equal	equal	equal	equal	subequal
pocketing between digits of hind feet (Dig-pok)	no	no	no	no	no
dorsal pattern faded (DP-fad)	yes	yes	slightly	yes	no
distinct dark blotches on top of head present (HD-mrk)	no	no	no	no	no
dark body bands with lightened centers (BB-Cntr)	yes	yes	yes	yes	no
dark body markings in light interspaces (BB-intr)	no	no	no	no	no
dark dorsal bands thin or ~ same width as light interspaces (BB-wd)	wider	wider	wider	wider	wider
light interspaces reduced to a narrow thin white band	no	no	no	no	no
dark dorsal bands bordered by prominently white tubercles (WHT-tub)	yes	yes	yes	yes	yes
dark caudal bands wider than light caudal bands (DCB-wd)	yes	yes	yes	yes	yes
light caudal bands bearing dark marking in adults (WCB-mrk)	yes	yes	yes	yes	yes
Morphometric characters (mm)
SVL	71	74	78.9	73.7	56.7
TL	85	82	88	/	81
TW	6.4	6.1	6.3	6.4	4.5
FL	10.8	12.8	13	13.2	9
TBL	14.9	15.5	15.7	15.7	11.7
AG	32.3	33.4	43.9	33.3	26.2
HL	21.2	21.6	22.6	21.6	16.6
HW	14.1	14.5	14.4	14.6	10.8
HD	8.6	8.7	9	9.2	6.9
ED	4.9	5	5	4.9	4.2
EE	5.6	6	6.1	4.9	4.2
SN	8.3	8.2	8.8	8.3	6.7
EN	6.3	6.3	6.6	6.2	4.9
IO	4.6	5.2	4.5	4.1	3.1
EL	2.2	1.6	2	2.1	1.8
IN	2.3	2.4	2.3	2.2	2

Table 7.

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Data for the referred series Sampeu from the Phnom Sampeu and Phnom Khpoh populations, Battambang Province, Cambodia.

Locality	Phnom Sampeu	Phnom Sampeu	Phnom Sampeu	Phnom Sampeu	Phnom Sampeu	Phnom Sampeu	Phnom Sampeu	Phnom Khpoh	Phnom Khpoh	Phnom Khpoh
specimen	LSUHC 15113	LSUHC 15115	LSUHC 15116	LSUHC 15118	LSUHC 15114	LSUHC 15117	LSUHC 15120	LSUHC 15230	LSUHC 15229	LSUHC 15231
sex	m	m	f	m	m	m	f	f	f	m
Meristic characters
supralabials (SL)	12	11	10	11	11	10	10	11	10	11
infralabials (IL)	10	9	9	10	10	11	11	10	9	9
paravertebral tubercles (PVT)	30	30	31	31	29	28	30	28	28	30
longitudinal rows of tubercles (LRT)	15	15	17	16	17	15	17	16	18	17
longitudinal rows of ventral scales (VS)	36	39	37	34	39	41	37	37	42	41
expanded subdigital lamellae on 4th toe (E4LT)	6	7	6	6	6	6	6	6	7	7
unmodified subdigital lamellae on 4th toe (U4LT)	12	13	12	14	13	12	13	13	13	13
total subdigital lamellae on 4th toe (T4TL)	18	20	18	20	19	18	19	19	20	20
enlarged femoral scales (R,L)	13+11	13+13	16+13	15+13	11+13	12+12	15+12	15+11	16+10	14+12
total of enlarged femoral scales (TFS)	24	26	29	28	24	24	27	26	26	26
femroal pores (FP)	0	0	8 dimples	0	0	0	8 weak dimples	0	0	0
enlarged precolacal scales (PS)	8	8	8	8	8	8	8	10	9	9
precloacal pores (PP)	8	8	8 small	8	8	8	8 small	10 dimples	9 dimples	9 dimples
post-precloacal scale rows (PPS)	2	3	2	3	2	3	3	3	3	3
postcloacal tubercles (PCT)	2	2	1	1	1	2	1	3	3	1
body dark bands (BB)	4	4	4	4	4	4	4	4	4 on sacrum	4
dark caudal bands (DCB)	10	11	10	/	/	/	/	10	/	/
light caudal bands (LCB)	9	10	10	/	/	/	/	10	/	/
Categorical characters
body tubercles greatly reduced (Tub-red)	no	no	no	no	no	no	no	no	no	no
body tubercles moderately keeled (Tub-kld)	yes	yes	yes	yes	yes	yes	yes	yes	yes	yes
caudal tubercles extend beyond base of tail (CT-ext)	yes	yes	yes	yes	yes	yes	yes	yes	yes	yes
subcaudals expanded (SubC-exp)	yes	yes	yes	yes	yes	yes	yes	yes	yes	yes
subccaudals extend up onto lateral side of tail (SubC-lat)	no	no	no	no	no	no	no	no	no	no
enlarged femoral and precloacal scales continuous (FS-PS)	yes	yes	yes	yes	yes	yes	yes	no	no	no
proximal femoral < 1/2 size of distal femorals (FS-sz)	equal	equal	yes	yes	equal	yes	yes	yes	yes	yes
pocketing between digits of hind feet (Dig-pok)	no	no	no	no	no	no	no	no	no	no
dorsal pattern faded (DP-fad)	yes	yes	yes	yes	yes	yes	yes	no	no	no
distinct dark pigmented blotches on top of head present (HD-mrk)	no	no	no	no	no	no	no	no	faint	no
dark body bands with lightened centers (BB-Cntr)	yes	yes	yes	yes	yes	yes	yes	yes	yes	yes
dark body markings in light interspaces (BB-intr)	no	no	no	no	no	no	no	slightly	slightly	slightly
dark dorsal bands thin or ~ same width as light interspaces (BB-wd)	equal	equal	equal	equal	slightly wider	slightly wider	slightly wider	wider	equal	wider
light interspaces reduced to a narrow thin white band (INT-red)	no	no	no	no	no	no	no	no	no	no
dark dorsal bands bordered by prominently colored white tubercles (WHT-tub)	yes	yes	yes	yes	yes	yes	yes	yes	yes	yes
dark caudal bands wider than light caudal bands (DCB-wd)	yes	yes	yes	yes	yes	yes	yes	yes	yes	yes
light caudal bands bearing dark marking in adults (WCB-mrk)	yes	yes	yes	yes	yes	yes	yes	faint	faint	faint
Morphometric characters (mm)
SVL	72.4	70.7	80.0	71.2	71.8	67.1	56.4	80.0	78.5	53.5
TaiL	100.0	88.0	104.0	75.0	88.0	75.0	73.0	81.0	77.0	58.0
TaiW	6.5	5.3	6.2	5.8	6.2	5.5	4.8	6.3	6.3	4.6
FL	12.2	11.9	11.6	12.4	11.4	10.4	8.8	13.3	12.8	8.2
TBL	15.2	13.2	14.6	14.2	13.6	12.7	11.0	15.4	15.5	10.2
AG	33.6	32.6	37	32.7	32.9	30	25.5	36.5	36.6	22.7
HL	21.2	20.1	21.7	20.1	20.6	19.2	16.5	22.3	22.8	15.7
HW	14.5	13.7	15.4	14.1	14.0	12.5	10.9	16.0	15.7	10.3
HD	8.4	7.6	9.0	8.7	8.6	8.6	6.5	8.9	9.7	6.7
ED	4.9	4.7	5.2	4.4	4.7	4.6	3.9	5.3	5.5	3.8
EE	5.1	5.4	6.1	5.6	5.4	5.6	4.5	6.4	5.9	4.2
SN	8.0	7.9	8.5	7.8	8.2	7.6	6.5	8.7	8.7	6.1
EN	6.1	5.9	6.3	6.0	6.0	5.6	4.8	6.3	6.4	4.2
IO	4.9	4.7	4.6	4.4	5.0	5.0	4.2	6.2	5.1	3.5
EL	1.4	2.0	2	1.7	1.6	1.9	1.4	2.3	1.7	1.6
IN	2.4	2.0	2.3	2.2	2.4	2.4	1.9	2.7	2.4	1.9

In meristics, Cyrtodactylus kampingpoiensis sp. nov. differs significantly from the P. Sampeu population in having more rows of longitudinal tubercles (LRT: 19–21 vs 15–17). It differs significantly from the P. Khpoh population and the P. Sampeu population in having a greater number of paravertebral tubercles (PVT 30–37 vs 28–30 and 28–31, respectively) (Tables 3–5). It differs significantly morphometrically from the P. Sampeu population in having a significantly shorter ear-eye distance (EE) and a wider and longer head (HL and HW, respectively) and from the P. Sampeu and P. Khpoh populations in having a longer snout (SN) and longer hind limbs (TBL) (Table 3). Categorically it differs from the P. Sampeu population in having discontinuous versus continuous enlarged femorals and precloacals (FS-PS) and there are no dark markings in the light interspaces between the dark body bands (BB-intr) whereas faint markings occur in the P. Khpoh population (Figs 7, 9, 10; Suppl. material 3).

Natural history.

All specimens of the type series were collected on P. Kamping Poi from 1030–2000 hrs. Phnom Kamping Poi is a long rectangularly shaped karstic hill ca 6.5 km in length and 1.6 wide and its widest point near its southeastern margin (Fig. 1). Phnom Kamping Poi reaches ca 242 m in elevation and is covered with drought-deciduous karst vegetation (Fig. 8). The hillsides are covered with karstic boulders of varying size which constitutes the prime microhabitat for C. kampingpoiensis sp. nov. although some specimens were found in a wide, open cave near the entrance. All age classes from hatchlings (LSUHC 15198; 39 mm SVL) to adults (LSUHC 15206, 15209, 15212; 82 mm SVL) were found and LSUHC 15207 (SVL 77.9 mm) was gravid with two eggs. Although geckos were found on all substrates, they were most commonly found on karst in all planes of orientation with fewer specimens being found on the ground and the karst vegetation – often at the base of trees. One specimen (LSUHC 15204 [in the FFI collection]) was found deep within a cave on top of a large (2.5 m diameter) log. Other species found on P. Kamping Poi were the lizards Eutropis longicaudata (Hallowell), Dixonius siamensis (Boulenger), Gekko gecko (Linnaeus), Gehyra cf. lacerata (Taylor), Gehyra mutilata (Wiegmann) Hemidactylus frenatus Duméril & Bibron, and Subdoloseps bowringii (Günther), and the snakes Lycodon capucinus Boie and Indotyphlops cf. braminus (Daudin).

Figure 8. 

Habitat at Phnom Kamping Poi A, B optimal microhabitat of boulders of varying size within the karst vegetation C cave microhabitat inside Damrei Cave D typical deciduous karst vegetation.

All specimens from P. Banan (Figs 9, 10) were collected near Rum Say Sok from 1930–2000 hrs. Phnom Banan is an irregularly shaped elongate karstic hill ca 6.6 km in length and 1.6 km wide at its widest point near its the eastern margin (Fig. 1). It reaches ca 203 m in elevation and is covered with drought-deciduous karst vegetation (Fig. 11). All age classes from hatchlings (LSUHC 15177 [in FFI collection]; 35.0 mm SVL) to adults (LSUHC 15168 [in FFI collection]; SVL 82.0 m) were found, indicating that mid-March is within the reproductive season for this population. Although geckos were occasionally found on the ground (most hatchings and juveniles), they were most commonly found on the karst surfaces in all planes of orientation with fewer specimens being found on karst vegetation (only at the base of trees less than 0.5 m above the ground or on fallen logs). Other species found on P. Banan were the frogs Polypedates cf. leucomystax (Gravenhorst); Glyphoglossus guttulatus (Blyth); Kaloula pulchra Gray; Microhyla mukhlesuri Hasan, Islam, Kuramoto, Kurabayashi & Sumida; the lizards Gehyra mutilata (Wiegmann); Subdoloseps bowringii (Günther); and Eutropis macularia (Blyth); and the snakes Oligodon cf. kampucheaensis Neang, Grismer & Daltry; and a Chrysopelea ornata (Shaw) found dead on a nearby road.

Figure 9. 

The Phnom Banan population A adult male LSUHC 15173 B adult female LSUHC 15168 in FFI collection C adult female LSUHC 15170 D juvenile LSUHC 15176 E hatchling LSUHC 15175.

Figure 10. 

Specimens from the Phnom Banan population.

Figure 11. 

Habitat at Phnom Banan A hillside microhabtiat B Cliff face microhabitat C deciduous karst vegetation following summer rains. Photographs by Sothearen Thi.

All specimens were from P. Sampeu (Figs 12, 13) from were collected from 1930–2050 hrs. Phnom Sampeu is a small oval karstic hill ca 1.8 km in length and 0.6 km across the center (Fig. 1). It reaches 102 m in elevation and is covered with drought-deciduous karst vegetation. The hillsides are covered with karstic boulders of varying size as well as shear vertical faces where specimens were most commonly found (Fig. 14). Geckos, however, were also found on the ground and on karst vegetation, usually at the base of trees. All age classes from hatchlings (LSUHC 15126; 35 mm SVL) to adults (LSUHC 151116; 80 mm SVL) were observed, indicating mid-March is within the reproductive season of the P. Sampeu population. Specimen LSUHC 15113 had eaten a large Huntsman Spider. Other species found on P. Sampeu were the frogs Duttaphrynus cf. melanostictus (Schneider), Kaloula pulchra Gray, and Polypedates cf. leucomystax (Gravenhorst), the lizards Dixonius siamensis (Boulenger), and the snakes Lycodon capucinus Boie and Trimeresurus cf. albolabris Gray.

Figure 12. 

A, C The Phnom Sampeu population A adult male LSUHC 15113 B juvenile LSUHC 15119 in FFI collection C hatchling LSUHC 15123 D–F the Phnom Khpoh population D gravid adult female LSUHC 15229 E juvenile female LSUHC 15231 F hatchling LSUHC 15233 in FFI collection.

Figure 13. 

Comparison of specimens from the Phnom Khpoh and Phnom Sampeu populations.

Figure 14. 

Habitat at Phnom Sampeu A vertical face microhabitat of a low karstic ridge B boulder microhabitat within open karst vegetation C vertical microhabitat D typical deciduous karst vegetation where LSUHC 15118 was found on the base of the tree in the foreground.

Phnom Khpoh is overall a somewhat circularly shaped karstic hill with deeply incised margins 8.6 km to the west of P. Sampeu. It is 5.0 km by 4.5 km in size with three major peaks – the western and eastern peaks reaching ca 250 m in elevation and a southern peak reaching ca 242 m in elevation (Fig. 1). Like the other hills, P. Khpoh is covered with drought-deciduous karst vegetation, varying karst boulder microhabitats, and caves of varying length and depth (Fig. 15). The three specimens collected were found on karst and vegetation from 1900–2030 hrs at the opening of a small cave. A fourth specimen was found low on a cave wall ~ 4 m from the opening (Figs 13, 15). All age classes from hatchlings (LSUHC 15232 [in the FFI collection; 36 mm SVL]) to adults (LSUHC 15230; 80.0 mm SVL) were observed. LSUHC 15229 (SVL 78.5 mm) is gravid. Other species found on P. Khpoh were the frogs Duttaphrynus cf. melanostictus (Schneider), Kaloula pulchra Gray, and Polypedates cf. leucomystax (Gravenhorst), the lizards Dixonius siamensis (Boulenger) and Hemiphyllodactylus khpoh Grismer, Sinovas, Quah, Thi, Chourn, Chhin, Hun, Cobos, Geissler, Ching & Murdoch, and the snakes Lycodon capucinus Boie and Trimeresurus cf. albolabris Gray.

Figure 15. 

Habitat at Phnom Khpoh A typical boulder microhabitat within karst vegetation B open rocky microhabitat where specimens are rarely found C vertical opening into a cave microhabitat D cave microhabitat shared with Python bivittatus.

The description of Cyrtodactylus kampingpoiensis sp. nov. brings the total number of named Cyrtodactylus in Cambodia to 10. Much of this diversity was due to the partitioning of C. intermedius into seven species (Murdoch et al. 2019) followed by descriptions of additional species (see summary in Chhin et al. 2024). However, given that the neighboring countries of Thailand, Laos, and Vietnam have 55, 25, and 55 named species, respectively, is a clear indication that Cyrtodactylus diversity in Cambodia is underestimated, even given its smaller size than the former countries. This is due mostly to the fact that large areas of Cambodia remain completely unexplored – especially areas with habitats such as karst which are known to harbor an inordinately large number of specialized karst-dwelling Cyrtodactylus (Grismer et al. 2021b). Western Cambodia is just such an area. Our survey covered only seven karstic outcroppings in a small area (Fig. 1), but we observed five separate populations, at least four of which for now represent a new species. During a reconnaissance of the region between the cities of Battambang and Palin near the Thai-Cambodia border, a straight-line distance of only ca 62 km, we observed ~ 30 unexplored karstic areas ranging from small towers to mountain ranges (Fig. 16). There are also unexplored karstic areas in northern and southern Cambodia. The discovery of this new species and its considerable degree of morphological interpopulational variation underscores the necessity for further exploration in order to begin understanding the overall herpetological diversity of Cambodia in general, and western Cambodia in particular, where dozens of isolated karstic habitats still remain to be surveyed. Given the extremely localized distribution of these four new populations, each meets the IUCN criteria of distinct genetic lineages that are Critically Endangered.

Figure 16. 

A Karstic ridge of Phnom Banan looking east B west end of Phnom Banan looking north. A cave opening can be observed near the summit. Several unsurveyed karst hills cab be seen in the background on both photographs. Drone photographs taken by Phyroum Chourn.

These data sets were analyzed using separate PCAs because each provides a different type of information concerning the morphological evolution and adaptation of these populations as well as different diagnostic systems. Even though all populations plot separately in both PCAs (Figs 3, 4) this does not mean they are all significantly different from one another in both analyses. The PERMANOVA and DAPC of the morphometric data clearly demonstrate that body shape among these populations is far more conserved than the plots illustrate, being that only two population pairs differ significantly from one another (Fig. 3B, C). Whereas the PERMANOVA and DAPC for the meristic data indicate that nearly all the populations differ significantly from one another and all but one of these species pairs differed at the more rigorous p-adjusted level (Fig. 4B, C). This would suggest that body shape for living in a highly specialized karstic microhabitat is under stronger selection pressure and as such, is able to vary less whereas the number of scales on various parts of the body can vary randomly and may be under no selection at all. A number of studies have shown that karst-dwelling Cyrtodactylus are morphometrically different than species living on other substrates and may even differ among species living on different kinds of karst (e.g., Chomdej et al. 2020; Grismer et al. 2020a, b, 2021a, 2023b; 2024a; Kaatz et al. 2021; Termprayoon et al. 2023; Wood et al. 2020). Understanding these differences can aid in putting into place more effective and efficient conservation measures.

The advent of integrative taxonomy with its incorporation of molecular phylogenies, has been instrumental in recovering unrealized species diversity within species complexes. Molecular analyses offer a more fine-grained, less subjective approach to species delimitation that is divorced from a strictly morphological approach which diagnoses and delimits species simultaneously. An inherent error with conflating these independent operations (see Frost and Hillis 1990; Frost and Kluge 1994; Hillis 2019) is the assumption that “diagnosable” populations lack gene flow – usually the primary criterion for delimiting species (de Queiroz 2007). Commonly exacerbating this issue, are morphological descriptions lacking statistical defensibility (see Chan and Grismer 2022), leaving any assumption of interspecific gene flow weak, thus rendering the validity of the “new” species dubious, typical of late 19^th and early 20^th century taxonomy (see for example Pauwels et al. 2024; Sumontha et al. 2024). We are aware that species descriptions based on older museum material do not avail themselves to molecular data, which is all the more reason, when possible, to provide rigorous statistical diagnoses.

We elect to recognize Cyrtodactylus kampingpoiensis sp. nov. as a new geographically variable species. Even though the mitochondrial phylogeny may indicate there is no interpopulational gene flow – given that all the populations are monophyletic – and it aligns with their statistically significant and categorical diagnosability, the presence or absence of gene flow requires definitive confirmation with nuclear markers – preferably a genomic data set – especially when genetic divergence is low. Notably, these newly discovered populations are allopatric on separate karst formations separated by ca 5–20 km of uninhabitable terrane (mostly degraded savannah or paddy fields) indicating that migration among these formations is extremely unlikely, particularly true of karst-adapted geckos with low vagility. Furthermore, unlike oceanic islands where passive overwater dispersal can result in admixture similar to upland sky-island habitats and land bridge islands that form and reform with glacial cycling, separate karst formations do not reform. In general, the separate towers, cones, and hills become isolated through erosion, weathering, and changing water courses (Wang et al. 2019). Thus, species adapted to live only in karstic microhabitats are essentially restricted to these environs. This is not to say that a habitat generalist may not disperse into a karstic area and hybridize with a karst-dwelling species, but we see no evidence of this in Cyrtodactylus even where karst-dwelling species and species with different habitat preferences are syntopic on the periphery of the karst formation (e.g., C. astrum [karst] and C. macrotuberculatus [granite forest] in Peninsular Malaysia; Grismer 2011; Grismer et al. 2012). Nonetheless, until the absence of gene flow is unequivocally confirmed, we reluctantly elect to consider all four populations to be a single geographically variable species. We realize this decision may not be in the true spirit of “integrative taxonomy” (Padial et al. 2010) that aims to integrate all forms of relevant data – geogrpahy, geology, ecology, and natural history, and that perhaps genetic distance alone should not be the overriding factor. We are currently working on a genomic analysis of these populations.

The high levels of biodiversity and site-specific endemism in many karstic regions rival that of most other habitats, yet these regions are rapidly becoming some of the most imperiled ecosystems in Southeast Asia (Clements et al. 2006; Grismer et al. 2016 Luo et al. 2016; Quah et al. 2021). Southeast Asia harbors more karst habitat than anywhere else in the world (Day and Urich 2000) but unregulated and unsustainable quarrying practices, the primary threat to karst-dwelling species, continues to degrade the integrity of these landscapes in many developing countries where enormous monetary gains override issues of conservation. This only serves to amplify the biodiversity crisis in Southeast Asia, where the overall rate of habitat loss is the highest among the world’s tropical regions (Sodhi and Brook 2006). A number of studies have shown that there are far more karst-associated vertebrates in Southeast Asia than previously reported (e.g., Luo et al. 2016; Connette et al. 2017) and the discovery rate of new species of karst-adapted amphibians and reptiles shows no signs of leveling off. Continued field surveys in western Cambodia will no doubt add to this increasing rate. The report here, and that of Grismer et al. (2025) of the new species of site-restricted, karst-dwelling Hemiphyllodactylus khpoh from Battambang Province alone, underlines the conservation and management needs of these areas in Cambodia. Given the extremely localized distribution of these newly discovered species and populations, each meets the IUCN criteria of being Critically Endangered. Some of the karst formations harboring site-specific endemics are also religious retreats and protected places of Buddhist worship, and thus under some level of de facto protection from quarrying operations. However, dozens are not and, as such, they remain under the threat of unsustainable quarrying. Intensive biodiversity systematic surveys of all these formations is paramount to underpin measures of protection for this portion of Cambodia’s natural heritage.

The authors have declared that no competing interests exist.

No ethical statement was reported.

This research was funded by the European Union through the BCOMING project, Horizon Europe Project 101059483.

Conceptualization: P. Sinovas and S. Thi. Generation of DNA data: M.L. Murdoch. Phylogeny construction and statistics: L.L. Grismer. Original draft: L. L. Grismer. Review and Editing: all authors. Field work: All authors.

Evan S. H. Quah https://orcid.org/0000-0002-5357-1953

L. Lee Grismer https://orcid.org/0000-0001-8422-3698

Pablo Sinovas https://orcid.org/0000-0001-5396-8910

Phyroum Chourn https://orcid.org/0009-0005-8664-5321

Sophea Chhin https://orcid.org/0000-0002-5612-7532

Seiha Hun https://orcid.org/0009-0004-0147-1565

Anthony Cobos https://orcid.org/0009-0001-9215-4052

Peter Geissler https://orcid.org/0000-0001-7408-1358

Christian Ching https://orcid.org/0000-0001-6972-6836

Matthew L. Murdoch https://orcid.org/0000-0001-5914-6408

Sothearen Thi https://orcid.org/0009-0009-4827-1719

Jesse L. Grismer https://orcid.org/0000-0002-2542-5430

All of the data that support the findings of this study are available in the main text or Supplementary Information.