Research Article |
Corresponding author: David Prötzel ( david.proetzel@mail.de ) Academic editor: Johannes Penner
© 2018 David Prötzel, Shea M. Lambert, Ginah Tsiorisoa Andrianasolo, Carl R. Hutter, Kerry A. Cobb, Mark D. Scherz, Frank Glaw.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Prötzel D, Lambert SM, Andrianasolo GT, Hutter CR, Cobb KA, Scherz MD, Glaw F (2018) The smallest ‘true chameleon’ from Madagascar: a new, distinctly colored species of the Calumma boettgeri complex (Squamata, Chamaeleonidae). Zoosystematics and Evolution 94(2): 409-423. https://doi.org/10.3897/zse.94.27305
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On a recent expedition to eastern Madagascar, we discovered a distinct new species of the genus Calumma that we describe here using an integrative approach combining morphology, coloration, osteology and molecular genetics. Calumma roaloko sp. n. has a dermal rostral appendage and occipital lobes, and belongs to the C. boettgeri complex, within the Madagascar-endemic phenetic C. nasutum species group. It is readily distinguished from other species of the C. boettgeri complex by a characteristic two-toned body coloration and small body size with a snout-vent length of 45.6 mm in an adult male. The osteology of the skull, with a prominent maxilla and broad parietal, is similar to the closest related species, C. uetzi. Analysis of uncorrected genetic distances within the C. nasutum group using the mitochondrial gene ND2 shows a minimum pairwise distance of 11.98% to C. uetzi from the Sorata massif and Marojejy National Park >500 km north of the type locality of C. roaloko sp. n.. Given an apparently small range (potentially <300 km2), located entirely outside of any nationally-protected areas, we recommend this new species be classified as Endangered under criterion B1ab(iii) of the IUCN Red List. The discovery of clearly distinct species like C. roaloko sp. n. in an area of Madagascar that is comparatively thoroughly surveyed highlights the critical role of continued field surveys for understanding the true extent of Madagascar’s spectacular biodiversity.
Calumma roaloko sp. n., Integrative taxonomy, Micro-computed tomography, Osteology, Calumma nasutum group
The biota of Madagascar is recognized as exceptional, both in terms of endemism and density of species (
With currently 90 endemic species (
The small-bodied chameleons of the Calumma nasutum group, usually characterized by their dermal rostral appendages, are distributed across the forests of eastern and northern Madagascar. Within this group, the species C. boettgeri (Boulenger, 1888), C. guibei (Hillenius, 1959) and C. linotum (Müller, 1924) differ from the others by the possession of well-defined occipital lobes and are referred to as the C. boettgeri complex (
During fieldwork in a forest fragment within the Réserve de Ressources Naturelles du Corridor Ankeniheny-Zahamena just south of Andasibe-Mantadia National Park in 2015/2016, we discovered a small-bodied chameleon with distinct coloration belonging to the Calumma nasutum group. Integrating morphological, molecular, and osteological data, we describe this new species of the C. boettgeri complex.
We located specimens at night using targeted searches of arboreal habitats during the rainy season, using flashlights to locate sleeping individuals. Following euthanasia, we removed a portion of liver tissue and transferred it immediately into 95% ethanol for use in DNA extractions for genetic analyses. Specimens were fixed in 10% formalin (buffered to pH 7.0 with sodium phosphate), and transferred to 75% ethanol for long-term storage after approximately two weeks. We deposited the holotype and four paratypes at the University of Kansas Biodiversity Institute, Lawrence, KS (
Terms of morphological measurements taken on these specimens were adapted from previous studies (
Morphological measurements of Calumma roaloko sp. n. All measurements in mm. For abbreviations, see Materials and methods.
Final museum no. |
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original museum no. |
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field no. | SML 213 | SML 177 | SML 210 | SML 178 | SML 166 |
sex | adult male | adult female | subad. male | subad. male | subad. female |
type status | holotype | paratype | paratype | paratype | paratype |
altitude [m] | 1100 | 1100 | 1100 | 1100 | 1100 |
SVL | 45.6 | 44.5 | 37.6 | 38.6 | 40.0 |
TaL | 48.1 | 41.0 | 44.3 | 42.3 | 34.8 |
TL | 93.7 | 85.5 | 81.9 | 80.9 | 74.8 |
RTaSV | 105% | 92% | 118% | 110% | 87% |
LRA | 5.2 | 2.3 | 4.6 | 3.9 | 2.7 |
RRASV | 11.4% | 5.2% | 12.2% | 10.1% | 6.8% |
DRA | 2.6 | 1.9 | 2.3 | 2.7 | 1.6 |
RDRSV | 5.7% | 4.3% | 6.1% | 7.0% | 4.0% |
NDRA | 5 | 6 | 4 | 5 | 7 |
NSRA | 16 | 29 | 28 | 33 | 31 |
RNLRA | 3.1 | 12.6 | 6.1 | 8.5 | 11.5 |
RC | + | + | + | + | + |
LC | + | + | + | + | + |
TCL | – | – | – | – | – |
TCR | – | – | – | – | – |
PC | + | – | – | – | – |
OL | c | c | c | c | c |
OLND | 0.4 | 0.2 | 0.3 | 0.2 | 0.3 |
RODSV | 8.8% | 10.1% | 10.4% | 10.6% | 10.3% |
OLD | 4.0 | 4.5 | 3.9 | 4.1 | 4.1 |
ROLSV | 4.6% | 4.3% | 5.1% | 5.2% | 4.5% |
OLW | 2.1 | 1.9 | 1.9 | 2.0 | 1.8 |
ROWSV | 0.9% | 0.4% | 0.8% | 0.5% | 0.8% |
DSOL | 0.7 | 0.5 | 0.7 | 0.6 | 0.7 |
DSCT | 0.7 | 0.6 | 0.7 | 0.6 | 0.6 |
DC | 2 | 0 | 1 | 0 | 0 |
DSA | 0.7 | 0.5 | 0.5 | 0.4 | 0.4 |
NSA | 11 | 14 | 13 | 15 | 15 |
NSL | 13 | 13 | 13 | 13 | 13 |
NIL | 13 | 14 | 13 | 13 | 12 |
For internal morphology, micro-Computed Tomography (micro-CT) scans of the head were prepared for the male holotype
Micro-CT scans of skulls of Calumma roaloko sp. n. Male holotype
Osteological measurements based on micro–CT scans of the skulls of the male holotype and an adult female of Calumma roaloko sp. n. All measurements in mm. For abbreviations, see Materials and methods.
Collection no. |
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field no. | SML 213 | SML 177 |
sex | male | female |
type status | holotype | paratype |
NaL | 1.4 | 1.8 |
RNaL | 11.6% | 15.7% |
FWPf | 3.2 | 2.8 |
RFWPf | 26.4% | 24.3% |
FWPo | 4.5 | 4.1 |
RFWPo | 37.2% | 35.7% |
FWPa | 4.1 | 3.9 |
RFWPa | 33.9% | 33.9% |
PWPo | 4.7 | 4.0 |
RPWPo | 38.8% | 34.8% |
PWm | 2.7 | 2.2 |
RPWm | 22.3% | 19.1% |
PL | 5.1 | 4.9 |
RPL | 42.1% | 42.6% |
FL | 7.0 | 6.0 |
RFL | 57.9% | 52.2% |
SCL | 14.5 | 13.5 |
RSCL | 119.8% | 117.4% |
SkL | 12.1 | 11.5 |
We extracted genomic DNA from tissue samples at the
Maximum-likelihood tree of the Calumma boettgeri complex, based on 513 base-pairs of the mitochondrial ND2 gene. Nodal support values indicate the proportional support from 1000 rapid bootstrap replicates. Support values for intra-specific relationships are not shown. Outgroup (C. oshaughnessyi FGZC 4577) not shown for graphical reasons.
Prior to phylogenetic analysis of the ND2 gene, conducted using maximum-likelihood in RAxML 8.2.6 (
The electronic version of this article in Portable Document Format (PDF) will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new names contained in the electronic version are effectively published under that Code from the electronic edition alone. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix http://zoobank.org/. The LSID for this publication is: urn:lsid:zoobank.org:pub:2433A9DD-8AC1-4139-A639-E24053D5C33F. The online version of this work will be archived and made available from the following digital repositories: CLOCKSS and Zenodo.
The ND2 alignment contained 513 sites and a total of 235 variable sites, of which 177 were parsimony informative. The genetic analysis of the ND2 gene fragment (Fig.
Map showing the location of the known range of Calumma roaloko sp. n. in central-eastern Madagascar. Red stars indicate localities where C. roaloko sp. n. was found, gray “X” indicate localities surveyed but with no detection of the species. The map is a composite of Landsat 7 and SRTM (Shuttle Radar Topographic Mission;
Calumma roaloko sp. n. is a member of the phenetic C. nasutum species group (
Calumma roaloko sp. n. differs from C. fallax, C. gallus, C. nasutum, C. peyrierasi, C. vatosoa and C. vohibola of the C. nasutum group by the presence of occipital lobes; from C. boettgeri, C. gehringi, C. guibei, C. lefona, C. linotum and C. juliae in the generally smaller body size with a maximum SVL of 45.6 mm and a maximum TL of 93.7 mm (vs. a range of SVL maxima in the former species of 49.1–59.6 mm and TL maxima of 98.7–126.1 mm), and a straight-lined dorsal margin of the supralabial scales vs. serrated (character ‘en dents de scie’ in
From the most similar taxon Calumma uetzi, C. roaloko sp. n. differs in the absence of a dorsal crest or presence of at most two cones (vs. presence of 5–14 cones), absence of a temporal crest (vs. presence of 1–2 temporal tubercles), greater number of supralabial scales (13 vs. 10–12) and infralabial scales (12–14 vs. 11–12), a longer rostral appendage in adult males of 5.2 mm with large tubercle scales (vs. 3.8 mm, small and smooth tubercle scales; note: n = 1 each), and less heterogeneous scalation on the head with diameter of largest scale in temporal region of 0.6–0.7 mm (vs. 1.0–1.3 mm). The osteology of the skull is similar in both species; C. roaloko sp. n. differs from C. uetzi only in the absence of elevated protuberances at the anterior end of the maxilla that characterize the skull of male C. uetzi. Calumma roaloko sp. n. furthermore differs from all other species by distinct differences in the mitochondrial genes ND2 and COI and a unique two-colored life-coloration.
Adult male (Figs
Description based on a micro-CT scan (Fig.
The micro-CT scan uncovered a worm-like structure that lies curled and fractured in the throat and proceeds posteriorly into the chameleon’s body presumably via the esophagus. We suppose that this shows an endoparasite trying to leave the dying chameleon after the processing, but note that it is remarkably strongly mineralized.
The body of the holotype in preservative (Fig.
The four paratypes agree well with the holotype in most characters of morphology and osteology. However, all four paratypes have more tubercle scales on rostral appendage on right side (28–33 vs. 16) and all paratypes lack a parietal crest (vs. indistinctly present); dorsal crest absent in
Based on observations and photographs of the type specimens (Figs
The hemipenes of the three male specimens (the holotype
There are no available names that could be attributed to a species of the C. nasutum group with occipital lobes.
The specific epithet “roaloko” is a combination of the Malagasy words “roa” meaning “two” and “loko” meaning “color”, in reference to the characteristic two-toned body colorations of males (green and white) and females (brown and tan) of this species. The epithet is to be treated as an invariable noun in apposition.
The specific natural history of C. roaloko sp. n. is little-known, but assumed to be similar to other small-bodied Calumma. As with other C. nasutum group species, individuals of C. roaloko sp. n. were encountered sleeping at night on leaves (Fig.
Given current evidence, the distribution of C. roaloko sp. n. is potentially restricted to a small fragment (~300 km2) of mid-elevation rainforest that lies outside of nearby Analamazaotra Special Reserve and Andasibe-Mantadia National Park in central-eastern Madagascar (Fig.
The ~300 km2 fragment of mid-elevation rainforest from which C. roaloko sp. n. is known is managed by several local government councils, and has recently been established as a new protected area (Réserve de Ressources Naturelles du Corridor Ankeniheny-Zahamena) within the scope of the expansion of Madagascar’s national parks (
The discovery of C. roaloko adds to a growing understanding of the diversity of small-bodied Calumma in Madagascar (
Biogeographically the pattern of diversity in the C. nasutum group currently suggests a complex history, possibly involving several major dispersal events, especially within the C. boettgeri complex, with C. roaloko being sister to C. uetzi, a species found >500 km to the north in the Sorata massif and Marojejy NP (
With a total length of 93.7 mm and a body size of 45.6 mm in the largest specimen (the male holotype), C. roaloko represents the smallest member of the “true” chameleons, subfamily Chamaeleoninae, (excluding the small, mostly ground dwelling species of the subfamily Brookesiinae) on Madagascar, and one of the smallest members of the Chamaeleoninae in the world. Within the C. nasutum group C. uetzi (with maximum TL of 101.2 mm and maximum SVL of 45.7 mm) and C. vohibola (with maximum TL of 90.5 mm – resulting from a measuring error due to a cut-off tail for DNA analysis – and maximum SVL of 49.8 mm,
Interestingly, Calumma roaloko and its sister taxon C. uetzi are among the only species within the C. nasutum group with strong sexual dichromatism. Males of both species differ clearly from the females by a conspicuous display coloration that contrasts well from the green and brown overall background of their habitat as shown for some chameleon species of the genus Bradypodion (
The increase in species richness in chameleons may come not only from the splitting of currently recognized and often widespread species (e.g., in the African chameleon genus Kinyongia,
We thank the Malagasy authorities for granting permits used to conduct field research and collecting efforts (Research conducted under research permit Nº329/15/MEEMF/SG/DGF/DAPT/SCBT, specimens exported under CITES permit Nº065C-EA01/MG16, 26/01/2016). Field research was supported by Global Wildlife Conservation through Grant 5019-0096 to CRH and SML. Landsat data were made available from the U.S. Geological Survey. We are grateful to Miguel Vences for providing DNA sequences and to the organization MICET for logistical support. We also thank the Vondron’Olona Ifotony (V.O.I.) of Iaroka, Maroala, and Fandrefiala for granting permission to work in their forests. We thank all of our Malagasy guides, cooks and porters, with special thanks to guides Jean Aime Rajaonarivelo (‘Gagah’) and Regis. We are also grateful to Asa Conover, Devin Edmonds and Vincent Premel for their assistance in the field.
Genetic distances of ND2
Genetic distances of COI
Movie of 3D model of the skull
Movie of 3D model of the skull