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Evolutionary reduction of adult body size (miniaturization) has profound consequences for organismal biology and is an important subject of evolutionary research. Based on two individuals we describe a new, extremely miniaturized chameleon, which may be the world’s smallest reptile species. The male holotype of Brookesia nana sp. nov. has a snout–vent length of 13.5 mm (total length 21.6 mm) and has large, apparently fully developed hemipenes, making it apparently the smallest mature male amniote ever recorded. The female paratype measures 19.2 mm snout–vent length (total length 28.9 mm) and a micro-CT scan revealed developing eggs in the body cavity, likewise indicating sexual maturity. The new chameleon is only known from a degraded montane rainforest in northern Madagascar and might be threatened by extinction. Molecular phylogenetic analyses place it as sister to B. karchei, the largest species in the clade of miniaturized Brookesia species, for which we resurrect Evoluticauda Angel, 1942 as subgenus name. The genetic divergence of B. nana sp. nov. is rather strong (9.9‒14.9% to all other Evoluticauda species in the 16S rRNA gene). A comparative study of genital length in Malagasy chameleons revealed a tendency for the smallest chameleons to have the relatively largest hemipenes, which might be a consequence of a reversed sexual size dimorphism with males substantially smaller than females in the smallest species. The miniaturized males may need larger hemipenes to enable a better mechanical fit with female genitals during copulation. Comprehensive studies of female genitalia are needed to test this hypothesis and to better understand the evolution of genitalia in reptiles.

Chameleons (Chamaeleonidae) are known for their arboreal lifestyle, in which they make use of their prehensile tail. Yet, some species have a more terrestrial lifestyle, such as Brookesia and Rieppeleon species, as well as some chameleons of the genera Chamaeleo and Bradypodion. The main goal of this study was to identify the key anatomical features of the tail vertebral morphology associated with prehensile capacity. Both interspecific and intra-individual variation in skeletal tail morphology was investigated. For this, a 3D-shape analysis was performed on vertebral morphology using μCT-images of different species of prehensile and nonprehensile tailed chameleons. A difference in overall tail size and caudal vertebral morphology does exist between prehensile and nonprehensile taxa. Nonprehensile tailed species have a shorter tail with fewer vertebrae, a generally shorter neural spine and shorter transverse processes that are positioned more anteriorly (with respect to the vertebral center). The longer tails of prehensile species have more vertebrae as well as an increased length of the processes, likely providing a greater area for muscle attachment. At the intra-individual level, regional variation is observed with more robust proximal tail vertebrae having longer processes. The distal part has relatively longer vertebrae with shorter processes. Although longer, the small size and high number of the distal vertebrae allows the tail to coil around perches.

We describe a new species of dwarf chameleon from the Brookesiaminima species group. Brookesiatedisp. n. occurs above 1300 m above sea level on the Marojejy massif in northeastern Madagascar. It is genetically sister to B.peyrierasi, a species occurring in lowlands around the Baie de Antongil, but is genetically strongly divergent from that species in both nuclear (c-mos) and mitochondrial (16S, ND2) genes, and morphologically in its smaller size and distinctly different hemipenis. It is the second species of the B.minima species group from Marojejy National Park, but is not known to occur in syntopy with the other species, B.karchei, due to elevational segregation.

Chameleons (Chamaeleonidae) feature many adaptations to their arboreal lifestyle, including zygodactylous feet, a prehensile tail, and epidermal microstructures. In arboreal tree chameleons, the substrate-contacting site of the feet and tail is covered by microscopic hair-like structures (setae) of 6-20 mu m length. Their friction enhancing function has been shown in recent studies. Leaf chameleons and one representative of the tree chameleons (Chamaeleo namaquensis) secondarily have become ground-dwelling. Because leaf chameleons are paraphyletic, one could expect that in the three leaf chameleon genera Brookesia, Rhampholeon, and Rieppeleon and the tree chameleon Ch. namaquensis, epidermis has adapted independently to terrestrial locomotion. Using scanning electron microscopy, we investigated the substrate-contacting surfaces of the feet (subdigital) of 17 leaf chameleon species and five tree chameleon species that have not yet been examined. Additionally, surfaces not involved in locomotion, the flanks (dorsolateral), and scale interstices, were examined. Although the subdigital microstructures in leaf chameleons are more diverse than in tree chameleons, we found some features across the genera. The subdigital microornamentation of Rhampholeon spinosus consists of long thin setae and spines, comparable to those of tree chameleons. All other Rhampholeon species have spines or short but broad setae. Rh. spectrum had tooth-like structures instead of setae. Subdigital scales of Brookesia have either thorns or conical scale-tops in the center and feature honeycomb microstructures. In Rieppeleon, subdigital scales have a thorn. Scale surfaces are covered by honeycombs and short hair-like structures (spines). As subdigital scales with a thorn in the center and honeycomb microstructures were also found in the terrestrial tree chameleon Ch. namaquensis, one can assume that this geometry is a convergent adaptation to terrestrial locomotion. Despite the great number of genus-specific traits, the convergent evolution of honey-comb structures in Brookesia, Rieppeleon, and Ch. namaquensis and the high variability of spines and setae in Rhampholeon suggests a rapid adaptation of subdigital microornamentation in Chamaeleonidae. J. Morphol. 276:167-184, 2015. (c) 2014 Wiley Periodicals, Inc.

The comparative vertebral morphology of different chamaeleonid genera has been generally neglected and some aspects such as the comparative anatomy of the neck region remain poorly known. The atlas and axis of all chamaeleonid genera (Brookesia, Rieppeleon, Archaius, Rhampholeon, Nadzikambia, Bradypodion, Chamaeleo, Calumma, Furcifer, Kinyongia, and Trioceros) are studied here. Considerable morphological differences are revealed. Additionally, some taxa exhibit sexual dimorphism in the atlas and axis. An extremely long, divided posterodorsal process is present in males of the Trioceros johnstoni + Trioceros jacksonii clade. The solid and well-developed morphology of the posterodorsal process in males of this taxon could reflect its competitive behavior-males fight with their horns and attempt to dislodge one another from branches during encounters. An additional area of insertion for the cervical musculature may indicate an incremental cervical musculature mass and cross sectional area that can add extra support and stability to the head and assist during combat involving lateral pushing. This character is not present in females. Heterochronic processes have played a role in the evolution of chamaeleonids, as evidenced in many characters of the atlas-axis complex. A new hypothesis of an anterior shifting of synapophyses of the axis is erected and a new derived anatomical structure of the parietal of Chamaeleo calyptratus is described (the processus parietalis inferior). The presence of the processus parietalis inferior is associated with the evolution of the dorsally elevated parietal crest.

International audience; Rhabdias casiraghii n. sp. and R. kibiraensis n. sp. are described from Trioceros ellioti from Burundi and co-infection was observed in one of the host specimens. Distinctive characters between these species are, among others, the mouth and buccal capsule in front view, both of which are round in the former and laterally flattened in the latter species. Both species have a complete set of submedian head papillae (three in each submedian axis) as observed in the fourth stage larva of R. americanus from anurans. This primitive character opposes them to other species parasitic in Chamaeleonidae that have a single papilla per axis. The third species is the first described from the primitive chamaeleonid genus Brookesia; R. brygooi n. sp. from B. superciliaris from Madagascar can be distinguished from other Rhabdias in Chamaeleonidae by the small diameter of its mouth and buccal capsule. In this character, it resembles parasites from anurans. However, its infective larva has a rounded caudal extremity ornated with buds, as described in species of Rhabdias parasitic in Chamaeleonidae.

Information on the distribution and abundance of chameleons in Madagascar is required to develop conservation plans that integrate protected area management and sustainable use. We surveyed chameleons in eight sites in deciduous forest in Menabe, western Madagascar. Brookesia brygooi was the most frequently detected species, with a population density of 35 ha–1. Furcifer species were less common, with calculated densities of 7.2 ha–1 (F. labordi), 3.0 ha–1 (Furcifer sp.) and 1.3 ha–1 (F. oustaleti). Chameleon abundance varied according to altitude (B. brygooi) and no clear effect from logging was detected (all species). A lack of information on chameleon diurnal habitat requirements impedes a fuller assessment of the extent to which these species are tolerant to forest degradation. There were interspecific differences in the height of nocturnal perches and additional studies are needed to determine whether these are related to diurnal resource partitioning. Furcifer labordi and Furcifer sp. are of conservation concern because they are restricted to native forests in western Madagascar.

We reviewed the diet of nocturnal and diurnal chameleon predators on Madagascar from the available literature and identified 28 species. Diurnal raptors were the main predators of chameleons, although the literature was biased towards birds and probably underestimated predation by other animals, notably snakes. Eutriorchis astul' (Accipitridae) and Falco zoniventris (Falconidae) are the principal avian predators of Furcifer spp. and Calumma spp. and the Brachypteracias leptosomus (Brachypteraciidae) of Brookesia spp.

Abstract Madagascar's Brookesia dwarf chameleons are believed to require relatively intact forest for survival. Although they have featured in herpetological surveys, taxonomic reviews and trade assessments, very little is known about their microhabitat requirements or ecology. Over a 5 night period in a deciduous forest in western Madagascar we recorded the night roosting sites for three sympatric Brookesia species. We calculated the area and distance between successive night roost locations and described the characteristics of each roost. Distance between roosts and the area used were larger for B. brygooi and B. perarmata than B. exarmata. The distance between roosts was significantly greater for male than female B. brygooi and B. perarmata. Roost sites were generally in low vegetation (

As a result of our recent herpetological surveys conducted in Madagascar, important new collections of large occipital-lobed Calumma chameleons have been made. These collections allow the recognition of six new Calumma species, in addition to the six currently recognized species. Five of the new species were located at specific montane regions, and one is broadly distributed in mid-altitude forest within the interior of the island. As previously reported for Brookesia dwarf chameleons, species richness in this Calumma group is greatest in northern Madagascar. An identification key is given for all 12 species and a new description provided for Calumma brevicorne, a species that has been confused with several other species. Previously described taxonomic uncertainties associated with this species are resolved, including the problematic status of the Calumma brevicorne tsarafidyi holotype, which we consider to represent the nominate form and unlikely to have been collected at Tsarafidy. Sexual dimorphism (including hemipenis development), biogeography, and conservation issues are also discussed for the group. The Bemanevika Lakes region of Southwest Tsaratanana harbors a new Calumma species not currently recorded for any protected area, and forest of this region is suggested for inclusion within a future reserve.

In a recent paper Raxworthy & Nussbaum (1995) included the Madagascan dwarf chameleons Brookesia peyrierasi and B. tuberculata in the synonymy of B. minima, regarding known hemipenial differences between B. tuberculata and B. peyrierasi as artefacts. Our studies demonstrate that the hemipenis description of B. tuberculata by Brygoo & Domergue (1975) was largely correct and referred to unusual but real structures. The enormous differences in the hemipenis morphology of B. tuberculata (hemipenes of six specimens examined) and B. peyrierasi (hemipenes of five specimens examined) strongly suggest that they are not conspecific. Furthermore, morphological differences in the arrangement of head crests and expression of supraocular spines were noted between these two taxa and B. minima. The differentiation of the three taxa is furthermore corroborated by both univariate and multivariate analyses of morphometric data. We therefore propose to consider Brookesia minima, B. peyrierasi, and B. tuberculata as distinct species. This conclusion corroborates the validity of biogeographic regions as suggested from the analysis of distribution patterns of other Brookesia species.

The phylogenetic analysis of skull characteristics, along with data derived from lung and hemipenial morphology, does not support monophyly of the predominantly terrestrial chamaeleons, Brookesiinae (Brookesia plus Rhampholeon), as sister-taxon of a monophyletic clade of arboreal chamaeleons, the Chamaeleoninae. Instead, the phylogenetic interrelationships of chamaeleonine lizards are (Brookesia (“Rhampholeon” (Bradypodion, Chamaeleo)). The monophyly of the genus Rhampholeon is not demonstrated. The arboreal chamaeleons form a monophyletic group comprising two genera, Bradypodion and Chamaeleo. Bradypodion may be placed as sister-group of the genus Chamaeleo, but its phyletic position and monophyly cannot at present be corroborated. This study documents that there is a high degree of intraspecific variability in skull characters which may be due to paedomorphic reduction in the skull, especially of small species. Characters affected by a variable degree of ossification include the separation of a prefrontal fontanelle, the contact of the squamosal with the parietal, the size and position of the fenestra vestibuli, the degree of ossification in the basicranial rim of the fenestra, the extent of ossification of the pterygoid wing, and the shape of the skull roof as indicated by the bones forming the dorsal margin of the orbit. The closure of the lateral aperture of the occipital recess might indicate that miniaturization, and concomitant paedomorphosis, may have played a role in the initial phases of chamaeleonine phylogeny.

During fieldwork carried out in January‐February 1995 in the Strict Nature Reserve No. 9 ≪Tsingy de Bemaraha≫ (western Madagascar), a new species of dwarf chameleon (genus Brookesia) was discovered on the western slopes of the Antsingy forest. The inaccessible habitat of this new taxon consists of dense subhumid tropophile forest growing among the limestone blocks belonging to the huge karstic complex of the ≪Tsingy de Bemaraha≫ Massif, where Brookesia exarmata sp. nov. is sympatric with Brookesia perar‐mata, a taxon likely endemic to the Antsingy forest, and Brookesia brygooi. The new taxon, formally described in this paper, is a small‐sized chameleon whose total adult length ranges between 30 and 45 millimetres: this feature, together with the absence of a pelvic shield and the presence of a series of dorsolateral tubercles (or a few small dorsolateral spines), leads to Brookesia exarmata being ascribed to the Brookesia minima group. Brookesia tuberculata and Brookesia peyrierasi are ressurrect...

Brookesia brunoi sp. nov. (Figs. 1���3) Holotype. ZSM 888 / 2010 (ZCMV 12784), adult male, collected in Anja Reserve (see Fig. 1 in Crottini et al. 2011), Ambalavao 21 �� 51 '06.8'' S, 46 �� 50 ' 38.5 '' E, about 950 m a.s.l., Haute Matsiatra Region, Fianarantsoa province, southern central Madagascar, on 8 th December 2010 by Aur��lien Miralles and Fanomezana M. Ratsoavina. A 3 mm long piece of muscle of the right hind limb has been cut and preserved in 96 % EtOH for genetic analyses. Paratypes. ZSM 889 / 2010 (ZCMV 13022), adult gravid female, collected in Anja Reserve, Ambalavao 21 �� 51 ' 2.64 '' S, 46 �� 50 ' 33.80 '' E, 949 m a.s.l., Haute Matsiatra Region, Fianarantsoa province, southern central Madagascar, on 9 th December 2009 by Angelica Crottini, D. James Harris, Iker A. Irisarri, Alexandra Lima, Solohery Rasamison and Emile Rajeriarison. One mm of the tail tip has been cut and preserved in 96 % EtOH for genetic analyses. UADBA uncatalogued (ZCMV 12783), adult female, collected at the same locality and dates of the holotype and hosted in the collection of the Universit�� d'Antananarivo, D��partement de Biologie Animale. A piece of muscle has been cut and preserved in 96 % EtOH for genetic analyses. Diagnosis. A medium-sized Brookesia species characterized by (1) body without a dorsal ridge (keel) and (2) presence of a complete series of laterovertebral pointed tubercles, with (3) the posteriormost laterovertebral pointed tubercle modified into a diamond-shaped pelvic shield, (4) nine pairs of laterovertebral pointed tubercles, (5) absence of enlarged pointed tubercles around the vent, (6) presence of poorly defined laterovertebral tubercles along the entire tail. Brookesia brunoi differs from all other Brookesia species, except for Brookesia brygooi, B. decaryi, B. bonsi, B. valerieae, B. ambreensis, B. antakarana, B. griveaudi, and B. stumpffi by characters 1���3 as listed in the previous paragraph. Brookesia brunoi differs from B. valerieae by its smaller size (SVL up to 42.8 vs. 46���53 mm), a clearly defined pelvic shield (vs. poorly defined), and rounded supraocular cone projecting forward to the level of nostril (vs. pointed supraocular cone not projecting forward to the level of nostril). It differs from B. ambreensis by the number of laterovertebral pointed tubercles (9 vs. 11���12), presence of poorly defined laterovertebral tubercles on tail (vs. absence), clearly defined pelvic shield (vs. poorly defined); from B. antakarana by the number of laterovertebral pointed tubercles (9 vs. 12���13); and from B. griveaudi by its smaller size (TL up to 67.8 vs. 86���99 mm), presence of poorly defined laterovertebral tubercles on tail (vs. absence), and rounded supraocular cone (vs. pointed supraocular cone). Brookesia brunoi differs from B. stumpffi by its smaller size (TL up to 67.8 vs. 81���93 mm) and the presence of poorly defined laterovertebral tubercles on tail (vs. absence). The most similar species to B. brunoi are Brookesia brygooi, B. decaryi and B. bonsi. Together with the morphologically highly distinct B. perarmata, these four species form a monophyletic group. Within this clade, B. brunoi can be distinguished by a combination of characters, especially by its lower number of laterovertebral pointed tubercles (9 vs. 10 in B. brygooi, B. decaryi, and B. bonsi), absence of enlarged pointed tubercles around the vent (present in the other three species), the configuration of its cephalic crest, the lateral tail tubercles (poorly defined), and hemipenial morphology. A summary of these differences is provided in Table 2. Description of the holotype. Well preserved male with everted hemipenis and extruded tongue. Snout-vent length 40.3 mm; tail length 25.9 mm. Head with lateral, orbital and posterior ridges that form a cephalic crest; crest divided into three plains by a pair of longitudinal parasagittal ridges that start above the eyes converging at the posterior part of the crest resulting in a median plain (MP) on the top of the crest, surrounded by two triangular parietal plains (PP) on each side (see Fig. 1, C). Four pointed tubercles on each side of the posterior part of the crest, one at termination point of lateral ridge, one at termination point of parasagittal ridge, and two between the parasagittal and lateral ridges; two pairs of pointed tubercles on lateral surface of head: one supralabial pair, just above posterior angle of mouth, composed of two adjacent conical scales, the posteriormost the biggest, and one infralabial pair of tubercules below posterior angle of mouth. Each tubercle is composed by a single conical scale; orbital crest denticulated; two small and poorly marked inter-orbital ridges converging anteriorly, forming a ���Vshaped��� pattern; supra-ocular cone rounded and projects forward to level of nostril; supra-nasal cone does not reach as far forward as snout tip. Dorsal surface of body flat (without a dorsal ridge or keel); 9 pairs of laterovertebral pointed tubercles form a complete longitudinal line on body; posterior-most (9 th) enlarged pair of pointed laterovertebral tubercles project posteriorly and laterally to form a diamond-shaped pelvic shield above insertion point of hind limbs; l st - 8 th anterior pointed tubercles equally spaced and almost equal in size, perpendicular to body; no ventrolateral pointed tubercle just posterior to the vent at the tail base; reduced laterovertebral pointed tubercles on the tail. Hemipenis short and stout, in general shape similar to that of B. brygooi as described by Brygoo & Domergue (1971) under the name B. ebenaui. Because only a single male is available and it is uncertain whether the organs are fully everted and turgid, the exact shape of the hemipenis is difficult to discern. No clear differentiation between truncus and apex. The sulcus spermaticus is short and deep, surrounded by a bulging basal lobe ("bourrelet basal" in Brygoo & Domergue 1971) and the lobular truncus. No field of denticulated papillae as in B. brygooi (Brygoo & Domergue 1971) is recognizable. A distinct denticulated rotula ("auricule bi-cr��t�� of Brygoo & Domergue 1971; "denticulated lobe" of Raxworthy & Nussbaum 1995) is visible, much larger and more prominent than in B. brygooi where this rotula is usually smaller, less prominent and less strongly denticulated in the specimens we examined. In comparison to the hemipenes of B. brunoi and B. brygooi, the organ of B. decaryi (examined in ZSM 558 / 2001 and 560 / 2001) differs by a sulcus spermaticus that is continuous to the apex, absence of distinct rotulae or papillary fields, and presence of one very characteristic 1 mm long papilla-like protrusion of the apex, along with two much smaller such apical structures. Coloration. According to the photographs (Fig. 1), the background coloration in life of the holotype specimen is beige, with exception of the flanks, the lateral side of the head, the ocular region, the throat and an indistinctly edged vertebral line that are darker. Several dark marks are present on the body: a median transversal black bar joins the summit of each supraocular cone, a median black mark extending from the inferior side of the posterior crest to the inferior side of the first pair of laterovertebral pointed tubercle, a pair of hourglass-shaped black marks running above the parietal ridge of the crest, one pair of black dots between the third and the fourth pair of laterovertebral pointed tubercles, one pair of black dots between the fifth and the sixth pair of laterovertebral pointed tubercle, large black dots below the eighth and the ninth pair of laterovertebral pointed tubercle (see Fig. 1, B), and a dozen tiny black dots on the flanks. The iris is faded orange with a thin central whitish ring bordering the pupil. Six months after fixation, the coloration was relatively similar to that in life, with the exception of the flanks that were darker, the dorsal side of the head, which was lighter, and the absence of tiny black dots on the flanks. Variation. Based on the female paratype ZSM 889 / 2010: SVL 42.8 mm; tail length 24.9 mm + 1 mm (cut for molecular analyses). The color of the paratype after one year and a half in alcohol was similar to that at the time of collection. Ground color of head, body, tail and dorsal parts of limbs dorsally and laterally beige or brown, often becoming darker dorsally. The ground color is contrasting, with a mosaic of whitish, beige, and brown areas, which can be interpreted as leaf mimesis (or more in general as a mimesis to plant debris of the dry forest floor including also sticks and dried bark) (Fig. 3). The blotches on the back break up the background color providing discontinuity thus enhancing the overall cryptic appearance. No measurements are available for the female paratype specimen in the UADBA collection (ZCMV 12783). Distribution, conservation and IUCN Red List status. The new species is currently known only from the type locality within the Anja Reserve (see Fig. 1 in Crottini et al. 2011), although a recent Brookesia record from Ambovomb�� in the far south (Hofmann 2012), ca. 375 km air distance from the type locality, seems to refer to B. brunoi. Furthermore, it is possible that some literature records of B. decaryi, B. brygooi, B. ebenaui and B. stumpffi from the central areas of Madagascar might actually refer to B. brunoi indicating that further investigations are required to better understand its actual distribution. In the Ambalavao area we did not observe any mineral or precious stone extractions or collecting for the pettrade, but deforestation for agriculture, logging and cattle grazing is widely afflicting the area. Although relatively abundant in the Anja reserve (several individuals were observed in a few hours of active searching) it is possible that B. brunoi will qualify for inclusion in one of the threatened categories, depending on the actual size of its distribution area. However, due to the limited knowledge on this species we suggest to consider its conservation status as ���Data Deficient��� according to IUCN criteria (IUCN 2001). Habitat and habits. The holotype of B. brunoi and the UADBA paratype (ZCMV 12783) were found around 21:00 at a roosting height of about 30 cm from the leaf litter on small bushes in the forest of Anja Reserve. With the exception of the paratype specimen ZSM 889 / 2010, found dead in the jaws of a female Furcifer oustaleti, all other individuals sampled for the molecular analyses were found between 13:00 and 14:00 in the leaf litter (ACZC 1938, ACZC 1981, ACZC 1924) (Fig. 3), in shaded spots inside the forest fragment with closed canopy. Another individual was found at night around 21:00 at a roosting height of about 40 cm on small herbal bushes (ACZC 1934). No other Brookesia species were found around the type locality. Although inhabiting the humid leaf litter this new species is one of the few Brookesia species adapted to the semi-arid deciduous forest of Madagascar. The other species found in arid habitats of the west are B. bonsi, B. exarmata, B. decaryi, B. perarmata, B. brygooi and B. stumpffi. Other reptiles found in the forest of the Anja Reserve during our two visits (December 2009, December 2010) were: Paragehyra sp. aff. petiti, Paroedura sp. aff. bastardi, Phelsuma gouldi, Thamnosophis lateralis, Furcifer lateralis, Furcifer oustaleti, Oplurus quadrimaculatus, Madagascarophis meridionalis, Trachylepis vato, and Hemidactylus frenatus. Etymology. A. Crottini dedicates this new species to Bruno Grassi in recognition of his love and support during these years. The specific name is thus a patronym, but in addition recalls the cryptic brownish coloration of the newly described species in Italian language. Mitochondrial variation, differentiation and phylogenetic relationships. The molecular data confirms the attribution of B. brunoi to the B. decaryi group (Townsend et al. 2009). The analyzed specimens of B. brunoi are genetically very uniform and show an intraspecific uncorrected divergence of 0.3 %, while the analyzed specimens of B. brygooi and B. decaryi (including the sample from Tsingy the Bemaraha) are more heterogeneous and show intraspecific uncorrected divergence of 4.8 % and 7.7 % respectively, in the ND 2 gene fragment sequences. The genetic distance between B. brunoi and the three other species of the B. decaryi group (molecular data of B. bonsi are not available for this gene) ranges between 17.6 % (comparison between B. brunoi and B. brygooi) and 19.7 % (comparison between B. brunoi and B. perarmata), and the genetic distance between B. brunoi and B. decaryi is 19.1 %. Among the species of the analyzed species group the smallest genetic distance is observed between B. brygooi and B. decaryi (16.2 %) and the highest value between B. perarmata and B. decaryi (20.8 %). More details are provided in table 1. the pairwise distance calculation for B. perarmata, B. brunoi, B. brygooi and B. decaryi. nc: not calculated. The phylogenetic analyses resulted in a tree with largely unresolved basal relationships (Fig. 4) but with good support for the monophyly of the samples of the B. decaryi group, and for each species of this species group, hence clearly supporting the distinctness of B. brunoi from the other three analysed species. In the analysis of Townsend et al. (2009) B. bonsi was sister to B. decaryi based on DNA sequences of the ND 4 gene available from the study of Raxworthy et al. (2002), and these two species were sister to B. brygooi. We could not include B. bonsi in our analysis due to the lack of samples and ND 2 sequences for this species, but the combined evidence from this study and the tree of Townsend et al. (2009) suggests these species are related as follows: (perarmata (brunoi (brygooi (bonsi, decaryi)))). In our analysis, samples of B. brygooi are organised in two mitochondrial lineages. Samples from Ankarafantsika belong to one lineage, while the samples of B. brygooi from Isalo and Kirindy belong to a second lineage, with an uncorrected average inter-lineage divergence of 5.9 %. Furthermore, our analysis includes one sample from the Tsingy de Bemaraha that in our tree is sister to B. decaryi from the type locality Ankarafantsika, albeit with a high average divergence (11.3 %). Because our sampling does not include B. bonsi, the sister species of B. decaryi (Townsend et al. 2009), we cannot decide whether the Tsingy de Bemaraha sample is to be assigned to B. decaryi, to B. bonsi, or possibly to a third, undescribed species. Neither B. bonsi nor B. decaryi have so far been reported from the Tsingy de Bemaraha, and a more detailed molecular and morphological study of this population is therefore necessary. bonsi. The table also includes B. stumpffi which is the only other superficially similar Brookesia occurring in parts of western Madagascar. Data for B. bonsi were taken from Ramanantsoa (1979). B. brunoi B. brygooi B. decaryi B. bonsi B. stumpffi The description of B. brunoi sp. nov. adds a distinctive new species to the most basal subclade of the ���typical��� Brookesia clade that contains B. decaryi, B. brygooi, B. bonsi and B. perarmata. All these four species inhabit dry deciduous forest areas of the west. On the contrary, B. brunoi is so far known to inhabit only the dry forest of the Anja Reserve, in the central high plateau of Madagascar, at an elevation of ca. 950 m a.s.l.. This small area (ca. 30 hectares) close to the town of Ambalavao has been protected since 1999, and is managed by the local community. Recent herpetological surveys in this area found evidence for the existence of several local endemics (Crottini et al. 2011) and the present study represents a step further towards the knowledge and description of its endemic fauna. Based on its morphological characteristics, B. brunoi in addition to its peculiar distribution, has at least two morphological characters that seem to be unique when compared with the other Brookesia species of the B. decaryi group: 1) the number of the laterovertebral pointed tubercles; and 2) the absence of enlarged pointed tubercles around the vent. The molecular analyses suggest that this species group forms a well-supported monophyletic lineage with high genetic divergences observed between all four species (more than 16 % uncorrected pairwise sequence divergence in the ND 2), indicating a long divergent evolutionary history. It has been suspected that the genetic variability observed between the different populations of B. brygooi across the island reflects the existence of a species complex (Townsend et al. 2009). However, the comparatively high genetic variability within B. brygooi, coupled with the lack of obvious morphological differences among the individuals analyzed in this study (see Appendix) suggests that this divergence might more appropriately be considered as intra-specific variability., Published as part of Crottini, Angelica, Miralles, Aur��lien, Glaw, Frank, Harris, James, Lima, Alexandra & Vences, Miguel, 2012, Description of a new pygmy chameleon (Chamaeleonidae: Brookesia) from central Madagascar, pp. 63-74 in Zootaxa 3490 on pages 65-71, DOI: 10.5281/zenodo.208594, {"references":["Crottini, A., Gehring, P-. S., Glaw, F., Harris, D. J., Lima, A. & Vences, M. (2011) Deciphering the cryptic species diversity of dull-colored day gekos Phelsuma (Squamata: Gekkonidae) from Madagascar, with description of a new species. Zootaxa, 2982, 340 - 348.","Raxworthy, C. J. & Nussbaum, R. A. (1995) Systematics, speciation and biogeography of the dwarf chameleons (Brookesia, Reptilia, Squamata, Chamaeleontidae) of northern Madagascar. Journal of Zoology, 235, 525 - 558.","Hofmann, T. (2012) A notable record of the genus Brookesia (Gray, 1865) from the extreme South of Madagascar. Sauria, 34 (1), 51 - 53.","IUCN (2001) IUCN red list categories and criteria. Version 3.1. IUCN Species Survival Commission, IUCN, Gland, Switzerland and Cambridge, United Kingdom, 30 pp.","Townsend, T. M., Vieites, D. R, Glaw, F. & Vences, M. (2009) Testing species-level diversification hypotheses in Madagascar: the case of microendemic Brookesia leaf chameleons. Systematic Biology, 58 (6), 641 - 656.","Raxworthy, C. J., Forstner, M. R. J. & Nussbaum, R. A. (2002) Chameleon radiation by oceanic dispersal. Nature, 415, 784 - 787.","Ramanantsoa, G. A. (1979) Description de deux nouvelles especes de Brookesia (Reptilia, Squamata, Chamaeleonidae): B. legendrei et B. bonsi. Bulletin du Museum National d'Histoire Naturelle, Paris, 4 (1), 685 - 693."]}

Brookesia dwarf chameleons, endemic to Madagascar, were surveyed at the following localities in northern Madagascar (north of 16°S): Montagne d'Ambre, Ankarana, Manongarivo, Tsaratanana, Marojejy and Masoala. A total of 15 species occur in this region. Six new species are described and five new synonyms are identified. The genus Brookesia, the most speciose chamaeleontid genus in Madagascar, contains 23 species. Almost all the northern Brookesia species are restricted to rainforest and occupy a relatively narrow elevational range. Although the northern rainforests represent just one-third of the total rainforest and about 5% of the total island area, 65% of the Brookesia species occur in this region, and 52% are endemic to the northern rainforest. Five new biogeographic regions of the northern rainforest are identified based on centres of Brookesia endemicity: Montagne d'Ambre, Northwest, Tsaratanana, Northeast and East. Speciation is thought to have been facilitated in the north through geographic isolation, with the Tsaratanana mountain range and the dry forests south of Montagne d'Ambre forming barriers to dispersal, and the Tsaratanana mountains acting as a centre of isolation. The fragmented distribution of several Brookesia species of low altitude rainforest suggests a period in Madagascar's history when the climate was wetter and low altitude rainforest much more widespread.

Brookesia confidens sp. n. ZooBank LSID: urn:lsid:zoobank.org:act:CB2A9146-0161-42B7-A145-6DED579F1F21 Remark.—This species has been considered before as conspecific with Brookesia sp. ‘‘Montagne des Francais’’ [7]. Holotype.— ZSM 2150/2007 (FGZC 1196), adult male (hemipenes incompletely everted), collected on trail to the ‘‘Petit Tsingy and Grotte des Chauves-Souries’’, 12°57'25"S, 49°0 7'0 6"E, 90 m a.s.l., Ankarana National Park, Antsiranana Province, northern Madagascar, on 1 March 2007 by P. Bora, H. Enting, F. Glaw, A. Knoll, and J. Köhler. Paratypes.— UADBA uncatalogued (FGZC 1194–1195), two specimens; ZSM 2151/2007 (FGZC 1197), probably male, ZSM 2152/2007 (FGZC 1198), adult female, ZSM 2153/2007 (FGZC 1199), adult female, all with same data as holotype; UADBA uncatalogued (FGZC 1608), UADBA uncatalogued (FGZC 1610), ZSM 1511/2008 (FGZC 1607), adult female, ZSM 1512 /2008 (FGZC 1609), adult male, all collected at ‘‘ Petit Tsingy’ ’, Ankarana National Park, Antsiranana Province, northern Madagascar, on 12 February 2008 by N. D’Cruze, M. Franzen, F. Glaw and J. Köhler. Diagnosis.— A member of the Brookesia minima group based on small body size (SVL Brookesia confidens is distinguished from other members of the group as follows: from B. dentata by probably smaller adult body size (no measurements of male B. dentata available), and absence of a supranasal cone (vs. presence); from B. exarmata by the absence of a supranasal cone (vs. presence); from B. karchei by a smaller adult body size (female SVL 20.6–22.6 mm vs. 30.7 mm; no measurements of clearly identified males available for B. karchei), and absence of a supranasal cone (vs. presence); from B. minima by a very narrow hemipenis (vs. balloon-like); from B. peyrierasi by generally smaller adult body size(male SVL 18.3– 20.1 mm vs. 19.7–22.4 mm), the absence of a supranasal cone (vs. presence), absence of a supraocular cone (vs. presence), and very narrow ornamentless hemipenis (vs. massive, bilobed hemipenis with four spines per lobe); from B. ramanantsoai by a smaller adult body size (male SVL 18.3–20.1 mm vs. 21.7 mm), absence of a supranasal cone (vs. presence), absence of a supraocular cone (vs. presence in some specimens), and hemipenis very narrow (vs. balloon-like); from B. tuberculata by the absence of a supranasal cone (vs. presence), absence of a supraocular cone (vs. presence), and hemipenis very narrow with pustules on apex (vs. wider, with crown-like apical structure). The new species is most similar to B. tristis but differs from this species by indistinct and short parasagittal crests (vs. distinct) and by 13 dorsolateral pointed tubercles (vs. 11), and ornamentless apical region of hemipenis (vs. spine-like papillae on apex). For a distinction from B. desperata and B. micra, described below, see the diagnoses of these species. Referencing a fragment of the 16S rRNA gene, B. confidens shows an uncorrected pairwise divergence of 6.7% to its sister species B. tuberculata, and divergences>9% to all other species of the B. minima group. Description of holotype.— Adult male in good state of preservation (Fig. 7; Supporting Information S 1). Both hemipenes everted. Measurements in Table 1. Head with lateral crest starting at median level at the posterior edge of eye; prominent orbital crests with distinctly enlarged pointed tubercle at posterior level slightly below lateral crest, and a crest at the posterior edge of the head, that form a weakly developed dorsal helmet; a pair of very short straight parasagittal crests that transform posteriorly into a broad area of elevated scales which almost cover the entire posterior head surface; three pointed tubercles on each side of posterior helmet crest, a prominent one at termination point of lateral crest, an even larger one at the imaginary termination point of parasagittal crest, and a weakly developed one between parasagittal and lateral crests; one pointed tubercle on lateral surface of head, below lateral crest in temporal region; orbital crest denticulated; no supraocular cone recognizable; supranasal cone not clearly recognizable; head longer (5.3 mm) than wide; chin and throat without longitudinal rows of slightly enlarged tubercles. Dorsal surface of body without a vertebral ridge or keel; 13 dorsolateral pointed tubercles form a complete longitudinal line on the body; most posterior (13th) pointed dorsolateral tubercle being largest, above insertion point of hindlimb; pointed dorsolateral tubercles almost equally spaced, pointing out almost perpendicularly from body; slightly enlarged, pointed tubercles form curved transversal crests on either side of the vertebral line between 2nd and 10th dorsolateral pointed tubercles; slightly enlarged tubercles on dorsal surface of tail, without any reconizable pattern; no well-defined dorsal pelvic shield in sacral area; few irregularly scattered enlarged rounded ubercles on the lateral surface of body; venter without enlarged tubercles; scattered, soft-pointed tubercles on limbs; no pointed tubercles around cloaca; longitudinal row of slightly enlarged pointed tubercles lateral on tail, forming a longitudinal row from tail base to two-thirds of tail length; no enlarged tubercles on ventral surfaces of tail. After almost four years in ethanol, all surfaces pale grey to beige, with most pointed tubercles being brown; upper surface of head pale brown. Variation.— For morphological measurements and proportions see Table 1 and Supporting Information S1. In ZSM 2153/2007, the ground colour is dark brown with a whitish vertebral stripe that starts at posterior half of body and extends onto the tail which is entirely beige with scattered brownish spots. The posterior dorsal surface of head is grey. The body of ZSM 1512/2008 is light grey whereas the head is brown with a greyish transverse line between the nostrils. The tail base of the females is less thickened than in the males. In life, all individuals with dorsal colouration of head, dorsum and tail light grey or pale beige, lateral parts of the body brown, with indistinct greyish marbling lateroventrally and with few small dark brown spots around the flank tubercles; limbs almost uniformly dark brown. This pattern is likely a stress colouration. Genital morphology.— For this species, everted hemipenes are available only for the holotype (ZSM 2150/2007). On both sides, the organs are single tube-like structures of low diameter which decreases from the apex to the distal part of the truncus. The left hemipenis is not fully everted whereas the one on the right side (length 1.3 mm; width at the pedicel 0.6 mm, width at the apex 0.4 mm) might be fully everted, but this cannot be verified without examination of further material. There are no ornaments on the pedicel and truncus. The sulcus spermaticus is not recognizable on most of the truncus but becomes more distinct in the apical region. The apical region is characterized by a slight increase in diameter but not bilobed, and is covered by relatively large but poorly defined pustular papillae (Fig. 6). The hemipenis is devoid of any further prominent structures such as the spiny apical papillae of B. peyrierasi (which are visible also in incompletely everted organs). Etymology.— The species epithet is an adjective derived from the Latin ‘‘confidens’’ meaning ‘‘confident’’, ‘‘trusting’’. The known range of the species is supposedly a well protected nature reserve with apparently limited habitat destruction. Furthermore, this area might benefit from natural protection by the tsingy limestone formations which are difficult to access, thus giving hope for the species’ survival. Distribution.— Only known from a single locality within Ankarana National Park. Natural History.— Most individuals were found roosting at night on thin branches about 5–20 cm above the leaf litter in deciduous dry forest close to a small forest trail within a small area, surrounded by tsingy outcrops. At this locality, the species was relatively abundant, whereas it was not found at similar localities nearby, suggesting a patchy distribution and a preference for certain microhabitats. This hypothesis is also supported by the fact that earlier herpetological surveys in Ankarana [38,39], did not record any species of the Brookesia minima group. When stressed, individuals can quickly change colour and display a broad pale vertebral stripe contrasting with the darker flanks., Published as part of Frank Glaw, Jörn Köhler, Ted M. Townsend & Miguel Vences, 2012, Rivaling the World's Smallest Reptiles: Discovery of Miniaturized and Microendemic New Species of Leaf Chameleons (Brookesia) from Northern Madagascar, pp. 1-24 in PLoS ONE 7 (2) on pages 15-16, DOI: 10.1371/journal.pone.0031314, http://zenodo.org/record/894041, {"references":["7. Glaw F, Vences M (2007) A Field Guide to the Amphibians and Reptiles of Madagascar. Koln: Vences & Glaw."]}

Brookesia micra sp. n. ZooBank LSID: urn:lsid:zoobank.org:act:D1A239D6-93E8-4C34-A428-F79A2C8B6405 Remark.—This species has been considered before as Brookesia sp. ‘‘Nosy Hara’’ [7]. Holotype.— ZSM 2181/2007 (FGZC 1271), adult male (hemipenes everted), collected on Nosy Hara island, 12°14'40"S, 49°0 0'30"E, ca. 10–20 m a.s.l., Antsiranana Province, northern Madagascar, on 7 March 2007 by H. Enting, F. Glaw and J. Köhler. Paratypes.— ZSM 2180/2007 (FGZC 1270), ZSM 2182/2007 (FGZC 1275), juveniles, ZSM 2183/2007 (FGZC 1278), ZSM 2185/2007 (FGZC 1280), adult males (hemipenes everted), ZSM 2184/2007 (FGZC 1279), ZSM 2186–2187/2007 (FGZC 1281– 1282), adult females, all with same data as holotype; UADBA uncatalogued (FGZC 1830), male, ZSM 1507 /2008 (FGZC 1831), male (not examined morphologically), UADBA uncatalogued (FGZC 1833), female, ZSM 1509 /2008 (FGZC 1834), adult female, ZSM 1510 /2008 (FGZC 1832), juvenile, all collected at small stream on Nosy Hara, 12°14'59"S, 49°0 0'28"E, 12 m a.s.l., Antsiranana Province, northern Madagascar, on 22 February 2008 by F. Glaw and J. Köhler. Diagnosis.— A member of the Brookesia minima group based on small body size (SVL Brookesia micra is distinguished from all other members of the group by a shorter relative tail length (tail length/ SVL 0.37–0.49 versus 0.49–0.92), and by orange coloured tails in life in adults (vs. inconspicuous brownish colour). In addition, from B. confidens by a smaller adult male size (SVL 15.1– 15.3 mm vs. 18.3–20.1 mm), supranasal cone present (vs. absent), and hemipenis with comb-like apical structure (vs. narrow without ornaments); from B. dentata by probably smaller adult body size (no measurements of male B. dentata available); from B. exarmata by smaller adult body size (female SVL 18.7–19.9 vs. 25.7–26.5, no male measurements available for B. exarmata); from B. karchei by smaller size (female SVL 18.7–19.9 vs. 30.7, no male measurements available for B. karchei); supraocular cone absent (vs. present); from B. minima by presence of a pelvic spine (vs. absent or indistinct pelvic spine), and hemipenis with comb-like apical structure (vs. balloon-like without ornaments); from B. peyrierasi by a smaller adult male size (SVL 15.1–15.3 mm vs. 19.1–27.4 mm), and hemipenis with comb-like apical structure (vs. bilobed with four spines on each lobe); from B. ramanantsoai by a smaller adult male size (SVL 15.1–15.3 mm vs. 21.7 mm), supraocular cone absent (vs. present in some specimens), and hemipenis with comblike apical structure (vs. baloon-like without ornaments); from B. tristis by a smaller adult male size (SVL 15.1–15.3 mm vs. 18.0– 18.2 mm), and hemipenis with comb-like apical structure (vs. small papillae on apex not arranged comb-like); and from B. tuberculata by supraocular cone absent (vs. present), and hemipenis with comb-like apical structure (vs. crown-like structure). For a distinction from B. desperata, described below, see the diagnosis of this species. Referencing a fragment of the 16S rRNA gene, B. micra shows an uncorrected pairwise divergence of 6.8% to its sister clade (B. tristis + B. desperata), and divergences>7.2% to all other species of the B. minima group. Description of holotype.— Adult male in good state of preservation (Fig. 7; Supporting Information S 1). Both hemipenes everted. Measurements in Table 2. Head with lateral crest starting at median level at the posterior edge of eye; prominent orbital crests; a weak crest at the posterior edge of the head, that forms a weakly developed dorsal helmet; a pair of low indistinct parasagittal crests slightly converging before terminating at the lateral crest; three similar-sized pointed tubercles on each side of posterior crest, one at termination point of lateral crest, one at termination point of parasagittal crest, and a one between parasagittal and lateral crests; one pointed tubercle on lateral surface of head, below lateral crest in temporal region; orbital crest denticulated; no supraocular cone recognizable; supranasal cone small; head longer (4.5 mm) than wide; chin and throat without longitudinal rows of slightly enlarged tubercles. Dorsal surface of body without a vertebral ridge or keel; 11 dorsolateral pointed tubercles form a complete longitudinal line on the body; most posterior (11th) pointed dorsolateral tubercle being largest, above insertion point of hindlimb; pointed dorsolateral tubercles almost equally spaced, pointing out almost perpendicularly from body; slightly enlarged, rounded tubercles form curved transversal crests on either side of the vertebral line between 1 st and 10th dorsolateral pointed tubercles; slightly enlarged scales forming four parallel longitudinal rows on dorsal surface of tail; no well-defined dorsal pelvic shield in sacral area; scattered enlarged rounded tubercles on the lateral surface of body; venter without enlarged tubercles; no pointed tubercles around cloaca; no enlarged tubercles on lateral or ventral surfaces of tail. After almost four years in ethanol, back, flanks, dorsal surfaces of limbs and anterior part of tail uniformly dark grey-brown; posterior part of tail beige; neck grey; throat and venter light grey. Variation.— For morphological measurements and proportions see Table 2 and Supporting Information S1. In preservative, ZSM 2185/ 2007 and 2188 /2007 have a greyish dorsal surface of head and neck and a slightly lighter vertebral region. The throat is uniformly dark brown (ZSM 2187/ 2007 and 2184 /2007) or beige with brown spots (ZSM 2185/2007, 2188/2007, and 1509/2008). The tail base of the males is only slightly more thickened than in the females. In life, most individuals with dorsal colouration of head, dorsum and tail light grey, lateral parts of the tail more yellowish becoming orange posterior to the tail base. Lateral parts of the body brown, with few dark brown spots; limbs almost uniformly dark brown. This pattern is likely to refer to a stress colouration. When unstressed, most parts of the body dark brown, except a beige area on the head anterior to the eyes. Tail base dark brown, orange in the middle and yellowish posteriorly. Colour and pattern of juveniles resemble those of adults and they can also show the stress colouration, but tails are slightly less colourful than in adults and usually do not show bright orange colour. Genital morphology.— Everted hemipenes of this species are available for three specimens (holotype ZSM 2181/2007, paratypes ZSM 2183/2007, ZSM 2185/2007). Each hemipenis is an elongated, relatively wide tubular structure. Apex and truncus are devoid of any ornaments. In the holotype, hemipenis length is 2.4 mm and maximum hemipenis width is 0.9 mm. When fully everted, the most characteristic feature is the apex which is a flat surface distally forming a symmetrical comb of six large, rounded papillae, of which the two inner ones are largest, followed by the two intermediate ones, whereas the outer ones are merely two slightly elevated knobs, recognizable only in fully everted organs. In general, this comb of papillae is not visible in hemipenes that are not fully everted, different from the spines of, e.g., B. peyrierasi that typically are also visible in incompletely everted organs. All three examined specimens have one hemipenis fully everted, the second one only partially everted. The organs are largely transparent, and the central retractor muscles can be easily seen through the outer hemipenial integument (Fig. 6). Etymology.— The species epithet is a latinized derivation from the Greek word ‘‘MIKRóz’’ (mikros), meaning ‘‘tiny’’ or ‘‘small’’ and referring to the extremely diminutive body size. It is used as an invariable noun in apposition. Distribution.— Only known from two sites (see localities in type series) on the small island of Nosy Hara, northern Madagascar (e.g. Fig. 8 D). Remarkably, no Brookesia species was recorded during intensive herpetological surveys of Nosy Hara, nearby islands and the adjacent mainland [40], suggesting that B. micra might be difficult to record. Natural History.— B. micra was found during the day active on the ground in a mosaic of eroded limestone boulders and dry forest leaf litter, and at night roosting on branches in very low vegetation (ca. 5–10 cm above the ground). In contrast to B. tristis and B. confidens its occurrence was not remarkably patchy., Published as part of Frank Glaw, Jörn Köhler, Ted M. Townsend & Miguel Vences, 2012, Rivaling the World's Smallest Reptiles: Discovery of Miniaturized and Microendemic New Species of Leaf Chameleons (Brookesia) from Northern Madagascar, pp. 1-24 in PLoS ONE 7 (2) on pages 16-17, DOI: 10.1371/journal.pone.0031314, http://zenodo.org/record/894041, {"references":["7. Glaw F, Vences M (2007) A Field Guide to the Amphibians and Reptiles of Madagascar. Koln: Vences & Glaw.","40. Metcalf CJE, Hampson K, Gray A, Andrianirina R (2007) Herpetofaunal assemblages on seven offshore islands of northwestern Madagascar. Trop Zool 20: 151 - 161."]}

Brookesia micra sp. n. ZooBank LSID: urn:lsid:zoobank.org:act:D1A239D6-93E8-4C34-A428-F79A2C8B6405 Remark.—This species has been considered before as Brookesia sp. ‘‘Nosy Hara’’ [7]. Holotype.— ZSM 2181/2007 (FGZC 1271), adult male (hemipenes everted), collected on Nosy Hara island, 12°14'40"S, 49°0 0'30"E, ca. 10–20 m a.s.l., Antsiranana Province, northern Madagascar, on 7 March 2007 by H. Enting, F. Glaw and J. Köhler. Paratypes.— ZSM 2180/2007 (FGZC 1270), ZSM 2182/2007 (FGZC 1275), juveniles, ZSM 2183/2007 (FGZC 1278), ZSM 2185/2007 (FGZC 1280), adult males (hemipenes everted), ZSM 2184/2007 (FGZC 1279), ZSM 2186–2187/2007 (FGZC 1281– 1282), adult females, all with same data as holotype; UADBA uncatalogued (FGZC 1830), male, ZSM 1507 /2008 (FGZC 1831), male (not examined morphologically), UADBA uncatalogued (FGZC 1833), female, ZSM 1509 /2008 (FGZC 1834), adult female, ZSM 1510 /2008 (FGZC 1832), juvenile, all collected at small stream on Nosy Hara, 12°14'59"S, 49°0 0'28"E, 12 m a.s.l., Antsiranana Province, northern Madagascar, on 22 February 2008 by F. Glaw and J. Köhler. Diagnosis.— A member of the Brookesia minima group based on small body size (SVL Brookesia micra is distinguished from all other members of the group by a shorter relative tail length (tail length/ SVL 0.37–0.49 versus 0.49–0.92), and by orange coloured tails in life in adults (vs. inconspicuous brownish colour). In addition, from B. confidens by a smaller adult male size (SVL 15.1– 15.3 mm vs. 18.3–20.1 mm), supranasal cone present (vs. absent), and hemipenis with comb-like apical structure (vs. narrow without ornaments); from B. dentata by probably smaller adult body size (no measurements of male B. dentata available); from B. exarmata by smaller adult body size (female SVL 18.7–19.9 vs. 25.7–26.5, no male measurements available for B. exarmata); from B. karchei by smaller size (female SVL 18.7–19.9 vs. 30.7, no male measurements available for B. karchei); supraocular cone absent (vs. present); from B. minima by presence of a pelvic spine (vs. absent or indistinct pelvic spine), and hemipenis with comb-like apical structure (vs. balloon-like without ornaments); from B. peyrierasi by a smaller adult male size (SVL 15.1–15.3 mm vs. 19.1–27.4 mm), and hemipenis with comb-like apical structure (vs. bilobed with four spines on each lobe); from B. ramanantsoai by a smaller adult male size (SVL 15.1–15.3 mm vs. 21.7 mm), supraocular cone absent (vs. present in some specimens), and hemipenis with comblike apical structure (vs. baloon-like without ornaments); from B. tristis by a smaller adult male size (SVL 15.1–15.3 mm vs. 18.0– 18.2 mm), and hemipenis with comb-like apical structure (vs. small papillae on apex not arranged comb-like); and from B. tuberculata by supraocular cone absent (vs. present), and hemipenis with comb-like apical structure (vs. crown-like structure). For a distinction from B. desperata, described below, see the diagnosis of this species. Referencing a fragment of the 16S rRNA gene, B. micra shows an uncorrected pairwise divergence of 6.8% to its sister clade (B. tristis + B. desperata), and divergences>7.2% to all other species of the B. minima group. Description of holotype.— Adult male in good state of preservation (Fig. 7; Supporting Information S 1). Both hemipenes everted. Measurements in Table 2. Head with lateral crest starting at median level at the posterior edge of eye; prominent orbital crests; a weak crest at the posterior edge of the head, that forms a weakly developed dorsal helmet; a pair of low indistinct parasagittal crests slightly converging before terminating at the lateral crest; three similar-sized pointed tubercles on each side of posterior crest, one at termination point of lateral crest, one at termination point of parasagittal crest, and a one between parasagittal and lateral crests; one pointed tubercle on lateral surface of head, below lateral crest in temporal region; orbital crest denticulated; no supraocular cone recognizable; supranasal cone small; head longer (4.5 mm) than wide; chin and throat without longitudinal rows of slightly enlarged tubercles. Dorsal surface of body without a vertebral ridge or keel; 11 dorsolateral pointed tubercles form a complete longitudinal line on the body; most posterior (11th) pointed dorsolateral tubercle being largest, above insertion point of hindlimb; pointed dorsolateral tubercles almost equally spaced, pointing out almost perpendicularly from body; slightly enlarged, rounded tubercles form curved transversal crests on either side of the vertebral line between 1 st and 10th dorsolateral pointed tubercles; slightly enlarged scales forming four parallel longitudinal rows on dorsal surface of tail; no well-defined dorsal pelvic shield in sacral area; scattered enlarged rounded tubercles on the lateral surface of body; venter without enlarged tubercles; no pointed tubercles around cloaca; no enlarged tubercles on lateral or ventral surfaces of tail. After almost four years in ethanol, back, flanks, dorsal surfaces of limbs and anterior part of tail uniformly dark grey-brown; posterior part of tail beige; neck grey; throat and venter light grey. Variation.— For morphological measurements and proportions see Table 2 and Supporting Information S1. In preservative, ZSM 2185/ 2007 and 2188 /2007 have a greyish dorsal surface of head and neck and a slightly lighter vertebral region. The throat is uniformly dark brown (ZSM 2187/ 2007 and 2184 /2007) or beige with brown spots (ZSM 2185/2007, 2188/2007, and 1509/2008). The tail base of the males is only slightly more thickened than in the females. In life, most individuals with dorsal colouration of head, dorsum and tail light grey, lateral parts of the tail more yellowish becoming orange posterior to the tail base. Lateral parts of the body brown, with few dark brown spots; limbs almost uniformly dark brown. This pattern is likely to refer to a stress colouration. When unstressed, most parts of the body dark brown, except a beige area on the head anterior to the eyes. Tail base dark brown, orange in the middle and yellowish posteriorly. Colour and pattern of juveniles resemble those of adults and they can also show the stress colouration, but tails are slightly less colourful than in adults and usually do not show bright orange colour. Genital morphology.— Everted hemipenes of this species are available for three specimens (holotype ZSM 2181/2007, paratypes ZSM 2183/2007, ZSM 2185/2007). Each hemipenis is an elongated, relatively wide tubular structure. Apex and truncus are devoid of any ornaments. In the holotype, hemipenis length is 2.4 mm and maximum hemipenis width is 0.9 mm. When fully everted, the most characteristic feature is the apex which is a flat surface distally forming a symmetrical comb of six large, rounded papillae, of which the two inner ones are largest, followed by the two intermediate ones, whereas the outer ones are merely two slightly elevated knobs, recognizable only in fully everted organs. In general, this comb of papillae is not visible in hemipenes that are not fully everted, different from the spines of, e.g., B. peyrierasi that typically are also visible in incompletely everted organs. All three examined specimens have one hemipenis fully everted, the second one only partially everted. The organs are largely transparent, and the central retractor muscles can be easily seen through the outer hemipenial integument (Fig. 6). Etymology.— The species epithet is a latinized derivation from the Greek word ‘‘MIKRóz’’ (mikros), meaning ‘‘tiny’’ or ‘‘small’’ and referring to the extremely diminutive body size. It is used as an invariable noun in apposition. Distribution.— Only known from two sites (see localities in type series) on the small island of Nosy Hara, northern Madagascar (e.g. Fig. 8 D). Remarkably, no Brookesia species was recorded during intensive herpetological surveys of Nosy Hara, nearby islands and the adjacent mainland [40], suggesting that B. micra might be difficult to record. Natural History.— B. micra was found during the day active on the ground in a mosaic of eroded limestone boulders and dry forest leaf litter, and at night roosting on branches in very low vegetation (ca. 5–10 cm above the ground). In contrast to B. tristis and B. confidens its occurrence was not remarkably patchy.

Brookesia desperata sp. n. ZooBank LSID: urn:lsid:zoobank.org:act:C62B456C-CDA8- 4DDE-AE0A-66DF43C57F7E Remark.—This species has been considered before as Brookesia sp. aff. karchei ‘‘Ambre’’ [7] and as Brookesia sp. nov. [41]. Holotype.— ZSM 2170/2007 (FGZC 1250), adult male (hemipenes everted), collected at Forêt d’Ambre Special Reserve, ca. 5 km southwest of Sakaramy village, 12°28'0 0"S, 49°13'37" E, 470 m a.s.l., Antsiranana Province, northern Madagascar, on 12 March 2007 by F. Glaw, J. Köhler and A. Razafimanantsoa. Paratypes.— ZSM 2171/2007 (FGZC 1251), ZSM 2175/2007 (FGZC 1258), ZSM 2176/2007 (FGZC 1260), ZSM 2177/2007 (FGZC 1263), ZSM 2179/2007 (FGZC 1269), adult males (all with everted hemipenes), ZSM 2172–2174 /2007 (FGZC 1252– 1254), ZSM 2178/2007 (FGZC 1265), adult females, all with same data as holotype; ZSM 1506/2008 (FGZC 1880, female), and UADBA uncatalogued: FGZC 1700, FGZC 1879 (male), FGZC 3111–3112, collected at Forêt d’Ambre Special Reserve, ca. 5 km southwest of Sakaramy village, 12°28'S, 49°13'E, 550 m a.s.l., Antsiranana Province, northern Madagascar, on 27 February 2008 by N. D’Cruze, F. Glaw and J. Köhler; ZSM 791 /2009 (ZCMV 13040), adult male (not examined morphologically), collected at Forêt d’Ambre Special Reserve, 12°28'18.0"S, 49°13'56"E, 438 m a.s.l., Antsiranana Province, northern Madagascar, on 16 November 2009 by A. Crottini, S. Hauswaldt, A. Lima, F. M. Ratsoavina and E. Rajeriarison. Diagnosis.— A member of the Brookesia minima group based on small body size (SVL 25–30 mm) and molecular phylogenetic relationships. Brookesia desperata is distinguished from all other species in the group by the presence of three enlarged tubercles on lateral head surface (versus 0–2). In addition it differs as follows: from B. confidens by a larger adult body size (male SVL 25.0–26.7 vs. 18.3–20.1 mm), supranasal cone present (vs. absent), and hemipenis with two apical processes each with a distinct spine (vs. narrow hemipenis without apical ornaments); from B. dentata by presence of well-developed lateral spines on the tail (vs. absence); from B. exarmata by a larger adult body size (female SVL 27.3– 30.0 mm vs. 25.7–26.5 mm), and presence of well-developed lateral spines on the tail (vs. absence); from B. micra by a larger adult body size (male SVL 25.0–26.7 vs. 15.3–15.8 mm), presence of well-developed lateral spines on the tail (vs. absence), and hemipenis with two apical processes each with a distinct spine (vs. comb-like arranged papillae on apex); from B. minima by a larger adult body size (male SVL 25.0–26.7 vs. 15.0– 20.6 mm), presence of well-developed lateral spines on the tail (vs. indistinct), pelvic spine present (vs. absent or indistinct), and hemipenis with two apical processes each with a distinct spine (vs. balloon-like hemipenis without ornaments); from B. peyrierasi by a larger adult body size (male SVL 25.0–26.7 vs. 19.7–22.4 mm), presence of well-developed lateral spines on the tail (vs. indistinct), and hemipenis with two apical processes each with a distinct spine (vs. four spines on each lobe); from B. ramanantsoai by presence of welldeveloped lateral spines on the tail (vs. absence), and hemipenis with two apical processes each with a distinct spine (vs. balloon-like hemipenis without ornaments); from B. tristis by a larger adult body size (male SVL 25.0–26.7 vs. 18.0– 18.2 mm), and hemipenis with two apical processes each with a distinct spine (vs. small spinelike papillae on apex); and from B. tuberculata by a larger adult body size (male SVL 25.0–26.7 vs. 14.4–18.8 mm), presence of welldeveloped lateral spines on the tail (vs. absence), and hemipenis with two apical processes with a distinct spine (vs. a single crownlike structure on apex). B. desperata is most similar to B. karchei in body size, number of dorsolateral pointed tubercles (12–13) and distinct lateral tubercles on tail (Fig. 9 B). However, B. karchei differs from the new species by more pronounced supraocular and supranasal cones, more prominent and spiny posterior crest and only one enlarged tubercle at lateral side of head (three in B. desperata). Referencing a fragment of the 16S rRNA gene, B. desperata shows an uncorrected pairwise divergence of 6.4% to its sister species B. tristis, and divergences>6.6% to all other species of the B. minima group. Description of holotype.— Adult male in good state of preservation (Fig. 7; Supporting Information S 1). Both hemipenes everted. Measurements in Table 2. Head with low lateral crest starting at median level at the posterior edge of eye; prominent orbital crests with distinctly developed supraocular cone directed anteriorly, and a crest at the posterior edge of the head, that form a weakly developed dorsal helmet; a pair of slightly curved parasagittal crests that start above the eyes and begin to converge slightly before terminating at the posterior crest; between the parasagittal crests there is a second pair of short parallel longitudinal crests; three pointed tubercles on each side of posterior helmet crest, one at termination point of lateral crest, one at termination point of parasagittal crest, and one between parasagittal and lateral crests; three pointed tubercles on lateral surface of head, one below lateral crest in temporal region, one at posterior edge of orbital crest below lateral crest, and one between lower edge of eye slightly anterior to angle of jaws; orbital crest with few enlarged conical tubercles; no supraocular cone recognizable; supranasal cone does not project beyond snout tip; head longer (6.4 mm) than wide; chin and throat without longitudinal rows of slightly enlarged tubercles. Dorsal surface of body without a vertebral ridge or keel; 13 dorsolateral pointed tubercles form a complete longitudinal line on the body; most posterior (13th) pointed dorsolateral tubercle being distinctly largest, slightly posterior to insertion point of hindlimb, projecting backwards; pointed dorsolateral tubercles almost equally spaced, 8th to 12th slightly smaller than others, pointing out almost perpendicularly from body; slightly enlarged, rounded tubercles form weakly elevated curved transversal crests on either side of the vertebral line between 1 st and 12th dorsolateral pointed tubercles; slightly enlarged tubercles on dorsal surface of tail, forming two weakly recognizable longitudinal parallel lines; barely defined dorsal pelvic shield in sacral area; lateral surface of body with densely scattered enlarged conical tubercles; venter without enlarged tubercles; scattered, conical tubercles on limbs; no pointed tubercles around cloaca; longitudinal row of distinctly enlarged tubercles lateroventrally on tail, forming a longitudinal row from tail base to three fourth of tail length; no enlarged tubercles on ventral surfaces of tail. After almost four years in ethanol, flanks, dorsal surfaces of limbs and tail uniformly dark grey-brown; neck with distinct light grey spot; vertebral region mottled with brown and grey; throat grey, venter and ventral surfaces of tail grey-brown. Hemipenes whitish. Variation.— For morphological measurements and proportions see Table 2 and Supporting Information S1. In life, dorsal colouration of head, dorsum and tail light grey. Lateral parts of the body beige, brown or dark brown, with few dark brown spots; limbs almost uniformly dark brown. This pattern is likely to refer to a stress colouration (Fig. 9). When unstressed, most parts of the body beige, with a slightly lighter area on the head anterior to the eyes and in the vertebral region of the dorsum. In preservative, ZSM 2172/2007 is greyish with brown blotches and flecks on dorsum and flanks whereas the other paratypes are more or less uniformly dark brown. The tail base of the males is only slightly more thickened than in the females. Genital morphology.— Everted hemipenes were available from the holotype (ZSM 2170/2007) and from four paratypes (ZSM 2171/ 2007, 2176–2177 /2007, 2179/2007). Structures were characteristic and concordant among all five specimens and are described based on the holotype. Hemipenis length is 3.2 mm. Hemipenis width in the fully turgid state is about 2.3 mm. The retractor muscle is visible through the transparent hemipenis integument and bifurcates after about 1/2 of the hemipenis length. From the wide apex, two narrow tubular processes (0.5 mm in width) extend about 1.1 mm distally, with each end of the retractor muscle ending at the tip of these processes. The end of each of the tubes bears a distinct spine. The apex processes with their large spines are easily recognizable also in incompletely everted hemipenes (Fig. 6). When the hemipenis is everted but not turgid (in preserved specimens), the processes are very distinct and project much more distinctly from the apex. Etymology.— The species epithet is an adjective derived from the Latin ‘‘desperatus’’ meaning ‘‘desperate’’. Although the known range of the species is within a nature reserve established decades ago, its habitat is in truth barely protected and subject to numerous human-induced environmental problems resulting in severe habitat destruction [41], thus threatening the survival of the species. Distribution.— Only known from the southern edge of Forêt d’Ambre Special Reserve, northern Madagascar. Natural History.— All individuals were found roosting at night on small branches or leaves about 5–100 cm above the ground in disturbed rainforest. Individuals occurred in apparently high abundance and were also found at the border of forest clearings on banana plants. One female laid two large eggs when kept in a plastic bag. When stressed, individuals can quickly change colour and display a broad pale vertebral stripe contrasting with the darker flanks (Fig. 9 A)., Published as part of Frank Glaw, Jörn Köhler, Ted M. Townsend & Miguel Vences, 2012, Rivaling the World's Smallest Reptiles: Discovery of Miniaturized and Microendemic New Species of Leaf Chameleons (Brookesia) from Northern Madagascar, pp. 1-24 in PLoS ONE 7 (2) on pages 17-18, DOI: 10.1371/journal.pone.0031314, http://zenodo.org/record/894041, {"references":["7. Glaw F, Vences M (2007) A Field Guide to the Amphibians and Reptiles of Madagascar. Koln: Vences & Glaw.","41. D'Cruze N, Kohler J, Franzen M, Glaw F (2008) A conservation assessment of the amphibians and reptiles of the Foret d'Ambre Special Reserve, north Madagascar. Madagascar Conservation and Development 3: 44 - 54."]}

Brookesia tristis sp. n. ZooBank LSID: urn:lsid:zoobank.org:act:56F36D4D-1F94-49C4-AC30-6E894B6AA998 Remark.—This species has been considered before as Brookesia sp. ‘‘Montagne des Francais’’ [7] and as Brookesia sp. nov. [37]. Holotype.— ZSM 1704 /2004 (no field number), adult male (hemipenes everted), collected at Montagne des Français, 12°19'S, 49°20'E, ca. 150 m a.s.l., Antsiranana Province, northern Madagascar, on 23 February 2004 by F. Glaw, M. Puente, R. D. Randrianiaina and guides of the hotel ‘‘ King’s Lodge’ ’. Paratypes.— UADBA uncatalogued (FGZC 477–478), ZSM 357/2004 (FGZC 654) [not examined morphologically], and ZSM 876/2010 [no field number, not examined morphologically], adult males, ZSM 354 /2004 (FGZC 651), adult female, same data as holotype; ZSM 1705 /2004–1707/2004 (no field numbers), three adult females, most likely same data as holotype. UADBA uncataloged (FGZC 1187, 1189, 1191); ZSM 2146–2149 /2007 (FGZC 1188, 1190, 1192, 1193), adult females, all collected at Montagne des Français, near remains of French Fort, 12°19'S, 49°20'E, 250–300 m a.s.l., Antsiranana Province, northern Madagascar, on 27 February 2007 by P. Bora, H. Enting, F. Glaw, A. Knoll and J. Köhler; ZSM 2018 /2008 (FGZC 1733 = MgF 061) and UADBA uncatalogued (FGZC 1734 = MgF 061), collected at Montagne des Français, canyon 2 km [air distance] W of Andavakoera, 12°19,838'S, 49°20,941'E, 250 m a.s.l., on 24 January 2006 by E. Randriamalala; UADBA uncatalogued (FGZC 1657), female, ZSM 1505 /2008 (FGZC 1656), adult male (hemipenes everted), both collected ca. 1.5 km southwest of Andavakoera (‘‘ Frontier base camp’’), 12°19'59.2"S, 49°21'20.6"E; 140 m a.s.l., Montagne des Français, Antsiranana Province, northern Madagascar, on 16 February 2008 by M. Franzen, F. Glaw, J. Köhler and Z.T. Nagy. Diagnosis.— A member of the Brookesia minima group based on small body size (SVL Brookesia tristis is distinguished from other members of the group as follows: from B. dentata by probably smaller adult body size (no measurements of male B. dentata available), and presence (in most specimens) of lateral spines on the tail (vs. absence); from B. exarmata by presence (in most specimens) of lateral spines on the tail (vs. absence); from B. karchei by probably smaller adult body size (female SVL 18.0– 23.8 mm vs. 30.7 mm; no measurements of clearly identified males available for B. karchei), and absence of a supraocular cone (vs. presence); from B. minima by the presence of a supranasal cone, a pelvic spine, and (in most specimens) lateral spines on the tail (vs. absence), and hemipenis with small apical spine-like papillae (vs. balloon-like without ornaments); from B. peyrierasi by smaller adult body size (male SVL 18.0– 18.2 mm vs. 19.7–22.4 mm), presence (in most specimens) of lateral spines on the tail (vs. absence), and by a hemipenis with small apical spine-like papillae (vs. bilobed hemipenis with four large spines per lobe); from B. ramanantsoai by smaller adult body size (male SVL 18.0– 18.2 mm vs. 21.7 mm), absence of a supraocular cone (vs. presence in some specimens), presence (in most specimens) of lateral spines on the tail (vs. absence), and hemipenis with small apical spine-like papillae (vs. hemipenis balloon-like without ornaments); and from B. tuberculata by the absence of a supraocular cone (vs. presence), presence (in most specimens) of lateral spines on the tail (vs. absence), and hemipenis with small apical spine-like papillae (vs. hemipenis with apical crown-like structure). For a distinction from B. confidens, B. desperata, and B. micra, described below, see the diagnoses of these species. Referencing a fragment of the 16S rRNA gene, B. tristis shows an uncorrected pairwise divergence of 6.4% to its sister species B. desperata, and divergences>6.9% to all other species of the B. minima group. Description of holotype.— Adult male in good state of preservation (Fig. 7; Supporting Information S 1). Both hemipenes everted. Measurements in Table 1. Head with lateral crest starting at median level at the posterior edge of eye, prominent orbital crests, and a crest at the posterior edge of the head, that form a weakly developed dorsal helmet; a pair of curved parasagittal crests that start above the eyes and begin to converge before terminating at the posterior crest; between the parasagittal crests there is a second pair of parallel longitudinal crests; three pointed tubercles on each side of posterior helmet crest, one at termination point of lateral crest, one at termination point of parasagittal crest, and one between parasagittal and lateral crests; one pointed tubercle on lateral surface of head, below lateral crest in temporal region; orbital crest denticulated; no supraocular cone recognizable; supranasal cone does not project beyond snout tip; head longer (5.1 mm) than wide; chin and throat without longitudinal rows of slightly enlarged tubercles. Dorsal surface of body without a vertebral ridge or keel; 11 dorsolateral pointed tubercles form a complete longitudinal line on the body; 10th dorsolateral tubercle least pointed, small; most posterior (11th) pointed dorsolateral tubercle being largest, above insertion point of hindlimb, very slightly projecting backwards; pointed dorsolateral tubercles almost equally spaced, 3rd to 6th tubercle slightly larger than others, pointing out almost perpendicularly from body; slightly enlarged, rounded tubercles form curved transversal crests on either side of the vertebral line between 1 st and 10th dorsolateral pointed tubercles; dorsal surface of tail with slightly enlarged rounded tubercles forming separated rounded crests that continue from tailbase to two-thirds down the tail; no well-defined dorsal pelvic shield in sacral area; lateral surface of body with evenly spaced enlarged rounded tubercles, mainly arranged in four longitudinal rows; venter without enlarged tubercles; scattered, soft-pointed tubercles on limbs; no pointed tubercles around cloaca; longitudinal row of slightly enlarged pointed tubercles lateral on tail, forming a longitudinal row from tail base to tail tip; no enlarged tubercles on ventral surfaces of tail. In life, colouration of head, body, limbs and tail brown to beige. Faint brown bars radiate from eye to lateral surfaces of head; chin and throat brown; ventral surfaces of body and tail pale brown. After almost seven years in ethanol, all surfaces pale grey to beige, with most pointed tubercles being brown; upper surface of head pale brown. Variation.— For morphological measurements and proportions see Table 1 and Supporting Information S1. In preservative, most paratypes are dorsally darker than the holotype, ranging from brown to dark brown. Their ventral surface is greyish except for the region of the chest, which shows brown mottling. In life several specimens got a light grey vertebral stripe when kept together with conspecifics in a bag, suggesting that this might be a stress colouration. The life colouration of juveniles generally resembles that of adults. The tail base of the females is distinctly less thickened than in the males. In life, two adult specimens had a weight of 0.18 g and 0.206 g, respectively. Genital morphology.— Only two specimens with everted hemipenes were available for examination: ZSM 1505/2008 and ZSM 1704/2004 (holotype). In both specimens, one hemipenis is clearly incompletely everted whereas the second one appears fully everted, although this cannot be stated with complete certainty. In ZSM 1505/2008, hemipenis length is 2.5 mm and maximum hemipenis width is 1.7 mm when fully turgid. The hemipenis is an irregular tubular structure without any ornaments, which in fully turgid state is somewhat balloon-shaped, widest at the central part of the truncus and more narrow at the pedicel and apex. Near the apex, two very small lobe-like structures are visible, corresponding to attachments of the retractor muscle, and a few very small spinelike papillae. However, it is not clear whether the hemipenis could possibly become further everted and then may become more strongly bilobed terminally, with a more prominent exposure of the spine-like papillae. There are no ornaments on pedicel and truncus, and no recognizable ornamental structures on apex (although apex might not be fully everted; Fig. 6). Etymology.— The species epithet is an adjective derived from the Latin ‘‘tristis’’ meaning ‘‘doleful’’, ‘‘sad’’, ‘‘sorrowful’’, and refers to the fact that the entire known range of this species (Montagne des Français) suffers from severe deforestation and habitat destruction [37] despite recently being declared as a nature reserve. Distribution.— Only known from the Montagne des Français limestone massif. Natural History.— Most individuals of B. tristis were found roosting at night on small branches about 5–50 cm above the leaf litter within a limestone massif with deciduous dry forest. By far most specimens were found in a few small areas where the species was abundant, whereas it was rare in other, apparently similar habitats. One of the females laid two large eggs (5.9 6 3.7 mm resp. 5.8 6 3.5 mm diameter) in February. The two juveniles hatched 64 and 69 days after egg deposition at temperatures between 20– 26°C. They measured 14 mm in total length. Eight days after hatching the weight of the older juvenile was 0.03 g., Published as part of Frank Glaw, Jörn Köhler, Ted M. Townsend & Miguel Vences, 2012, Rivaling the World's Smallest Reptiles: Discovery of Miniaturized and Microendemic New Species of Leaf Chameleons (Brookesia) from Northern Madagascar, pp. 1-24 in PLoS ONE 7 (2) on pages 12-15, DOI: 10.1371/journal.pone.0031314, http://zenodo.org/record/894041, {"references":["7. Glaw F, Vences M (2007) A Field Guide to the Amphibians and Reptiles of Madagascar. Koln: Vences & Glaw.","37. D'Cruze N, Sabel J, Green K, Dawson J, Gardner C, et al. (2007) The first comprehensive survey of amphibians and reptiles at Montagne des Francais, Madagascar. Herp Cons Biol 2: 87 - 99."]}

Brookesia confidens sp. n. ZooBank LSID: urn:lsid:zoobank.org:act:CB2A9146-0161-42B7-A145-6DED579F1F21 Remark.—This species has been considered before as conspecific with Brookesia sp. ‘‘Montagne des Francais’’ [7]. Holotype.— ZSM 2150/2007 (FGZC 1196), adult male (hemipenes incompletely everted), collected on trail to the ‘‘Petit Tsingy and Grotte des Chauves-Souries’’, 12°57'25"S, 49°0 7'0 6"E, 90 m a.s.l., Ankarana National Park, Antsiranana Province, northern Madagascar, on 1 March 2007 by P. Bora, H. Enting, F. Glaw, A. Knoll, and J. Köhler. Paratypes.— UADBA uncatalogued (FGZC 1194–1195), two specimens; ZSM 2151/2007 (FGZC 1197), probably male, ZSM 2152/2007 (FGZC 1198), adult female, ZSM 2153/2007 (FGZC 1199), adult female, all with same data as holotype; UADBA uncatalogued (FGZC 1608), UADBA uncatalogued (FGZC 1610), ZSM 1511/2008 (FGZC 1607), adult female, ZSM 1512 /2008 (FGZC 1609), adult male, all collected at ‘‘ Petit Tsingy’ ’, Ankarana National Park, Antsiranana Province, northern Madagascar, on 12 February 2008 by N. D’Cruze, M. Franzen, F. Glaw and J. Köhler. Diagnosis.— A member of the Brookesia minima group based on small body size (SVL Brookesia confidens is distinguished from other members of the group as follows: from B. dentata by probably smaller adult body size (no measurements of male B. dentata available), and absence of a supranasal cone (vs. presence); from B. exarmata by the absence of a supranasal cone (vs. presence); from B. karchei by a smaller adult body size (female SVL 20.6–22.6 mm vs. 30.7 mm; no measurements of clearly identified males available for B. karchei), and absence of a supranasal cone (vs. presence); from B. minima by a very narrow hemipenis (vs. balloon-like); from B. peyrierasi by generally smaller adult body size(male SVL 18.3– 20.1 mm vs. 19.7–22.4 mm), the absence of a supranasal cone (vs. presence), absence of a supraocular cone (vs. presence), and very narrow ornamentless hemipenis (vs. massive, bilobed hemipenis with four spines per lobe); from B. ramanantsoai by a smaller adult body size (male SVL 18.3–20.1 mm vs. 21.7 mm), absence of a supranasal cone (vs. presence), absence of a supraocular cone (vs. presence in some specimens), and hemipenis very narrow (vs. balloon-like); from B. tuberculata by the absence of a supranasal cone (vs. presence), absence of a supraocular cone (vs. presence), and hemipenis very narrow with pustules on apex (vs. wider, with crown-like apical structure). The new species is most similar to B. tristis but differs from this species by indistinct and short parasagittal crests (vs. distinct) and by 13 dorsolateral pointed tubercles (vs. 11), and ornamentless apical region of hemipenis (vs. spine-like papillae on apex). For a distinction from B. desperata and B. micra, described below, see the diagnoses of these species. Referencing a fragment of the 16S rRNA gene, B. confidens shows an uncorrected pairwise divergence of 6.7% to its sister species B. tuberculata, and divergences>9% to all other species of the B. minima group. Description of holotype.— Adult male in good state of preservation (Fig. 7; Supporting Information S 1). Both hemipenes everted. Measurements in Table 1. Head with lateral crest starting at median level at the posterior edge of eye; prominent orbital crests with distinctly enlarged pointed tubercle at posterior level slightly below lateral crest, and a crest at the posterior edge of the head, that form a weakly developed dorsal helmet; a pair of very short straight parasagittal crests that transform posteriorly into a broad area of elevated scales which almost cover the entire posterior head surface; three pointed tubercles on each side of posterior helmet crest, a prominent one at termination point of lateral crest, an even larger one at the imaginary termination point of parasagittal crest, and a weakly developed one between parasagittal and lateral crests; one pointed tubercle on lateral surface of head, below lateral crest in temporal region; orbital crest denticulated; no supraocular cone recognizable; supranasal cone not clearly recognizable; head longer (5.3 mm) than wide; chin and throat without longitudinal rows of slightly enlarged tubercles. Dorsal surface of body without a vertebral ridge or keel; 13 dorsolateral pointed tubercles form a complete longitudinal line on the body; most posterior (13th) pointed dorsolateral tubercle being largest, above insertion point of hindlimb; pointed dorsolateral tubercles almost equally spaced, pointing out almost perpendicularly from body; slightly enlarged, pointed tubercles form curved transversal crests on either side of the vertebral line between 2nd and 10th dorsolateral pointed tubercles; slightly enlarged tubercles on dorsal surface of tail, without any reconizable pattern; no well-defined dorsal pelvic shield in sacral area; few irregularly scattered enlarged rounded ubercles on the lateral surface of body; venter without enlarged tubercles; scattered, soft-pointed tubercles on limbs; no pointed tubercles around cloaca; longitudinal row of slightly enlarged pointed tubercles lateral on tail, forming a longitudinal row from tail base to two-thirds of tail length; no enlarged tubercles on ventral surfaces of tail. After almost four years in ethanol, all surfaces pale grey to beige, with most pointed tubercles being brown; upper surface of head pale brown. Variation.— For morphological measurements and proportions see Table 1 and Supporting Information S1. In ZSM 2153/2007, the ground colour is dark brown with a whitish vertebral stripe that starts at posterior half of body and extends onto the tail which is entirely beige with scattered brownish spots. The posterior dorsal surface of head is grey. The body of ZSM 1512/2008 is light grey whereas the head is brown with a greyish transverse line between the nostrils. The tail base of the females is less thickened than in the males. In life, all individuals with dorsal colouration of head, dorsum and tail light grey or pale beige, lateral parts of the body brown, with indistinct greyish marbling lateroventrally and with few small dark brown spots around the flank tubercles; limbs almost uniformly dark brown. This pattern is likely a stress colouration. Genital morphology.— For this species, everted hemipenes are available only for the holotype (ZSM 2150/2007). On both sides, the organs are single tube-like structures of low diameter which decreases from the apex to the distal part of the truncus. The left hemipenis is not fully everted whereas the one on the right side (length 1.3 mm; width at the pedicel 0.6 mm, width at the apex 0.4 mm) might be fully everted, but this cannot be verified without examination of further material. There are no ornaments on the pedicel and truncus. The sulcus spermaticus is not recognizable on most of the truncus but becomes more distinct in the apical region. The apical region is characterized by a slight increase in diameter but not bilobed, and is covered by relatively large but poorly defined pustular papillae (Fig. 6). The hemipenis is devoid of any further prominent structures such as the spiny apical papillae of B. peyrierasi (which are visible also in incompletely everted organs). Etymology.— The species epithet is an adjective derived from the Latin ‘‘confidens’’ meaning ‘‘confident’’, ‘‘trusting’’. The known range of the species is supposedly a well protected nature reserve with apparently limited habitat destruction. Furthermore, this area might benefit from natural protection by the tsingy limestone formations which are difficult to access, thus giving hope for the species’ survival. Distribution.— Only known from a single locality within Ankarana National Park. Natural History.— Most individuals were found roosting at night on thin branches about 5–20 cm above the leaf litter in deciduous dry forest close to a small forest trail within a small area, surrounded by tsingy outcrops. At this locality, the species was relatively abundant, whereas it was not found at similar localities nearby, suggesting a patchy distribution and a preference for certain microhabitats. This hypothesis is also supported by the fact that earlier herpetological surveys in Ankarana [38,39], did not record any species of the Brookesia minima group. When stressed, individuals can quickly change colour and display a broad pale vertebral stripe contrasting with the darker flanks.

BackgroundOne clade of Malagasy leaf chameleons, the Brookesia minima group, is known to contain species that rank among the smallest amniotes in the world. We report on a previously unrecognized radiation of these miniaturized lizards comprising four new species described herein.Methodology/principal findingsThe newly discovered species appear to be restricted to single, mostly karstic, localities in extreme northern Madagascar: Brookesia confidens sp. n. from Ankarana, B. desperata sp. n. from Forêt d'Ambre, B. micra sp. n. from the islet Nosy Hara, and B. tristis sp. n. from Montagne des Français. Molecular phylogenetic analyses based on one mitochondrial and two nuclear genes of all nominal species in the B. minima group congruently support that the four new species, together with B. tuberculata from Montagne d'Ambre in northern Madagascar, form a strongly supported clade. This suggests that these species have diversified in geographical proximity in this small area. All species of the B. minima group, including the four newly described ones, are characterized by very deep genetic divergences of 18-32% in the ND2 gene and >6% in the 16S rRNA gene. Despite superficial similarities among all species of this group, their status as separate evolutionary lineages is also supported by moderate to strong differences in external morphology, and by clear differences in hemipenis structure.Conclusion/significanceThe newly discovered dwarf chameleon species represent striking cases of miniaturization and microendemism and suggest the possibility of a range size-body size relationship in Malagasy reptiles. The newly described Brookesia micra reaches a maximum snout-vent length in males of 16 mm, and its total length in both sexes is less than 30 mm, ranking it among the smallest amniote vertebrates in the world. With a distribution limited to a very small islet, this species may represent an extreme case of island dwarfism.

Chamaeleons are well known for their unique suite of morphological adaptations. Whereas most chamaeleons are arboreal and have long tails, which are used during arboreal acrobatic manoeuvres, Malagasy dwarf chamaeleons (Brookesia) are small terrestrial lizards with relatively short tails. Like other chamaeleons,Brookesiahave grasping feet and use these to hold on to narrow substrates. However, in contrast to other chamaeleons,Brookesiaplace the tail on the substrate when walking on broad substrates, thus improving stability. Using three-dimensional synchrotron X-ray phase-contrast imaging, we demonstrate a set of unique specializations in the tail associated with the use of the tail during locomotion. Additionally, our imaging demonstrates specializations of the inner ear that may allow these animals to detect small accelerations typical of their slow, terrestrial mode of locomotion. These data suggest that the evolution of a terrestrial lifestyle inBrookesiahas gone hand-in-hand with the evolution of a unique mode of locomotion and a suite of morphological adaptations allowing for stable locomotion on a wide array of substrates.

Madagascar's flora and fauna are remarkable both for their diversity and supraspecific endemism. Moreover, many taxa contain large numbers of species with limited distributions. Several hypotheses have been proposed to explain this high level of microendemism, including 1) riverine barrier, 2) mountain refuge, and 3) watershed contraction hypotheses, the latter 2 of which center on fragmentation due to climatic shifts associated with Pliocene/Pleistocene glaciations. The Malagasy leaf chameleon genus Brookesia is a speciose group with a high proportion of microendemic taxa, thus making it an excellent candidate to test these vicariance scenarios. We used mitochondrial and nuclear sequence data to construct a Brookesia phylogeny, and temporal concordance with Pliocene/Pleistocene speciation scenarios was tested by estimating divergence dates using a relaxed-clock Bayesian method. We strongly reject a role for Pliocene/Pleistocene climatic fluctuations in species-level diversification of Brookesia. We also used simulations to test the spatial predictions of the watershed contraction model in a phylogenetic context, independent of its temporal component, and found no statistical support for this model. The riverine barrier model is likewise a qualitatively poor fit to our data, but some relationships support a more ancient mountain refuge effect. We assessed support for the 3 hypotheses in a nonphylogenetic context by examining altitude and species richness and found a significant positive correlation between these variables. This is consistent with a mountain refuge effect but does not support the watershed contraction or riverine barrier models. Finally, we find repeated higher level east-west divergence patterns 1) between the 2 sister clades comprising the Brookesia minima group and 2) within the clade of larger leaf chameleons, which shows a basal divergence between western and eastern/northern sister clades. Our results highlight the central role of phylogeny in any meaningful tests of species-level diversification theories.

We surveyed the lowland rainforest of the Foret d’Ambre Special Reserve in north Madagascar for amphibians and reptiles. We recorded a total of 20 amphibian and 39 reptile species via opportunistic searching and pitfall trapping in the first published survey to focus on this area. Consequently most of the species found were new records for the area. Approximately half of the species (51%) were only found in relatively undisturbed areas of forest and 61% appear to be restricted to lowland rainforest below 900 m elevation. The most vulnerable elements of this herpetofauna are the three species that appear to be locally endemic to Foret d’Ambre (according to the current knowledge): Boophis baetkei, Brookesia sp. nov., and Rhombophryne sp. nov. An additional 25 species are considered regional endemics, 14 species are threatened according to the 2007 Red List of Threatened Species and 15 species are listed on the CITES appendices. This paper contributes to the current understanding of Malagasy patterns of biodiversity by documenting the composition, geographical and ecological distribution of the herpetofauna found at this site. Despite its protected status, currently the Reserve is not being managed sufficiently as it is subject to numerous human-induced environmental problems resulting in habitat destruction and should therefore be considered a high conservation management priority. Herein, we provide conservation and development recommendations for this highly diverse site of herpetological importance. Furthermore, we provide an updated and revised species list of the amphibians and reptiles of Montagne d’Ambre National Park and a species list for the private Fontenay Nature Park. RESUME Lors d’un inventaire des especes d’amphibiens et de reptiles de la Reserve Speciale de la Foret d’Ambre au nord de Madagascar, nous avons recenses un total de 20 especes d’amphibiens et de 39 especes de reptiles par le biais d’echantillonnages directs et avec des trous-pieges « pitfall traps », constituant ainsi le premier resultat d’inventaire publie sur cette zone. De sorte que toutes les especes repertoriees constituent de nouvelles donnees pour cette region. Environ une moitie des especes (51%) n’a ete trouvee que dans des zones forestieres peu perturbees et 61% se limitent a la foret pluviale de basse altitude (altitude inferieure a 900 m). Les elements les plus vulnerables de cette herpetofaune sont les trois especes qui semblent etre localement endemiques, a savoir Boophis baetkei , Brookesia sp. nov. et Rhombophryne sp. nov. Par ailleurs, 25 autres especes sont endemiques de la region, 14 especes sont inscrites sur la liste rouge IUCN 2007 des especes menacees et 15 sont inscrites dans les annexes de la CITES. Ce travail contribue a une meilleure comprehension des schemas de la biodiversite malgache en documentant la composition ainsi que la distribution geographique et ecologique de l’herpetofaune recensee sur ce site. Malgre son statut de protection, la reserve est soumise a de nombreuses perturbations d’origine anthropique entrainant une destruction de l’habitat et devrait a ce titre etre consideree comme une zone prioritaire en matiere de conservation. Nous formulons des recommandations pour la conservation et le developpement de ces sites qui presentent une diversite faunique importance. Nous fournissons egalement une liste mise a jour et revisee des amphibiens et des reptiles du Parc National de la Montagne d’Ambre, ainsi qu’une liste du parc prive ‘Fontenay Nature Park’.

A phylogenetic hypothesis for the lizard family Chamaeleonidae is generated from 1503 aligned base positions (883 parsimony-informative) of mitochondrial DNA for specimens representing 59 species (57 ingroup and two outgroup). Sequences are reported for a genomic segment encoding eight transfer RNAs, NADH dehydrogenase component 2 (ND2), and portions of NADH dehydrogenase component 1 (ND1) and cytochrome c oxidase subunit 1 (COI). Newly reported genomic rearrangements and duplications support the hypothesis that mitochondrial gene order and content are destabilized by phylogenetic loss of a functional origin for light-strand replication between the genes encoding tRNAAsn and tRNACys. A novel gene order characterizes all sampled Brookesia except B. nasus. Brookesia nasus, the apparent sister taxon of a clade formed by all other Brookesia, has the ancestral gene order but contains a large tandem duplication. An apparently noncoding 220 base pair insertion between the genes encoding ND2 and tRNATrp is reported for Bradypodion tavetanum. Phylogenetic analysis identifies nine clades whose ancestral lineages diverged early in chamaeleonid evolutionary history: (1) Brookesia (possibly excluding B. nasus), (2) Chamaeleo subgenus Chamaeleo (excluding C. namaquensis), (3) Chamaeleo subgenus Trioceros, (4) viviparous Bradypodion, (5) oviparous Bradypodion, (6) genus Furcifer (except F. balteatus), and ( 7 – 9 ) three distinct clades of Calumma. Chamaeleo namaquensis, Brookesia nasus, Furcifer balteatus, Rhampholeon brevicaudatus, and R. spectrum represent ancient lineages dating to approximately the same time. Multiple independent losses and a possible secondary gain of horns are inferred for Trioceros. Viviparity has at least two separate origins in chameleons, one in Bradypodion and one in Trioceros.

The taxonomic status of recently discovered populations of pygmy chameleons (Rhampholeon) from the northern Mozambique montane isolates of Mt. Chiperone, Mt. Mabu, Mt. Inago and Mt. Namuli are assessed, and compared with the closest geographical congeners, including Rhampholeon platyceps Gunther 1893 from Mt. Mulanje, and Rh. chapmanorum Tilbury 1992 from the Malawi Hills, both in southern Malawi. Relationships were examined using morphological features and a phylogenetic analysis incorporating two mitochondrial and one nuclear marker. The phylogeny showed that each montane isolate contained a distinct, well-supported clade of chameleons. Chameleons from the Mozambican montane isolates are within a monophyletic clade inclusive of species from southern Malawi (Rh. platyceps and Rh. chapmanorum). Although some relationships are unresolved, the southern Malawi and Mozambican isolates appear to share their most recent common ancestor with species from the Eastern Arc Mountains and Southern Highlands of Tanzania and Malawi (Rh. moyeri, Rh. uluguruesis, Rh. nchisiensis). Along with Rh. beraduccii and Rh. acuminatus, all are included in the subgenus Rhinodigitum. Sister to this larger clade are species from west/central Africa (Rh. temporalis, Rh. spectrum) and the Rh. marshalli-gorongosae complex from southwest Mozambique and adjacent Zimbabwe. Morphological and molecular results confirm that Brookesia platyceps carri Loveridge 1953 is a junior subjective synonym of Rhampholeon platyceps Gunther 1892. Historical records of Rh. platyceps from the Shire Highlands (Chiromo) and the Zomba Plateau, are incorrect and the species is now considered endemic to the Mulanje massif. All of the four newly discovered, isolated populations are genetically and morphologically distinct, and we take the opportunity to describe each as a new species. Rhampholeon (Rhinodigitum) maspictus sp. nov. is restricted to Mt. Mabu and distinguished by its large size, well-developed dorsal crenulations, and bright male breeding coloration; Rhampholeon (Rhinodigitum) nebulauctor sp. nov. is restricted to Mt. Chiperone and distinguished by its small size, weakly-developed dorsal crenulations, and a large rostral process in males; Rhampholeon (Rhinodigitum) tilburyi sp. nov. is restricted to Mt. Namuli and distinguished by its small size, weakly-developed dorsal crenulations, and prominent flexure of the snout in males; and Rhampholeon (Rhinodigitum) bruessoworum sp. nov. is restricted to Mt. Inago and distinguished by its small size, weakly-developed dorsal crenulations, large rostral process in males, and relatively long tail in both sexes.

Chameleons of the Madagascan endemic genus Brookesia Gray are small to extremely small reptiles with a mostly terrestrial lifestyle, and due to their low mobility and their camouflage they are nearly invisible in the leaf litter of Madagascar's forests. The genus Brookesia is widely distributed (except in the arid south and south-west) but many Brookesia species are restricted to a relatively small area of the island (Glaw & Vences 2007). Speciation has been most prolific in the northern parts of Madagascar, an area that currently host about two-thirds of the 27 nominal Brookesia species (Raxworthy & Nussbaum 1995; Glaw & Vences 2007; Townsend et al. 2009).

Many of Madagascar's endemic chameleon species have restricted ranges and require relatively intact forest for survival but information on their response to habitat degradation is generally lacking, especially in western deciduous forests. We assessed the abundance and density of endemic chameleons from nocturnal surveys of roosting animals in Parc National Tsingy de Bemaraha in western Madagascar. We found 444 Brookesia brygooi, 192 B. perarmata, 98 B. exarmata, 22 Furcifer nicosiai and two F. cf. petteri. The highest pooled density was for B. brygooi (53.2 ha-1), followed by B. perarmata (29.2 ha-1) and B. exarmata (18.7 ha-1). B. brygooi was ubiquitous, with the highest density and abundance in the disturbed Ankazomanga forest outside the Park, where the similarly sized B. perarmata was absent. B. perarmata, a species endemic to the Park, was abundant in three of the five survey sites, especially where there was a well-developed leaf litter and understorey. B. exarmata, a species endemic to the Bemaraha plateau, was associated with an open understorey and small protrusions of limestone karst. Spatial variation in the density of B. perarmata is an important consideration when interpreting monitoring data and this species is a conservation priority because of its restricted distribution and apparent sensitivity to forest degradation.

The chameleon genus Brookesia, with 26 species, is endemic to Madagascar. The main threats to Brookesia are habitat destruction and collection for trade. At the CITES Conference of Parties 12, held in Santiago, Chile in 2002, all but one Brookesia species were added to Appendix II, under which trade is controlled in order to avoid utilization being incompatible with species' survival. The Vulnerable B. perarmata was added to Appendix I, under which trade is permitted only in exceptional circumstances. Summary information for Brookesia is presented here, reviewing taxonomy, habitat preferences, available population estimates, relevant legislation, major threats, and trade levels. Brookesia live on or close to the ground, with roost sites mostly

Malagasy chameleons are threatened by the relentless pace of habitat destruction. Their great popularity amongst herpetoculturalists has also resulted in collection for international animal markets. Most previous fieldwork has focused on the compilation of much needed species inventories, but little attention has been given to estimating chameleon population densities. This lack of information prevents a reliable assessment of the effects ofhabitat loss and direct exploitation on wild populations. A simple and repeatable methodology for the monitoring of chameleon populations is therefore urgently required. The present study developed survey techniques based on distance sampling within an area of relatively undisturbed rain forest at Ranomafana National Park. In total 394 individuals from six species were recorded and population densities (± SE) of 27.2 ± 5.4/ha Brookesia spp. and 21.4±4.0/ha Calumma spp. were estimated using the computer program DISTANCE. The authors propose that monitoring of chameleons in areas subject to different threats should begin immediately using the methods outlined in this paper.

Six species of Brookesia dwarf chameleons were recorded during two rainforest expeditions in Madagascar. One of these is described as a new species Brookesia valerieae based on the arrangement of the nine pairs of dorso-lateral spines, the lack of a pelvic shield, and dorsal chevron markings. Observations were made on the defence and roosting behaviours shown by these six species. Aspects concerning the evolution and conservation in Madagascar are discussed for this poorly studied group of endemic Malagasy reptiles.

The phylogenetic relationships of the African mainland species of the genus Rhampholeon (formerly referred to the genus Brookesia) with respect to Brookesia superciliaris, Bradypodion pumilus, and the genus Chamaeleo are analysed. Rhampholeon constitutes a genus well separated from Brookesia and may itself be subdivided in two subgroups on the basis of characters derived from the nasal and parietal complexes. A final section investigates problems of cladistic analysis, viz. the relationship of parsimony to character weighting, and pattern versus process as complementary perspectives of character analysis and coding.

Comparing the species of Brookesia and Rhampholeon with Chamaeleo it is concluded that Brookesia + Rhampholeon form a monophyletic group, arising from a branch of Chamaeleo, probably most related to the group around Chamaeleo nasutus. The separation between Rhampholeon and Brookesia is confirmed.

The form of the skull of Chamaeleo nasutus, especially the broad, trigonal parietal, is strikingly similar to the skull of Rhampholeon. This supports the hypothesis (Hillenius, 1986) that of all chameleons the group of species around Ch. nasutus is the most closely related to Rhampholeon.

5 Corresponding author Abstract We describe a new Brookesia species from a forest fragment located 13 km south of Ambalavao in the southern part of Madagascar's central high plateau. Brookesia brunoi sp. nov. is one of the few arid-adapted Brookesia species inhabiting deciduous forests on the western slope of the central high plateau of the island (around 950 m a.s.l.). So far the species has only been observed in the private Anja Reserve. The species belongs to the Brookesia decaryi group formed by arid-adapt- ed Brookesia species of western Madagascar: B. bonsi Ramanantsoa, B. perarmata (Angel), B. brygooi Raxworthy & Nussbaum and B. decaryi Angel. Brookesia brunoi differs from the other four species of the group by a genetic divergence of more than 17.6% in the mitochondrial ND2 gene, and by a combination of morphological characters: (1) nine pairs of laterovertebral pointed tubercles, (2) absence of enlarged pointed tubercles around the vent, (3) presence of poorly defined laterovertebral tubercles along the entire tail, (4) by the configuration of its cephalic crest, and (5) hemipenial morphology. Based on our molecular phylogeny this species is sister to a clade containing B. brygooi, B. decaryi, and probably B. bonsi for which no ND2 sequences were available. Our molecular data also confirm the presence of a divergent mitochondrial lineage in the Tsingy de Bemaraha, which might be assigned to either B. bonsi or B. decaryi, and point to the need of more research on this population.