Acta Herpetologica 10(2): 111-120, 2015 ISSN 1827-9635 (print) © Firenze University Press ISSN 1827-9643 (online) www.fupress.com/ah DOI: 10.13128/Acta_Herpetol-17171 A new species of Chameleon (Sauria: Chamaeleonidae: Kinyongia) highlights the biological affinities between the Southern Highlands and Eastern Arc Mountains of Tanzania Michele Menegon1,*, Simon P. Loader2, Tim R.B. Davenport3, Kim M. Howell4, Colin R. Tilbury5, Sophy Machaga3, Krystal A. Tolley5,6 1 Tropical Biodiversity Section, Museo delle Scienze, Corso  del Lavoro e della Scienza 3, 38122 Trento, Italy. *Corresponding author. E-mail: michele.menegon@muse.it 2 Department of Life Sciences, University of Roehampton, Holybourne Avenue, Room 1053, London SW15 4JD United Kingdom 3 Wildlife Conservation Society (WCS), PO Box 1475, Mbeya & PO Box 922, Zanzibar, Tanzania 4 Department of Zoology & Wildlife Conservation, PO Box 35064, University of Dar es Salaam, Dar es Salaam, Tanzania 5 South African National Biodiversity Institute, Private Bag X7, Claremont, Cape Town, South Africa 6 Department of Botany & Zoology, University of Stellenbosch Private Bag X1, Matieland, 7602, Stellenbosch, South Africa Submitted on 2015, 10th October; revised on 2015, 25th October; accepted on 2015, 27th October Editor: Sebastiano Salvidio Abstract. A new species of chameleon is described from the Livingstone and Udzungwa Mountains of Tanzania. The new species is morphologically most similar to Kinyongia vanheygeni. Furthermore, a single, short rostral appendage shows the species similarity to other Eastern Arc endemic Kinyongia species (e.g. K. uthmoelleri, K. oxyrhina, K. mag- omberae and K. tenuis). Females of all these species lack any rostral ornamentation and are all very similar morpho- logically. Males of the new species, on which the morphological diagnosis is based, can be distinguished from other Kinyongia by a shorter rostral appendage that bifurcates at the tip. They are easily distinguished from K. vanheygeni, otherwise the most similar species, by differences in head scalation and the length and shape of the rostral appendage. The new species is associated with montane rainforest and is known from only four forest fragments of which two are in the Udzungwa and two in the Livingstone Mountains. Phylogenetically, the new species is sister to K. tenuis and K. magomberae, which together, form a clade that also contains K. oxyrhina. The disjunct distribution of the new species, in the Livingstone and Udzungwa mountains, stretches across the ‘Makambako Gap’ which is a putative biogeographi- cal barrier separating the distinct faunas of the Southern highlands and Eastern Arc Mountains. Evidence from this species however, points to potentially closer biological affinities between the Livingstone and Udzungwa mountains. Keywords. Southern Highlands, Tanzania, Eastern Afromontane, Biodiversity, Chamaeleonidae, East Africa, new species, reptiles. INTRODUCTION Exploration and subsequent research in the past decades have substantially improved our understand- ing of the biodiversity from the Eastern Afromontane Region (EAR), which is known for its high species rich- ness (Menegon and Davenport, 2008). Collectively the Eastern Arc Mountains (EAM) and Southern Highlands of Tanzania form a system of mountain blocks spanning from southern Kenya, through Tanzania and into Malawi (Lovett and Wasser, 1993). Because of the prevailing cli- matic influence from the warm Indian Ocean, the EAM receives high orographic rainfall providing a relatively stable climate. This climate stability is thought to have reduced extinction rates for forest endemic taxa (e.g., Tol- ley et al., 2011; Loader et al., 2014). This, coupled to ele- 112 Michele Menegon et alii vated speciation rates as a result of specialization due to ecotones between forest and savanna (Caro et al., 2013), has presumably resulted in high diversity and endemism across the EAM (Loader et al., 2015). Our understanding of more general evolution- ary patterns and processes for the EAM, including bio- geographic patterns has increased dramatically for some taxonomic groups (e.g., Tolley et al., 2011; Dimitrov et al., 2012; Loader et al., 2014). Despite this, a number of vertebrate species are discovered and described each year (e.g., Rovero et al., 2014), indicating that our knowledge is far from complete in this region. A prime example are chameleons, for which new species are steadily being described (Tolley and Menegon, 2013), or previously named taxa are elevated from synonymy (Tilbury and Emmrich 1996; Menegon et al., 2002; Mariaux and Til- bury, 2006; Tilbury et al., 2006; Mariaux et al., 2008, Til- bury and Tolley, 2009; Menegon et al., 2009; Greenbaum et al., 2012; Branch et al., 2014) and these contributions have subsequently been utilized for revealing broader evolutionary patterns (Tolley et al., 2011; Tolley et al., 2013; Ceccarelli et al., 2014). Essentially, the scientific focus on EAM region has led to a substantial increase in knowledge on the flora and fauna, but the biota of the Southern Highlands, which is separated from the more northern lying EAM by the dry, low-lying Makam- bako Gap (Lovett and Wasser, 1993), is relatively poorly known. Indeed, the Makambako gap is considered an important turn-over region (e.g., Rovero et al., 2014) and as a result, many biodiversity studies have instead focused on the EAM because of its known biological wealth (e.g. Newmark, 1998; Stanley et al., 1999). Chameleons in the genus Kinyongia (Tilbury et al., 2006) are a prominent group in the EAM because find- ings have contributed to a broader understanding of spe- cies richness, endemism, and biogeography (e.g., Tolley et al., 2011). Twelve of the 16 described species of Kinyon- gia occur on isolated massifs within the EAM, with the remainder found to the northwest in mountainous regions of the Albertine Rift in Democratic Republic of the Con- go, Uganda and Rwanda and on isolated volcanoes, such as Kilimanjaro, Meru, Mt. Kenya (Tilbury, 2010; Tolley et al., 2011; Greenbaum et al., 2012). Many species have small distributional ranges, and are usually found on the forested slopes of just one or a few isolated massifs. Their isolated and restricted distributions have provided evi- dence for a long history of persistence in EAM, and as well as allowed inferences as to the formation and mainte- nance of refugial areas (Tolley et al., 2011). New biological surveys in unexplored regions such as southern Tanzania continually reveal the presence of new species, including Matilda’s horned viper Atheris matil- dae (Menegon et al., 2011), the two chameleons Kinyon- gia vanheygeni (Necas, 2009) and K. magomberae (Men- egon et al. 2009), and Africa’s only new genus of monkey described in the last 80 years, the kipunji Rungwecebus kipunji (Davenport et al., 2006). Unexplored forests to the south of the EAM (e.g. Southern Highlands) are there- fore predicted to contain a host of species not yet known to science. In this study, we describe a new chameleon species in the genus Kinyongia (Fig. 1) that is found in Afrotemperate forest from both the Livingstone (South- ern Highlands) and Udzungwa (EAM) Mountains (Fig. 2). Using both morphological and molecular evidence, we determine the taxonomic placement and evolution- ary relationships for this new taxon. Both morphologi- cal characters and genetic markers were examined and compared to the other species of Kinyongia. Furthermore we examine the biogeographical implications of this new taxon given the phylogenetic hypothesis inferred from the data. MATERIALS AND METHODS Material examined The following specimens (Table 1) were examined from the herpetological collections of the Science Museum of Trento, Trento, Italy (MTSN and MUSE), the Department of Zoology & Wildlife Conservation of the University of Dar es Salaam, Dar es Salaam, Tanzania and the collection of the WCS’ Southern Highlands Conservation Project (SHCP), Mbeya, Tanzania: K. vanheygeni (MUSE 13523 and MUSE 13524 from Mt. Rung- we), K. tenuis (KMH 21325 and KMH 21304 from Nilo Forest Reserve, East Usambara Mts.), K. oxyrhina (KMH 28277 from Ukami Forest, Udzungwa Mts.; KMH 28302 from Nyumbanitu Forest, Udzungwa Mts., MTSN 8454 and MTSN 8412 from Nguru South Forest Reserve), K. tavetana (MTSN 8658 and MTSN 8661 from Kindoroko Forest Reserve, North Pare Mts.) Molecular Analysis To understand the phylogenetic placement the new Kinyon- gia species a phylogenetic analysis was carried out which included 10 individuals from the two mountain ranges, plus multiple representatives from 17 of 19 Kinyongia species from published datasets (Menegon et al., 2009; Tolley et al., 2011; Greenbaum et al., 2012). The resulting dataset consisted of 47 individuals, including the outgroup taxa (Bradypodion pumi- lum and B. melanocephalum). DNA extraction, PCR amplifica- tion, and cycle sequencing of two mitochondrial gene fragments (ND2 and 16S) were carried out following standard proce- dures using the following primers for ND2: L4437b and H5934 (Macey et al., 1997a, b), and 16S: L2510 and H3080 (Palumbi, 1996). Standard PCR and sequencing were followed for this gene fragment, with PCR annealing temperature at 57˚C. All 113A new species of chameleon from Tanzania new sequences were deposited in European Nucleotide Archive (Table 2). Bayesian inference was used to investigate optimal tree space using MrBayes 3.1.2 (Huelsenbeck and Ronquist, 2001) for the combined mitochondrial markers (321 characters, parti- tioned by marker: ND2, 856 bp; 16S, 465 bp), although 18 bases were excluded for 16S due to ambiguous alignment. To inves- tigate which evolutionary model best fit the data, jModeltest was used (Posada, 2008), and the AIC test indicated the same model for all markers was appropriate (GTR + G). Therefore, MrBayes was run specifying six rate categories with uniform priors for the gamma distribution for each of the partitions. To ensure the results were robust for both datasets, the MCMC was run twice in parallel for 10 million and 20 million generations (four chains in each run), with trees sampled every 1000 gen- erations. Burn-in was estimated as 1 million generations (1000 trees), as determined by examination the average standard devi- ation of split frequencies, the convergence diagnostic (PSRF val- ues ~ 1.0) as well as the log-probabilities and the values of each parameter for stabilization (Ronquist and Huelsenbeck, 2003). In addition, Tracer v1.4.1 (Rambaut and Drummond, 2007) was used to check that the effective sample size (ESS) of all param- eters was greater than 200 after burn-in. A 50% majority rule tree was constructed and nodes with ≥ 0.95 posterior probabil- ity considered supported. In addition to the Bayesian analysis, a maximum likeli- hood (ML) search was run for both datasets using RAxML HPC 7.2.8 (Stamatakis, 2006). The datasets were partitioned as in the Bayesian analysis, with a GTR+I+G model for all mark- ers and rapid bootstrapping halted automatically (Stamatakis et al., 2008) This analysis was run three times to ensure that inde- pendent ML searches produced the same topologies. We con- sidered nodes with a bootstrap value of ≥ 70% as supported in this analysis. Both Bayesian and likelihood analyses were run on the CIPRES Science Gateway (Miller et al., 2010; www.phylo. org/sub_sections/portal/). Finally, to provide a rough indica- Fig. 1. Kinyongia msuyae sp. nov. from Livingstone Mountains in life. Pictures showing (upper) adult Male, (lower left) close up of male head, (lower right) Adult female. 114 Michele Menegon et alii tion of the degree of divergence between species, uncorrected p-distances were estimated in MEGA 5.05 for 16S which had the most complete taxon sampling (Tamura et al., 2011). RESULTS Molecular Analysis The likelihood and Bayesian searches produced the same topology and supported nodes (Fig. 3). The phylogenetic analysis showed that the Kinyongia sam- pled from the Livingstone and the Udzungwa moun- tains form a well-supported clade. The uncorrected net sequence divergence (p-distance) between this clade and other closely related species of Kinyongia range from 1.4- 3.7% (Table 3), which is similar to within species values of other chameleons (e.g. Menegon et al., 2009; Tilbury and Tolley; 2009, Greenbaum et al., 2012, Tolley et al., 2012; Branch et al., 2014). Although sequence divergence between chameleons from Livingstone and Udzungwa (< 1.3% for the 16S marker) are close to the lowest val- ues found between some other Kinyongia (e.g. K. tenuis and K. magomberae; Table 3), additional samples from Udzungwa would be needed to obtain a more accurate estimate of sequence divergence between the mountain ranges. Given the genetic distinctiveness, we take the opportunity to describe the individuals from Livingstone and Udzungwa Mountains as a new species. Taxonomy Kinyongia msuyae sp. nov (Fig. 1; 3) Holotype: Adult male in the Science Museum of Trento, MTSN 9374 collected in Mdandu Forest Reserve, Livingstone Mountains in January 2011 by Michele Men- egon, Tim Davenport, Simon Loader, Sandra Dürren- berger, Sandra Rudolf and Sophy Machaga Type locality: Mdandu Forest Reserve, Livingstone Mountains 1900 m above sea level, Mbeya Region, South Eastern Tanzania (-9.769549621; 34.78832024) K. oxyrhina K. vanheygeni K. magomberae K. tenuis K. msuyae K. uthmoelleri other Kinyongia species K. cf. oxyrhina possible dispersal routes Makambako Gap Udzungwa Mts. Southern Highlands (Livingstone Mts.) Mahenge Mts. Kilombero Valley Fig. 2. Distribution of Kinyongia species in the Eastern Afromon- tane Region. Inset map shows enlargement of Udzungwa and Southern Highlands region of Tanzania with possible dispersal routes of montane associated species. Table 1. Biometrics of the holotype and paratypes. Continuous measurements given in mm. Total Length SVL TL Head length Head Width Casque Lenght Casque Eye Snouth Lenght Eye Diameter Eye-eye Gap Upper Labials Rostral process MTSN 9374 150.74 69.56 81.18 23.54 10.12 16.68 10.78 7.02 6.61 5.38 20 4.62 MTSN 9375 142.3 65.02 77.28 23.18 10.42 15.11 10.29 8.14 6.69 5.39 20 4.4 MTSN 9898 103.77 43.84 59.93 17.55 18.21 10.58 7.98 6.39 4.88 4.52 18 3.13 MTSN 9373 129.44 57.46 71.98 19.64 9.29 13 8.04 6.58 5.38 5.37 14 no MTSN 9377 103.5 52.69 50.81 15.75 7.77 10.11 6.12 5.99 4.53 4.41 14 no MTSN 7497 124.46 61.99 62.47 21.08 11.13 13.39 8.23 7.07 7.75 5.93 13 no MTSN 9378 97.36 40.2 57.16 13.31 6.59 7.79 6.21 4.37 4.61 3.6 14 no MUSE 13521 141. 89 68.35 73.54 22.57 11.03 14.99 10.03 7.56 7.06 5.41 19 3.63 MUSE 13522 145.53 64.25 81.28 22.37 10.31 15.66 10.36 7.14 6.39 5.01 17 4.87 115A new species of chameleon from Tanzania Paratype: MTSN 9375, adult male, MTSN 9373, MTSN 9377, adult females; MTSN 9378, juvenile, same data as holotype. MTSN 7497 collected in Sakara Nyumo Forest Reserve (Livingstone Mountains) in January 2011 by Michele Menegon, Tim Davenport, Simon Loader and Sophy Machaga; MTSN 8686 (adult male) collected in Kigogo Forest Reserve, Udzungwa Mountains in Febru- ary 2006 by Michele Menegon; Referred material: MUSE 13521 (Field number CAM 1013) collected in Kigogo Forest Reserve by Charles A. Msuya. MUSE 13522 (Field number KMH 28302), adult male, collected in Nyumbanitu Forest Reserve by Louis Hansen. Diagnosis: A small, elongated chameleon, lacking distinctive colours or pattern, with a tail longer than the snout-vent length. It has a short, bone-based rostral appendage formed by a converging, scaly elongation of the canthi rostrales, the areas bound by the two canthi is concave and covered in flattened scales. The tips of these elongations are free and they appear like a double-tipped CT498 CT500 MTSN 9373 MTSN 9374 MTSN 9375 MTSN 9377 MTSN 9378 MTSN 7497 ludewa MTSN 8686 Livingstone Mountains Udzungwa Mountains CAS 168917 K. tenuis CT103 K. tenuis MTSN 8218 K. magomberae MTSN 8835 K. magomberae CT192 K. oxyrhina CT193 K. oxyrhina CT490 K. vanheygeni SCHP/08/R/50 K. vanheygeni SCHP/08/R/91 K. vanheygeni CT189 K. uluguruensis CT191 K. uluguruensis CAS168852 K. matschiei CT105 K. matschiei CAS168921 K. vosseleri CT104 K. vosseleri CT110 K. multituberculata CT111 K. multituberculata BM29 K. boehmei JM2946 K. boehmei CT334 K. fischeri MTSN8490 K. fischeri CT151 K. uthmoelleri CT339 K. uthmoelleri CAS201593 K. adolfifriderici CAS201594 K. adolfifriderici EBG2390 K. gyrolepis EBG2391 K. gyrolepis CT345 K. carpenteri CT346 K. carpenteri CT350 K. xenorhina CT351 K. xenorhina CT209 K. excubitor CT106 B. melanocephalum KT62 B. pumilum 0.05 substitutions/site CT113 K. tavetana CT207 K. tavetana MTSN8661 K. tavetana K. msuyae Fig. 3. The best scoring maximum likelihood tree for Kinyongia, with nodes supported by maximum likelihood (bootstrap > 70%) and Bayesian (posterior probabilities > 0.95) analyses indicated by black circles. Grey circle indicate support with Bayesian posterior probabili- ties only. 116 Michele Menegon et alii Table 2. Museum, GenBank and European Nucleotide Archive accession numbers (16S, ND2) for Kinyongia used in this study (CAS = Cali- fornia Academy of Sciences; MSTN = Science Museum of Trento (formerly Museo Tridentino di Scienze Naturali); PEM = Port Elizabeth Museum (Bayworld). N/A: sequences not available. Species Locality ID Specimen 16S ND2 B. melanocephalum KwaZulu-Natal, South Africa CT016 N/A AY289813 HF570475 B. pumilum Western Cape, South Africa KT62 N/A AY756639 AY756689 K. adlofifriderici Bwindi N.P., Uganda CAS201593 CAS201593 DQ923820 EF014304 K. adlofifriderici Bwindi N.P., Uganda CAS201594 CAS201594 GQ221944 GQ221965 K. boehmei Taita Hills, Kenya BM29 N/A GQ221942 GQ221963 K. boehmei Taita Hills, Kenya JM2946 N/A GQ221948 GQ221969 K. carpenteri Rwenzori Mtns, Uganda CT345 PEM R16572 DQ923821 EF014305 K. carpenteri Rwenzori Mtns, Uganda CT346 PEM R16573 DQ923822 EF014306 K. excubitor Mount Kenya, Kenya CT209 PEM R16571 DQ923823 EF014307 K. fischeri Nguru Mountains, Tanzania CT334 PEM R16566 DQ923829 EF014313 K. fischeri Nguru Mountains, Tanzania MTSN 8490 MTSN 8490 GQ221951 GQ221971 K. gyrolepis Lendu Plateau, DRC UTEP 20341 UTEP20341 JN602059 JN602049 K. gyrolepis Lendu Plateau, DRC UTEP 20342 UTEP 20342 JN602055 JN602050 K. magomberae Udzungwa Mountains, Tanzania MTSN 8218 MTSN 8218 GQ221950 GQ221970 K. magomberae Magombera Forest, Tanzania MTSN 8492 MTSN 8492 GQ221952 GQ221972 K. matschiei East Usambara Mtns, Tanzania CAS 168852 CAS 168852 FR716605 FR716641 K. matschiei East Usambara Mtns, Tanzania CT105 N/A GQ221946 GQ221967 K. multituberculata West Usambara Mtns, Tanzania CT110 PEM R5735 DQ923824 EF014308 K. multituberculata West Usambara Mtns, Tanzania CT111 N/A GQ221947 GQ221968 K. msuyae Livingstone Mountains, Tanzania CT498 N/A LN997632 K. msuyae Livingstone Mountains, Tanzania CT500 N/A LN997633 LN997642 K. msuyae Livingstone Mountains, Tanzania Ludewa N/A LN997638 K. msuyae Livingstone Mountains, Tanzania MTSN7497 MTSN7497 LN997637 LN997643 K. msuyae Udzungwa Mountains, Tanzania MTSN8686 MTSN8686 LN997639 K. msuyae Livingstone Mountains, Tanzania MTSN9373 MTSN9373 LN997634 LN997644 K. msuyae Livingstone Mountains, Tanzania MTSN9374 MTSN9374 LN997635 LN997645 K. msuyae Livingstone Mountains, Tanzania MTSN9375 MTSN9375 LN997636 LN997646 K. msuyae Livingstone Mountains, Tanzania MTSN9377 MTSN9377 LN997647 K. msuyae Livingstone Mountains, Tanzania MTSN9378 MTSN9378 LN997648 K. oxyrhina Uluguru Mountains, Tanzania CT192 PEM R16569 DQ923831 EF014315 K. oxyrhina Uluguru Mountains, Tanzania CT193 PEM R16552 DQ923832 EF014316 K. tavetana Mount Kilimanjaro, Tanzania CT113 PEM R5736 DQ991233 FJ717801 K. tavetana Mount Meru, Tanzania CT207 PEM R16563 DQ923833 EF014317 K. tavetana North Pare Mountains, Tanzania MTSN 8661 MTSN 8661 FR716615 FR716649 K. tenuis East Usambara Mtns, Tanzania CAS 168917 CAS 168917 DQ923834 EF014318 K. tenuis East Usambara Mtns, Tanzania CT103 PEM R5731 DQ923835 EF014319 K. uluguruensis Uluguru Mountains, Tanzania CT189 PEM R16565 DQ923825 EF014309 K. uluguruensis Uluguru Mountains, Tanzania CT191 PEM R16557 DQ923826 EF014310 K. uthmoelleri South Pare Mtns, Tanzania CT151 PEM R16585 DQ923836 EF014320 K. uthmoelleri Mount Hanang, Tanzania CT339 N/A DQ923837 EF014321 K. vanheygeni Poroto Mountains, Tanzania CT490 LN997631 LN997649 K. vanheygeni Poroto Mountains, Tanzania SCHP-08-R-50 LN997640 LN997650 K. vanheygeni Poroto Mountains, Tanzania SCHP-08-R-91 LN997641 LN997651 K. vosseleri East Usambara Mtns, Tanzania CAS 168921 CAS 168921 GQ221943 GQ221964 K. vosseleri East Usambara Mtns, Tanzania CT104 N/A GQ221945 GQ221966 K. xenorhina Rwenzori Mtns, Uganda CT350 PEM R16570 DQ923838 EF014322 K. xenorhina Rwenzori Mtns, Uganda CT351 PEM R15568 DQ923839 EF014323 117A new species of chameleon from Tanzania short horn protruding over the snout by 3 to 5 mm. The appendage is plated with subequal rounded tubercules. Laterally, the appendage continues from the supra-orbital crest, formed by low peaked tubercles, becoming more serrated over the anterior rim, from where it continues forward as a scaly rostral short horn. In the males exam- ined it extends between 3 and 4 mm beyond the ante- rior margin of the rostral scale. Females lack any rostral appendage and have a lower casque. K. msuyae does resemble K. vanheygeni Necas, 2009 and, to a lesser extent, K. uthmoelleri (Müller, 1938) in size, general body and head shape and by possession of a single, bone-based rostral appendage in males. It differs from K. vanheygeni in the length of the rostral append- age being longer, formed by more than ten scales and pointing straight forward (less than ten scales and slight- ly pointing upward in K. vanheygeni), from K. uthmoe- lleri by having a horn-like longer rostral appendage (can- thal scales in K. uthmoelleri males meet to form a ‘rostral wall’, or protruding in form of a very short rostral pro- jection). Kinyongia msuyae can easily be distinguished from the other known Kinyongia species by the combination of the following characters: (1) presence of rostral process in males formed by the partial fusion of the canthi rostrales and protruding forward over the snout by 3 to 5 mm. (2) tail longer than SVL in both sexes, and (3) gular, ventral and dorsal crest absent. Description of the holotype Adult male. Total length 162.4 mm, SVL 73.7, Tail length 88.7. Casque elongated, posteriorly raised, covered by flattened polygonal scales, giving it a smooth appear- ance. Parietal crest formed by a series of low peaked tuberculated scales, temporal and orbital crests present. No occipital lobes. Nostril posteriorly directed, posi- tioned halfway between tip of snout and the anterior rim of the eye, and separated from upper labials by two to three rows of flattened scales. Canthi rostrales converge above and before the nostrils in forming a single, short, rostral appendage with two tips, giving the appearance of two very short horns protruding beyond the rostral scale by 4.5 mm. The rostral process is completely ossified and covered by sub-equal, convex scales, the superior edge is serrated. Upper labials 16, lower labials 15 on each side. The sides of gular region is lined with 6 shallow grooves on each side. There is no dorsal or gular crest, while the central part of the gular region has no groove. No signs of dorsal, or ventral crests. Scales on body flat and homo- geneous, arranged is small clusters, those on the upper part of the dorsum are more quadrangular and arranged in vertical rows. Scales on limbs sub-equal, rounded, and flattened. Tail longer than the snout/vent length, later- ally compressed, and covered by quadrangular scales arranged in vertical rows. Hemipenes: unenverted. Colour in preservative: The overall colour is whitish- grey with few paler areas Paratype variation: Paratypes show no relevant mor- phological variation compared to the holotype. Variation in scutellation and body proportions for the type series and referred material are shown in Table 2. Colour in life: K. msuyae is an overall brown to green chameleon, sometimes with broad pale transversal bands and scattered blue spots formed by single scales or clus- ters of several scales. Females have often a larger round spot of contrasting colour on the flanks (Fig. 1). The tip of the snout, rostral appendage and limbs and top of the casque are often brownish to grey. Distribution: Refer to Figure 2. Etymology: The species is named after and dedicated to Charles A. Msuya, a pioneer of Tanzanian herpetology, who collected the first known specimen attributable to this species and has spent most of his life studying Tan- zanian wildlife. DISCUSSION The phylogenetic analyses, sequence divergence estimates, and morphological assessment suggest that there is a previously unknown but distinctive species of Kinyongia from the Livingstone and Udzungwa moun- tains, which we describe as Kinyongia msuyae. This spe- cies is sister to chameleons found in the Eastern Arc Mountains (i.e. K. tenuis from Usambara Mountains, K. magomberae from Udzungwa Mountains, and K. oxy- rhina from Uluguru Mountains). There was no clear morphological differentiation between the population on the Udzungwa and that from Livingstone Mountains, despite these two mountain ranges being separated by Table 3. Sequence divergence (p-distances) for Kinyongia within species (on diagonal) and between selected species for 16S (lower matrix). Populations of K. msuyae from Livingstone and Udzungwa are given separately. N/A = not available. 1 2 3 4 5 1 K. msuyae (Livingstone) 0.0006         2 K. msuyae (Udzungwa) 0.0129 N/A       3 K. tenuis 0.0262 0.0292 0.0000     4 K. oxyrhina 0.0208 0.0235 0.0246 0.0000   5 K. magomberae 0.0269 0.0370 0.0137 0.0186 0.0000 118 Michele Menegon et alii the Makambako Gap (ca. 150 km apart). Sequence diver- gence between chameleons from these localities less than what is normally found between species. Population level differences may exist, but additional sampling would be required to confirm that hypothesis. The close relationship among populations has some important biogeographic implications. The Southern Highlands have long been regarded as isolated and not part of the Eastern Arc Mountains, with the Makambako gap considered inhospitable, preventing dispersal. How- ever, recent molecular data has started to alter this view. Phylogenetic analyses for the shrew, Myosorex, suggest that the Makambako Gap is of little consequence in the historical biogeography of the genus (Stanley and Ess- eltyn, 2010). Similarly, there is little morphological vari- ation among populations of the murid rodent Hylomys- cus arcimontensis on either side of the Makambako Gap (Carleton and Stanley, 2005), and the newly discovered kipunji monkey (Rungwecebus kipunji) has populations on both sides of the gap (Jones et al., 2005; Davenport et al., 2006). There are also some commonalities in the avi- fauna among populations on the Nyika Plateau, Mount Rungwe, and the southern Udzungwas, with no evidence of the Makambako Gap having a biogeographic influence (Stuart et al., 1993). Furthermore the Southern Highlands might have served as a dispersal route for amphibians, connecting the Udzungwa and the Mahenge Mts, the two southernmost mountain blocks of the Eastern Arc (Men- egon et al., 2011; Loader et al., 2014). Interestingly, from a biogeographic perspective, the most suitable dispersal route for forest endemics from Udzungwa and Mahenge mountains – both part of the Eastern Arc Mountains - does not appear to be the short- est straight line distance, which would require crossing the Kilombero Valley (an ancient, deep, wide valley). Instead, the continuous ridge of highlands connecting the southern Udzungwa, through the Southern Highlands/ Livingstone Mountains via the Makambako Gap and then northeast to the Mahenge Mountains may have remained more suitable over historical times, potentially with for- ested areas (see Fig. 2). The description of Kinyongia msuyae provides a tan- talizing piece of evidence suggesting strong biogeographi- cal affinities between the Southern Highlands (i.e. Liv- ingstone Mountains) and the Eastern Arc Mountains (i.e. Udzungwa Mountains). The Makambako Gap may not be a turn-over region of high significance between the EAM and the Southern Highlands, rejecting previous claims of its biogeographical importance as a barrier. Instead, it is likely that some taxa can or have crossed this bar- rier, or that the gap was formerly less dry, forming a cor- ridor between Udzungwa and the Southern Highlands. Our increased understanding of the Southern Highlands is revealing that the region is more species rich than had been supposed, possibly similar in scale to some of the Eastern Arc Mountain forests. ACKNOWLEDGEMENTS For advice, help with fieldwork, granting national and local permits for research and export in Tanzania, we thank (no particular order) Tanzania Commission for Science and Technology (COSTECH research per- mit RCA 2001-272; RCA 2007-153, RCA 2009-306-NA- 2009-201, 2011-239-NA-2011-82, 2006 and 2007-72-Na- 2006-19), Tanzania Wildlife Research Institute (TAWIRI), Wildlife Division. We are also grateful to many people and organiza- tions that provided assistance in the field, logistical sup- port and advice, including Sandra Dürrenberger, San- dra Rudolf, Noah Mpunga, Tanzania Forest Conserva- tion Group, and colleagues of the Wildlife Conservation Societies Southern Highlands Conservation Programme. Thanks to Nicholas Barbieri for the help in the lab. 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