Systematic revision of the Malagasy chameleons Calumma boettgeri and C. linotum (Squamata: Chamaeleonidae) Author Prötzel, David Author Ruthensteiner, Bernhard Author Scherz, Mark D. Author Glaw, Frank text Zootaxa 2015 4048 2 211 231 journal article 10.11646/zootaxa.4048.2.4 2bc19ab1-a510-408b-8fc3-e526ee3dc59a 1175-5326 233651 2CAB0746-175E-4FE1-B0D0-23DFF395A559 Morphology of Calumma boettgeri sensu lato External morphology . Measurements of important morphological parameters were taken from 23 specimens ( 11 males , 12 females ) from Nosy Be, six specimens (five males, one female) from Montagne d’Ambre, and another nine specimens without exact locality data ( Table 1 ). Because there was only one female from Montagne d’Ambre available, only males from both species were considered for comparison of body size, extremities and appendages. The specimens without localities were not included in mean value calculations but could be assigned according to their morphology to the Nosy Be morphotype (four specimens) and the Montagne d’Ambre morphotype (five specimens), respectively. TABLE 1 . Morphological measurements of Calumma boettgeri and C. linotum . Abbreviations: m, male; f, female; SVL, snout-vent length; TaL, tail length; TL, total length; , ratio of tail to snout-vent length; LRA, length of rostral appendage from snout tip; RRS, ratio of length of rostral appendage and snout-vent length; RAPSC, number of peripheral scales on rostral appendage; NPSCM, number of peripheral scales per mm on rostral appendage; RC, rostral crest present (+) or absent (-); NSL, number of supralabials; NIL, number of infralabials; OLD, lateral diameter of the occipital lobe; OLN, depth of the dorsal notch in occipital lobe; PC, parietal crest absent (-) or number of parietal cones; DC, dorsal crest absent (-) or number of dorsal cones; AP, axillary pits present (+) or absent (-); DSC, diameter of broadest scale on upper arm; RSB, ratio of broadest scale to snout-vent length; NSC, number of big scales on upper arm from lateral view; UAD, upper arm diameter; RAS, ratio of arm diameter to snout-vent length; all measurements in mm.

collection no.	species	locality	sex	SVL	TaL	TL	RST	LRA	RRS	RAPSC	NPSCM	RC	NSL
ZFMK 51389	C. boettgeri	Nosy Be	m	51.5	51.8	103.4	0.99	4.0	0.077	26	6.5	+	12
ZFMK 51520	C. boettgeri	Nosy Be	m	50.7	54.2	104.9	0.94	3.0	0.060	21	7.0	+	13
ZFMK 45987	C. boettgeri	Nosy Be	m	51.6	52.8	104.4	0.98	2.6	0.051	13	4.9	+	11
ZFMK 45988	C. boettgeri	Nosy Be	m	49.9	54.4	104.3	0.92	3.1	0.062	18	5.8	+	12
ZFMK 51518	C. boettgeri	Nosy Be	m	46.0	45.5	91.4	1.01	2.6	0.057	17	6.5	+	12
ZFMK 51519	C. boettgeri	Nosy Be	m	50.7	50.2	100.9	1.01	3.2	0.062	26	8.3	+	12
ZFMK 51521	C. boettgeri	Nosy Be	m	41.1	45.8	86.9	0.90	4.1	0.100	22	5.4	+	12
ZSM 36/1913	C. boettgeri	Nosy Be	m	41.4	44.0	85.4	0.94	2.5	0.059	15	6.1	+	12
ZSM 440/2000	C. boettgeri	Nosy Be	m	50.1	54.1	104.2	0.93	3.0	0.060	14	4.6	+	12
ZSM 444/2000	C. boettgeri	Nosy Be	m	51.9	55.0	106.9	0.94	2.9	0.057	17	5.8	+	12
SMF 16471	C. boettgeri	Nosy Be	m	50.3	54.3	104.6	0.93	3.6	0.072	15	3.6	+	13
ZFMK 51514	C. boettgeri	-	m	49.4	51.2	100.6	0.96	3.5	0.072	17	4.8	+	13
ZSM 866/1920	C. boettgeri	-	m	54.6	55.7	110.3	0.98	3.6	0.065	17	4.8	+	16
ZFMK 45985	C. boettgeri	Nosy Be	f	48.9	46.7	95.6	1.05	3.8	0.077	20	5.3	+	12
ZFMK 51516	C. boettgeri	Nosy Be	f	51.4	51.2	102.6	1.00	2.2	0.042	20	9.2	+	11
ZFMK 51517	C. boettgeri	Nosy Be	f	50.5	47.6	98.1	1.06	3.1	0.062	18	5.8	+	11
ZFMK 50615	C. boettgeri	Nosy Be	f	42.0	41.8	83.8	1.00	3.0	0.072	18	5.9	+	11
ZFMK 45983	C. boettgeri	Nosy Be	f	50.8	48.6	99.4	1.05	4.1	0.081	-	-	+	12
ZFMK 45986	C. boettgeri	Nosy Be	f	45.0	45.2	90.3	1.00	-	-	-	-	+	12
ZFMK 48226	C. boettgeri	Nosy Be	f	47.6	47.3	94.9	1.01	2.5	0.052	14	5.7	+	15

……continued on the next page …….continued on the next page Continued.

TABLE 1. (Continued)
collection no. species	locality	sex	SVL	TaL	TL	RST	LRA	RRS	RAPSC	NPSCM	RC	NSL
ZFMK 45984 C. boettgeri	Nosy Be	f	55.5	52.5	108.0	1.06	1.2	0.022	10	8.1	+	12
ZSM 227/2002 C. boettgeri	Nosy Be	f	48.8	52.8	101.6	0.92	3.2	0.065	15	4.7	+	12
ZSM 441/2000 C. boettgeri	Nosy Be	f	45.5	43.4	88.9	1.05	2.7	0.060	16	5.9	+	12
SMF 16471 C. boettgeri	Nosy Be	f	50.0	46.7	96.7	1.07	2.8	0.056	14	5.0	+	13
SMF 16472 C. boettgeri	Nosy Be	f	46.0	45.7	91.7	1.01	3.0	0.065	14	4.7	+	12
ZSM 865/1920 C. boettgeri	-	f	49.4	46.7	96.1	1.06	3.5	0.070	20	5.8	+	15
ZFMK 51515 C. boettgeri	-	f	51.3	48.2	99.5	1.06	2.9	0.057	17	5.8	+	13
MCZ 5988 Ch. macrorhinus	‘Madagascar’	f	48.1	44.9	93.1	1.07	2.8	0.058	17	6.1	+	17

ZSM 236/2004	C. linotum	M. d'Ambre	m	55.4	63.0	118.4	0.88	4.3	0.077	16	3.7	+	13
ZSM 1683/2012	C. linotum	M. d'Ambre	m	53.0	56.3	109.3	0.94	4.7	0.089	cut	-	+	13
ZSM 2073/2007	C. linotum	M. d'Ambre	m	59.6	64.8	124.4	0.92	4.5	0.075	18	4.0	+	-
ZSM 2072/2007	C. linotum	M. d'Ambre	m	53.7	59.2	112.8	0.91	4.3	0.080	13	3.0	+	13
ZFMK 52308	C. linotum	Joffreville	m	36.2	37.9	74.1	0.96	2.5	0.068	15	6.1	+	-
ZSM 873/1920/3	C. linotum	-	m	54.6	58.1	112.6	0.94	4.4	0.081	14	3.2	+	12
ZSM 873/1920/1	C. linotum	-	m	52.4	cut	52.4	-	4.3	0.081	16	3.8	+	12
ZFMK 36630	C. linotum	-	m	55.6	61.9	117.5	-	4.7	0.085	15	3.2	+	12
ZSM 551/2001	C. linotum	Andampy	f	50.6	50.7	101.3	1.00	2.0	0.040	9	4.5	+	13
ZSM 622/1999	C. linotum	-	f	54.5	54.9	109.4	0.99	4.1	0.075	15	3.7	+	13
ZSM 873/1920/2	C. linotum	-	f	42.7	47.4	90.1	0.90	3.1	0.072	15	4.9	+	12
ZSM 21/1923	C. linotum	‘Madagascar’	m	56.1	70.0	126.1	0.80	4.3	0.077	15	3.5	+	14

collection no. species locality sex NIL OLD OLN PC DC AP DSC RSB NSC UAD RAS ……continued on the next page collection no. species locality sex NIL OLD OLN PC DC AP DSC RSB NSC UAD RAS SMF 16471 C. boettgeri Nosy Be f 10 4.2 0.0 - 0 - 0.4 0.007 9 2.2 0.044 SMF 16472 C. boettgeri Nosy Be f 13 3.5 0.1 - 11 - 0.4 0.009 10 1.8 0.039 ZSM 865/1920 C. boettgeri - f 13 2.8 0.1 - 0 - 0.3 0.006 9 1.9 0.038 ZFMK 51515 C. boettgeri - f 13 3.3 0.7 - 0 - 0.3 0.006 12 2.1 0.041 MCZ 5988 Ch . macrorhinus ‘Madagascar’ f 6.1 3.2 0.1 - 0 - 0.3 0.007 8 2.3 0.047 Individuals from Montagne d’Ambre show clear morphological differences from Nosy Be specimens ( Fig. 7 ). The adult males examined from Montagne d’Ambre are larger than those from Nosy Be (mean values of TL 107.8 mm in Montagne d’Ambre vs. 99.3 mm in Nosy Be, Table 2 ), their rostral appendage is longer related to the snoutvent length (RRS 0.078 vs. 0.065) and the ratio of arm diameter to snout-vent length is larger (UAD/SVL 0.053 vs. 0.042).

ZFMK 51389	C. boettgeri	Nosy Be	m	12	3.7	0.3	-	19 -	0.4	0.008	7	2.0	0.038
ZFMK 51520	C. boettgeri	Nosy Be	m	13	3.1	0.3	-	2 -	0.4	0.007	10	2.2	0.044
ZFMK 45987	C. boettgeri	Nosy Be	m	11	4.7	0.4	-	0 -	0.5	0.010	9	2.1	0.040
ZFMK 45988	C. boettgeri	Nosy Be	m	11	3.0	0.4	-	12 -	0.2	0.004	10	2.0	0.041
ZFMK 51518	C. boettgeri	Nosy Be	m	13	3.4	0.3	-	0 -	0.4	0.008	8	2.2	0.048
ZFMK 51519	C. boettgeri	Nosy Be	m	11	4.3	0.6	-	18 -	0.3	0.006	9	2.0	0.039
ZFMK 51521	C. boettgeri	Nosy Be	m	11	3.8	0.6	-	5 -	0.3	0.007	9	1.5	0.036
ZSM 36/1913	C. boettgeri	Nosy Be	m	12	3.2	0.0	-	0 -	0.4	0.008	12	1.8	0.043
ZSM 440/2000	C. boettgeri	Nosy Be	m	12	4.8	0.3	-	28 -	0.4	0.008	14	2.5	0.050
ZSM 444/2000	C. boettgeri	Nosy Be	m	12	3.7	0.5	-	15 -	0.4	0.008	11	2.3	0.045
SMF 16471	C. boettgeri	Nosy Be	m	13	4.0	0.0	-	11 -	0.4	0.009	9	2.0	0.040
ZFMK 51514	C. boettgeri	-	m	13	3.7	0.3	-	14 -	0.5	0.009	11	2.2	0.044
ZSM 866/1920	C. boettgeri	-	m	15	3.7	0.3	-	20 -	0.4	0.008	8	2.0	0.036
ZFMK 45985	C. boettgeri	Nosy Be	f	13	4.6	0.5	-	0 -	0.4	0.007	10	1.8	0.036
ZFMK 51516	C. boettgeri	Nosy Be	f	11	3.8	0.5	-	0 -	0.3	0.007	12	2.2	0.042
ZFMK 51517	C. boettgeri	Nosy Be	f	11	2.7	0.5	-	0 -	0.4	0.007	14	2.2	0.044
ZFMK 50615	C. boettgeri	Nosy Be	f	12	3.4	0.4	-	0 -	0.4	0.008	8	1.5	0.036
ZFMK 45983	C. boettgeri	Nosy Be	f	12	3.5	0.0	-	0 -	0.3	0.006	11	2.1	0.040
ZFMK 45986	C. boettgeri	Nosy Be	f	11	3.0	0.5	-	3 -	0.3	0.007	9	2.0	0.044
ZFMK 48226	C. boettgeri	Nosy Be	f	14	2.7	0.3	-	0 -	0.2	0.004	-	1.9	0.039
ZFMK 45984	C. boettgeri	Nosy Be	f	11	2.8	0.4	-	0 -	0.3	0.005	11	2.0	0.036
ZSM 227/2002	C. boettgeri	Nosy Be	f	12	3.3	0.1	-	0 -	0.3	0.006	11	2.0	0.041
ZSM 441/2000	C. boettgeri	Nosy Be	f	12	3.9	0.5	-	0 -	0.4	0.008	9	2.2	0.048

TABLE 1. (Continued)

ZSM 236/2004	C. linotum	M. d'Ambre	m	13	4.5	0.0	4 10 -	0.7	0.012	18	2.9	0.052
ZSM 1683/2012	C. linotum	M. d'Ambre	m	13	4.7	0.1	3 8 -	0.7	0.014	20	2.9	0.055
ZSM 2073/2007	C. linotum	M. d'Ambre	m	12	4.2	0.0	3 13 -	0.6	0.010	22	3.1	0.052
ZSM 2072/2007	C. linotum	M. d'Ambre	m	12	3.8	0.2	4 12 -	0.8	0.015	21	3.1	0.058
ZFMK 52308	C. linotum	Joffreville	m	-	2.8	0.0	3 9 -	0.4	0.011	20	1.7	0.048
ZSM 873/1920/3	C. linotum	-	m	14	4.1	0.2	6 5 -	0.7	0.013	19	3.0	0.054
ZSM 873/1920/1	C. linotum	-	m	12	4.8	0.1	5 4 -	0.8	0.015	19	3.0	0.058
ZFMK 36630	C. linotum	-	m	12	3.7	0.3	4 0 -	0.7	0.012	23	3.0	0.054
ZSM 551/2001	C. linotum	Andampy	f	13	5.1	0.0	(5) 6 -	0.6	0.011	16	2.8	0.055
ZSM 622/1999	C. linotum	-	f	13	4.1	0.2	(4) 0 -	0.6	0.011	22	2.9	0.054
ZSM 873/1920/2	C. linotum	-	f	12	3.9	0.0	6 0 -	0.5	0.013	17	2.8	0.065
ZSM 21/1923	C. linotum	‘Madagascar’	m	14	4.7	0.1	(5) 0 -	0.7	0.013	22	3.0	0.054

TABLE 2 . Mean values and standard deviations (SD) of morphological measurements of Calumma boettgeri , n (males) = 11, n (females) = 12 and C. linotum males (n =5); all measurements in mm. For abbreviations see Table 1. species C. boettgeri C. boettgeri C. linotum locality Nosy Be Nosy Be M. d'Ambre sex m f m

mean value SD	mean value	SD	mean value	SD
SVL	48.5 4.2	48.6	3.8	51.6	9.0
TaL	50.8 4.2	47.7	3.7	56.2	10.8
TL	99.3 8.1	96.3	7.2	107.8	19.7
RST	0.96 0.04	1.02	0.04	0.92	0.03
LRA	3.1 0.5	2.9	0.9	4.1	0.9
RRS	0.065 0.014	0.059	0.018	0.078	0.008
RAPSC	18.9 4.7	16.4	3.4	15.5	2.1
NPSCM	6.1 1.0	6.3	1.5	4.2	1.3
NSL	12 0.5	12	1	13	0
NIL	11.8 0.8	11.9	0.9	12.5	0.6
OLD	3.8 0.6	3.7	0.4	4.0	0.7
DSC	0.36 0.08	0.32	0.05	0.64	0.15
RSB	0.0075 0.0015	0.0066	0.0012	0.0124	0.0019
NSC	9.9 2.0	10.6	1.8	20.2	1.5
UAD	2.1 0.3	2.0	0.2	2.7	0.6
RAS	0.042 0.005	0.041	0.004	0.053	0.004

Both populations show heterogeneous scalation, especially at the extremities and the rostral appendage. The enlarged rounded tubercles on the limbs are distinctly larger in animals from Montagne d’Ambre; the mean diameter of the largest tubercle of the upper arm is 0.64 mm compared to 0.36 mm ( Nosy Be). Additionally the number of the enlarged tubercles on the upper arm of males from Montagne d’Ambre is approximately twice that of males from Nosy Be, with a mean of 20.2 compared to 9.9, respectively, and the tubercles are not bordering each other on Nosy Be individuals. This character is also confirmed from a juvenile of SVL 26.9 mm (ZFMK 48227, Nosy Be) and a juvenile of SVL 36.2 mm (ZFMK 52308, Joffreville). Another morphological difference between both populations can be found in the pholidosis of the rostral appendage. Although the rostral appendage is significantly smaller in males from Nosy Be (see above), they have more peripheral scales on it, with a mean of 18.9 compared to 15.5 in Montagne d’Ambre males. In relation to the size of the appendage, this means 6.1 scales per mm compared with 4.1 scales per mm in Montagne d’Ambre. In summary, specimens from Montagne d’Ambre show a more heterogeneous scalation with broader tubercles on extremities and the rostral appendage. This is true of females as well (note that only one female with locality data was examined). Montagne d’Ambre individuals differ also in both sexes from Nosy Be specimens in the presence of a parietal crest which is best visible in the micro-CT scan (see below). The other morphological features either were highly variable or did not differ between the populations. For example, the number of dorsal cones was 0–28 in Nosy Be, and 9–13 in Montagne d’Ambre. Likewise the edges of the occipital lobes vary between the specimens. They range between un-notched and clearly notched (up to 0.7 mm ) in Nosy Be, and are either not or only slightly notched (up to 0.3 mm ) in Montagne d’Ambre. All specimens have a rostral crest and none have axillary pits. Colouration. Males also show great differences between the populations in colouration (see Fig. 2 A, B). Males from Montagne d’Ambre were more colourful, with a true blue rostral appendage and greenish turquoise extremities ( Fig. 2 A). The colour of the legs is induced only by the coloured tubercle scales. The body is pale green or light brown with two dark brown spots and (occasionally) a beige lateral stripe on each side that stops at the base of the tail. The tail is the same colour as the body and (in stress colouration) possesses black annulations. The head is also greenish or brown with a dark stripe from the snout crossing the eyes to the occipital lobes. The skin around the mouth and the throat can be white. The colour description is based on a total of seven pictures of the Montagne d’Ambre form, referred to as C. boettgeri in Schmidt et al. (2010) , C. boettgeri (picture 1c) in Glaw & Vences (2007) Calumma sp. in Nečas (2004) and C. boettgeri in Garbutt et al. (2001) . The body and head of females is brown; the rostral appendage can be coloured bright blue, see picture of C. boettgeri in Schmidt et al. (2010) . FIGURE 2 . (A) Baseline colouration in life of male Calumma linotum from Montagne d’Ambre during day; (B) Baseline colouration in life of male C. boettgeri from Nosy Be during day; (C) female C. linotum from Montagne d’Ambre at night; (D) female C. boettgeri from Nosy Be at night. Photos taken by FG. The body colouration of males from Nosy Be in contrast is yellowish or greenish brown with little dark brown rosettes, when stressed ( Fig. 2 B). The legs are brown with little blue or green spots resulting from the tubercles. The colouration of the head is similar to the body colouration. The rostral appendage differs clearly from Montagne d’Ambre with the absence of any striking colour and is the same brown colour as the casque. Females are uniformly light or greenish brown coloured. Compare also a total of seven pictures of Calumma boettgeri in Hyde Roberts & Daly (2014) , in Glaw & Vences (2007: 191) picture 1a and 1b, in Nečas (2004) and in Henkel & Schmidt (1995) . Osteology of the skull based on micro-CT scans. Micro-CT scans of heads of two males and two females from Nosy Be and from Montagne d’Ambre exposed additional differences between the two forms. Specimens from Montagne d’Ambre ( Fig. 4 , D and E) bear tubercles on the parietal in both sexes. These form a little parietal crest in the middle with three to four tubercles, laterally followed by two tubercles on each side. The frontal is also irregularly spotted with tubercles. The parietal and frontal of animals from Nosy Be in contrast are smooth ( Fig. 4 , A and B). As in all species of the genus Calumma , the nasal bones are paired ( Rieppel & Crumly 1997 ). These are broader in our specimens from Montagne d’Ambre (mean NW 0.35 mm vs. 0.24 mm in Nosy Be; mean NW/NL 0.18 vs. 0.14, Table 3 , Fig. 4 ) and the anterior tip of the frontal bone does not exceed more than a half of the naris. In skulls from Nosy Be it does exceed this point, and the frontal meets the premaxilla, as described for C. nasutum ( Rieppel & Crumly 1997 ) . The parietal also varies between the two localities. In Nosy Be samples, the parietal tapers more tightly. Its diameter is at the tightest area on average 0.61 mm (vs. 1.06 mm ) and 11% of the largest diameter of the parietal (vs. 22%, Table 3 , Fig. 4 ). The parietal in Montagne d’Ambre samples appears wider and more compact. However, the form of the parietal is variable within localities and cannot be used as a diagnostic character. Although chameleons are sexually dimorphic animals, differences between sexes in skull structure were not proven ( Table 3 ). TABLE 3 . Osteological measurements of important characters of the skull for differentiation between Calumma boettgeri und C. linotum . Notes: m, male; f, female; NL, nasal length; NW, nasal width; RNWL, ratio of nasal width to length; PL, largest diameter; PS, parietal smallest diameter; RPSL, ratio of parietal smallest to largest diameter; PC, parietal crest absent (-) or number of tubercles.

collection no.	species	locality	sex	NL	NW	RNWL	PL	PS	RPSL	PC
ZSM 440/2000	C. boettgeri	Nosy Be	m	2.1	0.3	0.14	4.7	0.5	0.11	-
ZSM 441/2000	C. boettgeri	Nosy Be	f	2.2	0.2	0.09	3.9	0.5	0.13	-
ZSM 444/2000	C. boettgeri	Nosy Be	m	1.8	0.3	0.17	4.7	0.3	0.07	-
ZSM 227/2002	C. boettgeri	Nosy Be	f	1.3	0.2	0.15	4.1	0.6	0.15	-
mean value	1.85	0.25	0.14	4.35	0.49	0.11
SD	0.40	0.06	0.03	0.41	0.11	0.03
MCZ 5988	Ch. macrorhinus	‘Madagascar’	f	2.0	0.2	0.10	3.9	1.1	0.28	1
ZSM 2072/2007	C. linotum	M. d'Ambre	m	2.5	0.4	0.16	4.5	1.5	0.33	4
ZSM 2073/2007	C. linotum	M. d'Ambre	m	2.1	0.4	0.19	4.6	0.6	0.13	3
ZSM 873/1920/2	C. linotum	M. d'Ambre	f	2.3	0.3	0.15	4.2	0.9	0.21	4
ZSM 551/2001	C. linotum	Andampy	f	1.4	0.3	0.23	4.3	0.9	0.21	3
mean value	2.07	0.36	0.18	4.40	0.97	0.22	3.50
SD	0.49	0.04	0.04	0.18	0.38	0.08	0.58
ZSM 21/1923	C. linotum	‘Madagascar’	m	2.7	0.3	0.11	4.6	1.4	0.30	4

Hemipenial morphology based on micro-CT scans. The scans of hemipenes of specimens from each population enable a detailed view of their structure. The hemipenes are illustrated in sulcal and asulcal view with the apex on top ( Fig. 5 ). Both populations show large and deep calyces with smooth ridges on the asulcal side of the truncus. The apex is ornamented with two pairs of long pointed papillae and two pairs of rotulae. The papillae rise from the sulcal side of the apex and are curved to the asulcal side. They can be completely everted ( Fig. 5 , C) or retracted in the apex ( Fig. 5 , D). One pair of rotulae is placed on the asulcal side (the smaller one) and one pair on the sulcal side. Here some differences between the populations are recognizable; in Nosy Be (n = 2) the rotulae are slightly more denticulated, with 6–11 tips on asulcal side and 14–16 tips on sulcal side, compared to Montagne d’Ambre (n = 3) with 6–8 tips on the rotulae of the asulcal side and 11–14 tips on both rotulae on the sulcal side.