Description of ten tadpoles in the genus Boophis from Madagascar Author Glos, Julian Author Vences, Miguel text Zootaxa 2010 2694 1 25 journal article 10.5281/zenodo.199617 ddecc0b0-c3f3-46d5-b042-6bdc334ea2f7 1175-5326 199617 Boophis luciae Glaw, Köhler , de la Riva, Vieites & Vences The following description refers to one tadpole in developmental stage 25 (ZSM 0634/2008, field number ZCMV 3686, TL 20.8 mm , BL 7.8 mm ; Table 1 ), from Ranomafana National Park. The 16S rDNA sequence of this specimen was 97.7% identical to a reference sequence of an adult Boophis luciae . (accession AJ315913 ) from Vohidrazana. FIGURE 10. Drawings of preserved tadpole specimen of Boophis luciae (ZSM 0634/2008) from Ranomafana National Park (developmental stage 25). (a) dorsal view; (b) lateral view; (c) oral disk. In dorsal view, body ovoid, maximal width between the proximal 1/2 and 3/4 of the body (SBW 60% of BL), wide rounded snout. In lateral view, body depressed (BW 115% of BH), maximal body height between the proximal 1/2 and 3/4 of the body (SBH 68 % of BL), rounded snout. Eyes large (ED 14% of BL), not visible from ventral view, positioned dorsally and oriented laterally, situated between the proximal 3/10 and 4/10 of the body, wide distance between eyes (IOD 75% of BW). Large elliptical nares (NS 5% of BL), marked with marginal rim, positioned dorsally and oriented anterodorsally, situated nearer to eye than to snout (RN 128 % of NP), moderately wide distance between nares ( IND 45% of IOD), black spot on the back of the nare present, ornamentation absent. Moderately long sinistral spiracle (SP 11% of BL), directed posterodorsally, visible from dorsal and ventral views and perceptible from lateral view, inner wall free from body and aperture opens laterally instead of posteriorly, round opening, situated between the proximal 1/2 and 3/4 of the body (SS 66% of BL), located below the level of the point where the axis of the tail myotomes contact the body. Long medial vent tube (VL 10% of BL), not attached to the ventral fin. Tail of moderate length (TAL 162% of BL), maximal tail height lower than body height (MTH 84% of BH), tail height at midtail lower than body height and maximal tail height (THM 67% of BH and 79% of MTH), tail height at the beginning of the tail lower than body height (TH 82% of BH). Moderately developed caudal musculature (TMW 59% of BW, TMH 70% of BH and 83% of MTH, TMHM 100% of THM and 79% of MTH). Tail muscle reaches tail tip. Very low fins. Dorsal and ventral fins originate on the midtail, progresses almost parallel to the margin of the tail muscle up to the tail tip. Lateral tail vein visible on the proximal 1/2 of the tail, myosepta visible all along the tail, tail tip narrowly rounded. Very wide oral disk (ODW 80% of BW), positioned and oriented ventrally, not emarginated, maximal width in the middle. Oral disk not visible from dorsal view, upper labium is a continuation of snout. Single row of marginal papillae interrupted by moderately wide dorsal gap (DG 54% of ODW), total number of marginal papillae 129. Two hundred and thirty submarginal papillae positioned laterally and ventrally. Small conical papillae with protuberance, rounded tip, not visible from dorsal view, longest marginal and submarginal papillae measured 0.08 mm and 0.09 mm , respectively. Jaw sheaths fully keratinized with rounded serrations, narrow (JL 31% of ODW), medial convexity absent. Lower jaw sheath U-shaped, and ribbed (giving the appearance of vertical bars). LTRF 7(5–7)/3(1). Single row of keratodonts per ridge. Wide A1 (60% of ODW). Density of keratodonts on A1 119/mm (total 208). Short, discernible keratodont ( 0.14 mm ). The keratodonts in the rows A1, A2, and A3 are small and of equal size. For the others, the size decreases towards the margins. Coloration in preservative. Dorsally, body and tail brownish, not uniform. The region between the eyes and parts of the intestine are very dark. Dorsolaterally same color as dorsally and ventrolaterally, the part below the eye is almost whitish and the snout is transparent with a brown spot, intestine is dark. Caudal musculature is whitish with some dark round spots of different sizes. Pigmented stripes along the tail, tail fins are whitish. Ventrally, intestinal coil is not visible. Variation. Based on six tadpoles at stages 25–37, all from Ranomafana National Park (ZCMV 3619, 3631) of which two are DNA voucher specimens ( Table 6 ). All the specimens are deposited in the UADBA collection. TL and BL are 23–29 mm and 7–13 mm , respectively. BW 94–107% of BH; ED 9–13% of BL; RN 46–83% of NP; IND 45–61% of IOD; SS 54–63% of BL; TMH 95–100% of MTH; TMW 50–71% of BW; MTH 61–77% of BH; ODW 42–79% of BW; LTRF 7(5–7)/3. TABLE 6. Morphometric measurements (all in mm) of additional DNA voucher specimens of Boophis vittatus and B. luciae . For abbreviations, see Material and Methods.

Species Field number	B. vittatus FGZC 2237	B. vittatus FGZC 2251	B. vittatus FGZC 2276	B. vittatus FGZC 2276	B. luciae ZCMV 3619	B. luciae ZCMV 3631
BL BW	6.8 3.3	5.7 2.5	6.3 2.5	5.0 2.5	6.7 4.7	12.3 6.3
BH	2.7	2.2	2.0	2.0	4.7	6.5
ED RN	1.0 1.8	0.7 1.0	0.7 1.2	0.7 1.3	1.3 1.8	1.7 1.0
NP	1.7	1.5	1.0	1.2	2.3	2.2
IND IOD	1.3 3.0	1.3 2.0	1.0 2.0	1.0 2.0	2.0 4.0	2.7 4.3
SS	4.8	4.0	---	---	3.7	7.0
TMH TMW	2.0 2.2	1.3 1.3	1.5 1.3	1.3 1.3	3.3 3.3	4.0 3.3
MTH	2.0	---	1.5	1.5	3.5	4.0
BH TAL	2.7 11.8	2.2 ---	2.0 9.7	2.0 8.3	4.7 18.0	6.5 15.3
ODW	2.3	2.2	2.3	2.2	3.7	2.7
Discussion

Data presented herein confirm that, in general, tadpoles of species classified in the same species group of Boophis have relatively similar morphologies. This is true for tadpoles described herein belonging to the B. luteus group ( B. andreonei , B. anjanaharibeensis , B. elenae which all agree with other tadpoles in the same group), and in the B. albipunctatus group ( B. luciae which largely agrees with B. sibilans ). However, in the B. microtympanum group, the tadpoles of B. rhodoscelis are quite different from the more specialized tadpoles of B. microtympanum . FIGURE 11. Photographs of preserved DNA voucher specimen tadpoles (as in Figs. 1–10) in lateral and dorsal views. Not to scale. Two species for which larval morphological data are provided herein have been studied before in this respect: Boophis microtympanum by Blommers-Schlösser (1979b) and B. andreonei by Randrianiaina et al. (2009) . For B. microtympanum , the previously published descriptions in general agree with our data but were not very detailed, so that the possibility of comparison is limited. For B. andreonei , the previous description refers to a tadpole in developmental stage 25 from the site Maromalo in Manongarivo Special Reserve (close to the type locality of the species, Benavony). In general, the tadpoles from Marojejy agree with those from Manongarivo (e.g., in LTRF, ventral position and non-emarginated shape of oral disk). Differences are found in the lower numbers of marginal and submarginal papillae in Manongarivo (65 vs. 99, and 14 vs. 24), and lower numbers of keratodonts in A1 (108 vs. 143). This might be explained by the lower development stage of the Manongarivo specimen (25 vs. 37). The tadpole of Boophis luciae can be compared with the previously described tadpole of B. sibilans (see Raharivololoniaina et al. 2006 ). Both descriptions refer to specimens in stage 25. These two tadpoles closely agree in most morphological characters such as LTRF, position and form of oral disk, body proportions, and number of keratodonts in A1. Differences are observed in the number of oral papillae (130 vs. 303) and keratodont density on A1 (70/mm vs. 119/mm). These differences might partly be caused by different analysis methods (a digital measuring device and high-quality stereo microscope used only in the description of B luciae herein). However, it is unlikely that the difference in the number of papillae is only a counting error, and might indicate a stronger adaptation to strong currents in B . luciae , as the presence of high numbers of papillae is most common in tadpoles that attach to rocky surfaces in fast-moving streams ( Altig & McDiarmid 1999 ). TABLE 7. Comparison of morphological and ecological key characters of the tadpoles of ten species of Boophis as described herein. Ranges of ratios in this table are based on all DNA voucher specimens but not on additional specimens in the same series. LTRF, numbers of papillae and keratodonts are based only on the specimens used for description (see Table 1 ), LTRF variation on all other DNA voucher specimens.

Species	B. andreonei	B. anjanaharibeensis B. axelmeyeri	B. elenae	B. englaenderi
Species group	B. luteus	B. luteus B. goudoti	B. luteus	B. luteus
Breeding habitat	Stream	Stream Stream	Stream	Stream
BL (mm)	4.30–15.8	8.9 10.6–14	11.7–15.5	6.8–12
TAL/BL (%)	154–261	142 149–174	125–161	144–161
BW/BH (%)	109–167	124 115–130	75–149	112–138
TMH/BH (%)	56–79	67 65–82	63–75	69–81
TMH/MTH (%)	66–142	81 67–124	107–133	106–118
TMW/BW (%)	29–58	65 51–52	36–75	50–61
MTH/BH (%)	55–113	82 97–112	63–75	69–88
ODW/BW (%)	40–55	55 37–40	36–67	52–73
OD orientation	Ventral	Ventral Ventral	Ventral	Ventral
LTRF	6(5–6)/3(1)	5(4–5)/3(1) 4(3–4)/3(1)	3(2–3)/3(1)	6(4–6)/(3)
LTRF variations	3(2–3)/3(1) 4(3–4)/3(1) 5(4–5)/3(1)	5(4–5)/3(1) 4(3–4)/3(1) 5(4–5)/3(1)	2(1–2)/3(1) 3(2–3)/3(1)	6(5–6)/(3) 6(4–6)/(3) 7(5–7)/(3)

Number of keratodonts 143 in A1	108	132	78	236
Marginal papilae rows Single	Single	Single	Single	Double
Marginal papillae 99	85	68	56	128
Submarginal papilae 24	19	16	18	33

continued.

Species B. microtympanum	B. rhodoscelis	B. vittatus	B. roseipalmatus	B. luciae
Species group B. microtympanum	B. microtympanum	B. majori	B. goudoti	B. albipunctatus
Breeding habitat Brook	Brook	Brook	Brook	Brook
BL (mm) 8.5–23.8	15.3	4.9–6.8	10.5–13	6.7–12.3
TAL/BL (%) 128–204	141	130–173	153–219	124–270
BW/BH (%) 106–143	127	115–129	125–130	98–115
TMH/BH (%) 62–80	70	62–79	55–69	62–71
TMH/MTH (%) 67–129	59	92–110	69–120	83–120
TMW/BW (%) 35–64	51	53–65	37–50	53–71
MTH/BH (%) 64–127	118	75– 86	55–100	62–84
ODW/BW (%) 33–53	48	68–87	37–48	42–80
OD orientation Ventral	Ventral	Ventral	Ventral	Ventral
LTRF 6(5–6)/3(1)	5(4–5)/3(1)	7(4–7)/(3)	6(5–6)/3(1)	7(3–7)/(3)
LTRF variations 4(3–4)/3(1) 5(4–5)/3(1) 6(5–6)/3(1)	5(4–5)/3(1)	7(5–7)/(3) 6(3–6)/(3) 7(3–7)/(3)	4(3–4)/3(1) 5(4–5)/3(1) 6(5–6)/3(1)	7(3–7)/(3)
Number of keratodonts 287 in A1 Marginal papilae rows Single	160 Single	85 Single	188 Single	208 Single
Marginal papillae 70	105	113	72	230
Submarginal papilae 38	39	184	9	129

Tadpoles of Boophis roseipalmatus from Marojejy (described in this study) and its sister species, B . madagascariensis from Andasibe (tadpoles described by Raharivololoniaina et al. 2006 ), are similar in some characters: emargination and absence of ventral gap in oral disk, identical keratodont formulae, and similar numbers of keratodonts on A1 (150 vs. 164). However, they appear to differ by some essential characters, such as a possibly slightly stronger caudal musculature (and reduced fins) (TMH/MTH ratio 69–120 vs. 100), ventral vs. antero-ventral orientation of the oral disk, and slightly lower numbers of oral papillae (81 vs. 95) in B. roseipalmatus . While one of the major differences (caudal fin) may be caused by a different state of preservation of the examined specimens, others could represent real differences between these species, although their general similarity in the tadpole stage is obvious. Tadpoles of Boophis elenae from Ranomafana National Park (described in this study) and of B . sandrae from Andasibe (described by Raharivololoniaina et al. 2006 as candidate species B. sp. aff. elenae ) have some common morphological features, such as a ventral orientation and lack of emargination of the oral disk and general body proportions. However, there seems to be a difference in the keratodont formula, with B. elenae having two (vs. three) interrupted anterior keratodont rows and a lower number of oral papillae (74 vs. 164) and keratodonts on A1 (78 vs. 194). Although these differences may partly be influenced by different developmental stages (25 vs. 31) it is likely that they do represent real differences. In fact, despite their morphological similarity as adults (see Glaw & Vences 2007 ), B. elenae and B. sandrae are actually no sister species ( Vieites et al. 2009 ), and B. sandrae is more closely related to B. anjanaharibeensis which by external morphology (coloration) can be distinguished rather easily in the adult stage ( Glaw & Vences 2007 ). However, in all of these comparisons, it needs to be taken into account that the detailed data on numbers of papillae and keratodonts have only been assessed for a single individual per species or population. Since these variables seem to be valuable to distinguish tadpoles, additional morphometric studies are needed to assess their variability and thereby their taxonomic value.