BIHAREAN BIOLOGIST 9 (2): 117-120 ©Biharean Biologist, Oradea, Romania, 2015 Article No.: 151305 http://biozoojournals.ro/bihbiol/index.html

Morphology and Oral disc structure of the tadpole of alticola (Boulenger, 1882) from Cachar district (Assam, India)

Dulumoni TAMULY* and Mithra DEY

Department of Ecology and Environmental Science, Assam University, Silchar-788011, Assam, India. *Corresponding author, D. Tamuly, E-mail: [email protected]

Received: 12. November 2014 / Accepted: 13. February 2015 / Available online: 15. November 2015 / Printed: December 2015

Abstract. External morphology and the oral disc of tadpoles from Rosekandy Tea Estate of Cachar district, Assam are being described in the present paper. The mouthpart structures were examined using scanning electron microscope (SEM) at Gosner stage 26. Oral disc is composed of anterior (upper) labium and posterior (lower) labium. The upper jaw sheath is straight while the lower jaw sheath is V-shaped. Tooth rows are uniserial. Denticle formula is 5(3-5)/ 7(1) for stage 26 and 7(3-7)/7(1) for stage 38.The number of tooth rows increases during ontogeny.

Key words: SEM, oral disc, keratodont formula, tadpole, Clinotarsus alticola, Cachar district.

Introduction ferent measurements includes BL, TL, BD, BW, T, TH, BTMH, IO, IN, SO and SN. Abbreviations and definitions are in accordance with Clinotarsus alticola (Boulenger, 1882) (earlier alticola Altig & McDiarmid (1999). Tadpole descriptions and oral structure Boulenger, 1882) has been reported from Meghalaya under of the tadpoles were studied using light microscope based on Gosner stage 38. The oral structures of the selected tadpoles (stage 26) were the name Hylorana Pipiens (Jerdon, 1870) and then again by studied using Scanning Electron Microscopy. Description of oral Sahu & Khare (1980) and Sailo (2011). The morphology and disc and labial tooth row formula (LTRF) is in accordance with Altig buccopharyngeal anatomy of tadpoles of Rana (Nasirana) al- (1970). For Scanning Electron Microscopy the specimen were pre- ticola has been described from Thailand (Grosjean et al. pared as follows-The mouthparts of the tadpoles were fixed in 2.5 % 2003). This species is distributed in different latitude, longi- glutaraldehyde solution (prepared in 0.1 M Sodium Cacodylate tude and elevation of Assam, India up to Southern Thailand. buffer) for four hours. The pH of fixative and buffer was maintained The tadpoles of C. alticola in the present study were collected at 7.4. The samples were then dehydrated through ascending acetone grades and drying was done in Tetra methyl silane (Dey et al. 1989). from Rosekandy Tea Estate located in Cachar district of Ba- A thin conductive coating of gold was applied to the samples using a rak Valley, Assam. JFC 1100 (Jeol) ion sputter and the coated samples were examined Tadpole morphology is an important feature that may with the aid of a JSM -6360 (Jeol) scanning electron microscope at an play role in determining its position in a pond and the kind accelerating voltage of 20KV. of food they consume. Total length of the tadpole, tail fin, Abbreviations: BL-Body length, TL-Total length, BD-Body position of mouth, labial teeth rows, jaw sheath, are impor- depth, BW-Body width, T-Tail length, TH-Tail height, BTMH- Basal tant features for species identification. Tail fin shape is re- tail musculature height, IO- Inter-orbital distance, IN-Inter-narial lated to the water column, whether they occupy deep or distance, SO-Snout-orbit distance, SN-Snout-naris distance. shallow water pools, slow or fast flowing water body etc.

The structure of mouthparts provides a very useful charac- Results teristic feature in the discussion of taxonomical problems and identification of the tadpoles (Altig 1970, Altig & Pace Tadpole description 1974, Lee 1976). The mouthpart of the tadpole differs from External morphology (stage 38): The following description is species to species and it also depends on the habitat of the based on Gosner stage 38 tadpole of C. alticola. From dorsal species. The keratodont row also show wide variation in view, body oval or elliptic, posterior part of the body is shape, size and number among different species. Keratodont wider than the anterior part; snout broadly rounded. Inter- rows are absent in some species. The shape of beak, papilla- orbital distance (7.71mm) larger than snout-orbit (5.3mm) tion of labial fringes, pattern of dentition and development distance. In lateral view, body is depressed; snout rounded. of tooth rows vary among different anuran species (Smith Eyes positioned dorsolaterally. Naris oval in size, placed 1924, Daniel 1975). The oral papillae are believed to serve as nearer to the snout. In ventral view, intestine is not visible. chemosensory and tactile receptors (McDairmid & Altig Morphometric measurements of different developmental 1999). Position of oral disc of the tadpole reflects its feeding stages are presented in Table 1. Spiracle sinistral, aperture ecology, food preferences and feeding methodology em- oriented posterodorsally, extended as a short tube, fused to ployed for food acquisitions (Altig & Johnston 1989, Khan & body wall. Vent tube is dextral. Tail length 40.69 mm with Malik 1987). moderately high tail fin, dorsal fin is higher than the ventral fin. Tail musculature greatly narrowing to the tail tip, tail tip

rounded. V-shaped myomeres apparent. A pair of parotoid Materials and methods gland present behind the eyes, which is large and broadly The tadpoles of Clinotarsus alticola were collected from permanent oval in size. Supracaudal (an elongated strip at starting of water body inside the Rosekandy Tea Estate Cachar District dorsal fin) and infracaudal gland (beginning after origin of (24°42′29″–24º41′31″ N, 92°41′52″–92º42′39″ E). The larvae were fixed ventral fin) is also present in separate regions. in 10% formalin and staged following Gosner (1960). Morphometric Coloration (stage 38): The tadpoles are light olive to dark measurements of the tadpoles were taken using vernier calliper. Dif- brown black in colour. Anterior half of the dorsal side is 118 D. Tamuly & M. Dey

Table 1. Morphometric measurements (in mm) of C. alticola tadpoles in different developmental stages. N=10 (X±SD; range in parenthe- sis), [BL-Body length, TL-Total length, BD-Body depth, BW-Body width, T-Tail length, TH-Tail height, BTMH- Basal tail musculature height, IO- Inter-orbital distance, IN-Inter-narial distance, SO- Snout-orbit distance, SN Snout-naris distance]

Parameters 26 30 34 38 BL 8.1±0.45 17.95±0.42 25.49±1.82 28.63±0.61 (7-8.6) (17-18.5) (22-27.1) (27-29) TL 25.1±0.99 39.65±0.69 62.15±2.98 69.22±2.01 (23-26.2) (38.5-40.5) (57-65) (65-71.2) BD 4.42±0.33 5.91±0.65 8.85±0.25 9.43±0.66 (4-4.8) (5-6.9) (8.5-9.2) (8-10.1) BW 5.25±0.28 9.39±0.49 13.85±0.25 14.64±0.38 (4.9-5.6) (8.5-10) (13.5-14.1) (14-15.1)

T 17.03±0.57 21.81±0.68 30.47±0.84 40.69±1.56 (16-17.8) (21.1-22.8) (29-31.5) (38-42.2) Figure 1. Oral structure of stage 38 of C. alticola LTRF 7(3-7)/7(1) TH 8.54±0.65 11.35±0.21 12.47±0.80 14.07±0.53

(7.8-9) (11-11.5) (11.5-13.5) (13-14.8) BTMH 5.83±0.62 6.9±0.33 8.03±0.09 7.21±0.52 (5-6.5) (6.5-7.4) (7.9-8.2) (6.5-7.9) IO 3.33±0.19 4.81±0.35 6.99±0.33 7.71±0.44 (3-3.6) (4.2-5.2) (6.5-7.5) (7-8.2) IN 2.05±0.23 3.34±0.20 3.89±0.19 3.83±0.25 (1.8-2.4) (3-3.6) (3.5-4) (3.5-4.2) SO 2.37±0.17 3.15±0.24 5.11±0.11 5.3±0.16 (2.1-2.6) (2.7-3.5) (5-5.3) (5-5.5) SN 1.71±0.25 2.17±0.20 2.6±0.20 3.33±0.15 (1.3-1.9) (1.9-2.4) (2.4-2.9) (3-3.5)

lighter than the posterior half. Caudal muscle is lighter with black pigments and marked with two to three ocelli placed in a row at the tail base with orange halo. Oral structure: The oral structure of the tadpole of C. al- Figure 2. Scanning electron micrograph of oral structure of stage 26 st th ticola was studied with the help of light microscope (Olym- of C. alticola tadpole. A1, A2, A3, A4, A5 =1 -5 anterior tooth rows; P1, P2, P3, P4, P5, P6, P7 = 1st – 7th posterior tooth rows pus CX 41) and Scanning electron microscope. At stage 38, the oral structure of the tadpole is characterized with two uninterrupted teeth row and five interrupted teeth row on the upper labium. The lower labium consists of one inter- rupted teeth row followed by six uninterrupted teeth row. The margin of the mouth parts consists of marginal papillae border with a wide dorsal gap. Marginal teeth (accessory la- bial tooth row) are observed at this stage. The rows are short and present at junction of anterior and posterior labia. The LTRF is 7(3-7)/7(1) (Fig. 1). Scanning electron microscopic studies of C. alticola (stage 26) tadpoles showed that, the oral disc is composed of ante- rior (upper) labium and posterior (lower) labium. The upper lip have two continuous and three discontinuous rows and the lower lip have one discontinuous and six continuous la- bial rows and LTRF formula is 5(3-5)/ 7(1) (Fig. 2). The ante- Figure 3. Scanning electron micrograph of mouth (M) of C. alticola tadpole with upper jaw sheath (UJS) and lower jaw sheath (LJS) rior labial rows are designated as A1, A2, A3, A4, and A5 and posterior labial rows are designated as P1, P2, P3, P4, P5, P6 and P7. The margin of the mouth parts consist of a papillary Discussion (marginal papillae MP) border with a wide dorsal gap. The submarginal papillae (SM) are present in lateral parts of the Total length of the tadpoles in the present study in stage 38 upper and lower labia. Both the papillae (MP and SM) have was 69.22 mm (mean) which fits to that reported by Annan- a rounded tip. The suprarostrodont is straight while the in- dale (1912) i.e. 57 mm but the stage was not mentioned. Or- frarostrodont is V-shaped and both appeared to be serrated ton (1953) classified tadpoles on the basis of their and pointed. The mouth is present in between upper and mode of feeding. Ranoid tadpoles of genus Rana are type IV lower jaw sheath. The arrangement pattern of the tooth rows with well developed oral disc and its associated hard parts. are uniserial and tooth is keratinized and each keratinized External morphology is important for identification of some labial tooth consists of three regions; cusp, neck and base. species. The size of the tadpole may determine the moving Morphology and Oral disc structure of the tadpole of C. alticola 119

Figure 4. Scanning electron micrograph showing labial teeth

of C. alticola tadpole with B= Base, N= Neck, C= Cusp. Figure 7. Scanning electron micrograph showing marginal papillae (MP) and submarginal papillae (SM) of C. alticola tadpole.

This species can be identified by its prominent external mor- phological characters. Oral structure helps in identification of species as well as understanding the feeding ecology. Denticle formula is one of the strong characters for identifi- cation; though some of the tadpoles have more than one denticle formula (Inthara et al. 2005). The present study es- tablished the oral structure of C. alticola tadpole at stage 26 and 38 which is more or less similar to the findings of the earlier workers. The number of keratodont rows changes during ontogeny. Smith (1916) reported the denticle formula of C. alticola is 9(2-9)/ 8, where Sahu & Khare (1980) reported the LRTF is 2:5+5/1+1:6 from the north-eastern tadpoles.

Figure 5. Scanning electron micrograph of the Grosjean et al. (2003) described LRTF of Rana (Nasirana) alti- suprarostrodont (Sr) of C. alticola tadpole cola at stage 38 was 2:7+7/ 1+1:2. Inthara et al. (2005) also re- ported the LRTF of Rana alticola to be 2:5+5/1+1:6 from Thai-

land. Sailo (2011) reported in Rana alticola the highest num- ber of teeth rows was 2:5+5/1+1:7. The present study reports keratodont formula of C. alticola as 5(3-5)/7(1) (stage 26) and 7(3-7)/7(1) (stage 38).which shows that the number of tooth rows increases during ontogeny. James et al. (2000) found in Rana curtipes, where the number of teeth rows changed with growth. Ontogenic variation is found to be common in tad- poles with high number of keratodont rows. The differences in keratodont formula observed may be a geographical variation. Ontogenic variation is also common in species having large number of keratodont rows and has been also observed in tadpoles of Hoplobatrachus and Leptobrachium echinatum (Grosjean et al. 2003). In the present study tadpoles were collected from two

Figure 6. Scanning electron micrograph of different pools in undisturbed secondary forest area of the infrarostrodont (Ir) of C. alticola tadpole Rosekandy tea estate, however these pools were not con- nected to any lotic systems as reported by others (Grosjean

et al. 2003). They were conspicuous on the water surface in capacity of the tadpole. Labial tooth row number and jaw the pools and found in aggregation (Tamuly & Dey 2013). sheath also indicate the feeding pattern of the tadpole. The main characteristics of C. alticola are its big size and two to three ocelli place in a row at the tail base with orange halo, which is unique among ranid tadpoles. The caudal ocelli and Acknowledgments. The authors are grateful to SAIF, NEHU, glands could be interpreted as means of defence (Grosjean et Shillong, for the use of Scanning electron microscope (SEM) and to al. 2003). Glands secreting noxious substances are known to the University Grants Commission, India, for the award of a UGC- be efficient against predators (Liem 1961). The ocellus is AUS research fellowship. The authors are also grateful to the thought to mimic the eye of the larger (Altig & Department of Ecology and Environmental Science, Assam University, Silchar where the work was carried out. Channing 1993) and misdirect predator attacks. No other ranid species have that similar feature (Grosjean et al. 2003). 120 D. Tamuly & M. Dey

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