Iranian Journal of Fisheries Sciences 18(3) 575-582 2019 DOI: 10.22092/ijfs.2018.116671

First distribution record of regular echinoids (Echinodermata; Echinoidea) from Chennai Coast, South India

Radhika Rajasree S.R.1*; Gobala Krishnan M.1; Karthih M.G.1; Aranganathan L.1

Received: May 2017 Accepted: July 2017

1-Centre for Ocean Research, Sathyabama Institute of Science and Technology, Chennai-600119, Tamilnadu, India. *Corresponding author's Email: [email protected]

Keywords: , , Molecular phylogeny, South India

Introduction number of regular sea urchin Sea urchins have recently been such as Salmaciella oligopora, attracting considerable research interest radiata and Prinocidaris as sources of a number of highly baculosa along the continental shelf of valuable bioactive compounds that Chennai, Kovalam and Marakkanam possess antitumour, antiviral, coasts, Tamilnadu, South India. Among

Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 antimicrobial and anticoagulant these S.oligopora species has been properties that hold great promise for reported for the first time from Indian use in pharmacological applications waters.We identified and examined the (Ryoyama, 1974; Wardlaw and Unkles, morphological features of S.oligopora 1978; Okigbo et al., 2008). This and conducted molecular identification necessitated a thorough stock (18S ribosomal gene sequence). assessment study of regular echinoids in Indian coastal waters to promote Materials and methods viable culture practices. The Indian Sample collection and identification coastal waters harbour over 50 species The live specimens of sea urchin of sea urchins of which 14 have been species S.oligopora, A.radiata and P. found to be edible (James, 1983; baculosa were collected using mini Kaliaperumal and James, 1993). In this trawl nets with mesh size of 50mm at paper, an attempt has been made to depths of 10–45m from three areas provide an updated systematic account along the Chennai-Pondichery Coast, and distribution status which includes South East Coast of India viz., Kovalam the first record in Indian seas of a (12.°48’37.61’N,80°53’48.57.′E) 576 Radhika Rajasree et al., First distribution record of regular echinoids… Marakkanam (12°5'16.25"N DNA amplification and sequencing 80°15'20.80"E) and Pondicherry Genomic DNA was extracted from the (11°46'23.38’N 79°57'26.86’E) (Fig. gonadal tissue of the specimens by the 1). The live were stocked in Phenol Chloroform method (CAGL plastic containers filled with aerated standardized protocol for fish tissue seawater and were transported to the extraction and Quiagen kit (Anup et al., Marine Biotechnology Laboratory, 2014). Polymerase chain reaction Centre for Ocean Research, (PCR) for amplification of the partial Sathyabama Institute of Science and 18S rRNA gene was performed using a Technology, Chennai. In the laboratory, test sample along with positive and the specimens were photographed using negative control (Forward primer: Nikon (D5300) and the specimens were 5'-CAGCAGCCGCGGTAATTCC-3' washed with 0.5% Ampicillin solution and Reverse primer: to remove pathogenic bacteria before 5'-CCCGTGTTGAGTCAAATTAAGC-3'). immersing in UV- sterilized filtered Sequencing of the amplified product sea water stocked in closed glass was done (both forward & reverse aquarium tanks of 10L capacity. The direction) following the CAGL- animals were fed ad libitum with green standardized protocol using Genetic macro algae Chaetomorpha antennina, Analyzer ABI-3500. Additionally, Ulva rigida and Enteromorpha sp. The another combination of 18S rRNA sea urchin specimens were identified primers was tested and comparative based on the descriptions of Mortensen sequence analysis was done for (1943) and Clark (1916). The important probable species identification. morphological characters such as oral test and aboral view, morphometric Molecular analyses measurements of test and spine, The phylogenetic tree was drawn using Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 colouration, genital plates etc., were the Maximum Likelihood (ML) photographed with a fully automated method. For the resulting tree and Inverted Fluorescence microscope random resampling of the sequences, (Leica DM16000B) and automated bootstrapping method was performed. stereo zoom microscope (Leica The phylogenetic tree representing a M205A). The skeleton samples were consensus of 1000 trees was obtained. later dried in open air and zooidal Similarities were calculated from partial measurements as well as skeletal parts 18S rDNA sequences of were photographed using Field Temnopleuridae family, with the Emission Scanning Electron exclusion of ambiguous nucleotides Microscope (FESEM-SUPRA 55- using MEGA ver 7.0 (Tamura and CARL ZEISS, GERMANY) available Stecher, 2016). in the Centre for Nano Science and Nanotechnology, Sathyabama Institute Results and discussion of Science and Technology, India. Taxonomy Phylum Echinodermata Iranian Journal of Fisheries Sciences 18(3) 2019 577

Class Echinoidea Leske, 1778 Subclass Bronn, 1860 S.oligopora (H.L. Clark, 1916) Order Camarodonta Jackson, 1912 Description Infraorder Echinidea Kroh and Smith, Test is small in size, slightly pale brown 2010 with olive green tinge (Fig. 2A) strong, Family Temnopleuridae A. Agassiz, hemispherical or discoid in shape and a 1872 distinctly flattened ventral side (Fig. Genus Salmaciella Mortensen, 1942 2B). The margin of oral side slightly Salmaciella oligopora (H.L. Clark, sunken towards the peristome (Fig. 3C). 1916; Mortensen, 1942) Five genital plates have developed as madreporites sites. Ambulacrum almost Key features of family Temnopleuridae half as broad as interambulacrum (Fig. A. Agassiz, 1872 3F). The margin of Interambulacral Test not sculptured (or rarely so) but plates was formed as a zig–zag line on with pits on horizontal sutures or at the interracial suture (Fig. 3E). The angles of coronal plates; latter united by number of ambulacral plates in a dowelling. Tubercles usually column is from 45 to 56. crenulated; test with distinct pits, troughs or pores the angles of the Size sutures are extensively sculptured. Horizontal test diameter, 23.4- 40.1 mm; test height, 10.1-13.2 mm; Key to the genus of Salmaciella peristome, 7.1-11.2 mm. Mortensen, 1942 Sub conical test: Anal opening Spine and colour eccentrically arranged towards In S.oligopora the primary spines reach

Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 periproctal edge, ambulacral plates up to a length of 0.3–1.3cm. The long reduced on alternate plates aborally, ambital spines were widened distally very distinctive angular plates on the with distinct shovel like ends (Fig. 3A oral side. The primary spines are & 3B). The miliary spines and banded in dark and light greenish pedicellariae are white, the secondary tints.Test greyish brown spines have alternate white and maroon ...... S.erythracis bands. Equatorial spines short in size, widened distally with distinct ends. Spine have Biology bands of white with maroon, brown and This species occurs on the continental light olive. Test: color pale fawn to shelf along the upper limit of the light olive green……………...... S. intertidal zone. This specimen has been oligopora selected as the holotype. This species (Clark and Courtman-Stock, 1976; walks rapidly on its oral spines and can Schultz, 2010) travel 42cm in one minute (Miskelly, 2002).

578 Radhika Rajasree et al., First distribution record of regular echinoids… Distribution the interambulacral plates (Fig. 2E). Australia (Queensland), New South Periproct flat or low conical. Wales, Tasmania, Victoria, South Australia, and Western Australia and Color Philippines. Southeast coast of India Colour varies from red to black, but the (Chennai coast). red cross is always clearly visible.

Taxonomy Spine Phylum Echinodermata Red, orange band with spine (Fig. Class Echinoidea Leske, 1778 2D), Primary spines black, finely ridged Order and long, easily breakable, filled with Family Gray, 185 loose mesh work. Genus Astropyga Gray, 1825 Leske, 1778 Biology Key features of family Diadematidae A.radiata has been observed in shallow peters, 1855 sea grass beds, sandy shelves (De Beer, Spine usually hollow, long, cylindrical, 1990). In the coral region just below the thin and breakable, with a finely thorny low tide level on the shallow reef or ridged surface. Peristome is without (Herring, 1972). plates and with soft skin.Test often somewhat flattened, rather flexible, Distribution sometimes rigid. Well–known tropical Indo west central pacific (including the Red Sea) species Genus Astropyga Gray, 1825 (Rowe and Gates, 1995; Vanden Description Berghe, 2000), south east coast of Genital plates conspicuously elongated India. Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 into the interambulacra between the aborally bulging ambulacra, test very Taxonomy much flexible. Phylum Echinodermata Class Echinoidea Leske, 1778 Astropyga radiata Leske, 1778 Subclass Cidaroidea Taxonomic description Order Cidaroida Test diameter is between 110mm- Family Cidaridae Gray, 1825 125mm, flattened or slightly concave Genus Prionocidaris A. Agassiz, 1863 on the aboral side. Test with intense red Prionocidaris baculosa (Lamarck, bands bordered by fluorescent blue 1816) spots (Fig. 2C); anus red-brown with intermittent whitish red spots. The Key features of family Cidaridae Gray, spines are up to 20mm-40mm, long and 1825 are grouped in five vertical clusters in High interambulacral plates with between which are V-shaped areas with massive, blunt primary spines, widely no spines corresponding to separated from the each other, usually Iranian Journal of Fisheries Sciences 18(3) 2019 579

surrounded by much smaller, often closely covered with small tubercles, spatulate, secondary spines. Ambulacral naked zigzag line along. and interambulacral plates cover the peristome up to the mouth. Spine Brown with green band, Length up to Key characters of genus of 110mm-120mm (Fig. 2G), Primary Prionocidaris A. Agassiz, 1863 spine are Very variable; shape from Ocular plate in contact with the almost cylindrical to spindle – like, periproct; large globiferous tapering distally or to point more or less pedicellariae with numerous fringes on widened, banded, oral primaries simple, stalk; primary spines with serrated mostly straight cut. edges. The genus has spots and stripes on the collar and neck of the primary Color spines and its transverse oval and The denuded test has dark purple or confluent areoles. brown apical plates and ambulacra (Fig. 2I), irregularly banded, collar always Prionocidaris baculosa (Lamarck, with purple spots (Fig. 2H). 1816) Taxonomic description Distribution Test This species was common in the Indian Slightly flattened above and below, Ocean from the Red Sea to Mauritius ambitus round or slightly pentagonal, and Ceylon, also in the Philippines and flattend ventral side (Fig. 2F). Indonesian waters and from Japan to Horizontal diameter of the test may the west coast of Australia from the reach more than 50mm. Oral side shore to a depth of about 200m. Also

Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 slightly sunken towards peristome (Fig. distributed in SE Arabia, West India, 3H). Pakistan, Maldives area, Bay of Bengal, East Indies, China and south Japan Ambulacra (Clark and Rowe, 1971). Marginal series of tubercles very regular, small and continuous; inside Biology the marginal series, as a rule, two P.baculosa can cover itself in sea-grass double series of much smaller tubercles. and algal debris or lagoon areas with Plate conjugate pore pairs in single rocky outcrops and some isolated coral almost vertical series. (Fig. 3G). heads. Primary tubercles crenulated but not perforated (Fig. 3I). DNA sequence features The sequences of our Indian S. Interambulacral oligopora specimens were initially Scrobicular tubercles prominent, compared with GenBank data of 3 outside the circular a few tubercles Salmaciella species such as Genbank id about half that size, median space AF279211, AF279189, AF279163 and 580 Radhika Rajasree et al., First distribution record of regular echinoids… showed 99% similarity with S. identified under taxonomical key oligopora species. In total, 636 base guides: 1971 monographic work of pairs (bp) of the 18S rRNA gene Clark and Rowe, From sequence were obtained from the S. the Kiunga Marine National Reserve, oligopora. This sequence data have Kenya and Echinoidea: with pentameral been submitted to the Gen bank account symmetry. Schultz (2015) based on the of ID (KX838956). The study reported collection and field observations, the by Jeffery, 2003 reveals that the partial present study of the first distributional gene and 1773-bp 18S rRNA gene records for the regular echinoids identities always agreed at the genus species from the south coast of India, level, and 99% of assignments were the Chennai, resulted in recognitions of 3 Salmacis belli species assignments by species belonging to 3 families and 3 the partial gene method. genera. S.oligopora, which had not been previously reported from India, Phylogenetic tree has shown new distributional records In the phylogenetic tree, Maximum from the Indian waters. The previous likelihood (ML) method demonstrated description of S.oligopora species the presence of non-discriminatory distributed exclusively along New phylogentic branches (Fig. 4). We South Wales, Australian coast (Clarke, confirmed the identification of the 1916). Miskelly (2002) described the specimen as S. oligopora after species as endemic to Tasmania and comparison and analyzis with 18 described S. oligopora as a rapid mover closely related species of the in comparison with other species which Temnopleuridae family obtained from can travel 42 centimeters per minute on the Genbank. Totally nine different its oral spines. Though Mortensen genus groups of Temnopleuridae family (1942) described the species Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 were analysed. The phylogenetic tree Salmaciella and described in detail construction suggests that our specimen about 409 echnoid Taxa in his article has a very close resemblance to S. (1943), there are no descriptive records sphaeroides. The results were of S.oligopora other than Clark’s compared to groups of deuterostomes monogrpahy (1916) and we located the and was supported by a high value animals between 10m to 30m depths obtained by bootstrapping (71.2%). along the continental shelf. Recently Next, using the same alignment, we Schultz (2015) reported the occurrence constructed another phylogenetic tree of S.oligopora in the Philippines coast. by the maximum-likelihood method According to him, the animals live in using the fast DNAml program sheltered bays in the open bottom. (Felsenstein , 1985). A.radiata, and P.baculosa species recorded as first distribution echinoids Discussion species from the Chennai coastal Identification of echinoids species from waters, occur in depths of more than the south east coast of India has been 30m of the continental shelf region Iranian Journal of Fisheries Sciences 18(3) 2019 581

from Kovalam and Marakkanam coasts, Clark, A.M. and Rowe, F.E.W., 1971. Chennai, Tamil Nadu South India. A. Monograph of shallow-water indo- radiata is highly venomous and can west Pacific echinoderms. Printed by pierce through a wet-suit. Very order of the Trustees of the Museum: sensitive to light and have the ability to 238 P. shoot venom loaded spines at a short Clark, A. and Courtman Stock, J., distance and sometimes hitch lift on the 1976. The echinoderms of Southern back of crabs Africa. Publ. No. 766. British (www.whatsthatfish.com/fish/red- Museum of Natural History London. urchin/330).\Prionocidaris\baculosa ca 277 P. n be found hiding in the sea grass beds De Beer, M., 1990. Distribution Thalassodendron cilliatum (Forskal, patterns of regular sea urchins 1757). (Echinodermata: Echinoidea) across the Spermonde Shelf, SW Sulawesi Conclusions (Indonesia). in: De Ridder C, Dubois This is the first record of the regular P, Lahaye MC, Jangoux M, echinoid species A.radiata, P.baculosa eds, Research. and S.oligopora recorded for the first Rotterdam: Balkema. pp. 165-169 time from Indian waters, filling a gap of Felsenstein, J., 1985. Confidence limits its distribution in the East Coast of on phylogenies: an approach using India. It is clear that despite the efforts, the bootstrap. Evolution, 39, 783– knowledge and basic ecological 791. information about echinoderms and Forskål, P., 1775. Descriptiones infaunal communities in the Indian Seas animalium, avium, amphibiorum, and the region is still incomplete. piscium, insectorum, vermium; quae

Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 in Itinere Orientali observavit Petrus References Forskål. Post mortem auctoris edidit Mandal, A., Varkey, M., Sobhanan, Carsten Niebuhr. Adjuncta est S.P., Kottayil Mani, A., materia medica kahirina atque tabula Gopalakrishnan, A. and maris Rubri geographica. Havniae: Kumaran, G., 2014. Molecular ex Officina Mölleri. markers reveal only two mud crab Herring, P.J., 1972. Observations on secies of genus Scylla (Brachyura: the distribution and feeding habits of Portunidae) in Indian Coastal some littoral echinoids from Waters. Biochem Genet, 52, 338– Zanzibar. Journal of Natural 354. History, 6, 169-175. Clark, H.L., 1916. Report on the sea- James, D.B., 1983. Research on Indian lilies, starfishes, brittle-stars and sea- echinoderms - a review. Journal of urchins obtained by the F.I.S. the Marine Biological Association of Handbook of zoology Andreas India, 25 (1 & 2), 91-108. Schmidt-Rhaesa Jeffery, C.H., 2003. Phylogeny and (Ed.) Echinodermata. evolution of developmental mode in 582 Radhika Rajasree et al., First distribution record of regular echinoids… temnopleurid echinoids. Molecular 3rd ed. Hemidingen, Germany: Phylogenetics and Evolution, 28, 99 Scientific Publication. 484P. –118. Schultz, H.A.G., 2015. Handbook of Kaliaperumal,N. and James D.B., zoology: Echinodermata, Vol. 1: 1993. Hand book on aquafarming: Echinoidea: with pentameral seaweed, sea urchin and sea- symmetry., 386 P. cucumber.MPEDA.11:9-22. Tamura, K. Kumar, S. and Stecher, Miskelly., 2002. Sea urchins of G., 2016. MEGA7: molecular Australia and the Indo-Pacific. evolutionary genetics analysis Capricornica Publications, Sydney. version 7.0 for bigger datasets. Mortensen, T., 1942. Echinoderms Molecular Biology and from the Iranian Gulf. Asteroidea, Evolution, 33, 1870-1874. Ophiuroidea, and Echinoidea. Vanden Berghe, E., 2000. Danish Scientific Investigations in Annotated checklist of the Iran. Part, 2, 55-112. echinoderms from the Kiunga Mortensen, T., 1943. A monograph of Marine National Reserve, Kenya: 1. the Echinoidea. III, 2. Camarodonta. Echinoidea and I. Orthopsidæ, Glyphocyphidæ, Holothuroidea. Journal of East Temnopleuridæ and Toxopneustidæ, African Natural History, 89(1), 1-36. C. A. Reitzel, Copenhagen: pp. 146- Wardlaw, A.C. and Unkles, S.E., 148 1978. Bactericidal activity of Okigbo, R.N., Eme, U.E. and celomic fluid from sea-urchin Ogbogu, S., 2008. Biodiversity and Echinus esculentus. Journal of conservation of medicinal and Invertebrate Pathology, 32(1), 25- aromatic plants in Africa. 34. Microbiology and Molecular Downloaded from jifro.ir at 4:21 +0330 on Monday September 27th 2021 Biology Reviews Journal, 3(6), 127-

134. Rowe, F.W.E. and Gates, X., 1995. Echinodermata. In Zoological catalogue of Australia, Echinodermata. (CSIRO Publishing: Melbourne, Australia). Vol. 33: Melbourne: CSIRO Australia. 510 P. Ryoyama, K., 1974. Studies on the biological properties of coelomic fluid of the sea urchin. II. Naturally occurring hemagglutinin in sea urchin. Biological Bulletin, 14, 410- 414. Schultz, H., 2010. Sea urchins, a guide to worldwide shallow Water Species.