Iranian Journal of Fisheries Sciences 19(6) 3098-3111 2020 DOI: 10.22092/ijfs.2020.122939

Research Article Spawning and larval rearing of red (L. Agassiz and Desor, 1846;Echinodermata: Echinoidea)

Gobala Krishnan M.1; Radhika Rajasree S.R.2*; Karthih M.G.1; Aranganathan L.1

Received: February 2019 Accepted: May 2019

Abstract Gonads of sea urchin attract consumers due to its high nutritional value than any other seafood delicacies. Aquaculturists are also very keen on developing larval culture methods for large-scale cultivation. The present investigation systematically examined the larval rearing, development, survival and growth rate of Salmacis bicolor fed with various microalgal diets under laboratory condition. Fertilization rate was estimated as 95%. The blastula and gastrula stages attained at 8.25 h and 23.10 h post-fertilization. The 4 - armed pluteus larvae were formed with two well - developed post-oral arms at 44.20 h following post-fertilization. The 8 - armed pluteus attained at 9 days post fertilization. The competent larva with complete rudiment growth was developed on 25th days post - fertilization. Monodiet algal feed - Chaetoceros calcitrans and Dunaliella salina resulted medium (50.6 ± 2.7%) and low survival rate (36.8 ± 1.7%) of S. bicolor larvae. However, combination algal feed – Isochrysis galbana and Chaetoceros calcitrans has promoted high survival rate (68.3 ± 2.5%) which was significantly different between the mono and combination diet. From the observations of the study, combination diet could be adopted as an effective feed measure to promote the production of nutritionally valuable roes of S. bicolor.

Keywords: Sea urchin, Salmacis bicolor, Spawning, Embryo, Larvae, Microalgae

1-Centre for Ocean Research, Sathyabama Institute of Science and Technology, Chennai 600119, Tamilnadu, India. 2-Department of Fish Processing Technology, Kerala University of Fisheries and Ocean Studies, Cocin-682506, Kerala, India. *Corresponding author's Email:[email protected]

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Introduction bicolor and reported the early Echinoids aquaculture is being carried development and metamorphosis of the out as a commercial activity to harvest tropical echinoid from Chennai Coast. the gonads or roe which is a delicacy in Saravanan et al. (2017) have studied the Asian and Mediterranean countries. A temporal variation of gonadal maturity sea urchins roe is being consumed by of S. virgulata and Temnopleurus humans since pre-historic times in toreumaticus from Mandapam region of many countries around the world and Indian South-East Coast. Moreover, are considered a high nutritional Rahman et al. (2012) reported that valuable food source (Chen, 2013). spawning, embryonic and larval High exploitation rate has endangered development of the tropical sea urchin, this species in many countries that in Salmacis sphaeroides from Malaysian turn created interest towards coast. Earlier investigations on the development of sea urchin larval influence of dietary treatments on larval cultivation and rearing techniques survival and development of various (Kelly et al., 2000; Bustos and Olave, echinoids species were reported 2001). Development of sea urchin (Meidel et al., 1999; Kelly et al., 2000; aquaculture has greatly advanced by the Jimmy et al., 2003; Liu et al., 2007). enhancement of ranching techniques, Sea urchin releases large quantities of which directly targets gonadal growth eggs and sperm into the water for and reproduction by the nutritional fertilization and the larva of this enhancement. organism play an important role in the Aquaculture of sea urchins have marine food chain (Mishra et al., 2012). focused on larval feeding (George et Spawning and larval life span can be al., 2004; Liu et al., 2007), culture determined by the timing of larval methods for both larva and adults settlement. (Kelly et al., 2000). Adaption of the Twenty species of edible sea urchins recirculation system for larval growth are widely consumed globally. Indian and development in Paracentrotus coastal waters include over 150 species lividus progressing from fertilization to of sea urchins of which 14 have been harvest method was first developed by found to be edible (James, 1985; Le Gall and Bucaille (1989). The Kaliaperumal and James, 1993; system provides features beginning Venkataraman and Wafer, 2005). from spawning, larval and algal Edible sea urchins are selected based on cultures, and a separate rearing system three criteria: i) accessibility of the for the settlement and species, ii) palatability, and iii) development of mature adults for historical and culture practices needed harvesting. to be taken into consideration Aiyar (1935) has initially documented (Lawrence, 2007). the anatomy of the sea urchin S.

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Phytoplanktons are the major food In this paper, we made first report on source of larvae. Culturing the effect of various dietary microalgae practices of phytoplanktons support on growth, larval development and massive production of sea urchin larvae survival of sea urchin larvae S. bicolor (Sakai et al., 2004). Combinational diet from Tamil Nadu, South India. It also of I. galbana and C. gracilis were provides a possible solution to improve found to be useful in the mass production output of commercial production of competent larvae of hatchery. To this end, we compared sea (Carcamo, 2004). urchin larvae growth performance when Combination of various algal diet of fed different live microalgae that are Cricosphaera elongata, Pleurochrysis easy to culture and readily accepted by carterae and Dunaliella tertiolecta as the larvae. Furthermore, this work will well as Pleurocrysis elongata and D. help to further understanding the tertiolecta resulted in the successful physiological requirements of growth of P. lividus and P miliaris echinoplutei and enable optimization of larvae (Kelly et al., 2000; Stefanoet al., S. bicolor larval rearing protocols to 2012). increase hatchery profitability. The S. bicolor, a species of echinoid present study aimed at the influence of belonging to the family micro algal strains as monodiet: C. Temnopleuroidae, is a regular echinoid. Calcitrans, D. salina and combination Morphological features include 0.5 to 4 diet: C. calcitrans and I. galbana cm spines in length with white and red towards the morphological development color bands with long tube feet; circular and larval survival rate of S. bicolor test, dense covering of short spine, and grown under laboratory conditions. spine around peristome markedly flattened; spine, red around base with Materials and methods attractive yellow banding towards distal Geographical location ends. S. bicolor is a relatively well- The animal was collected from known tropical Indo - West Pacific Chennai, East Coast of Tamil Nadu species (Clark and Row, 1971; state, India (Lat. 13° 8'43.86’N; Richmond, 1997). Pearce (2010) Long. 80°21'42.43’E) (Fig. 1). The reported a scientific research aim to region covers shallow subtidal and bridge the gap between supply and continental slope area with typically demand of aquaculture products. covered muddy bottoms and rock Caboni et al. (2013) reported sea urchin materials. adults excellent quality gonads could be available for the market throughout the whole year, while juveniles bred in controlled conditions could be employed in restocking activities.

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Figure 1: Location of study sites, South East Coast of India. The map shows the collection site of S. bicolor from Chennai Coast, Tamil Nadu.

Live samples of S. bicolor were Ulva rigida and Enteromorpha sp. The collected by using the fishing net from a experimental study was carried out for depth of 15-20 m from Chennai Coast duration of 25 days. and transferred to the Marine Biotechnology lab in aerated plastic Larval rearing containers. After 2 to 4 days of acclimatization, Adult sea urchins were selected for Animal maintenance induced spawning according to their For maintenance in aquaria, UV- test diameter size (63 ± 3.4 mm) and sterilized 5-µm filtered seawater weight (54 ± 1.6 g). Adult sea urchins stocked in closed glass aquarium tanks were induced spawning by injection of 10 L capacity with ambient with 0.5 M KCl into the haemocoel via temperature (28 ± 2°C) was used. The the peristomal membrane (Fig. 2a). The were fed ad libitum with green injected animals (9 males and 6 macroalgae: Chaetomorpha antennina, females) were transffered to individual Iranian Journal of Fisheries Sciences 19(6) 2020 3102 vessels contained 5-µm filtered concentration of 1500 cells ml-1 were seawater. After 2-3 min, the animals provided separately, according to the started releasing the eggs and method described by Liu et al. (2007) spermatozoa into the water column in and Stefano et al. (2012). plastic bowls and extruded gametes All treatments were carried out in were collected by pipette. Totally, 15 triplicates. Water was exchanged individual culture jars were maintained. between 2-3 days intervals and the Eggs were washed twice with 5-µm larvae were monitored for the filtered seawater and high quality development of new arms or other gametes were selected for fertilization. structures and morphological features The suspension of eggs was fertilized such as lengths of the antero-lateral, with 2 ml volume of sperm which was postoral, somatic, posterodorsal and added in the water column and after posterolateral arms. The timing was gamete fusion, the fertilization rate was determined based on the morphological estimated to be 95%. Hatching rate was changes in the larval development. determined to be (88.5±2.3%). After the first week, larval cultures Embryos and larvae were maintained in were stirred with 50 rpm agitator paddle the 1 L glass beaker with water system similar to the method described temperature: 26–28˚C, salinity: 30–33 by Strathmann (1987). ppt; light density: 100–1000 lux without All the developmental stages of aeration. Initially, the count of embryos embryos and larvae were observed was maintained greater than 6 per ml under a phase contrast microscope followed by less than 1 per ml in the (Olympus CX41). All morphometric two arm stage. Microalgae cultures measurements of embryo and larvae at were grown in 150 L polyethylene bags different stages were monitored after in UV – sterilized filtered seawater fixation with 4% formalin solution prior enriched with Walne’s medium. to microscopic examination. Microalgae were harvested in the exponential growth phase and fed to the Statistical analysis larvae during 3rd day. Larval survival Tukey’s test and one-way ANOVA was rate was assessed volumetrically every performed to identify the significant three days interval. When the larvae difference among the dietary treatments reached the 2-arm stage, mono algal (Graph pad prism 7.0 software feed – C. calcitrans and D. salina, as package). Results with p<0.05 were well as 50% mixture of combinational considered statistically significant. The feed I. galbana and C. calcitrans, were experiment was carried in triplicates provided separately with the and expressed in mean ± SD. concentration of 500 cells/ml-1. When the larvae reached the 6 – arm stage, increased microalgal strains

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Results After 04.30 h post-fertilization, The mean diameter of the unfertilized occurred without micromere division eggs of S. bicolor (90.12- 96.45 µm ) and the embryos formed morula stage and fertilized egg (96.58 ± 2.38 μm) (Fig. 2j). In blastula, cells in one side was recorded. The fully developed eggs slightly more columnar than those on are transparent, spherical in shape and the other side. This pole is blastoporal light green in color (Fig. 2b). The end (Fig. 2k). The vegetal plate vitelline membrane of the egg was thickened and cilia were formed on the raised after 20-40 sec of sperm entry perimeter 08.25 h after fertilization, and the fertilization membrane began to before hatching stage. The gastrulation form (Fig. 2c). The fertilized egg with commencement was indicated by a few formation of fertilized membrane was of the cells from those pole pushing found within 10 mins after the inwards. This grow forwards rapidly, insemination (Fig. 2d). The 2-cell, 4- almost to the anterior end and this cell, 8-cell, 16-cell and 32-cell stages process was completed in about 23.10 h were achieved in 15 min, 30 min, 01.15 (Fig. 2l). h, 02.25 h and 03.20 h, respectively (Figs. 2e - i).

Figure 2: Induced spawning of S. bicolor by using 0.5 M KCl solution (a), Egg (b), Fertilization egg (c), Fertilized eggs with complete formation of fertilization membrane (d), 2 - cell stage (e), 4 – cell stage (f), 8 - cell stage (g), 16 - cell stage (h), 32 - cell stage (i), Morula stage (j), Blastula stage (k), Gastrula stage (l). Scale bar = 50 µM.

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The prism stage was observed 29.30 h constantly with arms directly upwards. after fertilization (Fig. 3a). The two - At this stage, competent larva with arm pluteus stage was observed 36.40 h complete rudiment growth was after fertilization (Fig. 3b). At this recorded (Fig. 3f). On 15th day, larva stage, opening of the mouth was fully matured with hydrocoel ring and observed. The larvae has become grew on amnion rudiment. On 19th day, humped with a presence of esophagus, old larvae form ear - shaped postero- stomach and a short intestine. Muscles lateral process supported by branches of the esophagus began to contract; the from the post-transverse rod which stomach grew in diameter, while its developed well and became prominent. epithelium became thinner. The 4 - arm The left side commences to bulge out pluteus larvae was observed 44.20 h considerably, the stomach being pushed after fertilization with two well - to the right. Sometimes variation seems developed postoral arms (Fig. 3c). In to be more pronounced in the post-oral this stage, larvae attained (423.76 µm - and postero-dorsal arms. Yellow 526. 17 µm) length. The well-defined pigment bodies are found in opening in the lower half of the larvae considerable numbers in relation to the represented the anus. postero-dorsal and post-oral arms (Fig. The 6 - arm pluteus larval stage was 3g). From 18th to 21st day, larvae shown formed on 4th day after fertilization the rapid growth of echinus rudiment (Fig. 3d). At this stage, postoral arm with enlargement of mouth and active further elongated and anterolateral arms larvae started feeding voraciously (Fig. were supported by well-formed skeletal 3h and 3i). On 25th day, competent rods. The 8 - arm pluteus larval stage larvae with complete rudiment growth was formed on the 9th day after were settled down. At this stage, larval fertilization. In this stage, postero- tissues accumulated on the aboral dorsal arms formed clearly as shown surface of the rudiment forming a and grew rapidly (Fig. 3e). No globoid structure. additional arms developed after 8 - arm Significant difference in larval survival pluteus larval stage. At this stage, larva was identified among the diet attained a maximum length of (830.43 treatments (p<0.05). During 2-5th day of µm - 890.43 µm) after 10th day of post fertilization, there was no fertilization. On 13th day, rudiment of significant difference (p<0.05) in the dorsal arch and two anterior coeloms larval survival between the dietary has formed followed by complete treatments. However, significant development of the larval skeleton, differences (p<0.05) in the growth of arch, and formation of first pedicellaria competent larvae was identified from stage. During 15th day , the pluteus 11th to 25th days of post fertilization larvae possess three pairs of well- between the mono and combination developed arms and kept swimming diets.

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Figure 3: Larval developmental stages of S. bicolor. Prism stage (a), 2 - arm pluteus larvae (b), 4 - arm pluteus (c), 6 – arm pluteus (d), 8 – arm pluteus (e), Competent larvae with complete rudiment growth development (f), Yellow pigment tips of left postoral arm (g), Rapid growth of echinus rudiment (h), Enlargement of mouth and started feeding voraciously (i). Scale bar = 50 µM.

Overall statistical analysis of the dietary monodiet. Moderate level was observed response showed the highest survival in larvae fed with C. calcitrans (50.6 ± rate of the larvae with significant 2.7%) and low level with D. salina difference (p<0.05) in combinational (36.8 ± 1.7%) at competent stage, diet of I. galbana and C. calcitrans (68. respectively. The statistical significance ± 2.5%) (Fig. 4). In contrast,variations firmly supports the promotional in the larval survival rate were observed influence of the combination diet. with significant difference (p<0.05) in the Iranian Journal of Fisheries Sciences 19(6) 2020 3106

Figure 4: Survival rate of S. bicolor larvae fed combination diet of I. galbana and C. calcitrans and monodiet of C. calcitrans and D. salina. Data are expressed as mean ± SD (n=3). Letters above bars indicate the results of Tukey’s test with different letters showing significant (P<0.05) difference between the three algal diets treatments. (Error bars shows standard deviation).

Discussion study will be useful for better The results of the present study understanding of the larvae culture. systematically documented the From the above observations, embroyolgy, larval development, combination diet - I. galbana and C. survivability and various microalgae calcitrans could be adopted as an diets which has influenced faster effective feed measure to promote the competency in the larval development. production of nutritionally valuable Preliminary study on larval roes of S. bicolor. This could serve as development and metamorphosis of the attractive nutritional source because same species was reported in which the when food is abundant with high growth parameters - optimal lab quality, growth is directed toward non- condition and survivial rate was not ephemeral larval structures which are reported (Aiyar, 1935). However, our carried through metamorphosis level. study was much concerned and varied Early stages of embryology and larval from the earlier study in several aspects development have been reported for a such as larval rearing protocol, growth number of bathyal and abyssal response and survival rate of the larvae invertebrates (Young et al., 1993; under the influence of different Mortensen, 1921). Competency of S microalgal feeds and also the study bicolor was attained after 25 days. provides better microscopic Similar observations in S. sphaeroides observations of the larvae captured at larvae (complete rudiment growth after different phases. The results of this 35 days) (Rahman et al., 2012). 3107 Gobala Krishnan et al., Spawning and larval rearing of red sea urchin Salmacis bicolor...

Cleavage and development of embryo has taken place by the end of 15th day. and larvae of S. bicolor were similar to All arms were broad at the base and those reported in various echinoids could be seen to open and shut. species - Clypeaster rosaceus and S Mortensen (1921) also reported sphaeroides (Emlet, 1986; Rahman et similarity at this stage on the 15th day in al., 2012). Mortensen (1921) described Mesopilia globulus. Microalgal genera - feeding larval forms of echinoids which Tetraselmis, Isochrysis, and generally showed clear morphological Chaetoceros include species preferred similarity to the temnopleurids family for feeding marine invertebrates in mass such as Temnopleurus toreuinaticus, T. cultures (Duerr et al.,1998; Brown, sculptum and Mespilia globulus. 2002). S. bicolor larvae fed with D. McEdward (1984) reported that salina possess low survival rate (36.7 ± morphometric changes in sea urchin 3.2%) that failed to reach competence larvae Dendraster excentricus undergo and this could be due to the low during the course of larval development nutritional value of this species. An have important functional consequences increased survival rate of S. bicolor as the shape is related to feeding (68.3 ± 2.5 %) was observed the capability and metabolic activity. significant difference in the On the 13th day, posterior coelom of S. combination diet of I. galbana and C. bicolor larvae extended backward but calcitrans. Survival up to 82.3 ± 9.0% the two layers are in contact throughout was achieved in Loxechinus albus by their extent. At this stage, madreporic providing a combined diet of C. vesicle well developed. Similar calcitrans and I. galbana (Carcamo et phenomena in Echinus al., 2015). Stefano et al. (2012) microtuberculatus and E. miliaris was reported that survival rate at the observed (Metschnikoff, 1869; Murti, competence of P. lividus larvae (82.8 ± 1932). S. bicolor larvae found yellow 10.6%) was achieved in D. tertiolecta pigmented presented on postero-dorsal diet. Bustos et al. (1992) suggested that and post-oral arms tips clearly. Also, larvae of L. albus may be fed Stomopneustes variolaris larvae have exclusively on Chaetoceros species as a found deep red pigmented tips on monodiet, as in the production of larvae posterolateral arms (Richard, 2009). of Strongylocentrotus intermedius and The body rods of S. bicolor larvae S. nudus and Glyptocidaris crenularis attained at 2.5 days age were similar to (Hagen, 1996; Chang and Wang, 2004; the findings in T. torematicus and S. Sakai et al., 2004). The apparently high virgulata larvae in 29 hours old age nutritional value of C. calcitrans may (Mortensen, 1921; Tennent, 1929). S. be attributable to high levels of 20:5n-3 bicolor anterior ciliated epaulettes and 20:4n-6 PUFA (Delaporte et al., began to appear on the 12th day, 2003). complete separation from the main band

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Rearing of S. bicolor larvae in lab MoES/36/OOIS/Extra/39/2014. We conditions was enhanced with a would like to thank the management of combination diet of C. calcitrans and I. Sathyabama Institute of Science and galbana to increase its survival rate . Technology for providing necessary The present data clearly show increased laboratory facilities. production output and a shorter production cycle when combination diet References of C. calcitrans and I. galbana is used Aiyar, R.G., 1935. Early development as larval diet providing production cost and metamorphosis of the tropical reduction and increased revenue thus echinoid Salmacis bicolor increasing hatchery profitability. S. (Agassiz). Proceedings Indian bicolor is an ideal candidate species for Academy of Science, IB, 714 – 728. aquaculture. The findings would Doi: 10.1007/BF03050571. immensely be helpful towards the Brown, M.R., 2002. Nutritional value development of induced breeding, of microalgae for aquaculture. In: commercial larval production for Cruz-Sua´rez, L.E., Ricque-Marie D, achieving high aquaculture potential of Tapia-Salazar M, Gaxiola-Corte´s sea urchins under MG, Simoes N. (Eds.), Avances en captive rearing condition. In future, NutricionAcuı´cola. Memorias del development of sea urchin aquaculture VI Simposium Internacional de will rely on positive interactions of Nutricio´nAcuı´cola. 3–6 September commercial fisheries and aquaculturists 2002. Cancu´n, Quintana Roo, towards mutual developmental activity Me´xico, pp. 281–292. for growth management and Bustos, E. and Olave, S., 2001. commercial production of larvae. As Manual: El cultivodelerizo the market demand for sea urchin roes Loxechinus albus. Divisio´ n de has been continuously increasing all Acuicultura, Instituto de over the globe, commercial culturing FomentoPesquero, Chile, 23P. activities to enhance productivity needs Bustos, E., Olave, S., Troncoso, R. to be addressed. The present report will and Godoy, C., 1992. be highly beneficial to understand the Investigacibnrepoblamiento de biological and environmental recursosbentdnicos Area Piloto IV parameters required for successful Regibn.Etapa IV. 5. Investigaciones culturing of other echinoids species. en erizo Loxechinus albus (Molina, 1782).CORFO-IFOP (SGI-IFOP Acknowledgments 92/8). 189P. We gratefully acknowledge the support Carboni, S., Hughes A.D., Atack, T., provided by Ministry of Earth Science, Tocher, D.R. and Migaud, H., Government of India, by funding the 2013. Influence of broodstock diet Research Grant Number: on somatic growth, fecundity, gonad

3109 Gobala Krishnan et al., Spawning and larval rearing of red sea urchin Salmacis bicolor...

carotenoids and larval survival of sea Delaporte, M., Soudant, P., Moal, J., urchin. Aquaculture Research, 46, Lambert, C., Que re, C., Miner, 969 - 976. Doi:10.1111/are.12256. Choquet, G., Paillard, C.and Carcamo, P.F., 2004. Massive Samain, J., 2003. Effect of a production of larvae and seeds of the monospecific algal diet on immune sea urchin Loxechinus albus. In: functions in two bivalve species Lawrence JM. Guzmán, O. (Eds.), Crassostrea gigas and Ruditapes Sea Urchins: Fisheries and Ecology. philippinarum. Journal of DEStech Publications Inc., Experimental Marine Biology and Lancaster, pp. 299–306. Ecology, 206, 3053 – 3064. Doi: Carcamo, P.F., Candia, A.I. and 10.1242/jeb.00518 Chaparro, O.R., 2015. Larval Duerr, E.O., Molna, A. and Sato, V., development and metamorphosis in 1998. Cultured microalgae as the sea urchin Loxechinusalbus aquaculture feeds. Journal of Marine (Echinodermata: Echinoidea): Biotechnology, 7, 65-70. Doi: Effects of diet type and feeding 10.1007/BF02161209. frequency. Aquaculture, 249, 375 – Emlet, R.B., 1986. Facultative 386.Doi:10.1016/j.aquaculture.2005. planktotrophy in the tropical 03.026 echinoid Clypeaster rosaceus Chang, Y. and Wang, Z., 2004. (Linnaeus) and a comparison with Production of seeds of the sea urchin obligate planktotrophy in Clypeaster Glyptocidaris crenularis. In: sub depressus (Gray) Lawrence, J.M., Guzma´n, O. (Eds.), (Clypeasteroida: Echinoidea). Sea Urchins: Fisheries and Ecology. Journal of Experimental Marine DEStech Publications Inc., Biology and Ecology, 95(2), 183 – Lancaster, pp. 331–338. 202. Doi: 10.1016/0022 - Chen, Y.C., Chen, T.Y., Chiou, T.K. 0981(86)90202-9. and Hwang, D.F., 2013. Seasonal George, S.B., Lawrence, J.M. and variation on general composition, Lawrence, A.L., 2004. Complete free amino acids and fatty acids in larval development of the sea urchin the gonad of Taiwan’s sea urchins Lytechinus variegatus fed an Tripneustes gratilla. Journal of artificial feed. Aquaculture, 242, 217 Marine Sciences Technology, 21 (6), - 228. Doi: 10.1016/j.aquaculture. 723-732.Doi:10.6119/JMST-013- 2004.06.024. 0429-1 Hagen, N.T., 1996. Echinoculture: Clark, A.M. and Rowe, F.E.W., 1971. from fishery enhancement to closed Monograph of Shallow-water Indo- cycle cultivation. World West Pacific . Printed Aquaculture, 27, 6 – 19. Doi: by order of the Trustees of the 11250/2384545 Museum, 238P.

Iranian Journal of Fisheries Sciences 19(6) 2020 3110

James, D.B., 1985. Echinoderm Fauna type on growth and fatty-acid of the Proposed National Marine composition of sea urchin larvae, I. Park in the Gulf of Mannar. Proc. Paracentrotus lividus (Lamark, Symp. Endangered Marine Animals 1816) (Echinodermata). and Marine Paries, Stefano pp.403- Aquaculture, 264, 247-262. Doi: 404. 10.1016/j.aquaculture.2006.12.021 Jimmy, R.A., Kelly, M.S. and McEdward, L.R., 1984. Morphometric Beaumont, A.R., 2003. The effect and metabolic analysis of the growth of diet type and quantity on the and form of an echinopluteus. development of common sea urchin Journal of Experimental Marine larvae Echinus esculentus. Biology and Ecology, 82, 259–287. Aquaculture, 220, 261 – 275. Doi: Doi:10.1016/00220981(84)90109-6. 10.1016/S0044-8486(02)00193-X. Meidel, S.K, Scheibling, R.E. and Kaliaperumal, N. and James, D.B., Metaxas, A., 1999. Relative 1993. Hand Book on Aqua farming: importance of parental and larval Seaweed, Sea Urchinand Sea- nutrition on larval development and Cucumber. Manual. MPEDA, metamorphosis of the sea urchin Cochin. Strongylocentrotus droebachiensis. Kelly, M.S., Hunter, A.J., Scholfield, Journal of Experimental Marine C.L. and Mckenzie, J.D., 2000. Biology and Ecology, 240, 161– 178. Morphology and survivorship of Doi:10.1016/S0022-0981(99)00046- larval Psammechinus miliaris 5 Gmelin Echinodermata : Echinoidea Metschnikoff, E., 1869. Studienuber / in response to varying food die Entwicklung der quantity and quality.Aquaculture, Echinoderinenand Nennertinen 183, 223–240. Doi: 10.1016/S0044- Mesta.de Acad. Imper. de St. 8486(99)00296-3. Petersb.14, Ser. 7, No. 8. Lawrence, J.M., 2007. Edible sea Mishra, A., Mishra, J.K. and Mohan, urchins; biology and ecology, P.M., 2012. Distribution pattern of Volume 38, 2nd Edition. ISBN: Echinoderm larvae with special 9780080465586. reference to the larvae of sea urchins Le Gall, P. and Bucaille, 1989. Sea in the Andaman Sea. Journal urchins production by inland of Coast Environmental, 3(1), 81- farming. Aquaculture - a 88. Doi: 10603/195355. biotechnology in progress. European Mortensen, T.A.,1921. Studies of the Aquaculture society, breden, Development and Larval Forms of belgium, pp. 53–59. Echinoderms. G.E. C.Gad, Liu, H., Kelly, M.S., Cook, E.J., Copenhagen. 260P. Black, K., Orr, H., Zhu, J.X. and Murti, N.N., 1932. The Development Dong, S.L., 2007. The effect of diet and Function of the Heart and

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Pericardium in Echinodermata, Proc. on gonadal maturity of two species Roy. Soc. London. (B), 109. of sea urchins. Indian Journal Fish, Pearce, C., 2010. Sea-Urchin 64, 190 -193. Doi: Aquaculture. Bulletin of the 10.21077/ijf.2017.64.specialissue.76 Aquaculture Association of Canada 265-28. 108–1. Stefano, C., Julien,V., Mariachiara, Rahman, M.A., Yusoff, F.M., Arshad, C., Douglas, R., Tocher, N. and A., Shamsudin, M.N. and Amin, Herve, M., 2012. Effects of dietary S.M.N., 2012. Embryonic, larval microalgae on growth, survival and development and early Juvenile of fatty acid composition of sea urchin the tropical sea urchin, Salmacis Paracentrotus lividus throughout sphaeroides (Echinodermata: larval development. Aquaculture, Echinoidea). The Scientific World 324, 325, 250 – 258. Doi: Journal, pp.1–9. Doi: 10.1016/j.aquaculture.2011.10.037 10.1100/2012/938482 Strathmann, M.F., 1987. Richard, B. and Emlet., 2009. The Reproduction and development of Bilaterally Asymmetrical Larval marine invertebrates of the northern Form of Stomopneustes variolaris. Pacific coast: data and methods for Journal of Biological bulletin, 216, the study of eggs, embryos, and 163 – 174. Doi: 10.2307/25470738. larvae. Seattle: University of Richmond, M.D., 1997. Washington Press, 684P. Echinodermata. In: A Guide to the Tennent, D.H., 1929. Early Seashores of Eastern Africa and the Development and Larval Forms of Western Indian Ocean Islands, (ed.) three Echinoidsof the Torres Straits M.D. Richmond, The SEA Trust, region, Papers from the Tortugas Zanzibar, pp. 290-321. Lab. Carnegie Lasts. Wash., 1929, Sakai, Y., Tajima, K.I. and 26. Agatsuma, Y., 2004. Mass Venkataraman, K. and Wafer, M., production of seed of the Japanese 2005. Coastal and marine edible seaurchins Strongylocentrotus biodiversity of India. Indian Journal intermedius and Strongylocentrotus of Marine Sciences, 34 (1), 54-75. nudus. In: Lawrence, J.M., Guzma´n, Doi: 10.1016/B978-0-12-801948 - O. (Eds.), Sea urchins: Fisheries and 1.00019-7 Ecology, DEStech Publications, Young, C.M., Tyler, P., Emson, R.H. Lancaster, pp. 287– 298. and Gage, J.D., 1993. Perception Saravanan, R., Jawahar, P., Francis, and selection of macrophyte detrital T., Ahilan, B., Santhakumar, R. falls by the bathyal echinoid and Gopakumar, G., 2017. Stylocidaris lineata. Journal of Deep Echinoid landings at Mandapam, Sea Research, 40,1475-1486. Doi: South-east Coast of India with a note 10.1016/0967-0637(93)90124-L.