Int. J. Biosci. 2019 International Journal of Biosciences | IJB | ISSN: 2220-6655 (Print) 2222-5234 (Online) http://www.innspub.net Vol. 14, No. 4, p. 350-359, 2019 RESEARCH PAPER OPEN ACCESS Bioactive compounds proximate composition and cytotoxicity of Echinoidea (Sea Urchin): Basis for future policy preservation and conservation Genevieve U Garcia*1,2, Nenita D Palmes1, Oliva P Canencia1 1 Department of Science Education, University of Science and Technology of Southern Philippines, Cagayan de Oro City, Philippines ]2 Department of Education, Division of Cagayan de Oro City, Philippines Key words: Echinoidea, Proximate composition, Cytotoxicity, Preservation and conservation, Philippines http://dx.doi.org/10.12692/ijb/14.4.350-359 Article published on April 30, 2019 Abstract The presence of Echinoidea in marine ecosystem at the inter-tidal zone of the Philippines is abundant. The proximate composition and cytotoxicity of the combined spines, tests and gonads of the selected three species of sea urchin namely: Diadema setusom, Tripnuestes gratilla and Astropyga radiata greatly vary in their proximate composition. Specifically, the three-sample species are potential source of nutrients and minerals. The cytotoxic activity using brine shrimp lethality test differ in selected sea urchins. The test for cytotoxicity showed that LC50 is ˃1000 ppm which is non-toxic. The results confirmed samples non-toxicity since the species are edible and local folks eat and sell them as source of livelihood. Preservation and conservation of sea urchin species are necessary for it is one of the most significant marine invertebrates used as bioindicator and as source for livelihood and commercialization. * Corresponding Author: Genevieve U Garcia [email protected] 350 Garcia et al. Int. J. Biosci. 2019 Introduction Sea urchins are not only important for their ecological The marine environment, which contains a vast array of roles in the environment but are found to be very organisms with unique biological properties, is one of beneficial to humans as well. Gonads of sea urchin the most underutilized biological resources (Ibañez, also named as roe are highly in demand and Herrero, Mendiola, & Castro-puyana, 2012). The economically important food because of its high Philippines is the center of sea urchin diversity in an in- nutritional value (Jinadasa, Zoysa, Jayasinghe, & depth study from intertidal to abyss according to the Edirisinghe, 2016). The roe of the sea urchin is Society for Integrative and Comparative Biology of the considered to be a prized delicacy due to its tasty University of California. As a biodiversity hotspot, the quality in Asian and Mediterranean countries. Also, in Philippines currently holds the gold medal in species Western Hemisphere countries such as Barbados and richness (Mooi & Munguia, 2014). Chile (Rahman et al., 2014). Gross chemical composition information of marine Due to the rich biodiversity of this country, organisms contributes largely to the importance of Philippines, though there is general diminishing of the species. Chemical analyses of the different some species, this study is timely before sea urchins compositions like proteins, lipids, carbohydrates, become extinct. Specifically, this study aims to nitrogen and phosphorus are significant as main determine the proximate composition and components of living matter (Barbarino & Lourenço, cytotoxicity of selected species of sea urchins namely 2009; Diniz et al., 2014). The total chemical the Diadema setosum, Astryopyga radiata and composition of an organism can be influenced by Tripnuestes gratilla using the combined spines, tests many factors such as physical characteristics, habitat and gonads. Furthermore, the study serves as the and life cycle, in addition to the environmental basis for drafting future policies on preservation and characteristics (Diniz, Barbarino, Oiano-neto, conservation of the sea urchin species. Pacheco, & Lourenço, 2014). The UN General Assembly last May 2017 made a The biological compounds of the sea urchin gonads, resolution for transforming our world through the tests, spines and amniotic liquid have been studied in “2030 Agenda for Sustainable Development”. This different species. The big quantity of polysaccharides, agenda consists of 17 goals. The present study is lipids and proteins are found in the ovary and testis aligned to the following goals: “Goal 2 is to … improve (Tenuzzo, Carata, Mariano, & Dini, 2017). The study on nutrition”, “Goal 3 is to ensure healthy lives and biological composition of edible urchin Stomopneustes promote well-being for all ages”, “Goal 13 is to take variolaris determined the proximate composition urgent action to combat climate change and its through moisture and ash, protein, lipid, carbohydrate, impacts”, and lastly, “Goal 14 is to conserve and fatty acid profile, carotenoid and vitamin analysis, and sustainably use the oceans, seas and marine resources essential trace metals (Jinadasa et al., 2016). for sustainable development” (UNESCO, 2017). The use of the brine shrimp A. salina has ecological In the Philippines, there are 70 species of sea urchin importance in marine environments, as these located in the different parts of the archipelago (Mooi & organisms are a symbol of the zooplankton Munguia, 2014). A higher density of T. gratilla species community and essential on the ecology of seashores. has been found in Northern Mindanao and in Southern The nuplii is very reactive to toxicants than the adults Mindanao (Pates, Responte, Mag-aso, & Dagoc, 2015). and microscopial measures are easy to perform using In addition, T. gratilla is the most known sea urchin that the brine shrimp A. salina (Lopes, Fernández, became a major source of livelihood. The populace of the Martins, & Vasconcelos, 2010). Asian Pacific Region has been using sea urchin for improving quality life and cure for many diseases In the Philippine context based on the published (Rahman, Arshad, & Yusoff, 2014). literature and studies cited, it dealt only on physical 351 Garcia et al. Int. J. Biosci. 2019 growth, taxonomy and management. It shows that of the species based on the abundance and maturity. there were no published studies on proximate The total body weight of each specimen and the composition and toxicity of the sea urchins in the average horizontal test diameter were measured Philippines which is the concern of the present study. perpendicularly without spines using a vernier The present study explored on this topic since sea calliper (Jinadasa et al., 2016). urchin is also one of the determinants of sea water pollution. It is therefore timely to undergo this study Preliminary identification was done based on the field for it might vanish in the future due to rapid human guide developed by Schoppe (2000) and the World population growth that results to water pollution. This Registry of Marine Species (WoRMS) using urgency calls for this study since we might lose the morphological characteristics. Final identification species without knowing the extent of its contribution was done at the Marine Center, Xavier University. For to humanity. further validation, photographs of the specimens were sent to the Institute of Environmental and Marine It is in these reasons that this study aims to determine Science, Siliman University. the proximate composition and cytotoxicity of the three species of sea urchin namely the Diadema setosum, The collected mature and fresh sea urchins were Astropyga radiata and Tripnuestes gratilla using the thoroughly washed with seawater to remove combined spines, tests and gonads from Vinapor and sediments. Selected species of sea urchins were Goso-on, Carmen Agusan del Norte, Philippines. Diadema setosum, Tripnuestes gratilla and Astropyga radiata as presented in Fig. 2, 3 and 4 Materials and methods respectively. The amniotic liquid was removed by Sampling Area making small hole in the shell, it was kept in a Ziplock The study was conducted in Goso-on and Vinapor and placed in a styropor box with ice for preservation Carmen, Agusan del Norte, Philippines. The samples during transport. The selected, clean and fresh were collected from the intertidal zone up to shallow samples were transported to USTP Chemistry waters of about 0-5m. The map of the collection site is laboratory within 24 hours and immediately stored in shown in Fig. 1. The Municipality of Carmen is a freezer to avoid decomposition. located in the province of Agusan del Norte of CARAGA Region. Fig. 2. The Diademata setosum. Fig. 1. Collection site in Barangay Gosoon and Vinapor, Carmen, Agusan del Norte. Sampling Scheme Purposive sampling scheme was adopted to where the sea urchins’ species were abundant based on the preliminary survey or reconnaissance of the area. The three sea urchins species were selected from the rest Fig. 3. The Tripnuestes gratilla. 352 Garcia et al. Int. J. Biosci. 2019 been oxidized. The following formula was used to calculate the ash content of the samples and expressed as the percentage by weight of the dry sample. 푀푎푠ℎ %퐴퐷 = x 100 (1) 푀 푑푟푦− 푀푚푙 Where: %AD = percentage of ash in the sample (dry basis) Mash = weight of ash sample Mdry = weight of the original dry sample Fig. 4. The Astropyga radiata. Mml= mass of the moisture lost (mass of the dry sample minus the mass of sample after moisture determination) Data Analysis Descriptive statistics was used in the presentation of b. Crude Protein the different parameters involved. Data gathered such Analysis of crude protein was done using the as total ash, crude lipid, crude protein, and total Kjeldahl’s method, which evaluate the total nitrogen carbohydrates for proximate composition was content of the sample after it has been digested in analysed using one-way analysis of variance (ANOVA) sulphuric acid with a mercury as a catalyst. The with Tukey Honest Significant Difference Mean Test simple method used was proposed by Chow et al. (Tukey HSD Mean Test) for pairwise comparison. The (1980). The following formulas were used to calculate differences with p-values of less than 0.05 (p<0.05) the ash content of the samples: level were statistically significant and data were 퐴푥퐵 %푁푠푎푚푝푙푒( 0.014) 푥 100 (2) expressed in mean ± SD.