Journal of Tationl and Complementary Meticne (2018) 341-351 Contents lists available at ScienceDirect e—> JTW Journal of Traditional and Complementary Medicine | journal homepage: http://www.elsevier.com/locate/jteme Review article Medicinal and health benefit effects of functional sea cucumbers Mons Ratih Pangestuti *", Zainal Arifin *” + serch Center for Oceanography, ndonesan Iu of Snes, Psi Pah, Anco Tina, oar Uta 14432 Indonesia Deputy of Ear Scenes. ndanesan state of Sees, ender Cat Sobrto No 0 Jakarta 12710 Indonesia ‘ete history ‘Sea cucumbers have long been used as food and traditional medicine in Asian counties with Stichopas ecelved 6 Ap 2017 hhermanni, Theleota ananas, Thelenota onex, Holothuria fuccova, and Acinopyga mauritiana as most Received in eis form Accepted 21 June 2017 Palate online 17 uly 2017 highiy-valued species. These organisms are potential Source of high value-added compounds with therapeutic properties such as rterpene glycosides, carotenoids, bioactive peptides, vitamins, minerals, Tatty acids collagens, gelatins, chondroitin sulfates, amino acid. In the recent ears, health benefit eects ‘of sea cucumbers have been validated through scentific research and have shown medicinal value such as wound healing, neuroprotective, anticumor, anticoagulant, antimicrobial, and antioxidant. These eae tanta ates eat potent development in vars oats an biomed fst hs Seal review we av prseted general view of major medial eat eet ees anton se Pay {cumbersome reson The star spec ad he potent apaon oa cae forte dered functional mater s wel thee nts sale are so disse. boars S201 comer or onda Boece exon Twan Universi. ductn and osingy svi Tainan te ts san open aes atu the Ce BY.N ND eense ih fae wea teensy nen) 1. imrodection The increasing number of scientific papers published in the last, few decades correlating functional materials derived from natural resources and some chronic diseases has shown the extraordinary possibilities of functional foods and nutraceuticals as well as biomedicine products to support, or even to improve, our healt beyond the provision of basic nutritional requirements.'? As a consequence, consumers interest in the relationship between health, diseases prevention, and well-being has grown substan- tially worldwide. The sources of functional foods, nutraceuticals and biomedicine products are exist in many reservoirs and may be {ound in terrestrial and marine environments. Terrestrial resources such as fruits, vegetables, cereals, probiotics, and. mushrooms: however, is by far more explored than the marine resources.” Even though the majority of those products in the marketplace are of terrestrial origin, marine organisms-based products are gaining. attention due to their unique features, which are not found in terrestrial-based resources.” * Corresponding author. [Emailed th anges. (anges Prot review under responsiblity of The Center for Food and Biomolecules, Navona Tawan Unversity utp do.org/1010163 me 20706007 BY-NND lene (ep eeatnecommonsorlcensesty nc nd) 407Ea Pong 2 Ain oul of Tonal and Complerentay Medicine 8 (208) 31-251 been reported, these economic value sea cucumbers which bring benefits to fishers for centuries were called as trepan or teripang.in Indonesia!” Not only, Indonesia, Malaysia and Philippines are also an important exporter of the sea cucumber ancl their products.’ Globally, The Southeast Asia represents the global market hot- spots for sea cucumbers trade due to their known mega biodiver- sity. Many of sea cucumber is gathered for human consumption and some are cultivated in aquaculture systems. Hence, the objectives ofthis article are first to present the re- sults obtained of a detailed bibliographical search about the composition of sea cucumbers from tropical regions especially Asia and secondly to discuss their biological activities and possibilities as new sources of functional ingredients. The information provided fn the various species of sea cucumbers does not refer in many cases to the same constituents since it has been taken from different research papers with different objectives. However, we believe the information provided can be useful to many research sroups considering a huge interest in the search for nutritional and ‘medicinal value of sea cucumbers. 2. Chemical composition of various sea cucumbers Functional ingredients from sea cucumbers have become an Increasingly interesting way to develop new foods as well as biomedicine products. Sea cucumbers are a source of high value- added compounds with health benefit effects to be used as func- tional ingredients. Bioactive’ peptides; vitamins, minerals. fatty acids, saponins, carotenoids, collagens, gelatins, chondroitin sul- fates, amino acids, fatty acids and other bioactive compounds are example of such sea cucumber derived functional ingredients that can be added at different stases of the food and biomedisine pro- duction process.” The sea cucumber species (Stichopus hermanni, Thelenota ananas, Thelenota anax, Holothuria fuscogilva, Holothuria leucospilota, Holothuria atra, Holotturia scabra and Actinopyga ‘mauritiana) described in this article has been selected considering edible species, medicinal effects, and low toxicity, The aforemen- tioned selected varieties are some of high-value sea cucumbers in Asia. Assuming that any new functional ingredient obtained from sea cucumbers could be used for further development of new products in food and pharmaceuticals industries. Another impor- tant factor is their nutritional value and potential as new sources of functional ingredients. 21. S. hermanni (local name in indonesia: gamat emas, garmat keacang, tikongkong) Sea cucumber 5. hermanni (curryfish, golden sea cucumbers) belongs tothe genus Stichopus: these species were formerly known as Stichopus variegarus. In Indonesia and Malaysia, sea cucumber ‘S. hermanni (Fig. 1) has long been used for the preparation of traditional medicinal products like gamat water and gamat ol ‘These species ate gaining much recognition among, consumers, ‘medical and biomedical researchers due to their potential health benefits. In Asian region communities, S. hermanni have been exploited for medicinal purposes; however such applications needs to be proven on a scientific basis using some clinical models. ‘As shown in Table I, S. hermanni contains high amount of pro- tein (47.00% + 0.36%) and low percentage of lipid (0:80% + 0.02%). This sea cucumber contain significant amount of sulfated glycos- aminoglycan. Glycosaminoglycan are long, unbranched poly- saccharides composed of repeating disaccharide units consisting of alternating uronic acids (D-glucuronic acid or L-iduronic acid) and amino sugars (o-galactosamine or D-glucosamine) Glycasamino- flyean are divided into non-sulfated and sulfated glycosamino- elycan. Sulfated glycosaminoglycan extracted from 5. hermanni Fi 1. Schou heron rom Lembeh tat, Indones possess various chemico-biological functions.” Compared to other parts such as internal organs and coelomic fluid; integument body wall of S. hermanni contain highest_glycosaminoglycan, both sulfated and non-sulfated Further, sullaled ghyeosaiioglicailfion integument has been demonstrated to accelerate wound healing process in rats!" More than 60% of wound heal area in rats was observed after daily treatment with sulfated glycosaminoglycan (20 iL of 1 ugimL) for 12 days. The healing activity of sulfated slycosaminoglycan was mediated through acceleration of wound contraction in wound healing phase | in addition, 40% of Stichopus hhermanii extract were able increase the numbet of lymphocytes during the healing process of traumatic ulcer on Wistar rat's oral MUCOUS! Most recently, Arundina et al, (2016) extracted 'S hermanni from Kalimantan, Indonesia and demonstrated their srowth stimulating effects in mesenchymal stem cells.” Mesen- «hymnal stem cells are self-enewing cells that have the capacity to Lierentiate into adipocytes, chondrocytes, myocytes, and osteo- biast. Following treatment with S. hermanni extract and osteogenic induction medium for 4 weeks, mesenchymal stem cells were differentiated into osteoblast. Collectively, it can be assumed that sea cucumber S. hermanni is able (© accelerate wound healing process. Further, these sea cucumber species can be used to prepare lotion ora topical ointment for wound healing management. Neuroprotection may defined as mechanisms and strategies used in order to protect neuronal cells against injury, apoptosis, dysfunction and or degeneration in the central nervous system (CNS).'In the CNS, there are two classes of cell, including neuron, and glia (microglia, astrocytes and the related Schwann cells and oligodendrocytes). Astrocytes plays an important structures that provide housekeeping functions necessary to maintain neuronal function, atively shape synaptic function, and act as neural pre- cursors in adult neurogenic regions. In addition, astrocytes also preserve the host integrity following injury. Recently, Patar etal (2012) prepared water extract of S, hermanni from Malaysia and showed their growth promoting effect to promote proliferation of spinal astrocytes." In pathological cases like spinal cord injury proliferating reactive astracytes are proven essential for early regeneration process, provide neuroprotective effects and preserve ‘motor function after acute injury. Further, it was demonstrated by GC-MS results that 37% ofthe total S. hermanni water extracts were comprised of amino acids (37%) followed by hydrocarbon (21%), ester compounds (168) the orther remaining compounds consisted of phenols, alcohol groups and unidentified compounds. The 2- carbamoy-3-methyiquinoaxaline was found to be the most abun- dant compounds in S. hermanni extracts.” Interestingly,Pung, 2 Ain | Jounal Talon and Complementary Medicine 8 (2018) 24-251 a6 ‘Tablet Proxima composition) fsa cacimbers (mean values standard deviation) spose Most Patan pa a ‘aboiyates et ‘Scape Renan ied) 1020 032 “4700-036 os0- 002 37902038 > Telonace ona ied) 5.10 +029 5520 038 190-001 25105030 » Thetonate oma (€e). 120-008 40702033, 9902027 520 +028 » ote scot (es) saason 3642456 1028 juss 679, 2 Holo eeepc (est) SiAl +060 4571020 40030 430.020 4496 «030 ™ Holo seabr esh) 8576 400, 48.43 £020 5155. 009 2262015, ‘885 £020, : ‘lath tad) 99000 sa20 2072 12s 000, Tiss saa ‘Actinopye aurtanaed) 150-03 es30 2085 tao = 002 1500 = 018 = P ‘quinoxaline derivatives has been reported to involved in reducing, neurological deficits and glia loss after spinal cord injury. These, quinoxaline may contribute to the neuroprotective effects of 5. herman. Based on several findings it may conclude that S. herman are valuable source of functional materials and could be introduced for the preparation of novel functional ingredients in food and biomedicine as a good approach for the treatment and or preven- tion_of many diseases. Furthermore, it can be suggested that have been observed in vitro. Therefore, further research studies are needed in order to investigate S. hermanni biological activities in vivo as well as human subject. 22, T-ananas (local Indonesian name: teripang nanas) and Tenax (local indonesian name: teripang babi, teripang conga, teripang. dduyung) ‘Tananas and T.anax are two sea cucumber species belong tothe Stichopodidae family which found in tropical waters. T ananas (Fig. 2) are known as pineapple sea cucumber or prickly redish ‘These species is considered as commercial sea cucumber species and one of the most popular edible sea cucumber species ‘consumed in China and Southeast Asian countries.” Due to intense ‘commercially exploitation population, these sea cucumber species declined by 80-90% in at least 50% of its range and listed as en- angered species by the International Union for Conservation of Nature. Medicinal value of T: ananas including antioxidant, antiin- flammatory, antitumor, antiproliferative, anticoagulant and ant viral effets have been established. i. 2. Telerta anaes fan Kap, doneti ‘Wu et al. (2010) have isolated novel fucosylated chondroitin, sulfate (Fig. 3) from the body wall ofthe sea cucumber T. ananas, which consisted of N-acetylgalactosamine (GalNAc), glucutanie acid (GicUA), fucose and ester sulfate with about 1:1:1:3.7, respectively. * Fucosylated chondroitin sulfate is a water-soluble depolymerized glycosaminoglycan isolated from echinoderm sea ‘cucumber: Physicochemical of the fucose branch ate differ ac- cording to the sea cucumber species. Anticoagulant activity of the fucosylated chondroitin sulfate from T. ananas as measured by the activated partial thromboplastin time assay varies in proportion to the molecular weight follows a logarithmic-ike function.” The molar ratio for types of fucose branch found in ananas is 25:22:53, {or 3-monosulfate, 4-monosulfate and 2,4-disullate, respectively. Fucose content and composition correlate with anticoagulant ac- tivities of fucosylated chondroitin sulfate: in addition to the composition. More recently, it was demonstrated that anticoagu: lant activity of fucosylated chondroitin sulfate from T. ananas was ‘mediated by inhibition of intrinsic tenase.”' However, fucosylated chondroitin sulfate from sea cucumber T. ananas also activated factor XII which further lead to hypotension when injected intra- vvenously in rats. Interestingly, the activation of factor XIl could be diminished by the low molecular weight fucosylated chondroitin sulfate; suggesting that molecular weight also plays an important role in anticoagulant effect of fucosylated chondroitin sulfate. Not only anticoagulant activity, low molecular weight fragment of {ucosylated chondroitin sulfate from sea cucumber T ananas which prepared by free radical depolymerization bas been demonstrated to inhibit virus HIV replication.” Fucosylated chongtoitin sulfate ‘was effective in blocking laboratory strain HIV-IIIB entry and replication, and inhibiting infection by clinic isolate HIV-Tyqgos and HIV-sre2 Fucosylated chondroitin sulfate might possess potential to be further developed as a novel HIV-1 entry inhibitor for reat- _ment of HIV/AIDS patients, particularly for those infected by T-20- resistant variants. However, further study to elucidate Fucosylated chondroitin sulfate structure and activity relationship will be required in the near future Pucoidan (Fig 3) is sulfated polysaccharide found in brown algae and sea cucumbers. In sea cucumbers, it was first isolated from Iudwigothurea grisea. Recently, low molecular weight fucoidan which composed of a novel tetrafucose repeating units has been isolated from sea cucumber T. ananas by enzymatic degradation. Fucoidan from T ananas was proven to possess a significant su- peroxide radical scavenging activity with an ICs value of T7AG + 0.14 ygiml."” The radical scavenging effect of fucoidan on superoxide radicals improved along with the increasing sulfate content. However, additional 2-O-sulphation in a specific residue increase the radical scavenging effect; suggesting that antioxidant activity of Fucoidan derived from 7. ananas depends on the sulfation pattern not simply on sulfate content. “Triterpene glycosides or also referred as saponins are substances consisting of a sugar moiety attached to a triterpene oF steroid aglycone, These substances are widely distributed in plants, marineun Pong 2 Ain oul of Tonal and Complerentay Medicine 8 (208) 31-251 Ey A (D) e- i ‘oso; ne, ) ~K Fg 2 Functional materal- dered rom sex cucumbers. Frosted hondrtin sults (A Facoidan (8) Holothrin A (Cd Hothrin A (). invertebrates and are characteristic secondary metabolites of echinoderms, octocorals, and sponges Two triterpene glycosides (stichoposide C and stichoposide D) have been isolated from the T.ananas and T. anax.” The structural differences between sticho- poside C and stichoposide D are sugar residue; where stichoposide C has quinovose, and stichoposide D has glucose as the second ‘monosaccharide unit. Stichoposice C showed potent anticancer activity in leukemia cells (HL-60) and mouse subcutaneous tumor cells (CT-26) by inducing apoptosis through the activation of both intrinsic and extrinxic pathway.”* Further, these compounds also showed antitumor activity in vivo which appears to be related to its membranotropic effects," Antifungal activity of stichoposide C from sea cucumber T. anax has also been reported.” Guo et al (2016) has investigated antiobesity effect of triterpene glycosides- enriched extracts from 10 sea cucumbers (including T: ananes). ‘Among them, T- ananas showed potent antiobesity effect through inhibition of pancreatic lipase activity.”® ‘Nutritional value of T. ananas and anax (Fig. 4) were charac~ terized by high protein with low lipid content (Table 1). Bicosa- pentaenoic acid (EPA, C20: Sn-3) was the primary n-3 polyunsaturated fatty acids (PUFA) in 7. ananas (3.92%) and T. anax (3.10%); however, docosahexaenoic acid (DHA, C22: 6n-3) was not detected in both species.”” EPA is the major n3-PUFA in sea cu- ‘umber. Consumption of EPA are associated with decreased risk of coronary heart disease, cancer and wound healing activity.” 23. H.fuscogiva (local Indonesian name: teripang susu) The popular name for H. fuscogiva (Fig. 5) is white teatfish.? FH. fuscogilva is one of the most prized of the commercially sea cu- cumber species in the world.” These species distributed throughout the Indo-Pacific and has a patchy distribution on reef Fig 4 Teenota an om Halmaber, ladon Fig 5. Malaria ue or Kup, IndonesiaPangea, 2 Ain | oun Talon and Complementary Medicine 8 (2018) 241-251 a slopes, seagrass, and lagoons at depth of 3~40 m. Due to their ease of capture, strong demand, improved fishing technologies: 1H. fuscogilva wild populations have been depleted by overfishing. In the recent years, stock enhancement and restocking of these spe- cies have been practiced successfully in Japan and Republic of Kiribati. For commercial purposes, once H, fuscogiva are caught, they usually are gutted, boiled and then dried. In China, HL. fuscogilva are highly valued for their reputed effects as an aphnrodisiac. The contents of saturated fatty acids, monounsaturated fatty acids (MUFA) and PUFA of H.fuscogia were 59.50 %, 32.19 %, 8.32%, respectively (Table 2)-H, fuscogilva have shown higher amounts of SFA and MUFA but lower contents of PUFA. H. fuscogilva extracts possess cytotoxic activity against Candida sp, Leishmania sp. and human supraclavicular lymph node metastasis cells (LoVo}.” Due to the high content of triterpene glycosides, itis assumed that biological activity of H.fuscogilva was due to the presence of ti terpene glycosides. In the nature, sea cucumbers are produced triterpene glycosides to protect them against predators, fungi and ‘ther protists. These cytotoxic effects of H.fuscagitva open the po- tential of H.fuscogiva as antinfective agents, 24, Holothuria atra (local Indonesian name: teripang susu) 1H, atra (Fig, 6) is commonly referred as black sea cucumber or Jollyfsh. Recently, their importance as a source of novel bioactive iz 6. Hotta ara rom Lembeh stat. Indonesia substances is growing rapidly and researchers have revealed that H1-atra originated compounds exhibit various biological activites. {As an example, phosphate-buffeted saline extract of H. atra from ‘Malaysia, exhibited significant antimicrobial activity and inhibited the growth of gram-negative and gram-positive bacteria. Inter- estingly, extracts obtained from the inner part of H. atva, showed ‘tabte2 Fatty ac pros 2) fsa acumber mean values standard deviation Faiyacits__Sihemanai__‘Trananas “Trane Hifscogva —_Hiewcorpiows —_Hsabra——_— Hata A rar 0. nd 003.001 nd na cio na oossoo1 nd 2507 = 004 C00 Ams0i6 2102004 47008 a son 170-2000 er0 4073 Yae2002 332002 133 2002 0372008 exo 0372001 024001 0292001 020001 FA, 4720-126 54702088 OLOO+ 18S —5850~126 sae cut 027001 008-001 0074001 OAR L001 42 «008 ant na Orss001— doreoo!' Oks oor 053 001 CsI ng 3700s) 3402012 3138021 3770120384007 orem ~—a29s0m 535.2015, Coin 1893 + 032 San20n2 436.012 045 £05 xing 5272021 isrs0o1 117-001 ting Sar 048 2554008 136s 001 MUTA 3170 128 ma0+056 3219-040 nan GosnsePA 1312009 St02009 = 12es0o1 —tass01) A200) O47 s0m2) 4142008 ind gaseoie — So02007 — Skss082 2322011 Pin 3254014 163009 1794007 25901 n3in8 O314014 ost sao ase s007 oss . . a References6 Pong 2 Ain oul of Tonal and Complerentay Medicine 8 (208) 31-251 stronger antimicrobial action compared to outer part extracts.” ‘The higher antimicrobial activity of inner part may correlate to the presences of microorganisms which are taken in together with the food substances. Antifungal activity of H. ata from Indonesia has also been reported. The antifungal activity of the above sea cucumber has been determined in Candida albicans by agar well diffusion assays. H, atra extract were not cytotoxic togingiva- derived mesenchymal stem cell in the concentration of < 0.5%: suggesting the extract safety and can be develop for further phase of clinical tral The use of human cells could increase the cor- relation hetween safety studies and clinical trials, an important benefit since conventional animal models of toxicity are not al- ways predictive of human responses. Extract of H.atra were also effective against the Malassezia furfur fungus that causes tinea versicolor.” More recently, antifungal activity of H. ata extracts against various fungal strains such as Trichoderma viride, Asper- sillus niger, Aspergillus flavis, C. albicans and Penicilium chrys- ‘genum has also been reported.” Antifungal activity of H.atra has opened the potency of sea cucumber as antifungal agent and could become the novel alternative solution as ora therapy of candia- asis and others Dhinakaran and his colleagues studied pharmacological effectof H, atra from the Indian ocean.” They demonstrated anti flammatory, analgesic activity, as well as antipyretic activity of H. tra extracts. H. ara extract showed potent inhibitory activity against tumor growth in human cervical cancer ces (Hea) as well as human breast cancer cells (MCF-7).” Extract from H. atra also Showed hepatoprotective activity against thioacetamide-induced liver fibrosis in rats. Subchronic administration of H. otra extract atthe 200 mg/kg to normal rats showed no toxie side effets on the host as evident by the insignificant changes inthe relative weights of the liver, heart kidneys, and spleen, favorable growth rate and survival rate (100%) ofthe animals.” Hepatoprotective and curative effects of the H. tra extract against 712-dimethylbenz [alanthra- cene (DMBA)-induced hepatorenal diseases in. rats has also re- ported.”” DMBA intake produced oxidative stress in liver of rats ‘The increased malondialdehyde concentration level suszests enhanced lipid peroxidation leading to tissue damage and failure of antioxidant defense mechanisms to prevent formation of excessive free radicals. Treatment with H. ata extract priar to or after DMBA intoxication significantly reversed these changes, suggesting that the mechanism of H. aira extract hepatoprotection may be due to its antioxidant effect. High-performance liquid chromatographic (HPLC) analysis of the H.atra extract revealed the presence of phenolic components, such as chlorogenic acid, pyrogallo, rutin, coumaric acd, catechin, and ascorbie acid.” Supporting HPLC analysis, GC-MS results showed thatthe presence of 59 compounds belongs tothe groups of flavonoids, phenolic, texpenoids, saponins, and alkaloids.” It was also reported that epidermal tissues, ovaries, viscera, gut contents Of H.atra contained carotenoid pigments, mycosporine-like amino acids (AAs) and other bioactive compouns such as gadusol. "The presence ofthe active phenolic and carotenoids compounds in the body wall of the H.atra may be due to phenolic and carotenoid-rich materials such as phytoplankton and particles derived from degrading seaweeds which are the main food sources for sea cu- cumbers Collectively. scientific evidence provides information that H. atra could be explored a5 a potential source of high-value bioactive metabolites and could be used in the food as well as pharmaceutical industry, 25. Holothuria leucospilota (local Indonesian name: getah, cera, Jepun, keeling, talengko) Sea cucumber H. leucospilota (Fiz. 7) is a tropical holothurian, Fig. 7 Holos knee. species that is widely distributed in shallow reef areas (eg. reef flats, shallow costal lagoons, seagrass beds) ofthe tropical and sub- tropical Indo-Pacific region, including the Red Sea.”” These sea cucumber species contain’ significant amount of carotenoids including f-carotene, f-echinenone, canthaxanthin, phoenicox- anthin, astaxanthin, lutein, zeaxanthin, diatoxanthin,alloxanthin, ketozeaxanthin, fucoxanthin, fucoxanthinol and idoxanthin.® Ca- rotenoids are linear polyenes that function as light energy har- vesters and antioxidants that inactivate reactive oxygen species (ROS) formed by exposure to light and ai.” Antioxidants may have a positive effect on human health as they can protect human body against damage by ROS, which attack macromolecules such as ‘membrane lipids, proteins and DNA, lead to many health disorders such as diabetes mellitus, neurodegenerative and inflammatory diseases with severe tissue injuries." It has been demonstrated that Holothuria leucospiota extract and their bioactive compounds hhave potential antioxidant activity. Antioxidant activity of H. leucospilota-derived carotenoids have been determined by various methods such as 1.1-diphenyi-2-pierylhydrazyl (DPPH), linoleic acid ree radical scavenging as well as f-carotene bleaching assays ‘Many bioactive metabolites have been characterized for the protection against some form of cancer cells. Sea cucumber H. leucospilota have also been considered as potential sources of cytotoxic compounds for such observations and much research hhave been conducted. Organic extract of H.leucospifota were found to be cytotoxic against Hela cells with ICsp values of 50 yim." Further, crude triterpene glycosides from H. leucospilota found to be cytotoxic against human lung carcinomana cells (A549) and skin ‘melanoma cells (BI6F10) with an IC3q values of 6 yl and 10 jg) ml. respectively "In addition, triterpene glycosides from the sea cucumber H. leucospilota has been isolated, namely leucospilota- side A, leucospilotaside B, leucospilotaside C. echinoside B, hol- othurin A, holotiurin 8, holothurin Ba" Itwas reported that sea cucumber triterpene glycosides exhibit pronounced anticancer ef- feets by direct interaction with tumor cells in the sub-cytotoxic range of concentration.” 26. Holothuria scabra (local Indonesian name: teripang pasir, uti, gosok, kariboa) 4H, scabra (Pg. 8)is also known as sandtish. Several numbers of studies have reported antimicrobial effect of H. scabra extracts Extracts of H,scabra were active against bacteria such as Aeromonas hrydropila, Escherichia coli, Enterococcus sp. Kibsilla pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Staphylococcus aureus and Vibrio harvey, and fih-borne mold, Aspergillus sp. Later, the Possible involvement of T-antigen binding lectin in HL scabraPung, 2 Ain | Jounal Talon and Complementary Medicine 8 (2018) 24-251 us Fie Hoth sobre antibacterial activity was demonstrated.” The expression of ‘defensive lectin is induced by bacterial challenge, wherein cell wall lycoconjugates of bacteria are involved in ‘lectin induction, Further, lectin also showed strong broad spectrum antibacterial activity, ‘Two new triterpene glycosides isolated from H. scabra collected from the South China Sea, scabraside A and B showed strong, cytotoxicity against HL-60, MOLT-4, AS49, and BEL-7402 cells. Scabraside A and B exhibit the same common structural features, ice, the presence of 12- and 17-hydroxy groups in the holostane- {ype triterpene aglycone with a 9 (11)-ene bond, but are different in the side chains ofthe triterpene aglycone.” Scabraside D which also a novel triterpene glycosides from H. scabra showed potent inhibitory activity against P-288, AS49, gastric cancer (MKN-28), snuman colon cancer (HCTII6), and MCF-7 cells with an ICsq values (0f0.96 umol/l, 172 pmol). 1.27 uno, 63 mol/l, and 1.80 umol/l, respectively. ” Subsequently, it was demonstrated that Scabraside induced disruption in mitochondria through the alteration of ‘Bax/Bel-2 ratio which stimulates the intrinsic apoptotic pathway. Disurption of mitochondria lead to release of cytochrome C and {urther stimulates caspase-9 and induced caspase-3 activation.” In addition, two other new triterpene glycosides, namely hol- ‘othurin A; and Ay (Fig. 3) were isolated from the methanol extract of H.scabra from Vietnam, Both saponins were found to be strongly ‘ytotoxic to human epidermoid carcinoma and human hepatoma (HepG2) cells with ICsp of 0.87 and 0.32 yigml. or holothurin As) and of L12 and 0.57 ygiml (For holothurin Aj), respectively.” [Aqueous extract of H. scabra were not active against in. human cervical cancer (C33 A) and A549 cells.” Several reviews suggested that the bioactivity of sea cucumber triterpene glycosides is a resultant of its strong membranolytic activity. The membranolytic activity is a function ofthe structural feature ofthe glycoside.” 2.7. A mauritiana (Indonesia local name: teripang buntal, eripang, ballang ul) ‘A. mauritiana commonly known as Surf Redfish, the body of this sea cucumber arched on the upper side and flat on the lower side ‘A. mauritiana is found in sandy ares, sea grasses, and sand lagoons in the coral reefs, These species i distributed up to 30 m deep, but abundantly found in between 5~10 m deep. A. mauritiana can grow toa length of 25 cm.” These species is amongst the most wide- spread holothurians, highly valued, in great demand and harvest in large number. Adults’ A. mauritiana are dried and processesed for their meat which is exported primarily to East Asia countries, The ‘meat ofA. mauritiana is high in protein, ow in fatand believed tobe aphrodisiac: ‘According to the study carried by Wen et al. (2010) on the chemical composition of composition of A. mauritiana, the major components is protein (63.30% =: 0.43%) with low percentage of lipid (140% + 002%). High protein content of A. mauritiana can be hhydrolized to obtain bioactive peptides.” The content of essential amino acids (EAA) in A. mauritiana was lower than non-essential amino acids (NEA) with the ratio of EAA: NEAA are (0.44 = 0.01.” Glycine is the most abundant amino acids in these species which constituted around 11 % of the total amino acids, followed by glutamic acd and proline. Glycine is used to nelp create muscle tissue and convert glucose into energy, itis also essential to ‘maintain healthy central nervous and digestive systems, and recently has been shown to provide protection from some types of cancer and antioxidant activities." It has been found also that slvcine reduced serum total cholesterol level. On the other side lysine: arginine ratio was lows it has been reported that this low ratio reduced concentrations of cholesterol in the serum and aorta and provide hypocholesterolemic effects. 3, Potential of sea cucumbers as functional ingredients in pharmaceuticals and foods industry ‘The latest trend in food and pharmaceutical sector has been the recovery of functional ingredients from marine resources. Itis well ‘known that consumption of marine foods rich in functional in- gredients beyond meeting basic nutritional needs, is also funda- mental for diseases risk reduction and health promotion (Fi. 9) The composition of different sea cucumbers described in the pre- vious section showed that these organisms can be interesting. natural sources of functional ingredients. In general, sea cucumber provide scientific evidences that sea cucumber contain multitude biologically active materials that provide health benefieffets such ‘Sea cucumber, from a nutritional point of view, is an ideal tonic food with medicinal value, as it contains a and 2 Jowerilevelof lipid:thanimostotherfoods."The main part of the described sea cucumber presents a ‘GIRS! The body wall of sea cucumber, which consists of insol- uble collagen, has been used as a nutrient supplement of hemato- sgenesis. The use of commercial enzymes to produce collagen from sea cucumbers appears to be a feasible process to convert an under- utilized species to a more useful product that contains a functional ingredient for the food and pharmaceutical industry. However, further studies are needed to isolate and identify the specific peptides and or amino acid sequences in sea cucumber collagen hydrolysates with functional activity for potential utilization in {ood and pharmaceutical production. Bioactive peptides are specific protein fragments that have a positive impact on a body's function ‘or condition and may ultimately influence human health. Bioactive peptides must be provided by a safe, reliable, and consistent oralus Pong 2 Ain oul of Tonal and Complerentay Medicine 8 (208) 31-251 Protection rom many diseases (ie neurodegenerative, diabetes melitus, cancer, hepatocenal diseases® inflammatory diseases) (ie S00. aH car) enti propagation of oxidative sess Protect macromoeces > [le'BNA prow, bach anton’ oe Fig. 9. Bologal acvies fsa cucumber derived oaive mar. ‘Another class of compounds of great interest that are common, in sea cucumbers described in this article is sulfated poly- saccharides. As an example, T. ananas contains fucoidan, and Inthe Further study, novel extraction and separation techniques, such as supercritical Op extraction, uitrasonic-aided extraction and membrane sepa- ration technology can be applied in development of sulfated poly- saceharides from sea cucumbers. In addition, PUFA probably the Substances that have attracted major interest since PUFA content in sea cucumber are varied. As an example, sea cucumber species from abyssal have shown higher amounts of SFA and MUFA but lower contents of PUFA compared to the tropical and temperate regions. Variations of fatty acid components among species of different sea cucumbers as well as from different regions might be linked to factors such as diet, natural habitat climatic conditions and post- hharvest processing regimes, especially the drying temperature. ‘The other compounds of great importance derived from sea ‘cucumber described in this work are triterpene glyco: i sive research has revealed that the sea cucumbers pounds exhibit wide spectrum of biological activities such as Linear withthe increased understanding of the health beneficial properties of sea eucumbers-derived compounds, considerable ef- forts have been exerted in discovering. more direct therapeutic- related food and pharmaceutical applications. but. despite high expectations, no commercially successful product ranges have yet been developed utilizing sea cucumber derived compounds tar- geting optimum health and nutrition for human. Up to now, there is ‘very few sea cucumbers-based products canbe seen covering food and pharmaceuticals market. However, s functional food product inthe market, sea cucumber is not up to the expectation. Fortii- cation of food products having higher consumer acceptance with sea cucumber health benefit effects and natural sources would provide an opportune approach to popularize sea cucumbers based products among consumers. Sea cucumber based-functonal foods present major challengesin the fod industry as they appear to bea ew and unfamiliar territory for product developers in marketing, and developing business strategies. Translating scientific advances Sand nutritional innovations into consumer products is a costly and complex process. Sound science must underlie the development, ‘marketing, and regulation of these new functional foods to gain success.” The ineeasing awareness of problems isthe skepticism of consumers of sea cucumbers. Hence, successful functional foods development is therefore a question of combining nutritional! medical insight, technological capabilites and a thorough under- standing of consumers. Importantly, when considering sea cucumbers sustainability fr foods and pharmaceuticals commercialization, these organisms are susceptible to overexplotation due to ther late maturity, density- dependent reproduction, and low rates of recruitment. Further- more, the high value of some species, the ease with which such shallow water forms can be harvested, and ther vulnerable nature due to their biology, population dynamics and habitat preferencesPangea, 2 Ain | oun Talon and Complementary Medicine 8 (2018) 241-251 20 ‘able Heavy metas content in ea cacumbe Ses cucumber peces ‘ean concentration of heney mee ie a ie oe c Mn ie ‘Sherman Ostman 003 mats 3500 make 3.00 make leone 810 male a Tana O24 mele 2a male 1522 a 134 ma aimee 565 mle = Taner 019 male ot als 295 male 095 mals a6 male AD mls 2 H pesca 69 make io mats 11.00 make 5700 maiko 130mahe 9.40 mths _ Hence 03 mele 005 gle 1027 mais 87 ale 155 mais 228 mele a ‘A meuriiena nd nd 5230 malt S110 alk nd 585 mas id all contribute to the overexploitation and collapse of fisheries that have been reported in several areas in Indonesia. During these yyeats, the shortage of sea cucumber seed was a bottleneck for ‘developing aquaculture. The potential area for sea farming of sea ‘cucumber in Indonesia is around 720.500 ha: however, only afew areas are used, eg, in East Java, West Nusa Tenggara, North Sula- ‘esi, Central Sulawesi, South-East Sulawesi, Molucca and Papua. Supporting aquaculture for sea cucumber based functional foods development, Research Center for Oceanography-LIPI currently developing research on sea cucumber aquaculture in Marine Bio- industry Implementation Unit, Research Center of Oceanography LIP Mataram* 4. Toxic materials in sea cucumbers ‘The ocean is vulnerable to human influences such as pollutions and toxic chemicals, Toxic chemicals released to the environment {rom point sources such as industrial and municipal discharges and from nonpoint sources such as agricultural runoff and atmospheric deposition have contaminated surface waters and their sediments and then flow to the ocean.’ One of the greatest problems with many of these contaminants is their tendency to build up in the bodies of aquatic organisms ia increasingly high levels. Uptake of a toxin from the water and from eating other contaminated organ- {sms s called bioaccumulation. Bioaccumulation of heavy metals in sea food poses a significant human health concern. Increased levels of heavy metals in seafood may constitute a food safety risk. Several cases of human disease. disorders, mal- function and malformation of organs due to metal toxicity have been reported. Heavy metals are accumulated in vital organs inthe ‘human body such asthe kidneys, bones and liver and are associated ‘with numerous serious health disorders. In addition, individual _metals exhibit specifi signs of their toxicity.” For example, Arsenic (As), is best known as a poison and can generate both acute and conic toxicity. Ingestion of As may induce peripheral vascular liseases and skin disease such as hyperkeratosis. Intake of lead (Pb) and cadmium (Cd) can cause kidney damage; with long-term ‘exposure to Cd may cause skeletal damage, which may result in fractures.” Therefore, safety of seafood is becoming of great concern in seafood consumption. n order to develop sea cucumber as functional foods, itis very important to estimate the potential land real risk for human health derived from sea cucumber con- sumption, heavy metals concentrations must be characterized. “Table 3 summarized heavy metals content in sea cucumber. Up tonow, there are no international (Codex Alimentarius 1995) as well as local standards (Ministry of Health, Republic of Indonesia) set as limits of Pb, Cd, 2n, Cu, Cr, Mn, As, Hg and Al specifically for Echinodermata, However, considering the ‘maximum limit for Pb in other marine products (Le-ish, molluscs, shellfish and crustaceans) is 0.3 mg/kg; Pb concentrations in some sea cucumber are acceptable. European Comission set the lowest ‘maximum limit set for Cd is 0.05 mg/kg for unspecified fish (EC 2008). Cd levels obtained in sea cucumbers presented inthis study ‘were below that value. In comparison with the level of concern set {or similar marine products, the concentration of heavy metals in sea cucumber were generally below the level af concern of inter- national standards; however, continuous care must be taken t0 ‘monitor the heavy metal levels in sea cucumber. 5. Conclusions ‘Sea cucumbers showed high potential as natural ingredients in foods as well as pharmaceutical industry; hence, these marine or- ganisms can be used maximally in the research areas of new drug. development. However, understanding the specific structures and bioactvities relationship of marine algal fibers is still a great challenge, further there is considerable gap in this area compared ‘with isolation rate of new compounds. Adequate clinical trials are needed in drug development of sea cucumber-derived bioactive ‘materials. More importantly, once their biological activities and health benefit effects are demonstrated, new aspects needs to be address such as culture of sea cucumber, production of functional ingredients at industrial scale, extraction and purification of func- tional ingredients, and scientifically demonstrated health properties. Conflict of interest ‘The authors declates no conflict of interest. Acknowledgement This research is supported by Thematic Research Program Research Center for Oceanography, Indonesian Institute of Sci cences-LIPI. The authors would like to thank Ismiliana Wirawati, Ucu Yanu Arbi, Indra bayu Vimono, and Pradina Purwati for providing, sea cucumber photographs. References 1, Pha ciuetes Ae Inthe search of ew funesonl food ingress ‘fom algae Trends Fd Se Tecmol 20081513138. 2 Frets AC. Rodrigues, Rocka Santos TA. Gomes AM, Duarte AC. Marine Dotechology advances towards applications in new facta Tos. Bo {ect fat 20122306) 1900" 1513 3, Pangesat Kim SK Bgl stvtes an health Bene es of natural Pamantscana Ritmo unease eas arieaarerase 4 Sram Y, Zhang W. Franco Ciscover of nove saponins rom the visera of the sea cucumber Holohura lesson Mar Dag. 201412(9)2653 2667 5. Parwat PTerpang Innes omposis jens dan seaah perkanan, sora 3005220191118 66 Wireat Pura. Rarely reported species of Indonesian sea cucumbers. ‘Mar Resour dons. 2012371923, 7. Dalla § Rahman MA. Yusot FNL Trends, prospects and ullzatons of sa ‘icumber Asher a Malaysia J Adv rc Enon Egy (JAAEE) 20163) tia 116 8. Kim SC Himaya 5. Ttepene gloss from see cucumbers ad thei bio- Topical actives, Ad Foo Nu ex 2012365:297- 319, 9. Zaha RM, Zaria ZAD,YusolN, Mustapha NM, Abdullah MNEL Sea cucumber (Stchopas hens) hased Iydzogel to went bara wounds at J Biomed ‘Mater es Part 8 App Bima 2011-98 1):30- 37.a0 Pung 2 Ain oul of Tada and Complementary Meine (208) 31-251 10. Masre SF Vip GW, Siaiuden Ghazal FC. Quantitative analysis of suphated ‘Syeasaminggljeas content of Maajlan sea cucumber Suchopus herman nd Suchopus vats. Nar ted Res, 2012267584589. 11 Mase sip GW, Szajudcen K Chal FC Wound healing activity of total sulfated syesaminglvean (GAG) from Stchopus vastus and tichopus he mann integument tissue as ne | Mol Me 2010364953. 12, Arundina Yl, SoesawatP, Daman DW, Manaran The ef of solden sea cucumber extract (Sichopus ermani) on the number of ya phocytes during the healing process of faumatie lee war fas Ol Focous Dent) (aj Reoktere Cig) 2015-48(2 100-103 18, Arundin I SdarataK,SetabudiH, Atani MD. Coden sea cucumbers (St ‘hopes herman) 3 growth factors of stem el me Dot Md ee, 20163 14 Pate A Jamal SMS, Jaa Abdullah JM. The effec of water extract of ‘en covumber Siehopes Vanegate on fat apna stocyee coll nes Ca ‘Newbie 2012:31) 15, Paar Apt amalullal Mss, Abdullah JM The ody wall rude exact of ‘Stehopus vargas promotes par of ace cnruse spinal or injury 19 ‘ats by impraiing motor function and eedaces itamedlry hemorrhage ‘Biomed es 20122238) 16, Yu, Kae G Chang, a. Stractreelciaton of xeon composed of 3 ove teafucose repeating unt fam sea cucumber Thelen ananas. Food (Chom 2014 196(0: 115-112 17, Wu Xu S. Zhao Kang i, Ding H, Physicochemical characteristics and ntengulant activites of low molecular weight factions by fee tad ‘epolymerission of 3 costed chondroitin suite fom sea cic ‘Theteate spans, Food Cen. 2010:1221) 716-723. 1m Wu Mo Kus Zhuo J. Kang H, Ding H. Feeadical depalymerzaton of ‘Syeasaminogyean ‘fom sea cucumber Thelema ananas by hyrogen ‘erode and copes fons. Carb Pom. 2010;303):1116~1124, 48, Huang N Wu MEV. Zhen CB, Zh Zhao |H, Zheng YT. The depalymeraed ucosyated chondrotin sulfate trom sea cucumber potently taht. HIV ‘replication via interfering with is en. Carboy es. 2013:360:64- 68. 20, Lio L Wu BM, Xu Le a Comparison af pysicoemeal carats apd Sagal activites of plysacchaids Hom thee Sea cicumber. Mar Drugs. 20131112)399-417. 21. Gap N LU. Xiao Cet a f-Eliminative depolymerization ofthe fooled hongrotn slate and anccoagulan actives of resulting Fagen. Caro Ina Ply 2015-12707 437 Si al pal Ste tele Aan elie ‘Yun 1, Park 5, shin SW, ta. Stichoposie C induces apoptess through {he generation of cami it feakemia and colorectal cancer cells an Shows Invivo aneurer activity. cn Cancer es 20121821) 3934-598, 24 Park eH Kien CG Stok VA Hak) Reatonstps betwen chem ‘rues and finctons of tsterpene pyeosides lated from sea cucumbers ‘rome chem. 2014:2(77. 25, Kohayash M, Hor M kan Ke aL Marin natural prods. XIE: dstbton (of lanostane-type tierpene ollgnlycsies In tn kinds of ldnawan Sea ‘icimbers. Cher Phar fl. 19 3%9)2282" 2287, 26, Gino lsGa07 Phang saponin enriched ea cocker extract exhib a9 ntbesty effect through inhbton of paneeati pase actiiy aod ope Iason of Lie signaling Pharm Bo 2016:548)1312~1325. ‘Wen Hu, Fans. Chemleal competion and aunitinal quay of ea ck umber. JS food gre 201090 124092474 RamoffiaC Batapene S Bel. Byrne M, Reproductive biology ofthe com- ‘mercial sea cucuner Hoiothura fascogina inthe solomon sands. Mar Be 2000:136665 1085-1056. ‘awrence A AIR Ahmed M. Khalifa, Page. Bloatvy 2s an options value ‘of ses cucumber inthe Egyptian Red Seu Consery Bil 20102041217 225, ‘idawan& Kaan, Azan Y, Fuad M,Serening for antibacterial agents in thre species of ea cucumbers rom coastal areas of Sabah. General Par acl Wes st 1985:26(7)1539~ 1543, ‘31, Pasha KReviant S- Antiungal fect of Scopus herman and Helos alta extrac and its cytotoxicity on gngiva-derived mesenchymal stem cel ‘ene jl Kedotean Ci) 2015:468) 218-23. 32, Mulyai A Alzwivah, Khtimah 5. Atveasancjamurekstakteripangdarah (Holothuria ata eager) terbadap pertumbuhan jamur Malaseia fur Penjebad pans. Pombione 20161. -33 Bhinslarin DL Lipton AP Aotitumor sod Antifungal Activities of Organi Ex ‘ocean Uni ena. 2013141). 185-18 34. Dhinataran Di, Lipton A. Pharmacological potentials of sea cucumber Hot ‘thar tra extracts fom the Tndan ocean. Aslan Bored. Pan aves, 35, Dhinalaran DL Lipton AP. Bioseive eompounds rom Holts aa of indian fccan Springer, 20143107. Esmat AY, Sal MM, Soliman AA, E-Masy KS. Rafa EA Biactve compounds antondane potential. and hepatororciveactviy of sea cucumber (Ha ‘churn era) again haces nomication mate uron, 201320 253267, Dakrry A Fahmy SR, Soliman AM, Mohamed AS. Amer SA. Protective and Ciratve ees of the se camber Hoth stra eatrat against DMBA- Induced epaterenal leases in ats BioMed es In. 2015, 2015. ‘38, hundaranayake W, Des Rocher A Roe of secondary metabolites and pigments BOR BOS 88 ® a7 inthe epidermal tues ripe ovaries, viscera gut contents and diet of he sea (hounber Holos ata Mar Bol 1908133116168, ‘38, Taquet Cau! S. Yasuda, subarsono, Nadaoka K Fist observation of lage grup of Molchralecspioa sea carer jens ata sey Manado (orth ulawes Indonesia. ece-de-mer 2011:31:30-34. 40, Matsuno. Tsushima M, Comparative Dechert studies of earoteois in sea ocumber. Comp Biochem Mysiolgy Pa B Bochmt Mot Bl. 1209;1118): 44, Westra Fangestut RK SIC Blloial acts and potential eat lenetts of sulted patsachanes dened Worm maine algae. Carly Polym 2010841) 14-21 42, Pangestt Re Muriasi T, Putra MY, ea. ree radical scavenging activity of Selected sea cucumber species om tararhdomBok,Indonest. J Tea dover 2) 4, Atunitat OF, Hath RO, Taher M, Dau JM, Mheda MEA, Za Bn wo {ntondant and aniprlieatve actives of three Malaysian Sea cucumber Species fur] St es. 209:373):376~ 387 “44 faharara Amin E VanfocanV. Concomitant use of sa cucumber organic fntact a ratinthecapy on roferaten snd apps ceva (Hea) cll line, 2aedan Jes Med Sc. 2016:18(2) 45, Sofa Parva K Baar Rrachian MA, As. Pattie mechanism for poptesi Inducing properties of erie sapein Isolate from sea cucumbe? (folomuralecorplot) a: an sotdant compound. mon) bate Med Se. dorsi) 180, 46, Sofa Mt Parivar K, aharara. Kerahian MA. Sl Hemolytic and etoronie ‘ropetes of sponia pure from Holos caspases ecimber op Biochem Ma ol. 20143013. 4. aharara Ail E, Mido! NSalek Abdo F. The cjtotoy af daar 2ie poteiatd by ea cucumber saponin ig fesstant B1GF10 melanoma cals {through apoptos indution. Avicenna] Med Biocecnel 2016:3):112. 48, Han HV YH iu BS, Wang XH. Fan OX Leucospltaside CA new sulfated ttespene glycoside from sea caumber Holes Teveespioc. Chi Chem Iau 2008:19(12) 1482" 1484 49, Han HV YH, LL etal A now tterpene gheosid from sea cucumber Hol- ‘thurs evospoia: Chin Chem Let 2007182) 161-164. $50. Han H, Zhang W, YEA, Lu BS, Pan MD, Wang XH. A nove! slated Holestane lyeosde fom sea cucumber Holothuris leucospiota Chem Bodies 2010:77).1764-176, St, Hh He Yanga ¥, Linge al Teterpene gycoids from sea camber Nolohuriaeacosplts chit] Nat Mec. 2008-73) 446—250. 52. AmininD, MenchunsayaEshaginESchenko A Avy 5, Kalin V. Ani ‘ancer activity of ea cicumbertterpenegycosiées. Mar drs. 2015:1303) tat, 53. Abraham). Nagaraan J Shanmugam S. Ancimicoial Substances of Potent ‘omadel portene fom Holohan Spe. 202 S54. Coma Nt, Cosa U, Khan MT antigen nding lectin with ance ‘acti from marine invertebrate sca cucumber (Holothura scab} possible invohement in diferent recognition of bacteria. j Inverter Paley ‘ove. 141- 145, 55. Han H.Y1Y. Xu La M.zhangH.Two new eytototterpene lyosies trom {the sea cacarber Hera scab, Pana Mes, 2003-75 15)1608- 1612 56, Hoa i ng iY Wang XH Pam NX. Tterpene lysis rom sca umber Hoothuria scab with corn att. Chin Herb Med 201243): 157, Assamasipaerk XK, Vania &,Chowwivattanakun ¢. Nobsthan wang Witayachumnanl Sebi D extracted frm Hota ‘scabra induces gpoptess and inhibits growth of human eholangicaeinoma ‘enogzats in mice. Asin Puc J Concer Pes. 2016:1725511- 317. ‘58 Dan NH Van Tha Van Kes, Huong LM Van ah Kim Tw now Itterpene glycosides from the vetamese sea CheumberHolohura sear ‘Arch Pharmac s- 200730(1) 13871301 58, Ahunibt OY, Hoshi RB, Taher M, Daud IM, thea MCA. Zli Bn vito Srtionane and antorleative actives of three Malaysian sea cucumber Species. fu St Res. 2000°373):576~ 357 (0, Kin SX Humaya's.Tterpene gvcesices rom sea cucumbers and thei bio- ogi sci Aa aod ur Re 201265207 319, ‘1, James D. Twenty sea cucumbers om seas around Inia, Nogn ALARM 2001:24142)4" 8. (2, Lovatell A Conand €Advoces in Se Cucumber Aqucatre and Manage ood Agicultre Og 2008. ‘5, angst yu Hinaya 5 Kin SK Optimization of hydrolysis conditions, ‘soliton and Wentcation of neroprtetive pepe derived rom seahorse Hippocampus tamaclats. mio As. 2015:45(2)'369- 31 ‘4, krpodoroa TA. Basiteyn VY. Bis NA, anova TS, Some macronutrient conten in myceia and cre broth of medina mushrooms ulated on Saran four tnt/ Mod Mushrooms 2012142) (65, ljamhan Tarp Lysine: rane rato oa protein nsencescolsteza ‘metabolism, Part t-stafles on sesame proein having low fsne: arginine fa. nan xp Bo 19735(1) 181223. ‘9, Pangstt Kim Ske. Horme once Peptide Sources: Cra Poms ond ‘te Petentl or New Trapt, New York: Wiley: 2013 (67. Zhong. Khan MA Shahi Compestonal characteristics and antioxidant ‘ropetes of fresh and processed sen camber (Cncumar andes) ) Ae Po cham. 2007°55 9): 11881182 (68 Gruner KG European consumers acceptance of functional foods. An Y Acad7, {Pangea 2 Ain | oun Taltionl and Complementary Medicine 8 (2018) 34-251 a Sei, 2010:11901: 165-173. Indiana Lr, jana The ulation of seagras and mactalge substrate for Settlement’ of stadfehHolotuns sabes lve. J Alla Indone. pos t30 6872, opt C, Arena G. Fore M, etal. Heavy metals concentrations in Fs and Shallfsh om easter Mediteranean Sea consumption avs. Food Che Tol 20135833 970 Mada! V, Mdaaa , udal A, Singh Mishra . lec of tie metas on Thum Heath: Ope Natracet J 2010;3'58-99. ‘Cot J, Hae Gr Khan Bet a Determination of tox ean metals a ch odermata and Chordata species fom South Korea Fed Add Contam Pat. 2014'74):295-90. Favezya YH, Januar, Susowati Chana F. Chemical compostion and {any Sd ofc of ime indonesian tea cucumber: Scien Bal May Pat Ftharves!otechnol 201551001727" Cmran NE-SE-S. Nutrtinal value of some Exyptian sea cucumbers. A] Bo teen 20131305), vai MY, lamin SM, Gide YEA. Ai SM, The proximate commpostion and the maton value of some sea ccumber specs intating the Sudanese ted sea Food Si Techn Manes 2015:41(11- 17) Fidewan 8 Hanta Mt Nurzaiah M.Novshuhadaa MS Manis ZF. re ty acids Composition nip extract of several sa ccumbers specs from Maly Int) asc chem Bonforma, 201443204 Haider Mt Sltana Jam Tara O, Azal WA stay on proximate compe ‘ton, amino aid patil, fe ac pale and Some miserlcontents af Species of seu cucumber. JAP) Ain Plat Sc. 201525(1) Wen HC Elemental composition of commercial sea cucumbers (hoothu- ans) Food Ad Contam. 2010: 3)246~252 ass Ml, Mar bin Sj Hussein MA, Hoque Z Detention o seven ea mets in igh specie of Sea Cucumber Set 20143281)