Red Sea Cucumber Research Articles

Simulated Digestion of Red Sea Cucumber (Parastichopus tremulus): A Study of Protein Quality and Antioxidant Activity

Sea cucumbers from unharvested areas, are underutilized resources which may have the potential to become a future food resource. The aim of this study was to evaluate protein quality and investigate the changes in antioxidant activity from frozen and freeze-dried red sea cucumber (Parastichopus tremulus) subjected to digestion, using an in vitro digestion model. P. tremulus constituted high moisture content (90%), comparable protein (4%) and ash (4%) content, and low lipid (1%) content. The biochemical components in freeze-dried samples were largely retained during the freeze-drying process. Frozen samples showed significantly higher (p < 0.05) antioxidant activity compared to freeze-dried samples (calculated on a dry weight basis). The quantity of essential amino acids was high (31%) and corresponds to the Food and Agriculture Organization of the United Nations reference protein. Frozen samples showed an increase in antioxidant activity during digestion measured by three different antioxidant assays. Freeze-dried samples only showed an increase in one of the antioxidant assays. Correlations (p < 0.05) were found between specific free amino acids and antioxidant activity. The amount of free amino acids exceeded the threshold to taste bitter, umami, sour, and sweet flavors. This study showed that P. tremulus constitutes good protein quality, performed antioxidant activity, and has the potential to be used as a flavor-enhancing ingredient.

Regio-specific lipid fingerprinting of edible sea cucumbers using LC/MS

Sea cucumbers are a rich source of bioactive compounds and are gaining popularity as nutrient-rich seafood. They are consumed as a whole organism in Pacific regions. However, limited data are available on the comparison of their lipid composition and nutritional value. In this study, untargeted liquid chromatography/mass spectrometry was applied to comprehensively profile lipids in the skin, meat, and intestinal contents of three color-distinct edible sea cucumbers. Multivariate principal component analysis revealed that the lipid composition of the intestinal contents of red, black, and blue sea cucumbers differs from that of skin, and meats. Polyunsaturated fatty acids (PUFAs) are abundant in the intestinal contents, followed by meats of sea cucumber. Lipid nutritional quality assessments based on fatty acid composition revealed a high P:S ratio, low index of atherogenicity, and high health promotion indices for the intestinal contents of red sea cucumber, suggesting its potential health benefits. In addition, hierarchical cluster analysis revealed that the intestinal contents of sea cucumbers were relatively high in PUFA-enriched phospholipids and lysophospholipids. Ceramides are abundant in black skin, blue meat, and red intestinal content samples. Overall, this study provides the first insights into a comprehensive regio-specific profile of the lipid content of sea cucumbers and their potential use as a source of lipid nutrients in food and nutraceuticals.

Seasonal and taxa impact on edible sea cucumber metabolome as analyzed via a multiplex approach of mass spectroscopy-based metabolomics

Sea cucumbers, belonging to class Holothuroidea, are marine invertebrates with substantial bioactive compounds and nutritive value. This study presents a multiplex metabolomics approach to dissect between winter and summer samples of two Red Sea cucumbers: Holothuria atra and Actinopyga crassa. LC-MS and GC-MS followed by multivariate data analyses were employed for metabolites profiling and seasonal biomarkers assignment for each species. A total of 314 metabolites were annotated in both species belonging to amino acids, sugars, sugar alcohols, acids, triacylglycerides (TAGs), phospholipids, lyso-phospholipids, sphingolipids, phosphatidylcholines, diacylglycerols, different glycolipids, and others, adding to the nutritive value of sea cucumber. GC-MS analysis revealed for amino acids i.e., alanine as the main polar primary metabolite class in sea cucumbers. LC-MS analysis targeting non-polar metabolites revealed TAGs as the main class followed by phospholipids and lyso-phospholipids. Multivariate orthogonal partial least square discriminant analysis (OPLS-DA) revealed that A. crassa is enriched in alanine and lactic acid than H. atra versus higher levels of pyroglutamic acid and ethanolamine and TAGs in the later. Summer A. crassa was distinguished from its winter specimen by its higher TAGs, glycine and glycerol levels versus enrichment of winter samples in lyso-phospholipids, isoleucine, and proline. Such metabolomic study revealed for A. crassa and H. atra richness in nutrient metabolites, alongside collection season effect on their amino acids and lipids profiles.

Effect of live-storage period and temperature on oxygen consumption rate in the cold-water sea cucumber Parastichopus tremulus

Sea cucumbers are of great significance in marine ecosystems all over the world. The red sea cucumber Parastichopus tremulus is also considered commercially interesting but very little information is available on its ecology and physiology. This study aimed at investigating this North Atlantic species` oxygen demand in relation to live-storage period and temperature in an attempt to deliver some physiological puzzle pieces useful in a potential aquacultural context. Oxygen consumption rate (OCR) was measured in P. tremulus stored in a flow-through system for up to one year, and in recirculation systems at different temperatures (4, 8 and 12°C) for up to four weeks. Throughout the experimental period, biometric data were collected from the different treatment groups and a condition index (CI) calculated. The results showed that long-term storage affected OCR and CI, which increased and decreased with time, respectively. Storage at 4°C resulted in significantly lower OCR and higher CI compared to 8 and 12°C after four weeks, suggesting that 4°C may be best for maintaining body mass during short-term storage, whereas storage at 8 and 12°C may require supplementary feeding to avoid biomass loss. Data presented in this study may be useful for a range of applications, for example in a prospective aquaculture setting, for optimizing live seafood storage, but also for impact assessment and management of wild stocks.

Direct assessment of giant red sea cucumber (Apostichopus californicus) sustainability through experimental fisheries

The giant red sea cucumber ( Apostichopus californicus) fishery in BC, Canada, is considered sustainable by managers but not by many harvesters. In 1997, the Kitasoo/Xai'xais First Nation began experimental fishing areas (EFAs) at Tolmie Channel and Laredo Inlet to test the effects of different exploitation rates on sea cucumber populations and determine what harvest levels are sustainable. These EFAs were annually surveyed by dive transects and subsequently harvested from 1998 to 2018. Bayesian surplus production models developed in 2011 indicated that the fishery was sustainable, after which the fishery changed from annual to 3-year rotational harvests. Here, we develop updated models to estimate the sustainability of the fishery using all currently available EFA data, including direct estimates of unfished biomass. Our models suggest that the fishery is likely to persist for 175 years at the current harvest rate annually or rotationally, though such long-term projections are likely inaccurate. We present methodology to generate scientific advice for the process of determining a maximum sustainable exploitation rate in the form of crash rate analysis.

The Effects of Breeding Environment and Food Type on the Survival and Growth of Red Sea Cucumber (Apostichopus japonicus) Larvae

Jwa, M.-S.; Kim, S.-C., and Hong, C.-Y., 2022. The effects of breeding environment and food type on the survival and growth of red sea cucumber (Apostichopus japonicus) larvae. Journal of Coastal Research, 38(3), 578–584. Coconut Creek (Florida), ISSN 0749-0208. This study aimed to investigate the effects of water temperature, salinity, and food type on the survival and growth of red sea cucumber (Apostichopus japonicus) larvae, over 14 days. Early Auricularia larvae (average length 475 ± 10.3 µm) were exposed to four different water temperatures (18, 22, 26, and 30 °C), four different salinities (25, 30, 35, and 40 psu), and four food types (Chaetoceros calcitrans, Pavlova lutheri, Isochrysis galbana, Rhodotorula+effective microorganisms (EM)). Subsequently, the survival rate, larval developmental stage (early Auricularia–Doliolaria), and the required time (days) to arrive at each stage were observed. The experiment was conducted at a private sea cucumber aquaculture farm in Jeju Special Self-Governing Province, and the results were statistically analyzed using SPSS version 22 (SPSS Inc., USA). This research team evaluated the average significance difference between groups, which was determined in p < 0.05, by applying a Duncan test due to variance heteroscedasticity. The results indicate the following optimum rearing conditions of red sea cucumber larvae: water temperature of 22 °C, salinity of 30–31 psu, with Rhodotorula+EM as food. The findings of this study could facilitate increasing seed production of the red sea cucumber A. japonicus.

6-Bromohypaphorine isolated from red sea cucumbers Apostichopus japonicus exhibits potent anticancer activity in A549 cancer cell line

Organisms such as Apostichopus japonicus are a group of marine invertebrates that produce a variety of alkaloids with bioactivities. Many of these alkaloids have been proven to be effective anticancer drugs. In this study, the anticancer activities of 6-bromohypaphorine (6-BHP) isolated from red sea cucumber, A. japonicus, were investigated. The anticancer effects of 6-BHP against tumor cell lines A549, HT-29, B16F10, MCF-7, and Hep3B were evaluated. The structure of 6-BHP was determined by mass spectroscopy (MS), as well as nuclear magnetic resonance spectroscopy (NMR). In the bioactivity assay, 6-BHP attenuated survival rate of A549 cell line with an IC50 value of (197.1 ± 7.4) μmol/L. Among the five cancer cell lines, 6-BHP showed the strongest anticancer activity against A549 than any other cell lines. The results of acridine orange-ethidium bromide and Hoechst 33342 staining showed that 6-bhp treatment mediated the formation of apoptotic body. In addition, 6-BHP down-regulated B-cell lymphoma-extra large (Bcl-xL), while up-regulated both Bcl-2-associated X protein (Bax) and caspase-3 in a dose- and time-dependent manner. We suggest that 6-BHP induces apoptosis via the mitochondria-mediated pathway. This is the first report on the isolation of 6-BHP from A. japonicus.

Immunomodulatory properties of enzymatic extract of Stichopus japonicus on murine splenocytes

Red sea cucumber Stichopus japonicus is an invertebrate animal inhabiting in coasts of Korea, China, and Japan. They are traditionally used for food and medicine and well known for their distinctive biologically and pharmacologically important compounds. We investigated the effect of amyloglucosidase (AMG) enzymatic extracts of S. japonicus (AESJ) on the proliferation and cytokine secretion of murine splenocytes stimulated with concanavalin A (Con A). AESJ enhanced the proliferation of splenocytes and the production of IL-2 (Th1 cytokine), IL-1β (Th1 promoting cytokine), and IL-4, IL-10 (Th2 type cytokines) when treated alone. However, under Con A stimulation, AESJ suppressed the proliferation of splenocytes, attenuated the secretion of IL-2, IL-4, IL-10, and enhanced IL-1β secretion. These results suggest that AESJ exhibits immunomodulatory effect by moderating the proliferation of splenocytes and the secretion of IL-2, IL-1β, IL-4, and IL-10 differently depending on the absence and presence of Con A stimulation. These data evidence the immunomodulatory potential of AESJ, which can be further developed into a functional food mediating homeostasis.

Controlled spawning and rearing of the sea cucumber, Parastichopus tremulus

AbstractThe red sea cucumber, Parastichopus tremulus, a cold water species with commercial potential, has recently attracted attention for wild harvest as well as for potential use in integrated multi‐trophic aquaculture in Scandinavian countries. Overharvesting has put natural stocks of sea cucumbers at risk in several countries. Our goal was to develop a rearing protocol for P. tremulus to enable sustainable production of this species. This study presents results from spawning and larval rearing conducted in both Norway (NO) and Sweden (SWE) during May–August 2019. We describe spawning induction and behavior, fertilization success, embryonic and auricularia larval development rate for this species under laboratory conditions. The larvae were fed a mixture of three species of live microalgae (SWE) and algal paste (NO). Larval development rate and survival were monitored at four different temperatures (7, 10, 13, and 16°C). Results showed faster development with increasing temperature. Daily food consumption rate was highest at the highest temperature. The combined effects of temperature and food availability on survival were investigated for the same four temperatures and three different feed concentrations. Only food availability affected the mortality rate, with the highest mortality in the low feeding regime of 1,000–2,000 cells ml−1 day−1.

Sulfated Triterpene Glycosides from the Saudi Red Sea Cucumber Holothuria atra with Antioxidant and Cytotoxic Activities

Sulfated Triterpene Glycosides from the Saudi Red Sea Cucumber Holothuria atra with Antioxidant and Cytotoxic Activities

Holospiniferoside: A New Antitumor Cerebroside from The Red Sea Cucumber Holothuria spinifera: In Vitro and In Silico Studies.

Chemical investigation of the methanolic extract of the Red Sea cucumber Holothuria spinifera led to the isolation of a new cerebroside, holospiniferoside (1), together with thymidine (2), methyl-α-d-glucopyranoside (3), a new triacylglycerol (4), and cholesterol (5). Their chemical structures were established by NMR and mass spectrometric analysis, including gas chromatography–mass spectrometry (GC–MS) and high-resolution mass spectrometry (HRMS). All the isolated compounds are reported in this species for the first time. Moreover, compound 1 exhibited promising in vitro antiproliferative effect on the human breast cancer cell line (MCF-7) with IC50 of 20.6 µM compared to the IC50 of 15.3 µM for the drug cisplatin. To predict the possible mechanism underlying the cytotoxicity of compound 1, a docking study was performed to elucidate its binding interactions with the active site of the protein Mdm2–p53. Compound 1 displayed an apoptotic activity via strong interaction with the active site of the target protein. This study highlights the importance of marine natural products in the design of new anticancer agents.

Inhibition of Lipid Accumulation in 3T3-L1 Adipocytes by Different Enzymatic Hydrolysates of Dried Red Sea Cucumber Stichopus japonicus

Inhibition of Lipid Accumulation in 3T3-L1 Adipocytes by Different Enzymatic Hydrolysates of Dried Red Sea Cucumber Stichopus japonicus

New Cytotoxic Cerebrosides from the Red Sea Cucumber Holothuria spinifera Supported by In-Silico Studies.

Bioactivity-guided fractionation of a methanolic extract of the Red Sea cucumber Holothuria spinifera and LC-HRESIMS-assisted dereplication resulted in the isolation of four compounds, three new cerebrosides, spiniferosides A (1), B (2), and C (3), and cholesterol sulfate (4). The chemical structures of the isolated compounds were established on the basis of their 1D NMR and HRMS spectral data. Metabolic profiling of the H. spinifera extract indicated the presence of diverse secondary metabolites, mostly hydroxy fatty acids, diterpenes, triterpenes, and cerebrosides. The isolated compounds were tested for their in vitro cytotoxicities against the breast adenocarcinoma MCF-7 cell line. Compounds 1, 2, 3, and 4 displayed promising cytotoxic activities against MCF-7 cells, with IC50 values of 13.83, 8.13, 8.27, and 35.56 µM, respectively, compared to that of the standard drug doxorubicin (IC50 8.64 µM). Additionally, docking studies were performed for compounds 1, 2, 3, and 4 to elucidate their binding interactions with the active site of the SET protein, an inhibitor of protein phosphatase 2A (PP2A), which could explain their cytotoxic activity. This study highlights the important role of these metabolites in the defense mechanism of the sea cucumber against fouling organisms and the potential uses of these active molecules in the design of new anticancer agents.

Characterization and anti-tumor activity of saponin-rich fractions of South Korean sea cucumbers (Apostichopus japonicus).

In this study, the saponin-rich fractions of five individual (two Red and three Black) sea cucumbers (Apostichopus japonicus) in South Korea were investigated for their antiproliferative effect against HL-60, B16F10, MCF-7, and Hep3B tumor cell lines. The red sea cucumber saponin-rich fraction (SSC) from Jeju Island (JRe) decreased the growth of HL-60 with an IC50 value of 23.55 ± 3.40μg/mL, which represented the strongest anticancer activity among the extracts. Further, SSC downregulated B-cell lymphoma extra-large (Bcl-xL), while upregulating, to different degrees, Bcl-2-associated X protein (Bax), caspase-9, caspase-3, PARP cleavage, and apoptotic bodies in cancer cells. Evidence for SSC inducing apoptosis via the mitochondria-mediated pathway was found. The contents of SSCs were determined using ultra high-performance liquid chromatography coupled with a quadrupole orbitrap mass spectrometry to comparatively evaluate the regional influence. In West Sea, the total SSC content of A. japonicus was 15.5mg/g, representing the highest content, while A. japonicus in the South Sea yielded the lowest content at 8mg/g. The major saponin constituent in SSC was identified as Holotoxin A1, which may the anti-tumor compound in A. japonicus.

Red Sea Cucumber (Stichopus japonicus) Suppresses Cancer Progression by Promoting the ROS-Me diated Inhibition of the MAPK Pathway

Red Sea Cucumber (Stichopus japonicus) Suppresses Cancer Progression by Promoting the ROS-Me diated Inhibition of the MAPK Pathway

Antioxidant and anti-melanogenic activities of ultrasonic extract from Stichopus japonicus

Objective: To investigate the antioxidant and anti-melanogenesis activities of an ultrasonic extract of red sea cucumber, Stichopus japonicus, collected from Jeju Island. Methods: Antioxidant activity experiments were assessed by an electron spin resonance system and a cellular model of immortalized human keratinocytes (HaCaT) to determine its radical scavenging activity and protective effects against 2,2’-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress. Anti-melanogenic activity of the ultrasonic extract of red sea cucumber was also examined using the melanoma cell model B16F10 and mushroom tyrosinase. Following the induction by α-melanocyte-stimulating hormone, the effects of the ultrasonic extract of red sea cucumber on intracellular tyrosinase activity, melanin content and the melanogenic protein expression of microphthalmia-associated transcription factor, tyrosinase, and tyrosinase-related proteins (TRP-1, and TRP-2) were examined. Results: The ultrasonic extract of red sea cucumber significantly scavenged 2,2-diphenyl-1-picrylhydrazyl and alkyl radicals [IC50: (0.924±0.035) and (0.327±0.006) mg/mL, respectively], as well as showed a protective effect against oxidative stress and attenuated generation of intracellular reactive oxygen species on AAPH- induced HaCaT cells, with no cytotoxicity (12.5-400 μg/mL). The ultrasonic extract of red sea cucumber also exhibited a tyrosinase inhibitory effect [IC50: (2.750±0.006) mg/mL]. On α-melanocyte-stimulating hormone-stimulated B16F10 melanoma cells, the ultrasonic extract of red sea cucumber (25-200 μg/mL) significantly inhibited not only melanin synthesis and tyrosinase activity, but also protein expressions of microphthalmia-associated transcriptional factor, tyrosinase, TRP-1, and TRP-2. Conclusions: The ultrasonic extract of red sea cucumber shows antioxidant and anti-melanogenic potential and may be a natural candidate for anti-aging as well as a whitening agent in the cosmeceuticals industry.

Integrated multi-trophic aquaculture of red drum (Sciaenops ocellatus) and sea cucumber (Holothuria scabra): Assessing bioremediation and life-cycle impacts

Environmental sustainability of aquaculture is a complex issue involving effects at local (e.g. benthic deterioration), regional (e.g. eutrophication) and global (e.g. catches for feed production) scales as a consequence of farming operations (e.g. waste emissions) and industrial processes involved in the product value chain. Integrating these effects using a holistic and multi-scale framework is essential to assess the environmental sustainability of innovative production systems such as Integrated Multi-Trophic Aquaculture (IMTA), in which organisms of different trophic levels are co-cultured on the same farm to minimize aquaculture waste. The environmental performances of theoretical production scenarios of red drum (Sciaenops ocellatus) sea cage monoculture and an open-water IMTA co-culturing of red drum and sea cucumber (Holothuria scabra) were assessed with mathematical models at local and global scales. First, the particulate waste bioremediation potential of sea cucumber production was estimated using an individual-based bioenergetic model. Second, environmental impacts of the monoculture and the IMTA systems were estimated and compared using life cycle assessment (LCA), calculated per kg of edible protein and t of product, including uncertainty analysis. Given the current limits to stocking density observed for sea cucumbers, its co-culture in sea cages suspended beneath finfish nets may decrease slightly (by 0.73%) farm net particulate waste load and benthic impact. The monoculture and IMTA showed little difference in impact because of the large difference in production scales of finfish and sea cucumber species. Removing 100% of finfish feces particulate waste requires cultivating sea cucumber at scale similar to that of finfish (1.3 kg of sea cucumber per kg of finfish). Nonetheless, LCA showed trends in IMTA performance: lower eutrophication impact and net primary production use but higher cumulative energy demand and climate change impacts, generating an impact transfer between categories. Intensification of sea cucumber culture could increase local and global environmental benefits, but further research is necessary to design rearing units that can optimize production and/or bioremediation and that can be practically integrated into existing finfish monoculture units. The methodology defined here can be a powerful tool to predict the magnitude of environmental benefits that can be expected from new and complex production systems and to show potential impact transfer between spatial scales. We recommend applying it to other IMTA systems and species associations and including socio-economic criteria to fully assess the sustainability of future seafood production systems.

The complete mitochondrial genomes from three body color variants of sea cucumbers, Apostichopus japonicus (Selenka, 1867)

To study the intraspecific variation of the sea cucumber, Apostichopus japonicus, we sequenced three complete mitochondrial genomes, especially from different body color variants (green, black, and red) of Apostichopus japonicus in Geomum-do, Republic of Korea. The full lengths of green, black, and red sea cucumber mitochondrial genomes are 16,106 bp, 16,096 bp, and 16,107 bp, respectively. They all contain 37 genes (13 protein-coding genes, 2 rRNAs, and 22 tRNAs), and have an average 38.0% of GC ratio. Two hundred thirty eight single nucleotide polymorphisms and 26 insertion and deletions were identified among three sea cucumber mitochondrial genomes. The phylogenetic analysis of A. japonicus with additional 14 available A. japonicus and two Parastichopus mitochondrial genomes were used as outgroups shows somewhat distinct three clades with body color patterns. Furthermore, there is no clear phylogenetic separation among Korean, Chinese, and Japanese individuals. These data will provide new insights and to facilitate the rapid growth of the sea cucumber aquaculture industry and its breeding programs.

Whitening Effect and Skin Regeneration Effect of Red Sea Cucumber Extract

Whitening Effect and Skin Regeneration Effect of Red Sea Cucumber Extract

염증성 사이토카인의 발현 억제를 통한 홍해삼 추출물의 항염증효과 및 아토피 개선효과

The red sea cucumber (RSC), Apostichopus japonicus, is a species of sea cucumber in the family Stichopodidae. It is widely distributed in China, Japan and Korea. Tumor necrosis factor-α (TNF-α) and Interferon (IFN-γ) induce inflammatory cytokines such as Thymus and activation-regulated chemokine (TARC/CCL17), Cutaneous T-cellattracting chemokine (CTACK/CCL27), interleukin-1β (IL-1β), IL-6 and IL-8. These cytokines are increased in atopic dermatitis condition. In this study, we investigated the antiinflammatory and skin regeneration effects by RCS. To investigate cytotoxic effects in HaCaT cells, we performed MTT assay after treatment with RSC extracted by 70% EtOH. As a result, RSC extracts did not have cytotoxic effect in HaCaT cells. Also, mRNA expression levels of CCL17, CCL27, IL-β, IL-6 and IL-8 genes induced by TNF-α/IFN-γ were decreased by RSC extracts. Moreover, we confirmed the release of CCL17 and CCL27 protein through ELISA assay. Wound healing assay and cell cycle assay were performed to identify cell regeneration effect of RSC extracts. These results demonstrated the prevention and treatment effects of RSC on skin inflammatory diseases such as atopic dermatitis.