Sea cucumbers are marine invertebrates of the phylum of Echinodermata that have been used in Asian traditional medicine since ancient times. This study was conducted to investigate the antioxidant and cytotoxic properties of aqueous and organic extracts from two sea cucumber species, Holothuria edulis Lesson (Holothuriidae) and Stichopus horrens Selenka (Stichopodidae). Antioxidant activities of the extracts were evaluated by DPPH· and β-carotene bleaching assays, while MTT and trypan blue exclusion assays were used to demonstrate the cytotoxic effects of the extracts against two human cancer cell lines, non-small cell lung cancer cells (A549) and esophageal cancer cells (TE1). The results showed that both aqueous and organic extracts of H. edulis were able to scavenge DPH radical (IC50 at 2.04 mg/ml and 8.73 mg/ml, respectively). Aqueous and organic extracts of S. horrens inhibited 79.62% and 46.66% of β-carotene oxidation by linoleate free radical. On the other hand, the organic extract of S. horrens exhibited the highest cytotoxic effects against A549 and TE1 cancer cells giving IC50 at 15.5 and 4.0 μg/ml, respectively. In conclusion, the present study revealed that H. edulis and S. horrens contain promising levels of antioxidant and cytotoxic natural products that might be used for cancer prevention and treatment.
The calcareous rings of two species of local sea cucumbers, Stichopus hermanni Semper and Holothuria atra Jaeger, were located, dissected, exposed and recorded. The calcareous rings of both species each composed of ten plates knit together by connective tissues. The radial plate of H. atra was a square with a notch at the anterior part whereas in S. hermanni it was squarish consisting of 4 ridges in the anterior part with a notch at the posterior part. The interradial plates of both species were smaller than the radial plates. A ridge was present at the anterior part of the interradial plate in H. atra. In S. hermanii, the ridge at the anterior part of the interradial was thin and prominent. Under the scanning electron microscope the calcareous rings from the two species exhibited a mosaic of small, numerous spicules bound to each other.
Atratoxin B1 which was extracted from a local sea cucumber, Holothuria atra is a potential antifungal agent against dermatophytes. Exposure of Microsporum canis mycelia inoculated into Sabouraud glucose agar medium to the 10 mg/ml atratoxin B1 solution, resulted in complete suppression of the fungal growth. Prolonged exposure to the atratoxin B] (72 hours) resulted with necrosis in a substantial portion of the existing hyphae. The main anomalies noted were the abnormally shaped hyphae and the alterations in its morphology and cytology.
Atratoksin B] yang diekstrak daripada timun laut tempatan, Holothuria atra, adalah suatu agen antikulat yang berpotensi terhadap dermatofit. Pendedahan miselium Microsporum canis yang diinokulat ke dalam medium agar glukosa Sabouraud kepada larutan atratoksin B] berkepekatan 10 mg/ml, mengakibatkan penindasan lengkap dalam pertumbuhannya. Pendedahan yang lebih lama kepada larutan atratoksin B] (72 jam) mengakibatkan nekrosis berlaku pada sebahagian besar hifanya. Keganjilan yang jelas berlaku adalah ketidaknormalan pada bentuk hifanya dan juga pengubahsuaian yang terjadi pada morfologi dan sitologinya.
Staphylococcus kloosii, an orange pigment-producing bacterium, was isolated from the respiratory tree of Holothuria (Mertensiothuria) leucospilota (Brandt 1835) from Teluk Nipah, Pangkor Island, Perak, Malaysia. This report is the first documentation of this Gram-positive strain, referred to as Strain 68 in Malaysia. A partial 16S ribosomal RNA gene sequence of the mesophilic strain has been registered with GenBank (National Center for Biotechnology Information, US National Library of Medicine) with accession number JX102547. Phylogenetic analysis using the neighbour-joining method further supported the identification of Strain 68 as S. kloosii. The circular strain produced orange pigments on tryptone glucose yeast extract agar (TGYEA) and in nutrient broth (NB) at approximately pH 7. The visible spectra of ethanolic and methanolic pigment extracts of the bacterial strain were considered identical with λmax at 426, 447 and 475 nm and λmax at 426, 445 and 473 nm, respectively. Both visible spectra resemble the visible spectra of lutein, which is a commercial carotenoid; however, further analyses are required to confirm the identity of this pigment. The methanolic extracts of the intracellular pigments comprised at least three pigment compounds: an orange pigment compound (major compound), a yellow pigment compound (the least polar) and a pink pigment compound (the most polar). These findings are the first documentation of the pigment composition of S. kloosii as no such record could be found to date.
Introduction: The aim of this study was to determine the effects of gamat extract on pain behaviour and Fos like immunoreactivity (FLI) expression in the ventral posterolateral thalamus using the acute pain model. Materials & Methods: Fourteen Sprague-Dawley male rats (220-300 gram) were given intraplantar injection of 0.05ml formalin (1%) followed by intraperitoneal administration of either 4 mg/kg gamat extracts (Holothuria spp.) or saline (control). Behavioural changes were observed and rats were sacrificed 2 hours post-formalin injection. Immunohistochemistry testing was done on the brain sections. FLI was examined using a light microscope attached to an image analyser. The behaviour and FLI data were analysed using repeated measure analysis of variance and independent t-test respectively. Significance level was taken as p
Holothuria (Mertensiothuria) leucospilota (Brandt 1835), white threads fish or locally known as bat puntil is currently considered as the most abundant sea cucumber species in Malaysia. This study aimed to generate the genetic profile of H. leucospilota from Malaysia and then to determine the phylogenetic relationship between H. leucospilota and other members of genus Holothuria using partial sequences of cytochrome c oxidase I (COI) mitochondrial DNA (mtDNA) gene. In this study, specimens of H. leucospilota were collected from Intan Besar Island, Langkawi, Kedah Darul Aman in the west coast of Peninsular Malaysia. Three main methods namely neighbour joining, maximum parsimony and maximum likelihood were used for the phylogenetic tree reconstruction. Tree topologies showed that H. leucospilota has its own monophyletic clade clearly distinct from the other species. The pairwise genetic distance calculated further supported these findings. In addition, the results also should that the COI mtDNA gene is capable to unravel the phylogenetic relationship of H. leucospilota.
This study aimed to resolve the taxonomic status of a morphologically undetermined sea cucumber species of order Apodida
from Malaysia (GenBank accession no.: FJ223867) using partial 16S mitochondrial rRNA gene sequences and subsequently
to determine the validity of morphological taxonomy of Holothuria species into its current subgenera. The undetermined
species clustered with all taxa of Holothuria in previous study. Phylogenetic analyses using maximum parsimony and
Bayesian methods suggest that the undetermined species was genetically closer to Holothuria (Lessonothuria) pardalis and
Holothuria (Acanthotrapeza) coluber; and its position in both phylogenetic trees further suggests its status as a Holothuria
taxon. Subgenera of Holothuria, Merthensiothuria and Metriatyla are monophyletic with strong bootstrap supports and
posterior probabilities of clades, thus strengthening their morphological taxonomies. Nonetheless, the non-monophyly of
subgenera of Halodeima, Microthele and Platyperona suggests a requirement for their taxonomic revisions using integrative
taxonomy. The status of Holothuria (Halodeima) edulis subgroups in the maximum parsimony and Bayesian trees is
indistinct and further taxonomic revisions are necessary. In terms of sister relationship, both phylogenetic trees suggest
that subgenus Holothuria is a sister taxon of subgenus Roweothuria while the other sister relationships were unclear due
to the undetermined species, paraphyly and polyphyly of a number of subgenera. Further studies with more specimens of
genus Holothuria from broader geographical locations and various mtDNA genes along with morphological approaches
may facilitate to provide better insights into the molecular phylogeny of subgenera of Holothuria.
This study aimed to determine phylogenetic relationship between and among selected species of sea cucumbers (Echinodermata: Holothuroidea) using 16S mitochondrial ribosomal RNA (rRNA) gene. Phylogenetic analyses of 37 partial sequences of 16S mitochondrial rRNA gene using three main methods namely neighbour joining (NJ), maximum parsimony (MP) and maximum likelihood (ML) showed the presence of five main genera of sea cucumbers: Molpadia from order Molpadiida and four genera of order Aspidochirotida namely Holothuria, Stichopus, Bohadschia and Actinopyga. All of the 17 species obtained from Malaysia distributed among the main genera except within Actinopyga. Interestingly, Holothuria excellens was out of Holothuria group causing Holothuria to be paraphyletic. High bootstrap value and consistent clustering made Molpadia, Stichopus, Bohadschia and Actinopyga monophyletic. The relationship of Actinopyga with the other genera was unclarified and Stichopus was sister to Molpadia. The latter finding caused the resolution at order level unclear. The pairwise genetic distance calculated using Kimura 2-parameter model further supported and verified findings from the phylogenetic trees. Further studies with more samples and different mitochondrial DNA genes need to be done to get a better view and verification on the molecular phylogeny of sea cucumbers.
Traditional Chinese Marine Medicine (TCMM) represents one of the medicinal resources for research and development of novel anticancer drugs. In this study, to investigate the presence of anticancer activity (AA) displayed by cold or hot nature of TCMM, we analyzed the association relationship and the distribution regularity of TCMMs with different nature (613 TCMMs originated from 1,091 species of marine organisms) via association rules mining and phylogenetic tree analysis. The screened association rules were collected from three taxonomy groups: (1) Bacteria superkingdom, Phaeophyceae class, Fucales order, Sargassaceae family, and Sargassum genus; (2) Viridiplantae kingdom, Streptophyta phylum, Malpighiales class, and Rhizophoraceae family; (3) Holothuroidea class, Aspidochirotida order, and Holothuria genus. Our analyses showed that TCMMs with closer taxonomic relationship were more likely to possess anticancer bioactivity. We found that the cluster pattern of marine organisms with reported AA tended to cluster with cold nature TCMMs. Moreover, TCMMs with salty-cold nature demonstrated properties for softening hard mass and removing stasis to treat cancers, and species within Metazoa or Viridiplantae kingdom of cold nature were more likely to contain AA properties. We propose that TCMMs from these marine groups may enable focused bioprospecting for discovery of novel anticancer drugs derived from marine bioresources.
Sea cucumbers, belonging to the class Holothuroidea, are marine invertebrates, habitually found in the benthic areas and deep seas across the world. They have high commercial value coupled with increasing global production and trade. Sea cucumbers, informally named as bêche-de-mer, or gamat, have long been used for food and folk medicine in the communities of Asia and Middle East. Nutritionally, sea cucumbers have an impressive profile of valuable nutrients such as Vitamin A, Vitamin B1 (thiamine), Vitamin B2 (riboflavin), Vitamin B3 (niacin), and minerals, especially calcium, magnesium, iron and zinc. A number of unique biological and pharmacological activities including anti-angiogenic, anticancer, anticoagulant, anti-hypertension, anti-inflammatory, antimicrobial, antioxidant, antithrombotic, antitumor and wound healing have been ascribed to various species of sea cucumbers. Therapeutic properties and medicinal benefits of sea cucumbers can be linked to the presence of a wide array of bioactives especially triterpene glycosides (saponins), chondroitin sulfates, glycosaminoglycan (GAGs), sulfated polysaccharides, sterols (glycosides and sulfates), phenolics, cerberosides, lectins, peptides, glycoprotein, glycosphingolipids and essential fatty acids. This review is mainly designed to cover the high-value components and bioactives as well as the multiple biological and therapeutic properties of sea cucumbers with regard to exploring their potential uses for functional foods and nutraceuticals.
This preliminary study aimed to identify a commercial gamat species, Stichopus horrens Selenka, 1867, and a timun laut species, Holothuria (Mertensiothuria) leucospilota (Brandt, 1835), from Pangkor Island, Perak, Malaysia, employing morphological techniques based on the shape of the ossicles and molecular techniques based on the cytochrome c oxidase I (COI) mitochondrial DNA (mtDNA) gene. In Malaysia, a gamat is defined as a sea cucumber species of the family Stichopodidae with medicinal value, and timun laut refers to non-gamat species. S. horrens is very popular on Pangkor Island as a main ingredient in the traditional production of air gamat and minyak gamat, while H. leucospilota is the most abundant species in Malaysia. In contrast to previous studies, internal body parts (the respiratory tree and gastrointestine) were examined in this study to obtain better inferences based on morphology. The results showed that there were no ossicles present in the gastrointestine of H. leucospilota, and this characteristic is suggested as a unique diagnostic marker for the timun laut species. In addition, the presence of Y-shaped rods in the respiratory tree of S. horrens subsequently supported the potential to use internal body parts to identify the gamat species. Phylogenetic analysis of the COI mtDNA gene of the sea cucumber specimens using the neighbour-joining method and maximum likelihood methods further confirmed the species status of H. leucospilota and S. horrens from Pangkor Island, Perak, Malaysia. The COI mtDNA gene sequences were registered with GenBank, National Center for Biotechnology Information (NCBI), US National Library of Medicine (GenBank accession no.: KC405565-KC405568). Although additional specimens from various localities will be required to produce more conclusive results, the current findings provide better insight into the importance of complementary approaches involving morphological and molecular techniques in the identification of the two Malaysian sea cucumber species.
Staphylococcus kloosii, an orange pigment-producing bacterium, was isolated from the respiratory tree of Holothuria (Mertensiothuria) leucospilota (Brandt 1835) from Teluk Nipah, Pangkor Island, Perak, Malaysia. This report is the first documentation of this Gram-positive strain, referred to as Strain 68 in Malaysia. A partial 16S ribosomal RNA gene sequence of the mesophilic strain has been registered with GenBank (National Center for Biotechnology Information, US National Library of Medicine) with accession number JX102547. Phylogenetic analysis using the neighbour-joining method further supported the identification of Strain 68 as S. kloosii. The circular strain produced orange pigments on tryptone glucose yeast extract agar (TGYEA) and in nutrient broth (NB) at approximately pH 7. The visible spectra of ethanolic and methanolic pigment extracts of the bacterial strain were considered identical with λmax at 426, 447 and 475 nm and λmax at 426, 445 and 473 nm, respectively. Both visible spectra resemble the visible spectra of lutein, which is a commercial carotenoid; however, further analyses are required to confirm the identity of this pigment. The methanolic extracts of the intracellular pigments comprised at least three pigment compounds: an orange pigment compound (major compound), a yellow pigment compound (the least polar) and a pink pigment compound (the most polar). These findings are the first documentation of the pigment composition of S. kloosii as no such record could be found to date.
A strategy to circumvent the problem of multidrug resistant pathogens is the discovery of anti-infectives targeting bacterial virulence or host immunity. Black sea cucumber (Holothuria atra) is a tropical sea cucumber species traditionally consumed as a remedy for many ailments. There is a paucity of knowledge on the anti-infective capacity of H. atra and the underlying mechanisms involved. The objective of this study is to utilize the Caenorhabditis elegans-P. aeruginosa infection model to elucidate the anti-infective properties of H. atra. A bioactive H. atra extract and subsequently its fraction were shown to have the capability of promoting the survival of C. elegans during a customarily lethal P. aeruginosa infection. The same entities also attenuate the production of elastase, protease, pyocyanin and biofilm in P. aeruginosa. The treatment of infected transgenic lys-7::GFP worms with this H. atra fraction restores the repressed expression of the defense enzyme lys-7, indicating an improved host immunity. QTOF-LCMS analysis revealed the presence of aspidospermatidine, an indole alkaloid, and inosine in this fraction. Collectively, our findings show that H. atra possesses anti-infective properties against P. aeruginosa infection, by inhibiting pathogen virulence and, eventually, reinstating host lys-7 expression.