Put O. Ang

Protective effects of seaweeds against liver injury caused by carbon tetrachloride in rats.

Three species of seaweeds collected from Tung Ping Chau, Hong Kong, were screened for their hepatoprotective activity using carbon tetrachloride (CCl4)-induced liver injury in the rat as a model of chemical hepatitis. A single oral dose of 1.25 ml/kg of CCl4 was able to produce significantly elevated levels of serum glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transminase (GOT). Administration of 150, 300 and 600 mg/kg of aqueous extracts from Myagropsis myagroides, Sargassum henslowianum and S. siliquastrum, respectively, significantly reduced the CCl4-induced acute elevation in the levels of GPT and GOT in rats. The same result was also seen in the histopathological study of liver tissue. The seaweed crude extracts probably acted to protect against CCl4-induced liver injury through their antioxidant properties.

Phylogeography of the marine macroalga Sargassum hemiphyllum (Phaeophyceae, Heterokontophyta) in northwestern Pacific.

Sargassum hemiphyllum is commonly found in Japan and Korea, with a variety, var. chinense, that is found distributed in the southern Chinese coast. We previously reported distinct genetic differentiation between the two taxa based on the PCR‐RFLP data of plastid RubiscoL‐S spacer. The present study aims at elucidating the phylogeographic pattern of S. hemiphyllum based on more markers in the nuclear and extranuclear genomes, with a view to reveal the occurrence of hybridization. The two allopatrically distributed taxa were found to be genetically distinct in nuclear ITS2, plastidial Rubisco (Rbc) and mitochondrial TrnW_I (Trn) spacers. Their divergence was postulated to be attributable to the vicariant event which resulted from the isolation of the Sea of Japan during the late Miocene (6.58–11.25 Mya). Divergence within both S. hemiphyllum and the chinense variety was observed based on Trn spacer, while the divergence in S. hemiphyllum was further confirmed in Rbc spacer. This divergence appears to correspond to the separation of the Japanese populations between the Sea of Japan and the Pacific that occurred around 0.92–2.88 Mya (the early Pleistocene). The presence of an ITS2 clone resembling var. chinense sequences in a Japanese population of S. hemiphyllum (JpNS) raises the possibility of the introgression of var. chinense individuals into S. hemiphyllum population. Compared to that between S. hemiphyllum and the chinense variety, hybridization among the Japanese and Korean populations of S. hemiphyllum is highly probable as all these individuals share a pool of nuclear ITS2 sequences, possibly attributable to incomplete concerted evolution of ITS2.

Changes in lipid profiles of rats fed with seaweed-based diets

Abstract The changes in lipid profiles of hypercholesterolemic rats fed with diets containing four Hong Kong seaweeds, Ulva sp., Hypnea charoides, Colpomenia sinuosa and Sargassum hemiphyllum , were evaluated. The cholesterol lowering potentials of the seaweeds were determined by comparing the levels of serum total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglyceride (TG) and hepatic cholesterol (HC) in the seaweed-based diet groups with those of the control group. All four seaweed-based diet groups showed similar trend of increase in HDL and triglyceride levels when compared to the control group. Although Hypnea charoides and Ulva sp. diet groups reduced the serum TC level (16% and 13.9%, respectively), these values exhibited no significant differences from those of the control. In addition, there were no significant differences of LDL level between control and H. charoides diet. However, H. charoides diet was the only seaweed diet that caused a depression of LDL level (10.7%). It seemed that H. charoides had a comparatively greater hypocholesterolemic potential than the others. Besides, all four seaweeds had no adverse effect on the growth of animals with respect to their body weight and the weights of internal organs except the C. sinuosa diet which caused a significant ( P

Antiviral activities of extracts from Hong Kong seaweeds.

We extracted six Hong Kong brown seaweed species with hot water for their antiviral properties. The cytotoxicity and antiviral activity of these extracts were tested by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenlytetrezolium bromide] method, cytopathic effect reduction assay, and plaque reduction assay. The antiviral effect was further determined by flow cytometric analysis. The results showed that most of these extracts inhibited the propagation of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) standard strains with very low cytotoxicity to the host cells. The extracts of Hydroclathrus clathratus and Lobophora variegata showed more potential anti-HSV activities than the extracts of the other four seaweeds. They also had moderate antirespiratory syncytial virus (RSV) activities but could not inhibit influenza A virus. Hydroclathrus clathratus was further extracted by diluted acid and alkali and the antiviral effects of the extracts were also detected. The result showed that the hot water extract contained the main carbohydrate components that exhibited the antiviral activities against various strains of HSV, including the acyclovir-resistant strain. HI-3, a compound fractionated from this hot water extract, showed a dose-dependent anti-HSV activity in flow cytometric analysis and plaque reduction assay.

Low genetic variability of Sargassum muticum (Phaeophyceae) revealed by a global analysis of native and introduced populations.

Sargassum muticum (Yendo) Fensholt is one of the most well‐known invasive species in the world. There have, however, been few genetic investigations on both its introduced and native populations. There are also some questions about the taxonomic status of this species. This study is the first to assess the genetic diversity of S. muticum on a global scale, by utilizing one marker each from the extranuclear genomes, namely, plastidial RUBISCO and mitochondrial TrnW_I spacers, as well as the nuclear internal transcribed spacer 2 (ITS2). Based on the markers investigated, both the invasive as well as the native populations of this species appeared very homogenous, when compared with other invasive and brown macroalgae. No variation in ITS2 and RUBISCO spacer was revealed in S. muticum populations, including those from its native ranges in Asia and the introduced ranges in Europe and North America. Two TrnW_I spacer haplotypes with a fixed two‐nucleotide difference were found between the populations of eastern Japan and the other 15 populations examined. This study confirms that there is no cryptic diversity in the introduced range of this species. All the materials collected globally are indeed S. muticum. Results depicting the distribution range of the two TrnW_I spacer haplotypes also support the earlier suggestion that the source of the introduced S. muticum populations is most likely western and central Japan (Seto Inland Sea), where the germlings of S. muticum were likely to have been transported with the Pacific oysters previously introduced for farming in Canada, UK, and France in earlier years.

Genetic structure analysis of natural Sargassum muticum (Fucales, Phaeophyta) populations using RAPD and ISSR markers

Sargassum muticum is important in maintaining the structure and function of littoral ecosystems, and is used in aquaculture and alginate production, however, little is known about its population genetic attributes. In this study, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure of four populations of S. muticum and one outgroup of S. fusiforme (Harv.) Setchell from Shandong peninsula of China. The selected 24 RAPD primers and 19 ISSR primers amplified 164 loci and 122 loci, respectively. Estimates of genetic diversity with different indicators (P%, percentage of polymorphic loci; H, the expected heterozygosity; I, Shannon’s information index) revealed low or moderate level of genetic variations within each S. muticum population, and a high level of genetic differentiations were determined with pairwise unbiased genetic distance (D) and fixation index (FST) among the populations. The Mantel test showed that two types of matrices of D and FST were highly correlated whether from RAPD (r = 0.9706, P = 0.009) or ISSR data (r = 0.9161, P = 0.009). Analysis of molecular variance (AMOVA) was conducted to apportion the variations among and within the S. muticum populations. It indicated that variations among populations were higher than those within populations, being 55.82% verse 44.18% by RAPD and 55.21% verse 44.79% by ISSR, respectively. Furthermore, the Mantel test suggested that genetic differentiations among populations were related to the geographical distances (r > 0.6), namely, conformed to the IBD (isolation by distance) model, as expected from UPGMA (unweighted pair group method with arithmetic averages) cluster analysis. On the whole, the high genetic structuring among the four S. muticum populations along the distant locations was clearly indicated in RAPD and ISSR analyses (r > 0.9, P < 0.05) in our study.

MORPHOLOGICAL AND GENETIC VARIATION IN THE POPULATIONS OF SARGASSUM HEMIPHYLLUM (PHAEOPHYCEAE) IN THE NORTHWESTERN PACIFIC(1).

Difficulty in species identification of Sargassum (Sargassaceae, Fucales) is partly attributed to the high polymorphism among its individuals and populations. This study aimed at assessing morphological and genetic variations in two varieties, var. hemiphyllum J. Agardh and var. chinense J. Agardh, of Sargassum hemiphyllum (Turner) C. Agardh, a widely distributed species in the northwestern Pacific. We investigated 26 measurable, five numerical, and 33 categorical morphological parameters associated with different branching levels of specimens from each of six localities within its distribution range using cluster analysis (CA) and principal coordinate analysis (PCoA). Leaf size of the primary and secondary branching levels and the vesicle size of the secondary branches of the specimens examined were determined to be the most important morphological parameters that were significantly different among populations. Change in leaf and vesicle length of individuals among the six populations followed a latitudinal gradient, with smaller leaves and vesicles associated with northern populations and larger ones in the southern populations. The possible influence of the gradual change in sea surface temperatures (SSTs) along this gradient in the northwestern Pacific on leaf and vesicle morphologies of this species was suggested. PCR‐RFLP analysis of the RUBISCO spacer in the chloroplast genome revealed two distinct and highly homogenous clades, a China clade and a Japan‐Korea clade, which corresponded to var. chinense and var. hemiphyllum, respectively. The formation of refugia along the “Paleo‐coast” in the East China Sea during glacial periods is suggested to have led to the vicariance of ancestral populations of S. hemiphyllum and thus to have promoted genetic differentiation. The massive freshwater outflow of the Yellow and Yangtze rivers may continue to act as a barrier, prolonging the allopatric distribution of the two varieties.

Homogeneous population of the brown alga Sargassum polycystum in Southeast Asia: possible role of recent expansion and asexual propagation.

Southeast Asia has been known as one of the biodiversity hotspots in the world. Repeated glacial cycles during Pleistocene were believed to cause isolation of marine taxa in refugia, resulting in diversification among lineages. Recently, ocean current was also found to be another factor affecting gene flow by restricting larval dispersal in animals. Macroalgae are unique in having mode of reproduction that differs from that of animals. Our study on the phylogeographical pattern of the brown macroalga Sargassum polycystum using nuclear Internal Transcribed Spacer 2 (ITS2), plastidal RuBisCO spacer (Rub spacer) and mitochondrial cytochrome oxidase subunit-III (Cox3) as molecular markers revealed genetic homogeneity across 27 sites in Southeast Asia and western Pacific, in sharp contrast to that revealed from most animal studies. Our data suggested that S. polycystum persisted in single refugium during Pleistocene in a panmixia pattern. Expansion occurred more recently after the Last Glacial Maximum and recolonization of the newly flooded Sunda Shelf could have involved asexual propagation of the species. High dispersal ability through floating fronds carrying developing germlings may also contribute to the low genetic diversity of the species.

Biosorption of di(2-ethylhexyl)phthalate by seaweed biomass

Samples of various Sargassum species were collected in the Hong Kong marine environment and used for studies on biosorption of di(2-ethylhexyl)phthalate (DEHP). Batch adsorption experiments were carried out to determine the removal capacity and removal efficiency of the biosorbents. The DEHP removal ability was similar among beached seaweed and three freshly collected Sargassum species. Different physico-chemical factors were evaluated in order to enhance the performance of the biosorbents. Under optimized conditions (25 mg biomass, initial pH 4, 25 °C, 40 mg L−1 DEHP), the mean removal capacity of beached seaweed and Sargassum siliquastrum was 5.68 and 6.54 mg g−1, respectively. Examination of the Langmuir and Freundlich adsorption isotherms showed that the biosorption phenomenon by these biosorbents could well be described by these models. Desorption of DEHP was also assessed with methanol, which showed the most satisfactory desorbing ability. Further study in multiple adsorption–desorption of DEHP by the biosorbents demonstrated the reusability of both beached seaweed and S. siliquastrum for biosorption of DEHP.

Gametogenesis, Embryogenesis, and Fertilization Ecology of Platygyra acuta in Marginal Nonreefal Coral Communities in Hong Kong

Understanding the reproductive biology of dominant coral species in subtropical nonreefal coral communities is critical in providing important information on the processes underlying the distribution limits of coral species and communities. This is the first study that investigates the reproduction cycle, gametogenesis, and fertilization ecology of Platygyra acuta. Results indicated that P. acuta is hermaphroditic and exhibits a single annual gametogenic cycle. Oogenic and spermatogenic cycle occurs for 6-7 months and for 2 months, respectively, prior to annual mass spawning event in May to June in Hong Kong. It took 18 hours for P. acuta to complete embryonic development, develop cilia, and start to rotate. High (>70%) fertilization success can be achieved under a broad range of sperm concentrations from 104 to 107 sperms mL−1. Fertilization success remained consistently high 6 h after spawning, indicating a prolonged viability of its gametes that is much longer than that recorded for other coral species. Significantly higher percentage of fertilization success was recorded in the first of the two consecutive nights of spawning, suggesting differences in the quality of the eggs and/or sperms between days of spawning. These results serve as important baseline information for better understanding of corals in marginal communities.