Antioxidant and hypolipidaemic properties of red seaweed,Gracilaria changii

The edible red seaweed, Gracilaria changii, was collected from the coastal area of Sarawak, Malaysia, and evaluated for its hypolipidaemic properties using high cholesterol/high fat (HF) induced male Sprague–Dawley rats. In the in vivo study, the HF diet group showed significantly higher total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), atherogenic index (AI) and body weight gain as compared to other treatment groups. At the end of treatment period, rats fed with a HF diet supplemented with 5 % freeze-dried G. changii powder had significantly reduced plasma TC (?39.19 %), LDL-C (?36.36 %), and triglycerides (TG) content (? 25.45 %). Meanwhile, 10 % seaweed powder significantly lowered the plasma TC, LDL-C and TG content by ?40.34, ?35.95 and ?30.91 % respectively, compared to the HF group. The AI of rats supplemented with 10 % seaweed powder was the lowest among the treatment groups and indicates a lowered risk for cardiovascular diseases. The plasma lipid peroxidation of the seaweed powder-fed groups was also significantly lower than the HF group, while the erythrocyte enzyme antioxidant activities of superoxide dismutase, catalase and glutathione peroxidase of the treatment groups were also improved. Diets supplemented with seaweed powder also decreased plasma aspartate aminotransferase and the alanine aminotransferase levels.     

Biosystematics of Gracilaria foliifera (Gigarthales, Rhodophyta)

Strains of Gracilaria foliifera (Forsskål) Børgesen isolated into culture from Ireland (where the species would appear to be introduced), England and the Atlantic coast of France were interfertile. Strains of G. foliifera from Texas and North Carolina, U.S.A. were similarly interfertile but did not cross with the European strains. The North Carolina strain was distinctly flattened in culture and superficially resembled G. foliifera from Europe whereas the Texas strain was terete or slightly flattened. Both of the American strains were interfertile with an isolate of G. tikvahiae McLa-chlan from the type locality of this species in northern Nova Scotia, Canada. It seems likely from these data that G. foliifera is not represented on the coast of North America and that G. tikvahiae is a highly morphologically variable species widely distributed from Nova Scotia south to the Caribbean. Examination of type material of Gracilaria foliifera and G. multipartita (Clemente) Harvey, together with phyto-geographical considerations, suggests that the species known as G. foliifera on eastem Atlantic masts should be known as G. rnultipartita , and that G. foliifera s. str . is probably confined to the Red, Arabian, and Indian Seas.     

Different regulation of haloperoxidation during agar oligosaccharide-activated defence mechanisms in two related red algae, Gracilaria sp. and Gracilaria chilensis

The related red seaweeds Gracilaria sp. from the eastern Mediterranean and Gracilaria chilensis from Chile were similar in their enzymatic inventory for halogenation. In both species, halogenation was dependent upon H(2)O(2) and thus driven by haloperoxidases. These could be inhibited with phosphate and reversibly inhibited with azide and were therefore apparently dependent upon vanadate. Both species generated in the first line bromoform and other brominated halocarbons. Gel electrophoresis under non-denaturating conditions demonstrated that both species expressed halogenating peroxidases. Elicitation of Gracilaria sp. with agar oligosaccharides resulted in marked increases in bromination, iodination, and chlorination. Production rates of volatile halocarbons and phenol red bromination both increased by a factor of eight, presumably due to increased availability for haloperoxidases of H(2)O(2) during the oxidative burst response. Elicitation of Gracilaria sp. also triggered a release of bromide ions through DIDS-sensitive anion channels, which allowed for some bromination in bromide-free medium. However, this effect was relatively limited. By contrast, agar oligosaccharide oxidation in G. chilensis did not increase halogenation. Obviously, agar oligosaccharide oxidation does not provide sufficient amounts of hypohalous acids for such increases, because it does not deliver H(2)O(2) at the active site of vanadium-dependent haloperoxidases. These results correlate with earlier findings that the agar oligosaccharide-elicited oxidative burst controls microorganisms while agar oligosaccharide oxidation does not.     

Seasonal variation of the agar quality and chemical composition of Gracilaria veleroae and Gracilaria vermiculophylla (Rhodophyceae,Gracilariaceae) from Baja California Sur,Mexico

Yield and physico‐chemical properties of agar from Gracilaria veleroae E.Y. Dawson and Gracilaria vermiculophylla (Ohmi) Papenfuss were studied and the chemical composition of the two seaweeds was determined. Samples were collected seasonally from summer 2003 to spring 2005. The agar yield did not vary significantly between seasons for both species. The lowest agar gel strength was obtained from G. veleroae (207.5 g cm^–2) in summer 2003 and the highest from G. vermiculophylla (793.1 g cm^–2) in winter 2004. Melting temperatures and hysteresis were higher in G. vermiculophylla, whereas gelling temperatures and 3,6‐anhydrogalactose content were higher for G. veleroae. Moisture, ash, crude fiber, and ether extract showed no significant seasonal variation for G. veleroae. The chemical composition of G. vermiculophylla showed significant seasonal variation. G. vermiculophylla possesses a better agar quality than G. veleroae and is a species that could be considered as a source of agar for commercial use.     

Growth,pigments, and biochemical composition of marine red alga Gracilaria crassa

The marine red alga Gracilaria crassa was investigated for its proximate composition, minerals, fatty acids, amino acids, and agar content to decipher its nutritional implications. The growth performance and pigments were studied under different combinations of temperature and salinity. On a dry weight basis the total lipid content was 1.30?±?0.05 %, protein was 5.18?±?0.64 %, carbohydrate was 42.0?±?1.2 %, ash was 43.18?±?1.15 %, and agar content was 21.52?±?0.73 %. Appreciable amounts of macro-, micro-nutrients (K?>?Na, Ca, Mg, and Fe), and essential amino acids (Ileu, His, Thr, Leu, and Lys) were found. Palmitic, stearic acid, and arachidonic acid were major fatty acids detected. The alga showed maximum daily growth rate (DGR %) 5.8?±?0.09 % at 25 °C, 35 ‰ salinity. The highest content of pigment R-phycoerythrin (444.7?±?1.9 μg g^?1 fresh weight (FW) basis) was obtained at 25 ‰ salinity at 35 °C while that of R-phycocyanin (476.3?±?2.3 μg g^?1 DW) at 30 ‰ salinity at 30 °C. This study revealed that this alga can be utilized as a potential source for food and feed. The data generated on best growth conditions will be very useful for farming of G. crassa in open sea. This alga could be used for production of natural colorants at defined control condition.     

Properties of Gracilaria agars

Data are presented concerning an investigation into the diversity, abundance, production and respiration of the oligochaete fauna in the eutrophic, polymictic Lake Zbechy and in a melioration canal in the Wielkopolska Region, an area of intensive agriculture. It was found that in the canal the average biomass of oligochaetes was about four times higher than in the lake. Oligochaetes expend 1.1–4 times more energy in respiration than in tissue production. Species diversity and species number are positively correlated, while the correlation between diversity and abundance is negative.     

Chromosomal and mutagenic study of the marine macroalga,Gracilaria tenuistipitata

A pressing need of algal aquaculture is the availability of strains with favourable characteristics. Since this goal cannot be achieved easily by traditional breeding methods for Gracilaria tenuistipitata, we developed mutagenesis and selection procedures that bypass the necessity for protoplast isolation and cultivation. A haploid gametophyte was identified through chromosome analysis and used for mutagenesis. Chromosomal analyses showed that the haploid plant contains several layers of mononucleated cells at the surface and many large multinucleated syncytiums in the internal region. Chromosomal division of all nuclei in a syncytium was in synchrony. Mononucleated cells were more sensitive to both UV and DNA insertion mutagenesis. UV treatment generated many cadmium (Cd) resistant mutants, but most mutants exhibited retarded growth rate. However, several stable Cd resistant mutants were obtained through reselection. Plasmid pCaMVCAT could be used for DNA insertion mutagenesis in G. tenuistipitata. Stable mutants with increased resistance to chloramphenicol were selected.     

World-wide use and importance of Gracilaria

The world's first source of agar, from the middle of the seventeenth century, was Gelidium from Japan, but by the beginning of the twentieth century demand for the phycocolloid exceeded of the supply of this alga. Since then Gracilaria has played an important role in the production of agar. Currently agars are obtained from five genera in three orders of red algae and marketed as ‘natural agar’ in squares or strips or as ‘industrial agar’ in powder form. The development of production processes through alkaline hydrolysis of sulphates has allowed a good quality food agar to be obtained from Gracilaria. This does not show the synergistic reaction with locust bean gum apparent with Gelidium agar. The term ‘agaroids’ is applied to Gracilaria agars produced without alkaline hydrolysis of sulphates, with greater sulphate content and much less gel strength. Unlike Gelidium, Gracilaria has to be processed in a short period of time and cannot be allowed to remain in storage for use during years of lower availability. Statistics of imports of agarophytes to Japan during the last 10 years give an indication of the state of the market. During this period there was a marked reduction in Gracilaria imports, mainly from Chile, but also the Philippines, Indonesia and South Africa, mainly due to the overall increase in the capacity of agar production in Gracilaria-producing countries.     

Effects of plant growth regulators on callus formation, growth and regeneration in axenic tissue cultures of Gracilaria tenuistipitata and Gracilaria perplexa (Gracilariales, Rhodophyta)

The effects of auxins and cytokinin on callus formation, growth and regeneration of Gracilaria tenuistipitata Chang et Xia and G. perplexa Byrne et Zuccarello (Gracilariales, Rhodophyta) are reported. Plant growth regulators (PGR) in concentrations ranging from 0.1 to 100.0 μmol of indole‐3‐acetic acid, 2,4‐dichlorophenoxyacetic acid (2,4‐D), and kinetin (K) were added to the ASP 12‐NTA solid medium (0.7% agar), and apical and intercalary segments (5 mm long) were inoculated as initial explants. K stimulated growth rates of intercalary segments of G. tenuistipitata in a linear relation, and 2,4‐D (1.0 μmol) and K (10.0 μmol) stimulated growth rates of apical and intercalary segments of G. perplexa, respectively. The simultaneous formation of apical, basal, and intermediate calluses is reported for the first time in axenic tissue cultures of red algae. With intercalary segments of G. tenuistipitata, basal callus induction rates were higher than those of apical and intermediate calluses in the majority of treatments, and auxins had stimulatory effects on the formation of all callus types. In apical segments of G. perplexa, intermediate callus formation was stimulated only by treatment with 1.0 μmol of K, while apical callus formation was stimulated by indole‐3‐acetic acid (1.0–10.0 μmol), 2,4‐D (10.0–100.0 μmol), or K (0.1 μmol). Intercalary segments of G. perplexa developed only intermediate calluses, and the majority of treatments with PGR stimulated higher rates than those presented by apical segments. Potential for regeneration (development of adventitious plantlets originated from callus cells) was higher in apical calluses than in basal and intermediate calluses developed in intercalary segments of G. tenuistipitata. Moreover, auxins and cytokinin were essential to the induction of regeneration in intermediate calluses, while specific concentrations stimulated regeneration from basal and apical calluses. Plant regeneration in G. perplexa was observed only after transferring calluses from solid to liquid medium, and the majority of treatments with PGR had stimulatory effects. Regenerating plants of G. perplexa developed tetrasporangia, and released tetraspores giving rise to adult gametophytes. Our results indicate that auxins and cytokinin have a regulatory role in the growth and morphogenesis in G. tenuistipitata and G. perplexa, and diversity of responses presented by both species is related to specific developmental systems.     

Yield, chemical composition and gel strength of agarocolloids of Gracilaria gracilis, Gracilariopsis longissima and the newly reported Gracilaria cf. vermiculophylla from Roscoff (Brittany, France)

Agarocolloids were extracted from field samples of Gracilaria gracilis, Gracilariopsis longissima and the newly reported Gracilaria cf. vermiculophylla harvested at different periods of the year near Roscoff (France). Native and alkali modified extracts were characterized by GLC, HPLC and FT-IR spectroscopy. The main components of agarocolloids isolated by freeze-thawing method, were 3,6-anhydrogalactose and galactose. In addition, minor components (6-O-methyl-galactose, 4-O-methyl-galactose and sulfate groups ranging from 4.4 up to 6.6% [w/w]) were detected. The highest rate of 6-O-methylgalactose was observed in agarocolloids from vermiculophylla (14 mole%). Sulfates were mainly branched on C4 of the D-galactose in gracilis and Gs. longissima agarocolloids. G. vermiculophylla agaroids isolated by EtOH and NaCl precipitations from the syneresis water were characterized by a high sulfation on C6 of galactose and a low sulfation on C2 of 3,6-anhydrogalactose. Native agarocolloid gel strengths from Gracilaria species were clearly higher than those of Gracilariopsis. Alkali treatments reduced the sulfate levels but increased slightly the gel strengths. An approximation of the polymer sizes carried out with colorimetric assays indicated that the polymer sizes were higher in G. gracilis than observed in Gs. longissima. This revised version was published online in June 2006 with corrections to the Cover Date.     

A review of the life history,reproduction and phenology of Gracilaria

The basic life history of the red alga Gracilaria is of the three-phase Polysiphonia type but a number of species show deviations. Plants can bear both gametangia and tetrasporangia, either on separate parts of the thallus or on the same. Explanations include the in situ germination of tetraspores (allowing gametophytic thalli to be epiphytic on tetrasporophytes), the coalescence of spores or developing discs (resulting in chimaeras), mitotic recombination during cell division in the mature diploid thallus (resulting in patches of diploid male and female cells on the tetrasporophyte), a mutation eliminating the repression of female expression allowing haploid male plants to be bisexual and initial failure of cell walls to form during the development of tetraspores. Polyploids can be produced from plants with diploid gametangia. The sexes and phases are usually morphologically identical but gametophytes or their parts may be smaller. The growth rates of the sexes may differ and diploid juveniles may survive better than haploid. Neither polyploidy nor hybridization results in superior growth. The sex ratio is probably 1:1 but females may appear to be more abundant. Diploid and haploid phases are usually either about equal or diploids predominate, often depending on the type of substratum. At high latitudes reproduction peaks in late summer whereas in the tropics it may be high all year. In temperate regions growth rate is fastest and biomass highest in late summer; in the tropics peak biomass is mainly in the winter. Spermatia are effective for only a few h. Spores vary in size around 25 \m, diploid ones usually being larger. Cystocarps or tetrasporangia in the field may not currently be releasing spores. In the laboratory spore release shows a diurnal rhythm, peaking during the night or day according to the species. All the above attributes are potentially important in planning and executing Gracilaria cultivation.     

Environmental variation and large-scale Gracilaria production

Temporal and spatial abiotic variation in seaweed farms should be anticipated to maximize production through alternative exploitation strategies. This study describes the basic assumptions and the most relevant data used to empirically develop a production model aimed at improving prediction and increasing production of Gracilaria farms in northern Chile. Continuous light and temperature recordings since 1986 have allowed us to relate abiotic variations with high production seasons of Gracilaria or with the presence of pests and epiphytes. Much of the interannual variations in light and temperature appear as part of a predictable pattern of change between ENSO (El Niño/Southern Oscillation) and inter-ENSO years. Production has been found to be a function of stock density and harvesting frequency, two parameters that can be effectively manipulated in the field. Thus, the range of climatic change now can be anticipated to some extent which, in turn, suggests the best farming strategy. During seasons or growth periods anticipated to be highly productive, farming activities are oriented to maintain high percentage removal of the stock through frequent harvesting. During seasons anticipated to be low in production, activities are oriented to prevent biomass losses due to the blooms of epiphytes and pests and to secure stocks to renew through planting the damaged parts of the beds after the blooms.     

Gracilaria species (Gracilariaceae, Rhodophyta) from southeastern Australia, including a new species, Gracilaria perplexa sp. nov.: Morphology, molecular relationships and agar content

Select species of the agarophyte Gracilaria were studied from southeastern Australia. The morphology and anatomy of species is described and molecular relations are inferred based on plastid and mitochon‐drial DNA sequence data. Agar yields and qualities are determined for each species. Gracilaria chilensis, found in Tasmania and Victoria, is morphologically and molecularly similar to G. chilensis from New Zealand and Chile and has low agar yields of 11–16%. Gracilaria cliftonii from Victoria, has high crude agar yield (52%) and is molecularly uniform. Gracilaria perplexa sp. nov., known only from Botany Bay, New South Wales, has an agar yield of 39%. The agar of G. perplexa is unusual in requiring the addition of 0.1 mol L^?1 NaCl for alcohol precipitation and is cold‐water (25°C) soluble because of the very high sulfate ester content. Molecular phylogeny shows that G. perplexa is closely related to Gracilaria preissiana from western Australia, but differs from the latter in its reduced branching and narrower more terete axes.     

Viability and dissemination of spermatia of Gracilaria verrucosa (Gracilariales,Rhodophyta)

The dissemination and viability of Gracilaria verrucosa spermatia were tested. Crosses were performed among three males and three females from Cape Gris Nez, northern France. Laboratory experiments show that spermatia have a mean fertile life of about five hours. Field studies show that spermatia are dispersed by stream and tidal currents and that fertilization can occur at least 80 m from a population.     

Polysaccharides from Gracilaria corticata: sulfation, chemical characterization and anti-HSV activities

In this study, we have analyzed water-extracted polysaccharides of Gracilaria corticata. The water extract (WE), a galactan-containing sub-fraction (F3) and their hyper sulfated derivatives (WES1, WES2, F3S1 and F3S2) had anti-HSV activity with inhibitory concentration 50% (IC50) from 1.1 to 27.4 microg/ml. Sub-fraction F3, which has a molecular mass of 30 kDa, consists of a backbone of beta-(1-->3) and alpha-(1-->4)-linked-galactopyranosyl residues. This linear galactan contained Gal2Xyl1, Gal2AnGal2, Gal4 and Me-Gal3AnGal2 as oligomeric building subunits. Sulfate group was located at C-4 of (1-->3)-linked galactopyranosyl residues of the native galactan, and appeared to be very important for the anti-herpetic activity.     

Agar polysaccharides from Gracilaria species (Rhodophyta, Gracilariaceae)

Yield, physical and chemical properties of agar from three agarophytes species (Gracilaria gracilis, G. dura and G. bursa-pastoris) were determined. The agar yield from the three species varied significantly (P<0.01). The highest yields of agar (34.8%) and the lowest (30%) were obtained from G. bursa-pastoris and G. gracilis, respectively. Highest gel strength (630+/-15 g cm(-2)) was obtained from agar extracted from G. gracilis and lowest from G. bursa-pastoris (26+/-3.6 g cm(-2)). The values of 3,6-anhydrogalactose were similar for G. gracilis and G. dura and there were no significant differences among the species. The sulfate contents varied significantly (P<0.01) and the higher value was obtained from G. bursa-pastoris. Among the three species, G. gracilis showed superior agar quality than the other two species, hence it can be considered a good potential source for industrial use.     

Integrated tank cultivation of salmonids and Gracilaria chilensis (Gracilariales,Rhodophyta)

In order to study the N and P balances in the Driss I reservoir, we measured concentrations in the water of these elements, their sedimentation rates, and their input by the river Inaouen and output through the dam. Supply and loss of N and P were calculated from samples collected every 48 hours and were for N 15 and 27. 5 mg m⁻² d⁻¹ and for P 33.6 and 1.1 mg m⁻² d⁻¹ respectively. The sedimentation rates, determined using sediment traps, were high, about 75 mg m⁻² d⁻¹ for N and 34 mg m⁻² d⁻¹ for P. Phosphate input came from point sources and was mainly in particulate form. A large fraction of the phosphate sedimented in the form of calcium bound phosphate, with some iron bound phosphate. Bioavailability was, however, low, as the particulate phosphate disappeared rapidly from the epilimnion because of the high sedimentation rate. The ratio Tot-N/Tot-P of the sedimented particulate matter varied between 0.05 and 7.74 and depended on primary production and watergate management.