The introduced alga Undaria pinnatifida (Laminariales,Alariaceae) in the lagoon of Venice

Since its appearance in 1992 in the lagoon of Venice, the brown algae Undaria pinnatifida (kelp) has gradually expanded along the banks of canals both at Chioggia and Venice, becoming the dominant species in the local algal community chiefly from February to July. In Chioggia Island, where another brown seaweed (Sargassum muticum) is present since 1992, the spreading of Undaria reaches a plateau. In Venice Island, instead, the colonisation process is still in progress. During 1999, the kelp has colonised the main canals (e.g. The Grand Canal) and, subsequently, the small inner ones. In order to evaluate the dynamics of substrate re-colonisation by Undaria, two mechanical eradications were carried out during (March) and after the fertile period (July) of the algae. Such experimental manipulations provided evidence of the fast re-colonisation potential of the algae mainly attributable to its efficient reproductive system. Eradication made during the fertile period, in fact, permitted the kelp development during the following year, while re-colonisation has started 2 years later where eradication was performed after the reproductive period. A significant decrease in the surface covered by other species has been observed both in shallow (Ulva rigida, Enteromorpha spp., Antithamnion pectinatum, Chondracanthus acicularis) and deeper areas (Rhodymenia ardissonei) during the period of maximum development of U. pinnatifida. On the basis of the results of this study, the following conclusions can be drawn: (1) the alga U. pinnatifida is continuously expanding in lagoon environments of Venice; (2) in order to limit its spreading, mechanical eradications would be done on a large spatial scale and before the zoospores release; (3) there is suggestive evidence of competition between Undaria and the remaining indigenous algae.     

Surveys of Undaria pinnatifida (Laminariales,Phaeophyta) in Golfo Nuevo,Argentina

In December 1992, some sporophytes of the Asian kelp Undaria pinnatiftda were found growing subtidally at 6 m depth below A. Storni Port, Puerto Madryn, Argentina. During the winter of 1994, the species expanded significantly from its original location. Sporelings appear in early autumn and attain their maximum size (1.65 ± 0.10 m) during winter and early spring, when most of them become fertile. The fronds are lost in summer, with only some holdfasts and sporophylls surviving, and these disappear by the end of summer. The occurrence of U. pinnatida in Golfo Nuevo is reportedly due to an accidental introduction by cargo ships or fishing vessels arriving from Asian ports.     

Cultivation and utilization of Undaria pinnatifida (wakame) as food

A brief review is presented concerning wakame, Undaria pinnatifida, one of the most popular seaweeds used for food in Japan. Although it has been cultivated since about 1940, full-scale cultivation occurred after 1955. As methods for providing ‘seed stock’ and of processing the harvested sporophytes progressed, the yield increased rapidly. The main areas of cultivation are in Japan (e.g. Sanriku, Naruto), Korea and China, while ‘wild’ U. pinnatifida has been introduced into France, New Zealand and Australia. The total world yield of wakame exceeds 500 000 t fresh weight. Cultivated and harvested Undaria is boiled and salted (thus becoming green) and refrigerated; in the factory, it is removed its foreign matter and salt and dried. After checking for quality, the product is packaged in forms convenient for cooking and eating.     

Antioxidant activities of sulfated polysaccharides from brown and red seaweeds

The in vitro antioxidant activities of the following six sulfated polysaccharides were investigated: iota, kappa and lambda carrageenans, which are widely used in the food industry, fucoidan (homofucan) from the edible seaweed Fucus vesiculosus and fucans (heterofucans) F0.5 and F1.1 from the seaweed Padina gymnospora. With respect to the inhibition of superoxide radical formation, fucoidan had an IC₅₀ (the half maximal inhibitory concentration) of 0.058 mg·mL⁻¹, while the IC₅₀ for the kappa, iota and lambda carrageenans were 0.112, 0.332 and 0.046 mg·mL⁻¹, respectively. All of the samples had an inhibitory effect on the formation of hydroxyl radicals. The results of peroxidation tests showed that fucoidan had an IC₅₀ of 1.250 mg·mL⁻¹ and that the kappa, iota and lambda carrageenans had an IC₅₀ of 2.753 and 2.338 and 0.323 mg·mL⁻¹, respectively. Fucan fractions showed low antioxidant activity relative to fucoidan. These results clearly indicate the beneficial effect of algal polysaccharides as antioxidants.     

Microscopic observations on tissue calcium distribution in the brown alga,Undaria pinnatifida (Harvey) Suringar

The distribution of Ca²⁺ in thallus tissues of the brown algaUndaria pinnatifida was investigated by microspectro-photometric analysis after fixation with Carnoy's reagent and staining with an alizarine sulfonate dye. Ca²⁺ accumulation in the young thallus was much higher in mucilage cells of the basal than the apical region; there was least in the pith of the basal region. A roughly uniform pattern was found in the stipe from the central part of the young thallus, while a highly irregular pattern occurred in both stipe and blade from the corresponding area of the mature thallus; the Ca²⁺ content was much lower than in the cortex, mucilage cells and vacuoles of the mature stipe than in the corresponding area of the young stipe. The contents of all regions of the mature blade were higher than those in the young blade; they were particularly high in mucilage cells.     

Study of antioxidant activities of sulfated polysaccharides from Laminaria japonica

The composition, molecular weight and in vitro antioxidant activity of various sulfated polysaccharides obtained by anion exchange chromatography, acid hydrolysis and radical process degradation of the crude sulfated polysaccharide extracted from Laminaria japonica were compared. The low sulfated F-A2, with a peak-molecular weight (Mp) of 5–15 kDa, 14.5% sulfated ester and 21.8% glucuronic acid, exhibited a very strong antioxidant activity on superoxide and hydroxyl radicals, with activity even higher than that of large molecular weight fractions F-A and F-B. However, highly sulfated fractions with a peak-molecular weight below 15 kDa had much lower antioxidant activities than other fractions. These results indicated that the sulfate group of the low molecular weight fractions represents a physical block for the reaction with oxygen radicals. The chemical properties and antioxidant activities of sulfated polysaccharide fractions obtained by radical process degradation of crude sulfated polysaccharide were quite different from those obtained by acid hydrolysates. By radical process degradation, the high molecular weight was decreased to give LM2 (Mp 8 kDa) and LM1 (Mp 1.5 kDa), with a yield of 40% and 15%, respectively. LM2 was enriched with fucose and sulfated ester, while containing low amounts of glucuronic acid. The antioxidant activity showed that LM2 was unable to scavenge either superoxide or hydroxyl radical, which suggested that radical process degradation targeted mainly ascopyllan-like species rich in glucuronic acid, while the fraction rich in sulfated l-fucose remained unchanged. However, LM1 with Mp 1.5 kDa still retained apparent scavenging ability for superoxide radical, although it contained no glucuronic acid and certain amounts of galactose and mannose as main neutral sugars. These result suggest that the antioxidant activity of sulfated polysaccharides is apparently related not only to molecular weight and sulfated ester content, as previously determined, but also to glucuronic acid and fucose content.     

Bioactive compounds from brown seaweeds: Phloroglucinol,fucoxanthin and fucoidan as promising therapeutic agents against breast cancer

Breast cancer is one of the most common cancers among women and its incidence tends to increase year by year. Chemotherapy is an effective treatment for many types of cancer, however its toxicity in normal cells and acquired tumor resistance to the drug used are considered as the main barriers. New strategies have been proposed to increase the success of anticancer drugs namely it combination with natural dietary compounds, decreasing drug dose administered and reducing its toxicity to normal cells. Seaweeds are rich in bioactive compounds and, in Traditional Chinese Medicine and Japanese folk medicine are used to “treat” tumors. Attending to the attractive biological effects of some seaweed several efforts have been made to isolate the bioactive compounds and explore its action mechanisms. Phloroglucinol, fucoxanthin and fucoidan are bioactive compounds present in brown seaweed showing chemopreventive and chemotherapeutic effects against cancer. Several mechanisms namely antioxidant, cell cycle arrest, induction of cell death and inhibition of metastasis and angiogenesis have been mentioned as responsible for it anticancer activity. Beside the promising biological effects of these compounds, synergistic effects with cytotoxic drugs have been less explored. This review focuses on the potential protective and therapeutic effect – mainly against breast cancer – of the bioactive compounds phloroglucinol, fucoxanthin and fucoidan present in the brown seaweeds. Current knowledge about interaction between each of these compounds and the conventional anticancer drugs and the further research opportunities are discussed.     

Regeneration from callus of Undaria pinnatifida (Harvey) Suringar (Laminariales,Phaeophyta)

Calli were formed on the explants of midrib, meristem and immature stipe parts from freshly collected Undaria pinnatifida sporophytes. Each part was sterilized by Betadine and ethanol, and was cut into explants. The explants were incubated on an agar medium at 10 hours light and 14 hours dark photoperiod under a photon flux density of 80 µmol m^–2 s^–1. Callus was formed best on the explants of meristem parts at a temperature of 13 °C on PESI medium. Calli were cut off from the explants and were transferred into a sterile liquid PESI medium in flasks. Callus was dark brown in colour and was composed of well-pigmented cells. The cells were loosely bound and were separated by low power sonication, and were easy to attach to vinylon strings. From the calli formed on the explants of meristem parts, entire fronds were regenerated, but from the calli formed on the explants of midrib parts, only thin layered laminae were regenerated. The calli formed on the explants of immature stipe parts did not exhibit any regeneration at all.     

Influence of mineralisation methods on the determination of the mineral content of the brown seaweed Undaria pinnatifida by atomic absorption spectrophotometry

The influence of 8 mineralisation (dry and wet ashings) procedures on the determination of the mineral content of the brown seaweed Undaria pinnatifida by atomic absorption spectrophotometry was studied. Two procedures are proposed for routine analysis: dry ashing using sample calcination at 450 °C, followed by ash treatment with hydrochloric acid and silica elimination by hydrofluoric acid, and wet digestion using hot nitric acid.     

Seasonal changes in growth rate, morphology and alginate content in Undaria pinnatifida at the northern limit in the Sea of Japan (Russia)

The growth, morphology, alginate yield and composition of Undaria pinnatifidawas studied from March to August in 2000 and 2001 at the northern limit of distribution of the species in Peter the Great Bay, the Sea of Japan (Russia). The changes in morphology, alginate yield and composition were caused by sporophyte growth and sporulation. The average rate of biomass change was 2–5% d⁻¹. The highest alginate content (51% d.wt) was obtained from blades, with lower values for sporophylls and midribs. An increase in alginate content was detected before sporulation. The conditions seem favourable for farming the alga in this region, with June the optimum month for harvesting. This revised version was published online in August 2006 with corrections to the Cover Date.     

Intraspecific Genetic Diversity of Undaria pinnatifida in Japan, based on the Mitochondrial cox3 Gene and the ITS1 of nrDNA

The intraspecific genetic diversity of the kelp Undaria pinnatifida (Harvey) Suringar (Laminariales, Phaeophyceae) was investigated using DNA sequences of the mitochondrial cytochrome oxidase subunit 3 (cox3) gene and internal transcribed spacer 1 (ITS1) of nuclear ribosomal DNA in plants collected from 21 localities along the Japanese coast between 2001 and 2003. Morphological variation was also examined and compared with the genetic diversity. Cox3 analyses of 106 plants revealed 9 haplotypes (I–IX) that differed from each other by 1–7 bp (all synonymous substitutions). Haplotype I was distributed in Hokkaido and the northern Pacific coast of Honshu, while haplotype III was found along the Sea of Japan coast of Honshu. Other types were found along the central and southern coast of Honshu. ITS1 analyses of 42 plants revealed 0–1.7% nucleotide differences, but plants from the Sea of Japan coast and northern Japan had similar sequences. The lower genetic differentiation along the Sea of Japan and northern coasts might be due to the recent establishment (after the middle of the last glacial period) of the Sea of Japan flora. The cox3 haplotype of cultivated plants was found in natural populations occurring close to cultivation sites (Naruto, Tokushima Pref., and Hokutan, Hyogo Pref.). This suggests that cultivated plants possibly escaped and spread or crossed with plants of natural populations. Morphological analyses of variation in 10 characters were conducted using 66 plants. The results showed no significant local variation owing to the wide variation in each population and did not support any forma previously described. No correlations between the morphological characters and cox3 haplotypes were detected.     

Effect of season on the composition of bioactive polysaccharides from the brown seaweed Saccharina longicruris

The structural features of laminarans and galactofucans extracted from the brown seaweed Saccharina longicruris were determined for four harvest periods (M05, A05, N05 and J06). Crude laminarans were purified and crude galactofucans were fractionated using DEAE Sepharose anion exchange chromatography with increasing levels of NaCl (0.5, 1 and 2 M). The results showed differences in terms of their monosaccharide compositions. Purified laminaran contained a high proportion of d-glucose, between 45.1% and 69.1%, with a higher amount in M05 and A05, while the amount of d-mannitol remained constant (less than 1.7%). Crude galactofucans from M05, A05, and N05 contained 19.9-21.5% of sulphates, where J06 had only 14.3%. The 2 M fractionated galactofucans contained a higher proportion of sulphate groups, from 27.1% to 36.9%, for each harvest period, while the 1 M fraction contained 9.2% to 15.9% of sulphates. An important variation in the amount of l-fucose and d-galactose was observed for crude and fractionated galactofucans. In M05, a higher content of l-fucose was observed for crude galactofucans compared to that observed for d-galactose (21.5% vs. 11.1%), whereas the opposite was found for A05 (18.5% vs. 36.6%), N05 (20.9% vs. 36.8%), and J06 (12.8% vs. 19.6%). Also, the 0.5 and 2 M fractions were similar to the crude galactofucans. A05, N05, and J06 contained lower amounts of l-fucose than d-galactose, while the M05 fractions showed the opposite behaviour. However, the 1 M fraction showed a higher amount of l-fucose than d-galactose for each harvest period. The next step will be to study the biological activity of the fractions and to attempt to relate this activity to the structure of the galactofucan and laminaran fractions.     

In vitro and in vivo immunomodulatory activity of sulfated polysaccharides from Enteromorpha prolifera

Water-soluble sulfated polysaccharides extracted from Enteromorpha prolifera and fractionated using ion-exchange chromatography (crude, F₁, F₂ and F₃ fractions) were investigated to determine their in vitro and in vivo immunomodulatory activities. The sulfated polysaccharides, especially the F₁ and F₂ fractions, stimulated a macrophage cell line, Raw 264.7, inducing considerable nitric oxide (NO) and various cytokine production via up-regulated mRNA expression. The in vivo experiment results show that the sulfated polysaccharides (the crude and F₂ fractions) significantly increased Con A-induced splenocyte proliferation, revealing their potential comitogenic activity. In addition, IFN-γ and IL-2 secretions were considerably increased by the F₂ fraction without altering the release of IL-4 and IL-5. This implies that the F₂ fraction can activate T cells by up-regulating Th-1 response and that Th-1 cells might be the main target cells of the F₂ fraction. These in vitro and in vivo results suggest that the sulfated polysaccharides are strong immunostimulators.     

Immunomodulation and antitumor activities of different-molecular-weight polysaccharides from Porphyridium cruentum

The extracellular polysaccharides (EPSs) isolated from Porphyridium cruentum were degraded by hermetical-microwave and H₂O₂ under ultrasonic waves. Six products were obtained with molecular weights of 6.53, 256, 606, 802.6, 903.3 and 1002 kDa. The antitumor and immunomodulatory activities of different-molecular-weight (MW) polysaccharides were evaluated by the S180-tumor-bearing mouse model in vivo and peritoneal macrophage activation in vitro. The degraded EPSs all showed clear immunomodulation to different extents. The MW of the EPSs had a notable effect on their activity. The 6.53-kDa fragment had the strongest immunoenhancing activity. Different doses of EPS all inhibited the growth of the implanted S180 tumor. The tumor inhibition index at high, middle and low doses was 53.3%, 47.5% and 40.5%, respectively. In addition, three different concentrations of EPS significantly increased lymphocyte proliferation, which indicated the unique mechanism of the antitumor effect of EPS.     

Epi-endophytic symbiosis between Laminariocolax aecidioides (Ectocarpales, Phaeophyceae) and Undaria pinnatifida (Laminariales, Phaeophyceae) growing on Argentinian coasts

The present is the first study on epi-endophytic algae on thalli of Undaria pinnatifida growing along Argentinian coasts. The main goal is to describe the nature and the morphology of this symbiosis. Individuals of Laminariocolax aecidioides were detected in both June and December 2004, growing on U. pinnatifida sporophytes. In nature, the epi-endophyte were macroscopically observed as dark zones that partially covered the hosts’ fronds. L. aecidioides vegetative thalli were irregularly branched uniseriate filaments. The life cycle is described from laboratory cultures started from Patagonian populations. Caryology revealed that the sporophytic diploid phase presented 16 chromosomes whereas the gametophytic haploid phase presented 8 chromosomes. Isolates made from thalli growing in the interior of infected hosts developed into filamentous, branched sporophytes that reproduced by both unispores and plurispores that were produced in unilocular and plurilocular sporangia, respectively. The results of this paper also allowed us to conclude that L. aecidioides uses the thalli of U. pinnatifida as a proper substrate. The penetration of endophitic filaments among the host′s cortical cells produced a lateral compression and, finally, their thalli development generated perforations in the host tissues. The effects of the epi-endophytic infection of L. aecidioides on U. pinnatifida are neither severe nor deleterious.     

Sulfated polysaccharides from marine green algae Ulva conglobata and their anticoagulant activity

Sulfated polysaccharides from the green algae Ulva conglobata were isolated and prepared by extraction in hot water, precipitation with ethanol and purification by ion-exchange and size-exclusion column chromatography. The characterizations of the sulfated polysaccharides were defined, and containing 23.04–35.20% sulfate ester groups, 10.82–14.91% uronic acid and 3.82–4.51% protein. Gas chromatography analysis shows that the sulfated polysaccharides from Ulva conglobata are mainly consisted of rhamnose with variable contents of glucose and fucose, trace amounts of xylose, glactose and mannose. The anticoagulant properties of the sulfated polysaccharides were compared with those of heparin by studying the activated partial thromboplastin time using normal human plasma. The sulfated polysaccharide from Ulva conglobata collected in Qingdao, China is the most potent among the sulfated polysaccharides tested. The mechanism of anticoagulant activity mediated by the sulfated polysaccharides is due to the direct inhibition of thrombin and the potentiation of heparin cofactor II.     

The expanding range of Undaria pinnatifida in southern New Zealand: distribution, dispersal mechanisms and the invasion of wave-exposed environments

Very few studies have addressed how the invasive kelp Undaria pinnatifida (Harvey) Suringar spreads beyond initial founding populations in harbours. Surveys of the harbours and accessible areas of open coast throughout southern New Zealand were conducted to determine how far U. pinnatifida populations had extended since initial incursions. Our findings clearly demonstrate that U. pinnatifida is capable of invading native communities and can establish reproductive populations in locations subjected to significant and consistent wave action. The extent of spread from source populations differs between harbours in which it has established. Dispersal is greatest in harbours with long established populations, those where populations have not been strategically managed, harbours with high water exchange with surrounding coastal waters, and where prevailing currents allow establishment of U. pinnatifida on suitable substrata close to harbour entrances. Dispersal along the open coast is primarily achieved by drifting adult sporophytes that are washed up in the rocky intertidal zone. Founding populations are most often found in the intertidal zone, primarily within rockpools. Subtidal transects and observations indicate that U. pinnatifida is well adapted to invade exposed coastlines and can establish within a broad range of niches in wave-exposed areas including rockpools, the low intertidal, shallow subtidal, Macrocystis pyrifera kelp forests, and in low light areas beyond the vertical extent of large native macroalgae. The current range of U. pinnatifida is much greater than expected and appears to be expanding. Due to its ability to grow in a broad range of environments and to form dense monospecific stands, U. pinnatifida has the potential to strongly modify almost all rocky subtidal and intertidal communities in temperate locations.     

The antitumor and antioxidative activities of polysaccharides isolated from Isaria farinosa B05

The water-soluble intra-polysaccharides WIPS1 and water-soluble extra-polysaccharides WEPS1 were isolated from Isaria farinosa B05 through ethanol precipitation and gel permeation chromatography (GPC). Their characteristics were determined by chemical analysis, gas chromatography, GPC and IR spectroscopy. The results show that WIPS1 contained 90.3% carbohydrate, 8.00% uronic acid, 7.15% protein and three kinds of monosaccharides including mannose, galactose and glucose with a molar ratio of 8.0:4.8:1.0. WEPS1 contained 93.4% carbohydrate, 8.06% uronic acid, 4.40% protein and three kinds of monosaccharides including mannose, galactose and glucose with a molar ratio of 21.6:4.7:1.0. WIPS1 and WEPS1 had a molecular weight of 42 and 208kDa, respectively. The in vivo tests in mice indicate that WIPS1 and WEPS1 had significant antitumor and antioxidative activities to some extent.