Supplementation with Ascophyllum nodosum extracts mitigates arsenic toxicity by modulating reactive oxygen species metabolism and reducing oxidative stress in rice.

Ascophyllum nodosum extract (ANE) is considered as an effective source of biostimulants that have the potential of ameliorating the negative impact of different abiotic stresses in plants. Considering the growth-promoting ability and other regulatory roles of ANE, the present investigation was executed to evaluate the role of ANE in conferring arsenic (As) tolerance in rice (Oryza sativa L. cv. BRRI dhan89). Rice seedlings (35-d-old) were exposed to two doses of sodium arsenate (As1 - 50 mg As kg-1 soil; As2 - 100 mg As kg-1 soil) at 25 days after transplanting through irrigation, whereas only water was applied to the control. Foliar application of 0.1% ANE was also supplemented under control as well as As-stressed conditions at 7 days intervals for 5 times. Arsenic-induced oxidative stress was evident through a sharp increase in lipid peroxidation, hydrogen peroxide, methylglyoxal, and electrolyte leakage in the As-treated plants. As a consequence, plant growth and biomass, leaf relative water content, as well as yield attributes were reduced noticeably. On the other hand, ANE supplemented plants accumulated enhanced levels of ascorbate and glutathione, their redox balance, and the activities of antioxidant and glyoxalase enzymes viz. ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, catalase, glutathione peroxidase, and activities of glyoxalase I and glyoxalase II, respectively. Furthermore, relative water content, plant growth, yield attributes and yield were increased in the As-treated rice plants with ANE supplementation. The results reflected that foliar spray with ANE alleviated As-induced oxidative stress in rice plants by modulating the antioxidative defense and glyoxalase system.

Insight into the physiological and biochemical mechanisms of biostimulating effect of Ascophyllum nodosum and Moringa oleifera extracts to minimize cadmium-induced oxidative stress in rice.

Cadmium (Cd) is a serious threat for environmental sustainability as it can be taken up quickly by plants and transported to the food chain of living organisms. It alters plants' metabolic and physiological activities and causes yield loss, thereby, enhancing plant tolerance to Cd stress is of utmost essential. Therefore, an experiment was executed to investigate the potential role of Ascophyllum nodosum extract (ANE) and moringa (Moringa oleifera) leaf extract (MLE) to confer Cd tolerance in rice (Oryza sativa cv. BRRI dhan89). Thirty-five-day-old seedling was subjected to Cd stress (50 mg kg-1 CdCl2) alone and in a combination of ANE (0.25%) or MLE (0.5%) in a semi-controlled net house. Exposure to Cd resulted in accelerated production of reactive oxygen species, enhanced lipid peroxidation, and disrupted antioxidant defense and glyoxalase system, thus retarded plant growth, biomass production, and yield attributes of rice. On the contrary, the supplementation of ANE or MLE enhanced the contents of ascorbate and glutathione, and the activities of antioxidant enzymes such as ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase, glutathione peroxidase, and catalase. Moreover, supplementation of ANE and MLE enhanced the activities of glyoxalase I and glyoxalase II which prevented the overgeneration of methylglyoxal in Cd stressed rice plants. Thus, because of ANE and MLE addition Cd-induced rice plants showed a noticeable declination in membrane lipid peroxidation, hydrogen peroxide generation, and electrolyte leakage, whereas improved water balance. Furthermore, the growth and yield attributes of Cd-affected rice plants were improved with the supplementation of ANE and MLE. All the studied parameters indicates the potential role of ANE and MLE in mitigating Cd stress in rice plants through improving the physiological attributes, modulating antioxidant defense and glyoxalase system.

Ascophyllum nodosum and Fucus vesiculosus Extracts Improved Lipid Metabolism and Inflammation in High-Energy Diet-Induced Hyperlipidemia Rats.

Ascophyllum nodosum and Fucus vesiculosus both contain unique polyphenols called phlorotannins. Phlorotannins reportedly possess various pharmacological activities. A previous study reported that the activity of phlorotannin is strongly correlated with the normalization of metabolic function, and phlorotannins are extremely promising nutrients for use in the treatment of metabolic syndrome. To date, no study has explored the antihyperlipidemic effects of phlorotannins from A. nodosum and F. vesiculosus in animal models. Therefore, in the present study, we investigated the effects of phlorotannins using a rat model of high-energy diet (HED)-induced hyperlipidemia. The results showed that the rats that were fed an HED and treated with phlorotannin-rich extract from A. nodosum and F. vesiculosus had significantly lower serum fasting blood sugar (FBS), aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triacylglyceride (TG) and free fatty acids (FFAs) levels and hepatic TG level and had higher serum insulin, high-density lipoprotein cholesterol (HDL-C) levels and lipase activity in their fat tissues than in the case with the rats that were fed the HED alone. A histopathological analysis revealed that phlorotannin-rich extract could significantly reduce the size of adipocytes around the epididymis. In addition, the rats treated with phlorotannin-rich extract had significantly lowered interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) levels and increased superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities than did those in the HED group. These results suggested that the phlorotannin-rich extract stimulated lipid metabolism and may have promoted lipase activity in rats with HED-induced hyperlipidemia. Our results indicated that A. nodosum and F. vesiculosus, marine algae typically used as health foods, have strong antihyperlipidemic effects and may, therefore, be useful for preventing atherosclerosis. These algae may be incorporated into antihyperlipidemia pharmaceuticals and functional foods.

Supplementation of Ascophyllum nodosum meal and monensin: Effects on diversity and relative abundance of ruminal bacterial taxa and the metabolism of iodine and arsenic in lactating dairy cows.

Previous research has shown that the brown seaweed Ascophyllum nodosum (ASCO) has antimicrobial and antioxidant properties and also increases milk I concentration. We aimed to investigate the effects of supplementing ASCO meal or monensin (MON) on ruminal fermentation, diversity and relative abundance of ruminal bacterial taxa, metabolism of I and As, and blood concentrations of thyroid hormones, antioxidant enzymes, and cortisol in lactating dairy cows. Five multiparous ruminally cannulated Jersey cows averaging (mean ± standard deviation) 102 ± 15 d in milk and 450 ± 33 kg of body weight at the beginning of the study were used in a Latin square design with 28-d periods (21 d for diet adaptation and 7 d for data and sample collection). Cows were fed ad libitum a basal diet containing (dry matter basis) 65% forage as haylage and corn silage and 35% concentrate and were randomly assigned to 1 of the following 5 dietary treatments: 0, 57, 113, or 170 g/d of ASCO meal, or 300 mg/d of MON. Supplements were placed directly into the rumen once daily after the morning feeding. Diets had no effect on ruminal pH and NH3-N concentration, which averaged 6.02 and 6.86 mg/dL, respectively. Total volatile fatty acid concentration decreased linearly in cows fed incremental amounts of ASCO meal. Supplementation with ASCO meal did not change the ruminal molar proportions of volatile fatty acids apart from butyrate, which responded quadratically with the lowest values observed at 56 and 113 g/d of ASCO supplementation. Compared with the control diet or diets containing ASCO meal, cows fed MON showed greater molar proportion of propionate. Diets did not affect the α diversity indices Shannon, Simpson, and Fisher for ruminal bacteria. However, feeding incremental levels of ASCO meal linearly decreased the relative abundance of Tenericutes in ruminal fluid. Monensin increased the relative abundance of the CAG:352 bacterial genus in ruminal fluid compared with the control diet. Linear increases in response to ASCO meal supplementation were observed for the concentrations and output of I in serum, milk, urine, and feces. Fecal excretion of As increased linearly in cows fed varying amounts of ASCO meal, but ASCO did not affect the concentration and secretion of As in milk. The plasma activities of the antioxidant enzymes and the serum concentrations of thyroid hormones did not change. In contrast, circulating cortisol decreased linearly in diets containing ASCO meal. The apparent total-tract digestibilities of dry matter, organic matter, and crude protein increased linearly with ASCO meal, but those of neutral and acid detergent fiber were not affected. In summary, feeding incremental amounts of ASCO meal decreased serum cortisol concentration, and increased I concentrations and output in serum, milk, feces, and urine.

Seasonal Variation of the Proximate Composition, Mineral Content, Fatty Acid Profiles and Other Phytochemical Constituents of Selected Brown Macroalgae.

The main objective was to determine the chemical, phytochemical, fatty acid and mineral profiles of three commercially relevant brown macroalgae (Laminaria digitata, Laminaria hyperborea and Ascophyllum nodosum) collected each season for two years off the west coast of Ireland. All the chemical, phytochemical, fatty acid and minerals analysed varied significantly depending on the macroalgal species, season and year of collection. Overall, the protein contents of macroalgae were negatively correlated with carbohydrate content. Protein (2-11%) was at its highest during winter and/or spring, decreasing to a minimum during summer and/or autumn. The three macroalgal species analysed in this study had clearly differentiated fatty acid profiles. The concentration of fatty acids was higher in A. nodosum compared with both Laminaria species. The mineral profile of the three macroalgal species was rich in essential metals, particularly Ca, Mg and P, while the levels of I were approximately 9- to 10-fold higher in both Laminaria spp. compared with A. nodosum. The levels of toxic metals (Cd, Hg and Pb) in all the macroalgal species studied were low in the current study; while the levels of total As were high (49-64 mg/kg DW macroalgae) compared with previous reports.

Suppressive effects of sulfated polysaccharide ascophyllan isolated from Ascophyllum nodosum on the production of NO and ROS in LPS-stimulated RAW264.7 cells.

In this study, we found that a sulfated polysaccharide isolated from the brown alga Ascophyllum nodosum, ascophyllan, showed suppressive effects on stimulated RAW264.7 cells. Ascophyllan significantly inhibited expression of inducible nitric oxide synthase mRNA and excessive production of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells in a dose-dependent manner without affecting the viability of RAW264.7 cells. Ascophyllan also reduced the elevated level of intracellular reactive oxygen species (ROS) in LPS-stimulated RAW264.7 cells. Furthermore, preincubation with ascophyllan resulted in concentration-dependent decrease in ROS production in phorbol 12-myristate-13-acetate-stimulated RAW264.7 cells. Our results suggest that ascophyllan can exhibit anti-inflammatory effects on stimulated macrophages mainly through the attenuation of NO and ROS productions.

Consumer acceptability and chemical composition of whole-wheat breads incorporated with brown seaweed (Ascophyllum nodosum) or red seaweed (Chondrus crispus).

BACKGROUND: Seaweeds have been eaten in the diets of coastal cultures for centuries; however, consumption of seaweeds has been limited in Western diets owing to undesirable sensory characteristics and lack of familiarity. Apart from healthful bioactive metabolites, seaweeds are good sources of fibre and minerals. They are nearly a complete protein and have a low fat content (mainly mono- or polyunsaturated). The objectives were (i) to investigate if the addition of brown seaweed, Ascophyllum nodosum, or red seaweed, Chondrus crispus, altered the chemical composition and sensory properties of whole-wheat bread; and (ii) to determine what percentage the addition of brown or red seaweed to whole-wheat bread is acceptable to consumers. The two seaweeds were incorporated into separate batches of whole-wheat bread by percentage weight flour at 0% (control), 2%, 4%, 6%, and 8%. RESULTS: The products containing the highest amounts of A. nodosum and C. crispus had the highest ash and total dietary fibre. A. nodosum and C. crispus breads were acceptable at 4% and 2% levels respectively. The attributes of no aftertaste, soft, and chewy drove consumer liking of the whole-wheat bread, whereas attributes dry, dense, strong aftertaste, and saltiness detracted from liking. CONCLUSION: This project's significance is to demonstrate the acceptability of seaweed in a Western population, which may lay the groundwork to encourage and promote the consumption of seaweed or to exemplify seaweed incorporation into foodstuffs. © 2020 Society of Chemical Industry.

Fucoidan isolated from Ascophyllum nodosum alleviates gut microbiota dysbiosis and colonic inflammation in antibiotic-treated mice.

Antibiotic treatment, as an important therapeutic intervention, can cause damage to the host microbiome and the intestinal mucosal barrier. In order to find a way to alleviate the side effects of antibiotics, the present study investigated the effects of fucoidan (ANP) isolated from Ascophyllum nodosum on gut microbiota dysbiosis and colonic inflammation induced by ciprofloxacin-metronidazole (CiMe) in C57BL/6J mice. Our results showed that dietary ANP prevented colon shortening, alleviated the colonic tissue damages, and partially reversed the alteration of gut microbiota by increasing the abundance of potentially beneficial bacteria, e.g., Ruminococcaceae_UCG_014 and Akkermansia and decreasing the abundance of harmful bacteria, e.g., Proteus and Enterococcus. ANP also suppressed the overproduction of TNF-α, IL-1ß, and IL-6 and promoted the expression of IL-10. In addition, ANP reversed the decreased production of short-chain fatty acids in CiMe-treated mice. Furthermore, correlation analysis indicated the presence of critical gut microbiota, which played important roles in reducing the inflammation-related indices. Thus, the present study suggests that fucoidan isolated from Ascophyllum nodosum is effective in providing protection against the negative effects of antibiotics on gut microbiota and colonic health.

Adsorption of indium by waste biomass of brown alga Ascophyllum nodosum.

The biosorption capacities of dried meal and a waste product from the processing for biostimulant extract of Ascophyllum nodosum were evaluated as candidates for low-cost, effective biomaterials for the recovery of indium(III). The use of indium has significantly grown in the last decade, because of its utilization in hi-tech. Two formats were evaluated as biosorbents: waste-biomass, a residue derived from the alkaline extraction of a commercial, biostimulant product, and natural-biomass which was harvested, dried and milled as a commercial, "kelp meal" product. Two systems have been evaluated: ideal system with indium only, and double metal-system with indium and iron, where two different levels of iron were investigated. For both systems, the indium biosorption by the brown algal biomass was found to be pH-dependent, with an optimum at pH3. In the ideal system, indium adsorption was higher (maximum adsorptions of 48 mg/g for the processed, waste biomass and 63 mg/g for the natural biomass), than in the double metal-system where the maximum adsorption was with iron at 0.07 g/L. Good values of indium adsorption were demonstrated in both the ideal and double systems: there was competition between the iron and indium ions for the binding sites available in the A. nodosum-derived materials. Data suggested that the processed, waste biomass of the algae, could be a good biosorbent for its indium absorption properties. This had the double advantages of both recovery of indium (high economic importance), and also definition of a virtuous circular economic innovative strategy, whereby a waste becomes a valuable resource.

Optimization of hyper-thermal acid hydrolysis and enzymatic saccharification of Ascophyllum nodosum for ethanol production with mannitol-adapted yeasts.

In this study, Ascophyllum nodosum was studied as a biomass for ethanol production. A. nodosum was degraded to monosaccharide by hyper-thermal (HT) acid hydrolysis and enzymatic saccharification and analyzed using response surface methodology (RSM) and the Michaelis-Menten equation. Maximum monosaccharide concentrations of 20.3 g/L glucose and 7.0 g/L mannitol were obtained from HT acid hydrolysis and enzymatic saccharification from 8%(w/v) of A. nodosum. Fermentation was conducted using Pichia stipitis and P. angophorae adapted to high mannitol concentrations. Neither non-adapted P. stipitis and P. angophorae nor adapted P. stipitis could ferment all mannitol in the A. nodosum hydrolysate. Adapted P. angophorae produced the highest ethanol concentration among various yeasts, with ethanol production reaching 13.6 g/L with an ethanol yield (YEtOH) of 0.50. Optimization of HT acid hydrolysis and enzymatic saccharification, in combination with the use of adapted yeast, could enhance overall A. nodosum ethanol fermentation yields.

Extracts from the edible seaweed, Ascophyllum nodosum, inhibit lipase activity in vitro: contributions of phenolic and polysaccharide components.

A polyphenol-rich extract (PRE) from the edible seaweed, Ascophyllum nodosum, inhibited pancreatic lipase activity in an oil-based turbidimetric assay with an IC50 of 200 µg gallic acid equivalents (GAE) perassay) [∼230 µg DW] whereas the known inhibitor, Orlistat, gave an IC50 at 0.4 µg per assay. A phlorotannin-enriched fraction (TRF) purified from the PRE was more potent with an IC50 = 60 µg GAE per assay (∼65 µg DW). When the assay was started by the addition of lipase, both Orlistat and TRF were much less effective which suggests that pre-incubation of enzyme and inhibitor improved inhibition. Based on phenol content, water extracts from Ascophyllum were more potent lipase inhibitors than PRE (IC50 ∼ 150 µg GAE per assay). However, this was equivalent to ∼580 µg DW and these extracts contained polysaccharides (e.g. alginate content = 110 µg mL-1) which may also contribute to inhibition. Indeed, a polysaccharide-enriched fraction obtained by ethanol precipitation gave an IC50 of 1000 µg DW which was equivalent to 130 µg GAE and 420 µg alginate per assay. Therefore a >3 fold increase in alginate content did not markedly improve inhibition. Re-precipitation increased alginate content and reduced polyphenol content but lipase inhibition was markedly reduced (i.e. IC50 at ∼1100 µg DW per assay, 700 µg alginate and 25 µg GAE). Purifying the polysaccharide fraction by ion exchange removed all phenolics but the IC50 increased to >2500 µg DW, equivalent to >1970 µg alginate per assay. In conclusion, polysaccharides and phlorotannins may inhibit lipase in an additive fashion, with phlorotannins apparently more effective in vitro. However, interactions between these components may be important when food products containing this edible seaweed are consumed.

Differences in Metal Accumulation in the Growing Shoot Tips and Remaining Shoot Tissue in Three Species of Brown Seaweeds.

In this study we compared the accumulation of eight metals and two metalloids in the growing tips and the remaining shoot tissue of three species of brown seaweeds commonly used in biomonitoring studies (Fucus ceranoides, Fucus spiralis and Ascophyllum nodosum). Regression analysis of the data obtained showed that there was no statistically significant difference in accumulation in numerous cases; although most of the relationships were significant, many of the coefficients of determination were low. However, the concentrations of Mn and Zn in the growing tips were closely related to the concentrations in the rest of the tissue in all three species, possibly due to redistribution of these elements. Interspecies differences in bioconcentration of the elements may be partly explained by differences in the relative growth rates.

Seasonal variation of chemical composition and biomethane production from the brown seaweed Ascophyllum nodosum.

Ascophyllum nodosum, an abundant Irish brown seaweed, shows significant seasonal variation in chemical composition and biogas production. The polyphenol content is shown to be a more important factor in biogas production than ash content. High polyphenol content in summer months adversely affected biogas production; suggesting two potential harvest dates, March and October. A. nodosum harvested in October showed a relatively low level of polyphenols (2% of TS) and ash (23% of volatile solids), and exhibited a specific methane yield of 215LCH4kgVS(-1), which was 44% of theoretical yield. The highest yield per wet weight of 47m(3)CH4t(-1) was achieved in October, which is 2.9 times higher than the lowest value (16m(3)CH4t(-1)), obtained in December. The gross energy yield of A. nodosum based on the optimal biogas production can achieve 116GJha(-1)yr(-1) in October.

Incremental amounts of Ascophyllum nodosum meal do not improve animal performance but do increase milk iodine output in early lactation dairy cows fed high-forage diets.

The objective of this study was to investigate the effects of incremental amounts of Ascophyllum nodosum meal (ANOD) on milk production, milk composition including fatty acids and I, blood metabolites, and nutrient intake and digestibility in early lactation dairy cows fed high-forage diets. Twelve multiparous Jersey cows averaging (mean±standard deviation) 40±21 d in milk and 464±35 kg of body weight and 4 primiparous Jersey cows averaging 75±37 d in milk and 384±17kg of body weight were randomly assigned to treatment sequences in a replicated 4×4 Latin square design. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Cows were fed a total mixed ration (64:36 forage-to-concentrate ratio) supplemented (as fed) with 0, 57, 113, or 170 g/d of ANOD. Milk yield as well as concentrations and yields of milk components (fat, protein, lactose, milk urea N) were not affected by increasing dietary amounts of ANOD. Concentration (from 178 to 1,370 µg/L) and yield (from 2.8 to 20.6 mg/d) of milk I increased linearly in cows fed incremental amounts of ANOD as a result of the high concentration of I (820 mg/kg of dry matter) in ANOD. Overall, only minor changes were observed in the proportion of milk fatty acids with ANOD supplementation. Quadratic trends were observed for dry matter intake and total-tract digestibilities of organic matter and neutral detergent fiber, whereas negative linear trends were observed for serum concentration of cortisol and crude protein digestibility with ANOD supplementation. Serum concentrations of triiodothyronine and thyroxine were not affected by ANOD supplementation and averaged 1.1 and 48.4 ng/mL, respectively. However, feeding increasing amounts of ANOD linearly reduced the plasma concentration of nonesterified fatty acids (from 164 to 132 mEq/L). Quadratic effects were found for the total-tract digestibility of ADF and urinary output of purine derivatives, suggesting that ANOD supplementation may stimulate growth of ruminal cellulolytic bacteria in a dose-dependent fashion. In general, feeding incremental amounts of ANOD to early lactation dairy cows dramatically increased milk I concentration and output with no effect on animal performance.

Ascophyllan purified from Ascophyllum nodosum induces Th1 and Tc1 immune responses by promoting dendritic cell maturation.

Marine-derived sulfated polysaccharides have been shown to possess certain anti-virus, anti-tumor, anti-inflammatory and anti-coagulant activities. However, the in vivo immunomodulatory effects of marine-derived pure compounds have been less well characterized. In this study, we investigated the effect of ascophyllan, a sulfated polysaccharide purified from Ascophyllum nodosum, on the maturation of mouse dendritic cells (DCs) in vitro and in vivo. Ascophyllan induced up-regulation of co-stimulatory molecules and production of pro-inflammatory cytokines in bone marrow-derived DCs (BMDCs). Moreover, in vivo administration of ascophyllan promotes up-regulation of CD40, CD80, CD86, MHC class I and MHC class II and production of IL-6, IL-12 and TNF-α in spleen cDCs. Interestingly, ascophyllan induced a higher degree of co-stimulatory molecule up-regulation and pro-inflammatory cytokine production than fucoidan, a marine-derived polysaccharide with well-defined effect for promoting DC maturation. Ascophyllan also promoted the generation of IFN-γ-producing Th1 and Tc1 cells in the presence of DCs in an IL-12-dependent manner. Finally, myeloid differentiation primary response 88 (MyD88) signaling pathway was essential for DC maturation induced by ascophyllan. Taken together, these results demonstrate that ascophyllan induces DC maturation, and consequently enhances Th1 and Tc1 responses in vivo. This knowledge could facilitate the development of novel therapeutic strategies to combat infectious diseases and cancer.

Tasco®: a product of Ascophyllum nodosum enhances immune response of Caenorhabditis elegans against Pseudomonas aeruginosa infection.

The effects of Tasco®, a product made from the brown seaweed (Ascophyllum nodosum) were tested for the ability to protect Caenorhabditis elegans against Pseudomonas aeruginosa infection. A water extract of Tasco® (TWE) reduced P. aeruginosa inflicted mortality in the nematode. The TWE, at a concentration of 300 µg/mL, offered the maximum protection and induced the expression of innate immune response genes viz.; zk6.7 (Lypases), lys-1 (Lysozyme), spp-1 (Saponin like protein), f28d1.3 (Thaumatin like protein), t20g5.7 (Matridin SK domain protein), abf-1 (Antibacterial protein) and f38a1.5 (Lectin family protein). Further, TWE treatment also affected a number of virulence components of the P. aeuroginosa and reduced its secreted virulence factors such as lipase, proteases and toxic metabolites; hydrogen cyanide and pyocyanin. Decreased virulence factors were associated with a significant reduction in expression of regulatory genes involved in quorum sensing, lasI, lasR, rhlI and rhlR. In conclusion, the TWE-treatment protected the C. elegans against P. aeruginosa infection by a combination of effects on the innate immunity of the worms and direct effects on the bacterial quorum sensing and virulence factors.

Impacts of ambient salinity and copper on brown algae: 2. Interactive effects on phenolic pool and assessment of metal binding capacity of phlorotannin.

The aim of this study was to establish in laboratory experiments a quantitative link between phenolic pool (production, composition and exudation) in Ascophyllum nodosum and Fucus vesiculosus and their potential to bind metals. Additionally, the copper binding capacity of purified phlorotannin was investigated. A reduction in salinity decreased total phenolic contents, altered phenolic composition by increasing proportion of cell-wall phenolics, and also increased phenolic exudation of the two seaweed species. After 15 days at a salinity of 5, the inhibition of photosynthesis observed previously for A. nodosum coincided with the high exudation of phenolic compounds into the surrounding water of the seaweed tips which resulted in a significant reduction of phenolic contents. Increased copper concentration also reduced total phenolic contents, changed phenolic composition (increase in proportion and level of cell-wall phenolics), and positively affected phenolic exudation of A. nodosum and F. vesiculosus. A decrease in salinity enhanced the copper toxicity and caused the earlier impact on the physiology of seaweed tips. An involvement of phlorotannins in copper binding is also demonstrated; purified phlorotannins from A. nodosum collected from a site with little anthropogenic activity contained all four metals tested. When placed in copper-enriched water, as for the seaweed material, copper contents of the phenolics increased, zinc and cadmium contents decreased, but no change in chromium content was observed. The use of cell-wall phenolic content as biomarker of copper contamination seems promising but needs further investigation.

Impacts of ambient salinity and copper on brown algae: 1. Interactive effects on photosynthesis, growth, and copper accumulation.

The effect of copper enrichment and salinity on growth, photosynthesis and copper accumulation of two temperate brown seaweeds, Ascophyllum nodosum and Fucus vesiculosus, was investigated in laboratory experiments. A significant negative impact of reduced salinity on photosynthetic activity and growth was observed for both species. After 15 days at a salinity of 5, photosynthesis of A. nodosum was entirely inhibited and growth ceased at a salinity of 15. Increased copper concentration negatively affected photosynthetic activity of A. nodosum and F. vesiculosus resulting in chlorosis and reduced seaweed growth; 5 mg L⁻¹ copper caused an inhibition of the photosynthesis and the degradation of seaweed tips. Under reduced salinity, copper toxicity was enhanced and caused an earlier impact on the physiology of seaweed tips. After exposure to copper and different salinities for 15 days, copper contents of seaweeds were closely related to copper concentration in the water; seaweed copper contents reached their maximum after 1 day of exposure; contents only increased again when additional, free copper was added to the water. At high water copper concentrations or low salinity, or a combination of both, copper content of A. nodosum decreased. By contrast, copper content of F. vesiculosus increased, suggesting that different binding sites or uptake mechanisms exist in the two species. The results suggest that when using brown seaweeds in biomonitoring in situ, any change in the environment will directly and significantly affect algal physiology and thus their metal binding capacity; the assessment of the physiological status of the algae in combination with the analysis of thallus metal content will enhance the reliability of the biomonitoring process.

Technetium-99 ((99)Tc) in annual growth segments of knotted wrack (Ascophyllum nodosum).

The distribution of technetium-99 ((99)Tc) in annual growth segments of the brown seaweed Ascophyllum nodosum (Fucales, Phaeophyceae) from the southwestern coast of Norway is examined in samples collected from January to November 2006. A twenty-fold increase in the (99)Tc-concentration from the youngest to the oldest growth segments was found. The concentrations ranged from 42 to 98Bq/kg dry weight (d.w.) and from 964 to 1000Bq/kg d.w. in growth segments formed in 2006 and 1996, respectively. In addition, a seasonal variation in the (99)Tc concentration was observed in the actively growing 2006-segments: concentrations decreased from 98Bq/kg d.w. in April to 54Bq/kg d.w. in June; there was a further reduction from June to August (42Bq/kg d.w.); and, finally there was an increase from August to November (93Bq/kg d.w.). In most of the segments formed between 2000 and 2005, there was a tendency of slightly decreasing (99)Tc-concentrations between June and November but this pattern was not observed for the older growth segments. In order to find an explanation for the non-homogenous distribution of (99)Tc within thalli of A. nodosum, different hypotheses are discussed. Uptake and elimination of (99)Tc appears to be most pronounced in the actively growing segments. To date, such non-homogenous distribution of (99)Tc within thalli of A. nodosum has not been taken into consideration, neither in connection with sample collection nor analysis. This paper shows that special protocols must be followed if A. nodosum is going to be used as a bioindicator for (99)Tc in the marine environment. A sampling strategy is proposed.

Optimization of Cu(II) biosorption onto Ascophyllum nodosum by factorial design methodology.

A Box-Behnken factorial design coupled with surface response methodology was used to evaluate the effects of temperature, pH and initial concentration in the Cu(II) sorption process onto the marine macro-algae Ascophyllum nodosum. The effect of the operating variables on metal uptake capacity was studied in a batch system and a mathematical model showing the influence of each variable and their interactions was obtained. Study ranges were 10-40 degrees C for temperature, 3.0-5.0 for pH and 50-150 mg L(-1) for initial Cu(II) concentration. Within these ranges, the biosorption capacity is slightly dependent on temperature but markedly increases with pH and initial concentration of Cu(II). The uptake capacities predicted by the model are in good agreement with the experimental values. Maximum biosorption capacity of Cu(II) by A. nodosum is 70 mg g(-1) and corresponds to the following values of those variables: temperature=40 degrees C, pH=5.0 and initial Cu(II) concentration=150 mg L(-1).