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The ubiquitous and species rich genus Ulva comprises entities of green macroalgae with variable morphologies. Ulva species are important from ecological and economic perspectives, but their identification is often problematic. Current knowledge on Ulva diversity has focused mainly on foliose individuals of temperate regions, but genetic and morphological data on tubular species are often insufficient and the species richness is ambiguous due to the lack of molecularly identified type vouchers. Together with a previous study, our study demonstrates that due to the crypticity of tubular entities of the genus Ulva present in the Atlantic-Baltic Sea transect, certain species remained undetected until recently whereas molecular evidence of other historically identified species is missing. An entity which appears to be a relatively frequent species in the Atlantic-Baltic Sea transect and which was probably mis-identified with other species in the past is here described as Ulva capillata sp. nov.. The description is based on molecular identification using tufA and rbcL sequences, and by comparing the species´ phylogenetic relationships, distribution and range margins in the Atlantic-Baltic Sea transect, as well as on morpho-anatomical characters, and early ontogenetic development. By comparisons with closely related and potentially morphologically overlapping species concepts we were able to identify the uniqueness of U. capillata. Therefore, the description of U. capillata as a new species within the genus Ulva is supported by a combination of molecular, morphological, and ontogentic evidence which confirm their uniqueness in comparison to other species concepts.Furthermore, our results strongly emphasize the importance and necessity to molecularly investigate especially tubular historic type vouchers within the genus Ulva to facilitate a clear species identification to omit continuing with taxonomic confusion and ongoing misapplication of names of e.g. cryptic species concepts within this important green algal genus.

The cultivation of kelp typically involves two stages, where an indoor hatchery phase proceeds the grow-out phase at-sea. The in situ adhesion of microscopic propagules onto specifically designed substrates using binders has been proposed as an alternative to conventional hatchery methods where juvenile seaweeds are cultured on seeded twine, aimed at saving resources while increasing productivity as it obviates the hatchery phase. Here we tested how well kelp (Saccharina latissima) gametophytes adhere to cultivation ropes using two binder types (agar and κ-carrageenan) and application treatment (separate or mixed application) under two ecologically relevant flow regimes (5 and 15 cm s− 1), and a control condition (0 cm s− 1) in a laboratory flume. Our findings indicate that the effectiveness of a binder to retain S. latissima gametophytes onto cultivation rope, measured by the sporophyte density, was comparable for all binder types in the high flow velocity (15 cm s− 1) treatments, including the non-binder control treatment. Sporophyte densities were highest in the low flow velocities (0 and 5 cm s− 1) in the absence of a binder compared to all other treatment combinations. In conclusion, our results highlight that the effectiveness of binder assisted seeding of kelp propagules did not differ between the binder and non-binder treatments and was unaffected by flow velocities for the binder treatments. These findings are important in the development of novel methods and further optimisation of existing binder-based methods aimed at retaining seaweed propagules onto cultivation rope.

The green seaweed Ulva is a model system to study seaweed-bacteria interactions, but the impact of environmental drivers on the dynamics of these interactions is little understood. In this study, we investigated the stability and variability of the seaweed-associated bacteria across the Atlantic-Baltic Sea salinity gradient. We characterized the bacterial communities of 15 Ulva sensu lato species along 2,000 km of coastline in a total of 481 samples. Our results demonstrate that the Ulva-associated bacterial composition was strongly structured by both salinity and host species (together explaining between 34% and 91% of the variation in the abundance of the different bacterial genera). The largest shift in the bacterial consortia coincided with the horohalinicum (5-8 PSU, known as the transition zone from freshwater to marine conditions). Low-salinity communities especially contained high relative abundances of Luteolibacter, Cyanobium, Pirellula, Lacihabitans and an uncultured Spirosomaceae, whereas high-salinity communities were predominantly enriched in Litorimonas, Leucothrix, Sulfurovum, Algibacter and Dokdonia. We identified a small taxonomic core community (consisting of Paracoccus, Sulfitobacter and an uncultured Rhodobacteraceae), which together contributed to 14% of the reads per sample, on average. Additional core taxa followed a gradient model, as more core taxa were shared between neighbouring salinity ranges than between ranges at opposite ends of the Atlantic-Baltic Sea gradient. Our results contradict earlier statements that Ulva-associated bacterial communities are taxonomically highly variable across individuals and largely stochastically defined. Characteristic bacterial communities associated with distinct salinity regions may therefore facilitate the host's adaptation across the environmental gradient. info:eu-repo/semantics/publishedVersion

Ulva fenestrata is an economically and ecologically important green algal species with a large potential in seaweed aquaculture due to its high productivity, wide environmental tolerance, as well as interesting functional and nutritional properties. Here, we performed a series of manipulative cultivation experiments in order to investigate the effects of irradiance (50, 100, and 160 μmol photons m−2 s−1), temperature (13 and 18 °C), nitrate (pCO2 (200, 400, and 2500 ppm) on the relative growth rate and biochemical composition (fatty acid, protein, phenolic, ash, and biochar content) in indoor tank cultivation of Swedish U. fenestrata. High irradiance and low temperature were optimal for the growth of this northern hemisphere U. fenestrata strain, but addition of nutrients or changes in pCO2 levels were not necessary to increase growth. Low irradiance resulted in the highest fatty acid, protein, and phenolic content, while low temperature had a negative effect on the fatty acid content but a positive effect on the protein content. Addition of nutrients (especially nitrate) increased the fatty acid, protein, and phenolic content. High nitrate levels decreased the total ash content of the seaweeds. The char content of the seaweeds did not change in response to any of the manipulated factors, and the only significant effect of changes in pCO2 was a negative relationship with phenolic content. We conclude that the optimal cultivation conditions for Swedish U. fenestrata are dependent on the desired biomass traits (biomass yield or biochemical composition).

Seaweed biomass has the potential to become an important raw material for bio-based production. The aim of this study was to screen the overall composition of several seaweed species on the Swedish west coast, including some scarcely studied species, to provide fundamentals for evaluation of biorefining potential and to benchmark with already potentially industrially relevant species and commercially important land-based biomasses. Twenty-two common seaweed species (green, red, brown) were collected and the carbohydrate, ash, protein, water and metal contents were measured. Carbohydrate content varied between 237 and 557 g kg−1dry weight (dw), making it the largest constituent, on a dry weight basis, of most species in the study. Ash, which is considered unwanted in biorefining, ranged between 118 and 419 g kg−1dw and was the largest constituent in several seaweeds, which were therefore considered unsuitable for biorefining. Protein content was most abundant in the red seaweeds but was generally low in all species (59–201 g kg−1dw). High contents of several unwanted metals for processing or human consumption were found (e.g. aluminium, arsenic, copper, chromium and nickel), which need to be considered when utilizing seaweeds for certain applications. Potential targets for further biorefinery development mostly include species already known for their potential (Saccharina latissima,Laminaria digitataandChondrus crispus) while some, such asHalidrys siliquosaandDilsea carnosa, have not been previously noted. However, more detailed studies are required to explore biorefinery processes for these seaweeds, as well as how to potentially cultivate them.

In recent years, the interest in using seaweed for the sustainable production of commodities has been increasing as seaweeds contain many potentially worthwhile compounds. Thus, the extraction and refining processes of interesting compounds from seaweeds is a hot research topic but has been found to have problems with profitability for novel applications. To increase the economic potential of refining seaweed biomass, the content of the compounds of interest should be maximized, which can potentially be achieved through optimization of cultivation conditions. In this study, we studied how the monosaccharide composition of the green seaweed species Ulva fenestrata is influenced by the abiotic factors; irradiance, temperature, nitrate, phosphate, and pCO2. It was evident that lower nitrate concentration and cultivation at elevated temperature increased monosaccharide contents. A 70% increase in iduronic acid and a 26% increase in rhamnose content were seen under elevated irradiance and temperature conditions, though the absolute differences in monosaccharide concentration were small. Irradiance and nitrate impacted the ratio between iduronic and rhamnose, which is an indicator of the ulvan structure. These results could potentially be utilized to coax the ulvan towards specific bioactivities, and thus have a considerable impact on a potential biorefinery centered around Ulva.

Seaweed is a promising sustainable source of vegan protein as its farming does not require arable land, pesticides/insecticides, nor freshwater supply. However, to be explored as a novel protein source the content and nutritional quality of protein in seaweed need to be improved. We assessed the influence of pH-shift processing on protein degree of hydrolysis (%DH), protein/peptide size distribution, accessibility, and cell bioavailability of Ulva fenestrata proteins after in vitro gastrointestinal digestion. pH-shift processing of Ulva, which concentrated its proteins 3.5-times, significantly improved the %DH from 27.7±2.6% to 35.7±2.1% and the amino acid accessibility from 56.9±4.1% to 72.7±0.6%. Due to the higher amino acid accessibility, the amount of most amino acids transported across the cell monolayers was higher in the protein extracts. Regarding bioavailability, both Ulva and protein extracts were as bioavailable as casein. The protein/peptide molecular size distribution after digestion did not disclose a clear association with bioavailability.

Climate change leads to multiple effects caused by simultaneous shifts in several physical factors which will interact with species and ecosystems in complex ways. In marine systems the effects of climate change include altered salinity, increased temperature, and elevated pCO2 which are currently affecting and will continue to affect marine species and ecosystems. Seaweeds are primary producers and foundation species in coastal ecosystems, which are particularly vulnerable to climate change. The brown seaweed Fucus vesiculosus (bladderwrack) is an important foundation species in nearshore ecosystems throughout its natural range in the North Atlantic Ocean and the Baltic Sea. This study investigates how individual and interactive effects of temperature, salinity, and pCO2 affect F. vesiculosus, using a fully crossed experimental design. We assessed the effects on F. vesiculosus in terms of growth, biochemical composition (phlorotannin content, C:N ratio, and ∂13C), and susceptibility to the specialized grazer Littorina obtusata. We observed that elevated pCO2 had a positive effect on seaweed growth in ambient temperature, but not in elevated temperature, while growth increased in low salinity at ambient but not high temperature, regardless of pCO2-level. In parallel to the statistically significant, but relatively small, positive effects on F. vesiculosus growth, we found that the seaweeds became much more susceptible to grazing in elevated pCO2 and reduced salinity, regardless of temperature. Furthermore, the ability of the seaweeds to induce chemical defenses (phlorotannins) was strongly reduced by all the climate stressors. Seaweeds exposed to ambient conditions more than doubled their phlorotannin content in the presence of grazers, while seaweeds exposed to any single or combined stress conditions showed only minor increases in phlorotannin content, or none at all. Despite the minor positive effects on seaweed growth, the results of this study imply that climate change can strongly affect the ability of fucoid seaweeds to induce chemical defenses and increase their susceptibility to grazers. This will likely lead to widespread consequences under future climate conditions, considering the important role of F. vesiculosus and other fucoids in many coastal ecosystems.

Globally, there is a need for novel vegetarian protein sources. We recently showed that the pH-shift process, using alkaline protein solubilization followed by isoelectric precipitation, is an efficient way to produce extracts with high protein concentrations from Ulva lactuca (>50% on a dry matter basis). However, the total protein yield was low, and to improve this, the effects of adding ulvan lyase, preincubating the seaweed homogenate at pH 8.5 and using different protein extraction temperatures (8 °C, RT and 40 °C), were evaluated in this study. Addition of ulvan lyase reduced protein solubility but increased the precipitation. Incubation at pH 8.5, without ulvan lyase added, significantly increased both protein solubility and precipitation at 8 °C and RT. Temperature per se had no effect on protein solubility, while protein precipitation increased with decreasing temperature. Highest protein yield (29%) was achieved when keeping the process at 8 °C with a preincubation step at pH 8.5 for 1 h. By these process modifications, the yield was 3.2 times higher than achieved by the control process (9.2%).

In the 1930s, outbreaks of the wasting disease pathogen Labyrinthula zosterae caused a severe reduction of the eelgrass Zostera marina meadows in the Atlantic Ocean. Many surviving populations were found in low-salinity environments, and low-salinity environments have therefore been hypothesized to act as a refuge for eelgrass against L. zosterae infection. Here, we investigated L. zosterae pathogen load and wasting disease symptoms in eelgrass over a similar to 970 km salinity gradient (6-25 PSU) along the Swedish coast. Furthermore, laboratory infection experiments and studies of inhibitory compounds were carried out to investigated whether resistance against the pathogen is correlated to differences in natural pathogen pressure among eelgrass populations. The degree of L. zosterae infection was positively correlated to salinity and the pathogen was absent in several of the eelgrass meadows in lower salinity (7-8 PSU). However, a low L. zosterae pathogen load was also found in some eelgrass populations in the lowest salinity (6 PSU). No correlation between resistance and pathogen pressure in situ was detected, and all eelgrass shoots produced chemical compounds that inhibited L. zosterae growth. These results imply that positive correlations between L. zosterae and salinity are not due to eelgrass resistance, but rather to the poor ability of L. zosterae to cope with low salinity. However, our results also indicate that some strains of L. zosterae may adapt to low salinity, and therefore there may also be a risk of wasting disease outbreaks in low-salinity eelgrass meadows, in contrast to what so far has been the general hypothesis.

We report (1) successful extraction and characterization of cellulose from northern hemisphere green macroalgae Ulva lactuca (Ulva fenestrata) collected along the Swedish west coast and cultivated indoors under controlled conditions, followed by (2) its utilization in the production of lignin-free cellulose nanofibrils (CNF). Cellulose was extracted by sequential treatment with ethanol, hydrogen peroxide, sodium hydroxide, and hydrochloric acid, yielding a cellulose-rich insoluble fraction. The extracted cellulose was disintegrated into CNF using a mechanical homogenization process without any further enzymatic pre-treatments. In addition, regenerated cellulose was prepared. XRD characterization of the CNF showed characteristic peaks for the cellulose I allomorph and confirmed that the nanofibrils were semicrystalline with a crystallinity index of 48%. Regenerated cellulose was mostly amorphous with an XRD pattern indicating the presence of the cellulose II allomorph. The cellulose fractions were essentially free from inorganic substances and thermally stable up to around 260 degrees C. Structural mapping with CP-MAS C-13-NMR sustains the cellulose content of CNF and regenerated cellulose, respectively, yet ion chromatography identified the presence of 10-15% xylose in the fractions. Optotracing was used as a novel and non-disruptive tool to selectively assess the polysaccharide composition of the cellulose fractions and produced CNF aiming to shed light on this hitherto non-resolved origin of xylose in Ulva cell wall matter. Fluorescence excitation and emission spectra of a panel of 4 oligothiophenes identified and verified the presence of cellulose and sustain the conclusion that the isolated fractions consist of cellulose intertwined with a small amount of a xylose-containing glucan copolymer. [GRAPHICS] . Funding Agencies|Royal Institute of Technology; Swedish Foundation for Strategic Research (SSF)Swedish Foundation for Strategic Research [RBP14-0045]; Erling-Persson Family Foundation; Getinge AB

The growing world population demands sustainably produced proteins and cultivation of seaweeds may become an important solution for this so-called protein-shift. To increase the biomass yield and protein content of seaweeds, nutrient-rich wastewaters originating from different processes and activities such as aquaculture, industrial food processing, and municipal wastewater management could be used. Our aim was to synthesize previous literature to assess the impact that different wastewaters have on seaweed growth and protein content, and to identify research gaps. We show, by using standardized and quantitative methods, that cultivation in wastewaters increases both growth and nitrogen content (often used as an approximation for protein) of seaweeds. We further show that all groups of seaweeds (green, brown, and red) benefit from cultivation in wastewater, that the effect is similar between ocean- and land-based seaweed cultivations, and that seaweed cultivation with wastewater is not limited to only fish aquaculture wastewaters. Our results clearly demonstrate that seaweeds can be cultivated in a wide range of wastewaters to generate protein-rich biomass with increased growth and biomass yields. These results can be valuable when searching for sustainably produced proteins for the growing human population.

peer-reviewed The demand for vegetable proteins increases globally and seaweeds are considered novel and promising protein sources. However, the tough polysaccharide-rich cell walls and the abundance of polyphenols reduce the extractability and digestibility of seaweed proteins. Therefore, food grade, scalable, and environmentally friendly protein extraction techniques are required. To date, little work has been carried out on developing such methods taking into consideration the structural differences between seaweed species. In this work, three different protein extraction methods were applied to three Swedish seaweeds (Porphyra umbilicalis, Ulva lactuca, and Saccharina latissima). These methods included (I) a traditional method using sonication in water and subsequent ammonium sulfate-induced protein precipitation, (II) the pH-shift protein extraction method using alkaline protein solubilization followed by isoelectric precipitation, and (III) the accelerated solvent extraction (ASE®) method where proteins are extracted after pre-removal of lipids and phlorotannins. The highest protein yields were achieved using the pH-shift method applied to P. umbilicalis (22.6 ± 7.3%) and S. latissima (25.1 ± 0.9%). The traditional method resulted in the greatest protein yield when applied to U. lactuca (19.6 ± 0.8%). However, the protein concentration in the produced extracts was highest for all three species using the pH-shift method (71.0 ± 3.7%, 51.2 ± 2.1%, and 40.7 ± 0.5% for P. umbilicalis, U. lactuca, and S. latissima, respectively). In addition, the pH-shift method was found to concentrate the fatty acids in U. lactuca and S. latissima by 2.2 and 1.6 times, respectively. The pH-shift method can therefore be considered a promising strategy for producing seaweed protein ingredients for use in food and feed.

Here we evaluated the levels of lipid oxidation products, fatty acids, ascorbic acid and colour of Porphyra and Ulva after oven-drying at 40 °C, and during subsequent storage for ≥370 days under light, semi-light and dark conditions. Part of the seaweed was pre-soaked in freshwater or pre-coated with a whey protein mixture. Controls consisted of freeze-dried seaweeds. Throughout storage there was a moderate development of the lipid oxidation-derived aldehydes, malondialdehyde, 4-hydroxy-trans-2-hexenal and 4-hydroxy-trans-2-nonenal, while there was a great loss of unsaturated fatty acids and ascorbic acid. Light storage and freeze-drying stimulated the fatty acid loss as well as pigment bleaching, seen as increased a*-values. For Ulva, the coating reduced malondialdehyde, 4-hydroxy-trans-2-hexenal and 4-hydroxy-trans-2-nonenal formation during drying and slightly prevented loss of polyunsaturated fatty acids during light storage. Pre-soaking in freshwater had no effect on the seaweed stability, although it reduced the ash content and thereby increased the relative content of ascorbic acid and fatty acids of the biomasses.

Hydrogels based on the polysaccharide ulvan from the green macroalgae Ulva fenestrata were synthesized and evaluated as an adsorbent for heavy metals ions and methylene blue. Ulvan was extracted from Ulva fenestrata using diluted hydrochloric acid and recovered by precipitation with EtOH. The extracted ulvan was converted into ulvan dialdehyde via periodate-oxidation and subsequently combined with gelatin yielding hydrogels. The hydrogels showed good water-uptake capacity with a maximum swelling degree of 2400 % in water and 900 % in PBS buffer. Adsorption tests of methylene blue showed a maximum adsorption capacity of 465 mg/g. The adsorption data of methylene blue followed the pseudo-second order kinetics and agreed with the Langmuir adsorption isotherm. The maximum adsorption capacity of heavy metal ions was 14 mg/g for Cu2+, 7 mg/g for Co2+and 6 mg/g for Ni2+and Zn2+ indicating that the hydrogels have a stronger affinity for Cu2+ than for Co2+, Ni2+, and Zn2+.

Shipworms (teredinids) are highly specialized marine bivalves that consume terrestrially derived wood. Changes in environmental variables may result in shipworms spreading into the Baltic Sea – which would have devastating consequences for maritime cultural heritage and submerged wooden structures. We investigated the distribution and abundance of the shipworms Teredo navalis and Psiloteredo megotara along the Swedish coast in 2006–2008, and compared our findings with data collected at partly the same locations in 1971–1973. Wooden test panels were submerged in near-surface waters at 18 harbours. The presence of shipworms was determined by X-ray radiography of each panel. Sea surface temperature and salinity data were analysed to investigate whether any changes in distribution were correlated to changes in environmental variables. We found that past and present distributions of T. navalis were similar – indicating that no range expansion of shipworms into the Baltic Sea has taken place the last 35 years. The abundance of T. navalis was similar between decades at all investigated sites except two (Arild and Barsebäckshamn), where abundances were higher in 2006–2008. The abundance of T. navalis varied along the coast and was positively correlated to mean sea surface salinity, but not to mean sea surface temperature (2006–2008 data). The distribution and abundance of P. megotara were similar during the two study periods with only single observations at a few sites. In conclusion, we found no evidence of range expansion of shipworms along the Swedish coast.

Summary 1. Allelopathy is an important non-resource interaction in terrestrial plant communities that may affect invasions by non-indigenous plants. The ‘novel weapons hypothesis’ (NWH) predicts that non-indigenous plants will become invasive if they have allelopathic compounds that assemblages in the new range are not adapted to. Recently, the non-indigenous, chemically rich macroalga Bonnemaisonia hamifera (Hariot) has become one of the most abundant filamentous red algae in Scandinavian waters. 2. We used B. hamifera to specifically test the aspect of the NWH that concerns invasion success based on novel allelochemicals in the invaded range. Allelopathic interactions were tested through effects on the growth rate of adult native macroalgae in co-cultures with B. hamifera and through the settlement success of native macroalgal propagules and microalgae on surfaces coated with 1,1,3,3-tetrabromo-2-heptanone. We also investigated whether 1,1,3,3-tetrabromo-2-heptanone can be transferred from B. hamifera to its native host algae, as a means of pre-emptive competition. 3. The settlement of native macroalgal propagules and microalgae was strongly inhibited on surfaces coated with 1,1,3,3-tetrabromo-2-heptanone at ecologically relevant concentrations, but there were no effects of adult B. hamifera on growth rates of adults of the six native naturally co-occurring species. The compound was shown to be transferred from B. hamifera to the surface of its native host algae at inhibitory concentrations in both laboratory and field experiments. 4. By inhibiting the settlement of propagules on its thallus and on surrounding surfaces, B. hamifera achieves a competitive advantage over native macroalgae, a finding that parallels previous reports on soil- and litter-mediated allelopathic interactions among vascular plants. Because competition for available substrata in marine benthic systems is intense, the ability to reserve space may be vital for B. hamifera’s successful invasion. This is the first example of an allelopathic compound that can be transferred by direct contact from an exotic to a native species, with an active and unaltered function. 5. Synthesis. Our results clearly show that the main secondary metabolite of the invasive red alga B. hamifera has strong allelopathic effects towards native competitors, suggesting that its novel chemical weapon is important for the highly successful invasion of new ranges.

Sessile invertebrates and algae are ubiquitous as hosts for a range of fouling organisms in marine benthic habitats. Fouling can sometimes impose negative fitness effects, and hosts may decrease the amount of fouling through production of chemical defenses. Waterborne chemical defenses may rapidly become diluted and should be released continuously in order to inhibit fouling; this process may be costly for the host. Therefore, it has been hypothesized that natural selection should favor the evolution of defense metabolites that stay on the host surface in aquatic environ- ments. We investigated chemical deterrence of fouling in the marine sessile invertebrate Halichon- dria panicea (breadcrumb sponge), through both field observations and a series of laboratory bio- assays. Natural populations of H. panicea had very few macrofouling organisms. Cyprid larvae of the barnacle Balanus improvisus were deterred from attaching to the substratum in the presence of water-soluble compounds that were rapidly exuded from the sponges both under laboratory condi- tions and in the field. Artificial substrata placed near H. panicea in the field received less fouling by barnacles than control substrata placed farther away from sponges. These results clearly show that H. panicea releases water-soluble compounds that deter attachment of B. improvisus. However, these compounds may not have evolved as a chemical defense against fouling in the sponges. H. panicea can release epidermal cell layers, and therefore the barnacles may have evolved an ability to recog- nize water-soluble compounds from the sponges in order to avoid attaching to unsuitable substrata.

Terrestrial herbivores can use anthelmintic properties of plant secondary metabolites to decrease parasite/parasitoid infections. Secondary metabolites with potential anthelmintic prop- erties have also been described from different seaweed species, but the effects of diet on parasitism in marine herbivores has not been studied previously. In the present study, a combination of labora- tory and field experiments was used to test the hypotheses that infection by digenean trematode par- asites affects the feeding pattern of herbivorous gastropods (Littorina littorea), and that the seaweed diet prevents parasitism in the snails. Heavily infected snails consumed less of the red seaweed Osmundea ramosissima than uninfected snails, but there was no difference in consumption when snails were offered the green seaweed Ulva lactuca. Healthy snails placed in cages with different food treatments and transplanted to different field sites became infected by trematodes, but there was no significant difference in parasite resistance between snails offered different foods. Further- more, there was no significant negative relationship between infection abundance and consumption when O. ramosissima was incorporated into agar-based artificial diets, indicating that the difference in feeding pattern found in the initial consumption experiment was due to the morphology, and not the chemistry of the seaweeds. In conclusion, these preliminary results do not indicate that para- sitized L. littorea feeds on O. ramosissima to prevent parasite infection. However, the hypothesis that algal chemical compounds prevent or decrease parasite infections in herbivores should be tested using additional candidate species before drawing general conclusions about the effect of diet on parasitism in marine systems.

1 Terrestrial plants can sense and respond to herbivory, which may lead to increased resistance towards further grazing if the responses have negative effects on the preference and/or performance of the herbivores. Marine plants (seaweeds) are exposed to a considerable grazing pressure by herbivores ranging from large, mobile fishes and sea urchins to small, sedentary crustaceans and molluscs. The number of investigations studying induced resistance in seaweeds has increased during the last decade, but empirical results are conflicting. 2 We performed a categorical meta-analysis to evaluate statistically the overall seaweed responses to damage or damage-related cues, and factors that may explain the observed variation in inducible seaweed resistance to herbivores. 3 We found a highly significant overall effect of damage on induced seaweed resistance to further herbivory. Division of the studies into different categories showed that brown and green, but not red, seaweeds induce significant resistance to further grazing in response to grazing by small crustaceans and gastropods, but not in response to large gastropods and sea urchins. The seaweeds showed stronger responses when exposed to damage for 11-20 days than in shorter or longer experiments. 4 Seaweeds are very important both as habitat and food for a wide range of marine animals. Our findings contribute importantly to the general ecological understanding of marine plant-herbivore interactions by showing that induced resistance in seaweeds is more common than previously assumed. Many recent marine investigations included in this study have not put emphasis on the ecological relevance and underlying mechanisms of the investigated plant-herbivore interactions. We suggest that the scientific value of future investigations concerning induced defences in marine algae would benefit from formulating more advanced and/or complex hypotheses including the genetic and biochemical mechanisms, cost and constraints of damage-induced civilian and defensive seaweed responses, as well as the effects of these responses on herbivores and other organisms/trophic levels, and on community structure and functioning.

Plants have different strategies to cope with herbivory, including induction of chemical defences and compensatory growth. The most favourable strategy for an individual plant may depend on the density at which the plants are growing and on the availability of nutrients, but this has not been tested previously for marine plant–herbivore interactions. We investigated the separate and interactive effects of plant density, nutrient availability, and herbivore grazing on the phlorotannin (polyphenolic) production in the brown seaweed Ascophyllum nodosum. Seaweed plants grown at low or high densities were exposed either to nutrient enrichment, herbivorous littorinid gastropods (Littorina obtusata), or a combination of nutrients and herbivores in an outdoor mesocosm experiment for 2 weeks. Seaweeds grown at a low density tended to have higher tissue nitrogen content compared to plants grown at a high density when exposed to elevated nutrient levels, indicating that there was a density dependent competition for nitrogen. Herbivore grazing induced a higher phlorotannin content in plants grown under ambient, but not enriched, nutrient levels, indicting either that phlorotannin plasticity is more costly when nutrients are abundant or that plants responded to herbivory by compensatory growth. However, there were no significant interactive or main effects of plant density on the seaweed phlorotannin content. The results indicate that plants in both high and low densities induce chemical defence, and that eutrophication may have indirect effects on marine plant–herbivore interactions through alterations of plant chemical defence allocation.

Terrestrial plants have long been known to induce resistance towards herbivores in response to direct grazing, and strong evidence of inter-plant information transfer through volatile signals has been reported recently. Still, little is known about information exchange in aquatic plant–herbivore interactions. In this study, 12 Swedish seaweed species were exposed either to direct grazing by a generalist crustacean herbivore (Idotea granulosa), or to waterborne signals produced by actively feeding herbivores for 1 week. In order to test for the presence of induced chemical resistance in the different seaweed species, the dried and homogenized seaweed tissues were incorporated into an agar matrix, and herbivores were allowed to choose between the resulting control and induced artificial diets in two different two-choice feeding trials. The herbivores were actively feeding from all seaweed species in the induction experiments, although the amount of seaweed tissue consumed differed significantly between species. A chemically based induced herbivore resistance was found in response to direct grazing in four of the tested seaweed species (two red, one brown, and one green seaweed species). Furthermore, four seaweeds (one red, two brown, and one green seaweed species) induced resistance towards further grazing in response to waterborne chemical signals. Several seaweed species responded differently when exposed to different herbivore-related cues, indicating that both cues and responses can be highly specific. The results show that herbivore-induced resistance is present in 7 of 12 of the tested Swedish seaweed species, but that different signals (i.e., direct grazing and waterborne cues) elicit complex responses in the seaweeds.

The allocation of resources to different life-history traits should represent the best compromise in fitness investment for organisms in their local environment. When resources are limiting, the investment in a specific trait must carry a cost that is expressed in trade-offs with other traits. In this study, the relative investment in the fitness-related traits, growth, reproduction and defence were compared at central and range-edge locations, using the seaweed Ascophyllum nodosum as a model system. Individual growth rates were similar at both sites, whereas edge populations showed a higher relative investment in reproduction (demonstrated by a higher reproductive allocation and extended reproductive periods) when compared to central populations that invested more in defence. These results show the capability of A. nodosum to differentially allocate resources for different traits under different habitat conditions, suggesting that reproduction and defence have different fitness values under the specific living conditions experienced at edge and central locations. However, ongoing climate change may threaten edge populations by increasing the selective pressure on specific traits, forcing these populations to lower the investment in other traits that are also potentially important for population fitness.

The life cycle of many sessile marine invertebrates includes a dispersive planktonic larval stage whose ability to find a suitable habitat in which to settle and transform into benthic adults is crucial to maximize fitness. To facilitate this process, invertebrate larvae commonly respond to habitat-related chemical cues to guide the search for an appropriate environment. Furthermore, small-scale hydrodynamic conditions affect dispersal of chemical cues, as well as swimming behavior of invertebrate larvae and encounter with potential habitats. Shipworms within the family Teredinidae are dependent on terrestrially derived wood in order to complete their life cycle, but very little is known about the cues and processes that promote settlement. We investigated the potential for remote detection of settling substrate via waterborne chemical cues in teredinid larvae through a combination of empirical field and laboratory flume experiments. Natural populations of teredinid larvae were significantly more abundant close to wooden structures enclosed in plankton net compared to empty control nets, clearly showing that shipworm larvae can sense and respond to chemical cues associated with suitable settling substrate in the field. However, the flume experiments, using ecologically relevant flow velocities, showed that the boundary layer around experimental wooden panels was thin and that the mean flow velocity exceeded larval swimming velocity approximately 5 mm (≈ 25 larval body lengths) from the panel surface. Therefore, we conclude that the scope for remote detection of waterborne cues is limited and that the likely explanation for the higher abundance of shipworm larvae associated with the wooden panels in the field is a response to a cue during or after attachment on, or very near, the substrate. Waterborne cues probably guide the larva in its decision to remain attached and settle, or to detach and continue swimming and drifting until the next encounter with a solid substrate.

Brown seaweeds can serve as habitat and food for a number of mesoherbivore species, but the effect of mesoherbivore grazing on the fitness of individual seaweed plants is not well documented. Here we investigate how mechanical wounding, which mimics the grazing damage made by the herbivorous gastropod Littorina obtusata, affects the biomass change, apical growth, and individual shoot strength of the fucoid seaweed species Ascophyllum nodosum both in natural field populations and in the laboratory. We did not detect a significant effect of wounding on apical shoot growth in the laboratory, but wounding decreased the plant biomass in half of the studied A. nodosum populations in the field. Furthermore, we found that the strength of individual A. nodosum shoots was significantly lower when compared with unwounded control shoots initially after wounding. However, after 11 days incubation in the laboratory, the strength of the wounded shoots had increased almost to the level of the control shoots, although there was still a significant difference in the force required to break the shoots. These results indicate that wounding can increase the tissue loss (and thereby decrease the fitness) of A. nodosum plants in natural populations, probably through weakening of individual shoots. However, A. nodosum appears to have a mechanism to strengthen the shoots after mechanical wounding, and therefore, the timing of damage (e.g. prior to a storm) probably determines the amount of plant tissue lost through wave-induced forces.

Optimal Defense Theory predicts that plants exposed to herbivory should allocate more resources to produce costly secondary metabolites in tissues with higher fitness values. To increase plant resistance, the secondary metabolites must have a negative impact on the preference and/or performance of herbivores. We tested the hypotheses that induction of secondary metabolites (phlorotannins) in a brown seaweed in response to grazing by herbivorous gastropods will differ between seaweed tissues with different fitness values (basal stipes and annual shoots), and that the subsequent change in food value will affect the fitness (growth and fecundity) of the gastropods. Induction of phlorotannins was significant in both tissue types but was more pronounced in basal stipes, which have a higher fitness value. Basal tissues also had significantly higher constitutive defense levels than did apical tissues. No effects of algal tissue type or grazing history on the growth rate of the gastropods were detected. However, the number of viable eggs was significantly lower for gastropods feeding on basal shoots, and there was a significantly lower proportion of viable eggs produced by gastropods that were offered previously grazed seaweed tissues. The results show that induced resistance, and its variation among different plant parts, can have significant negative effects on herbivore performance that may reduce future herbivore pressure and thus enhance plant fitness.

Many brown macroalgae contain substantial concentrations of polyphenolic compounds, known as phlorotannins. Previous studies have shown that much of the variation in phlorotannin concentration is correlated with taxonomy and biogeography, but little is known about patterns of variation within species and populations. In this study, we examined intraspecific variation in the phlorotannin content of the intertidal seaweed Ascophyllum nodosum, by testing for differences within as well as between genetic individuals (genets) and populations, and between algal tissues of similar age from juveniles and adults. The results show that variation in the phlorotannin content of annual shoots within primary shoots and genets contributes substantially to the total intraspecific variation. This pattern probably indicates that the phlorotannin content is a highly plastic trait that can be significantly influenced by small-scale variation in extrinsic factors. There were also significant differences in mean phlor...

Optimal defense theory (ODT) predicts that plants should have the highest defense levels in parts that have the highest value in terms of fitness. Assumptions about differences in fitness value among plant parts have previously been based on general reasoning, e.g., that reproductive tissue is more valuable than vegetative tissue since fitness ultimately depends on reproduction. Here we present a first example of how demographic elasticity (proportional sensitivity) analysis can be used as a tool to generate more objective and specific estimates of the fitness value of different plant parts, based on the entire life cycle. These estimates were further used to generate testable predictions about intraplant variation in defenses of the brown seaweed Ascophyllum nodosum (knotted wrack). The predictions were clearly supported by the results from both analyses of chemical defense levels (phlorotannins) and bioassays (feeding preference experiments), which showed that reproductive tissues had the lowest defense level of the three examined tissue types, in contrast to previous assumptions. We suggest that a stronger focus on variation in life history among species and populations, with demographic perturbation analysis as one important tool, could lead to a better understanding of intraplant variation in defense levels, in seaweeds as well as vascular plants and other modular organisms.

The habitat and feeding preference of two gastropod mesoherbivore species, Ansates (Helcion) pellucida and Lacuna vincta, inhabiting the kelp Laminaria hyperborea were investigated by studying the distribution, habitat choice and food preference of the herbivores between new and old kelp fronds. Nitrogen content and chemical defence (phlorotannin) level of the kelp fronds were measured in order to determine whether they were correlated with the feeding preference of the herbivores. The habitat choice of Lacuna vincta was correlated with the food preference of this species, while the habitat choice of A. pellucida probably was due to some other factor, e.g. competition or mortality. Ansates pellucida preferred to reside on the old fronds of Laminaria hyperborea, but consumed an equal amount of both tissue types. Lacuna vincta, however, preferred the new Laminaria hyperborea fronds both as habitat and food. There was a large variation in the phlorotannin content among the tissue samples of L. hyperborea. However, there were no overall statistically significant differences in tissue nitrogen or phlorotannin content between the new and the old L. hyperborea fronds.

Phlorotannins (polyphenolic compounds) are found exclusively in brown algae and are commonly regarded as an important component of algal defense against herbivores. The variation in phlorotannin concentration within and among individual seaweed specimens of the same species can be substantial, and seaweeds may respond to herbivore damage by an increased production of phlorotannins in order to prevent further herbivory. In the present study, we investigated the natural variation of phlorotannins in different parts of the kelp Laminariahyperborea in an observational field survey. We also performed a manipulative induction experiment in which we studied the effect of natural grazing by the patellid limpet Ansates (=Helcion) pellucida and the littorinid snail Lacunavincta, as well as artificial damage on the production of phlorotannins in L. hyperborea. There was a large variation in phlorotannin content both among and within L. hyperborea individuals and populations. The control plants in the induction experiment had a higher phlorotannin content compared to plants in natural populations, but grazing by A. pellucida or L. vincta, or artificial damage did not induce a higher phlorotannin concentration in the kelps. Instead, the phlorotannin content in wounded kelps was significantly lower than in the control plants, possibly due to removal of phlorotannin-rich outer layers of the kelp thallus, or to an induced compensatory growth response in the L. hyperborea plants. These results imply that L. hyperborea phlorotannins probably do not function as an inducible chemical defense against A. pellucida and L. vincta, instead grazing by these herbivores decreases the phlorotannin content of the kelps.

Factors determining the degree of dietary generalism versus specialism are central in ecology. Species that are generalists at the population level may in fact be composed of specialized individuals. The optimal diet theory assumes that individuals choose diets that maximize fitness, and individual specialization may occur if individuals' ability to locate, recognize, and handle different food types differ. We investigate if individuals of the marine herbivorous slug Elysia viridis, which co-occur at different densities on several green macroalgal species in the field, are specialized to different algal hosts. Individual slugs were collected from three original algal host species (Cladophora sericea, Cladophora rupestris and Codium fragile) in the field, and short-term habitat choice and consumption, as well as long-term growth (proxy for fitness), on four algal diet species (the original algal host species and Chaetomorpha melagonium) were studied in laboratory experiments. Nutritional (protein, nitrogen, and carbon content) and morphological (dry weight, and cell/utricle volume) algal traits were also measured to investigate if they correlated with the growth value of the different algal diets. E. viridis individuals tended to choose and consume algal species that were similar to their original algal host. Long-term growth of E. viridis, however, was mostly independent of original algal host, as all individuals reached a larger size on the non-host C. melagonium. E. viridis growth was positively correlated to algal cell/utricle volume but not to any of the other measured algal traits. Because E. viridis feeds by piercing individual algal cells, the results indicate that slugs may receive more cytoplasm, and thus more energy per unit time, on algal species with large cells/utricles. We conclude that E. viridis individuals are specialized on different hosts, but host choice in natural E. viridis populations is not determined by the energetic value of seaweed diets as predicted by the ODT.

Tannins, a large and diverse group of phenolic secondary metabolites, are common in terrestrial plants and marine brown algae. It is sometimes desirable to remove the tannins from plant or algal extracts, e.g., when isolating enzymes and nucleic acids, when using certain colorimetric methods to quantify the tannin content, or to create reliable controls when using tannins in experimental studies. Insoluble polyvinylpolypyrrolidone (PVPP) can be used to specifically remove tannins from solution. In the present study, we evaluated the effect of different factors (amount of PVPP, number of PVPP treatments, type of solvent, pH, and incubation time) on the PVPP removal of dissolved brown algal phlorotannins. Our results imply that there is a limited amount of phlorotannins that can bind to a given amount of PVPP, and that it is preferable to use low quantities of PVPP repeatedly, compared to using fewer treatments with a high amount of PVPP. Furthermore, we found no consistent effect on the removal of phlorotannins due to solvent type (acetone, methanol, distilled water or filtered seawater). There was a slight decrease in the amount of phlorotannins removed from extracts with increasing pH when repeatedly treated with PVPP. All phlorotannins were removed from extracts with pH ≤6.2, and 89% of the initial phlorotannin content was removed at pH 9.7. These results are compared with previous methodological studies on tannin removal with PVPP. Furthermore, the implications of phlorotannin removal in analytical and ecological investigations are discussed.

Physical damage caused by herbivores or artificial clipping is known to induce responses in secondary chemistry as well as increased resistance to further grazing in a large number of terrestrial vascular plants, but this has only rarely been described for marine algae. In the present study, it was found that a few weeks of grazing by the gastropod Littorina obtusata can induce substantially increased concentrations of phlorotannins in the intertidal brown seaweed Ascophyllum nodosum.In contrast, grazing by the isopod Idotea granulosa and simulations of herbivory through momentary and continuous clipping caused no significant changes in phlorotannin levels. This indicates a high degree of specificity in the elicitation of chemical responses to physical damage, something which previously has been shown for terrestrial vascular plants but not for marine algae. Ascophyllum plants that had been grazed by L. obtusata were also less susceptible than undamaged plants to further grazing by gastropods, but no such induced resistance was found in the experiments with I. granulosa. Feeding experiments with undamaged Ascophyllum plants and artificial food containing different levels of phlorotannins provided further support for the sensitivity of the feeding behavior of L. obtusata, and the insensitivity of I. granulosa, to intraspecific variation in the phlorotannin content of Ascophyllum. Since L. obtusata is specialized to live and feed on a few fucoid species, including A. nodosum, the results imply that phlo- rotannins have an important mediating role in the interactions between these macroalgae and L. obtusata. The experimentally induced increase of phlorotannins was consistent with results from phlorotannin analyses of Ascophyllum individuals from natural populations, where plants that had been heavily grazed by L. obtusata contained significantly higher levels of phlorotannins compared to undamaged plants. A field survey of the distribution and abundance of L. obtusata revealed that the density of the gastropod is highly variable at the same spatial scale as the phlorotannin content of Ascophyllum in the study area. These results suggest that grazing by L. obtusata can be an important factor in explaining natural phenotypic variation in the phlorotannin content of Ascophyllum. Together with the results from a few other studies on the interactions between meso- herbivores and marine algae, the results of this study support the previously proposed hypothesis that it is feeding by relatively small, less mobile herbivores that is most likely to cue for induced production of defense chemicals in seaweeds. More studies on such interactions may reveal that the apparent rarity of inducible chemical defenses in seaweeds is misleading.

Heavy metal resistance in plants and algae may involve an internal detoxification mechanism, where toxic metal ions are chelated to substances that are stored in special compartments within the cells. Phlorotannins (brown algal polyphenolics) are strong chelators to heavy metals in solution and are found in special compartments (physodes) within the cells, and have therefore been suggested to function as an internal detoxification mechanism in brown seaweeds. Phlorotannins are also believed to function as a chemical defence against herbivory and UV radiation, and the production of phlorotannins has been shown to be induced by increased intensities of these factors. In the present study, we investigated the possible role of phlorotannins as an inducible and internal detoxification mechanism in the brown seaweed Ascophyllum nodosum (L.) Le Jol., in response to increased copper (Cu) concentrations in the surrounding water. The phlorotannin content, tissue Cu content and growth of Cu-exposed seaweeds from areas with different levels of contamination were compared in an observational field study and 2 manipulative induction experiments. The phlorotannin content and growth of the seaweeds were not affected by high Cu concentrations, although A. nodosum plants exposed to high Cu levels accumulated high concentrations of Cu in their tissues. The accumulated Cu was not specifically associated with phlorotannins, since only a small part of the total tissue Cu content of the seaweeds was found in the extracted phlorotannin fractions. The results of the present study show that phlorotannin production in A. nodosum is not induced by high Cu concentrations and that, although plants accumulate high levels of Cu in the tissues, binding of Cu to phlorotannins is probably not an important internal detoxification mechanism in A. nodosum. Since plants were not negatively affected by elevated Cu levels in terms of growth, substances other than phlorotannins are probably more important in the metal resistance mechanism of A. nodosum.

Summary 1 The potential cost of polyphenolic (phlorotannin) production in the brown seaweed Ascophyllum nodosum was investigated by testing for phenotypic trade-offs between phlorotannin content and annual growth. Relationships between tissue nitrogen and phlorotannin content, as well as between tissue nitrogen content and annual growth, were also examined to evaluate the importance of the carbon–nutrient balance in explaining intraspecific variation in phlorotannins. 2 Annual shoots were collected from a total of 300 A. nodosum individuals in natural populations at three sites in each of two study areas, in 2 consecutive years. 3 Data were analysed using analyses of covariance (ancovas), which showed that there was a significant negative relationship between phlorotannins and growth in both study areas in both years. Shoots with a relatively high phlorotannin content (> 9% dry weight) had a mean reduction in growth that varied from 25% to 54%, compared with shoots with relatively low concentrations (

Many planktonic microalgae produce a range of toxins and may form harmful algal blooms. One hypothesis is that some toxins are allelopathic, suppressing the growth of competitors, and it has been suggested that allelopathy may be one important mechanism causing algal blooms. In a metaanalysis of recent experimental work, we looked for evidence that allelopathy may explain the initiation of algal blooms. With few exceptions, allelopathic effects were only significant at very high cell densities typical of blooms. We conclude that there is no experimental support for allelopathy at prebloom densities, throwing doubts on allelopathy as a mechanism in bloom formation. Most studies tested allelopathy using cell-free manipulations. With simple models we show that cell-free manipulations may underestimate allelopathy at low cell densities if effects are transmitted during cell–cell interactions. However, we suggest that the evolution of allelopathy under field conditions may be unlikely even if based on cell–cell interactions. The spatial dispersion of cells in turbulent flow will make it difficult for an allelopathic cell to receive an exclusive benefit, and a dispersion model shows that dividing cells are rapidly separated constraining clone selection. Instead, we propose that reported allelopathic effects may be nonadaptive side effects of predator–prey or casual parasitic cell–cell interactions.

Photosynthesis is a fundamental process sustaining heterotrophic organisms at all trophic levels. Some mixotrophs can retain functional chloroplasts from food (kleptoplasty), and it is hypothesized that carbon acquired through kleptoplasty may enhance trophic energy transfer through increased host growth efficiency. Sacoglossan sea slugs are the only known metazoans capable of kleptoplasty, but the relative fitness contributions of heterotrophy through grazing, and phototrophy via kleptoplasts, are not well understood. Fitness benefits (i.e. increased survival or growth) of kleptoplasty in sacoglossans are commonly studied in ecologically unrealistic conditions under extended periods of complete darkness and/or starvation. We compared the growth efficiency of the sacoglossan Elysia viridis with access to algal diets providing kleptoplasts of differing functionality under ecologically relevant light conditions. Individuals fed Codium fragile, which provide highly functional kleptoplasts, nearly doubled their growth efficiency under high compared to low light. In contrast, individuals fed Cladophora rupestris, which provided kleptoplasts of limited functionality, showed no difference in growth efficiency between light treatments. Slugs feeding on Codium, but not on Cladophora, showed higher relative electron transport rates (rETR) in high compared to low light. Furthermore, there were no differences in the consumption rates of the slugs between different light treatments, and only small differences in nutritional traits of algal diets, indicating that the increased growth efficiency of E. viridis feeding on Codium was due to retention of functional kleptoplasts. Our results show that functional kleptoplasts from Codium can provide sacoglossan sea slugs with fitness advantages through photosynthesis.

Herbivory on marine macroalgae (seaweeds) in temperate areas is often dominated by relatively small gastropods and crustaceans (mesoherbivores). The effects of these herbivores on the performance of adult seaweeds have so far been almost exclusively investigated under artificial laboratory conditions. Furthermore, several recent laboratory studies with mesoherbivores indicate that inducible chemical resistance may be as common in seaweeds as in vascular plants. However, in order to further explore and test the possible ecological significance of induced chemical resistance in temperate seaweeds, data are needed that address this issue in natural populations. We investigated the effect of grazing by littorinid herbivorous snails (Littorina spp.) on the individual net growth of the brown seaweed Ascophyllum nodosum in natural field populations. Furthermore, the capacity for induced resistance in the seaweeds was assessed by removing herbivores and assaying for relaxation of defences. We found that ambient densities of gastropod herbivores significantly reduced net growth by 45% in natural field populations of A. nodosum. Seaweeds previously exposed to grazing in the field were less consumed by gastropod herbivores in feeding bioassays. Furthermore, the concentration of phlorotannins (polyphenolics), which have been shown to deter gastropod herbivores, was higher in the seaweeds that were exposed to gastropod herbivores in the field. This field study corroborates earlier laboratory experiments and demonstrates that it is important to make sure that the lack of experimental field data on marine mesoherbivory does not lead to rash conclusions about the lack of significant effects of these herbivores on seaweed performance. The results strongly suggest that gastropods exert a significant selection pressure on the evolution of defensive traits in the seaweeds, and that brown seaweeds can respond to attacks by natural densities of these herbivores through increased chemical resistance to further grazing.