Temporal and spatial distribution of epibenthic dinoflagellates in the Kattegat-Skagerrak, NE Atlantic-Focus on Prorocentrum lima and Coolia monotis.

Epibenthic dinoflagellates occur globally and include many toxin-producing species of concern to human health and benthic ecosystem function. Such benthic harmful algal blooms (BHABs) have been well described from tropical and sub-tropical coastal environments, but assessments from north temperate waters, e.g., northern Europe, and polar regions are scarce. The present study addressed the biodiversity and distribution of potentially toxic epibenthic dinoflagellate populations along the west coast of Sweden (Kattegat-Skagerrak) by morphological and molecular criteria. Morphological analysis conducted by light- and electron-microscopy was then linked by DNA barcoding of the V4 region of 18S rRNA gene sequences to interpret taxonomic and phylogenetic relationships. The presence of two potentially toxigenic epibenthic dinoflagellates, Prorocentrum lima (Ehrenberg) F.Stein and Coolia monotis Meunier was confirmed, along with a description of their spatial and temporal distribution. For P. lima, one third of the cell abundance values exceeded official alarm thresholds for potentially toxic BHAB events (>1000 cells gr-1 of macroalgae fresh weight). The same species were recorded consecutively for two summers, but without significant temporal variation in cell densities. SEM analyses confirmed the presence of other benthic Prorocentrum species: P. fukuyoi complex, P. cf. foraminosum and P. cf. hoffmannianum. Analyses of the V4 region of the 18S rRNA gene also indicated the presence P. compressum, P. hoffmannianum, P. foraminosum, P. fukuyoi, and P. nanum. These findings provide the first biogeographical evidence of toxigenic benthic dinoflagellates along the west coast of Sweden, in the absence of ongoing monitoring to include epibenthic dinoflagellates. Harmful events due to the presence of Coolia at shellfish aquaculture sites along the Kattegat-Skagerrak are likely to be rather marginal because C. monotis is not known to be toxigenic. In any case, as a preliminary assessment, the results highlight the risk of diarrhetic shellfish poisoning (DSP) events caused by P. lima, which may affect the development and sustainability of shellfish aquaculture in the region.

Stranding events of drift organisms (Arribadas) in southern Brazil and the spread of invasive bryozoan in South America.

The accumulation of drift organisms (Arribadas, constituted by benthic microalgae, macroalgae and bryozoans) on Central Beach in Balneário Camboriú (SC) has drawn attention since the early 2000s. However, historical surveys suggest an ancient phenomenon that has been increasing in intensity, promoted by urbanization and the eutrophication of the bay in recent years, leading to changes in the taxonomic composition. Previously, these Arribadas were comprised of two species of benthic microalgae (Amphitetras antediluviana and Biddulphia biddulphiana) and the bryozoan Arbocuspis ramosa. However, since 2019, a substantial increase in biomass has been observed on the beach and the dominance of the macroalgae Bryopsis plumosa strongly suggests an increase in the load of organic matter and nutrients in the bay. Recently (2022) the presence of a new invasive bryozoan species (Amathia alternata) was detected, highlighting the need to continue investigating the Arribadas to monitor the ecological evolution of this process.

Microphytobenthos spatio-temporal dynamics across an intertidal gradient using Random Forest classification and Sentinel-2 imagery.

Microphytobenthos (MPB) provides important ecosystem functions and services, contributing significantly to the total primary production in shallow coastal ecosystems. However, determining the factors that regulate the seasonal changes of MPB and its distribution patterns at larger scales is hindered by the considerable spatial and temporal variability in these environments. Here, we studied the dynamics of intertidal MPB biomass, cover, and net growth rates in a south European tidal flat (Cadiz Bay, Spain) over a four-year period using the Normalized Difference Vegetation Index (NDVI) calculated from Sentinel-2 satellite images. Pixels dominated by different benthic communities (MPB, Zostera sp., Caulerpa sp. and green macroalgae) were identified at a 10-m resolution using a Random Forest (RF) machine learning classification algorithm. MPB dominated the intertidal zone. MPB cover did not show a clear seasonal pattern and was clearly higher in the middle of the intertidal range of sea level. Despite interannual variability, MPB biomass was always higher during winter, coinciding with observations from other low latitude intertidal flats with temperate climate, and in the upper-middle intertidal. Net rates of MPB biomass change, calculated from the differences in MPB NDVI over time, showed maximal net growth rates from autumn to winter and maximum loss rates during spring and summer, although with high variability. Our study demonstrates that RF algorithms allow mapping MPB and other intertidal communities from Sentinel-2 multispectral satellite imagery accurately obtaining invaluable information from large areas at very high spatio-temporal resolution. The dissimilarities observed in the patterns of MPB variables over time or sea level, indicate differences in their ecological regulation, still largely unknown both here and in other temperate climate intertidal flats. High resolution remote sensing can aid in their detailed and systematic study producing a more integrated view of these systems and contributing to their science-based management and conservation.

Succession pattern and phylotype analysis of microphytobenthic communities in a simulated oil spill seagrass mesocosm experiment.

Microphytobenthic communities play a significant role in nutrient modulation, sediment stabilization, and primary production in seagrass beds, which provide various ecosystem services. We hypothesized that microphytobenthic communities in sediments of chronically oil-exposed seagrass beds will exhibit increased resiliency to stressors associated with oil exposure as opposed to seagrass beds never exposed to oil spills. We prepared 14-liter seawater mesocosms, each containing a submersed macrophyte Ruppia maritima collected from the Chandeleur Islands, Louisiana, and Estero Bay, Florida. Mesocosms were initially exposed to 50% water-accommodated oil fractions (WAF) and subsequently diluted by 50% with daily artificial seawater exchanges over 8 days to simulate tidal dilution. High-throughput amplicon sequencing based on 23S rRNA gene targeting cyanobacteria and chloroplasts of eukaryotic microphytobenthos was conducted to assess the impact of oiling on microphytobenthic communities with additional assessment via microscopy. High-throughput sequencing in combination with traditional microscopic analysis provided a robust examination in which both methods roughly complemented each other. Distinct succession patterns were detected in benthic algal communities of chronically oil-exposed (Louisiana) versus unexposed (Florida) seagrass bed sediments. The impact of oiling in microphytobenthos across all samples showed that benthic diatoms dominated all algal communities with sample percentages ranging from 42 to 97%, followed by cyanobacteria (2 to 50%). It is noteworthy that drastic changes in microphytobenthic community structure in terms of the larger taxonomic level were not observed, rather change occurred at the phylotype level. These results were also confirmed by microscopy. Similarity percentages (SIMPER) analysis identified seven phylotypes (Cyanobacteria, Bacillariophyceae, and Mediophyceae) in the Louisiana samples and one phylotype (Bacillariophyceae) in the Florida samples that increased in relative sequence abundance after oil exposure. The detailed phylotype analysis identifying sentinel microphytobenthic indicators provides a base for future research on benthic microalgae response to ecosystem disturbance.

Heteroaggregates of Polystyrene Nanospheres and Organic Matter: Preparation, Characterization and Evaluation of Their Toxicity to Algae in Environmentally Relevant Conditions.

The environmental fate and behavior of nanoplastics (NPs) and their toxicity against aquatic organisms are under current investigation. In this work, relevant physicochemical characterizations were provided to analyze the ecotoxicological risk of NPs in the aquatic compartment. For this purpose, heteroaggregates of 50 nm polystyrene nanospheres and natural organic matter were prepared and characterized. The kinetic of aggregation was assimilated to a reaction-limited colloid aggregation mode and led to the formation of heteroaggregates in the range of 100-500 nm. Toxicities of these heteroaggregates and polystyrene nanospheres (50 and 350 nm) were assessed for a large range of concentrations using four benthic and one planktonic algal species, in regards to particle states in the media. Heteroaggregates and nanospheres were shown to be stable in the exposure media during the ecotoxity tests. The algal species exhibited very low sensitivity (growth and photosynthetic activity), with the noteworthy exception of the planktonic alga, whose growth increased by more than 150% with the heteroaggregates at 1 µg L-1. Despite the lack of a strong direct effect of the NPs, they may still impair the functioning of aquatic ecosystems by destabilizing the competitive interactions between species. Moreover, further work should assess the toxicity of NPs associated with other substances (adsorbed pollutants or additives) that could enhance the NP effects.

Examining the dynamic nature of epiphytic microalgae in the Florida Keys: What factors influence community composition?

The factors that influence the composition of marine epiphytic microalgal assemblages are poorly-understood. To address this short-coming, 93 samples were collected from four distinct regions in the Florida Keys National Marine Sanctuary (FKNMS) during winter and summer months to test the model that epiphytic microalgal communities are influenced by environmental gradients related to different sites, seasons, and host macrophyte species. One hundred and eighty-three morphotypes from 13 classes (7 phyla) were identified, dominated by 106 Bacillariophyta (77 identified to species equivalent or below), 37 Cyanophyta (13 identified to species equivalent or below), and 30 Dinophyta (21 identified to species equivalent or below). The largest proportion of variability in epiphytic communities was related to physico-chemical parameters (37%), followed by site location (ocean-versus bayside; 15%), seasonal differences (11%), and host macrophyte species (10%). Four physico-chemical variables were found to be most influential: wave height, temperature, ammonium concentration, and salinity. Only six out of 616 epiphyte - host comparisons exhibited significant differences in individual epiphyte taxon abundance between different host species (within site and season), further demonstrating that host-specificity was not strongly evident in this study. Overall, the results of this (sub)tropical study indicate that changing environmental characteristics between sites and seasons were the primary drivers influencing epiphyte community composition. Similar findings were found in an accompanying study of phytoplankton and other studies from temperate and (sub)polar regions, suggesting that common, underlying processes exist among these disparate environments.

Ecología funcional de las algas perifíticas en el Chocó colombiano: limitación de recursos, competencia y variables ambientales / Functional ecology of periphytic algae in the Colombian Chocó: resource limitations, competition, and environmental variables

Introducción: El conocimiento de las comunidades perifíticas es importante, dada su participación en la producción primaria de los ambientes fluviales; además, el perifiton es una valiosa comunidad bioindicadora. No obstante, hasta la fecha es poco lo que se conoce sobre la ecología funcional de esta comunidad en los ríos colombianos. Objetivo: En este trabajo se buscó caracterizar algunos aspectos funcionales de la comunidad de ficoperifiton de cuatro sistemas lóticos de la Reserva Natural La Planada, ubicada en el departamento de Nariño, Colombia. Métodos: Durante mayo de 2019, en cada río se establecieron entre uno y tres sitios de muestreo, en los cuales se midieron las características hidrológicas y fisicoquímicas y se determinó la composición taxonómica y algunos rasgos funcionales de las comunidades de algas perifíticas. Se calcularon la media ponderada comunitaria (MPC) y la varianza ponderada comunitaria (VPC) de estos rasgos y se exploró su relación con las variables abióticas mediante regresiones, correlaciones y análisis de correspondencia canónica (ACC). Resultados: Se registraron 59 géneros de algas, de los cuales a 47 se les midieron los rasgos funcionales. La MPC del rasgo exoesqueleto silíceo fue estadísticamente mayor en la quebrada El Tejón. La MPC y la VPC de los demás rasgos no varió entre ríos, pero en algunos de ellos la presencia de mucílago y la organización en filamentos tendió a ser mayor. La amplia fluctuación de la VPC de todos los rasgos funcionales parece señalar que estas comunidades ficoperifíticas no tienen limitaciones de recursos, la competencia es reducida y las especies tienden a ser funcionalmente diferentes. Las variables ambientales con mayor influencia fueron el caudal, la mineralización del agua, el pH y la transparencia. La presencia de mucílago se asoció con la dureza del agua y el predominio de algas filamentosas con la transparencia; el pH influyó positivamente la relación superficie/volumen (S/V). Conclusiones: Este estudio constituye una línea base que permitirá evaluar cambios en las comunidades de algas bentónicas ante posibles intervenciones y brindar pautas para eventuales acciones de restauración de los sistemas fluviales de esta región de importancia mundial por su elevada biodiversidad.

Introduction: Knowledge on the periphytic community is essential because of its central role in river primary production. Also, periphyton is a valuable bioindicator of the community. However, to the date, little is known about the functional ecology of these communities in Colombian rivers. Objective: This work sought to characterize functional aspects of the phycoperiphytic community of four lotic systems of the La Planada Natural Reserve, located in the Department of Nariño, Colombia. Methods: In May 2019, between one and three sampling sites were established in each river. Hydrological and physicochemical variables were measured. Periphyton was sample to determine the taxonomic composition of the benthic algae as well as some functional traits. From these traits, the community weighted mean (CWM) and community weighted variance (CWV) were calculated, and their relation with abiotic variables was explored using regressions, correlations, and a canonical correspondence analysis (CCA). Results: 59 genera of algae were recorded, from which 47 had its functional traits measured. The CWM of the siliceous exoskeleton trait was statistically higher in El Tejón creek. The other traits had similar CWM and CWV values in all rivers. However, the traits of the presence of mucilage and organization in filaments showed predominance in some streams. The fluctuating data of CWV in all rivers seem to indicate that these phycoperiphytic communities have no resource limitations, competition is low, and species tend to be functionally different. The environmental variables with the greatest influence were flow, water mineralization, pH, and water transparency. The hardness of the water and the presence of mucilage were associated, while the predominance of filamentous algae was related to transparency; pH positively influenced the surface/volume ratio (S/V). Conclusions: This study represents a baseline that will allow evaluating changes in the benthic algae communities in the face of possible interventions and providing guidelines for eventual actions to restore the river systems of this important region due to its high biodiversity.

Hydrological connectivity and herbivores control the autochthonous producers of coastal salt marshes.

Sea-level rise and overfishing could enhance the strength of hydrological connectivity and the presence of herbivores, resulting in salt marsh loss through physical stress and trophic cascade effects. Our aim was to estimate the effects of these two stresses on autochthonous producers. Survivorship and biomass of Suaeda salsa (S. salsa) were the lowest in areas with high hydrological connectivity, whereas the highest biomass was observed in the areas with moderate connectivity. The biomass of benthic microalgae was higher under low hydrological connectivity and no herbivores. The interactive effects between hydrological connectivity and herbivores on S. salsa but not on benthic microalgae were observed. Herbivores were somewhat (28%) important for the survival of initial S. salsa seedlings, while hydrological connectivity controlled (50%) the biomass of benthic microalgae. Our study highlights that, the autochthonous producers in coastal salt marshes may disappear due to strong hydrological connectivity and the excessive presence of herbivores.

Diversity and Toxicity of the Genus Coolia Meunier in Brazil, and Detection of 44-Methyl Gambierone in Coolia tropicalis.

Coolia is a genus of marine benthic dinoflagellates which is widely distributed in tropical and temperate zones. Toxicity has been reported in selected Coolia species, although the identity of causative compounds is still controversial. In this study, we investigated the taxonomical and toxicological aspects of Coolia species from Brazil. Since light- and electron microscopy-based morphology was not enough to distinguish small-celled species, ITS and LSU D1-D3 phylogenetic analyses were used for species definition. Cultures of Coolia palmyrensis and Coolia santacroce were established from samples collected along the northeastern Brazilian coast, the first record of both species in South Atlantic waters. Cultures of Coolia malayensis and Coolia tropicalis were also established and exhibited acute in vivo toxicity to adults of Artemia salina, while C. palmyrensis and C. santacroce were non-toxic. The presence of 30 yessotoxin analogues, 7 metabolites of Coolia and 44 Gambierdiscus metabolites was screened in 14 strains of Coolia. 44-methyl gambierone (formerly referred to as MTX3) and a new isomer of this compound were detected only in C. tropicalis, using both low- and high-resolution LC-MS/MS. To our knowledge, this is the first report of gambierone analogues in dinoflagellates other than Gambierdiscus; the role of C. tropicalis in ciguatera poisoning thus deserves to be considered in further investigations.

Spatiotemporal variability in microphytobenthic primary production across bare intertidal flat, saltmarsh, and mangrove forest of Asia and Australia.

The ecological role of intertidal microphytobenthos (MPB) is increasingly recognized in coastal production systems. MPB primary production (PP) measured in coastal wetlands of Korea, Cambodia, and Australia confirmed large variability at the global scale. Surprisingly, MPB biomass in mangrove forests almost doubled those measured in nearby bare tidal flats. However, MPB productivity (Pb) in vegetated habitats was significantly reduced (by ~50%) compared to that on bare tidal flats. Extensive measurements of MPB biomass, PP, and Pb across 12 Korean tidal flats revealed large spatiotemporal variations, suggesting complex sediment-MPB coupled dynamics. The key factors included sediment type, tide, bed elevation, irradiation, temperature, and vegetation. Winter MPB blooms and the elevated Pb seem to be unique characteristics of the Korean intertidal flats. The present study provides the baseline data of MPB PPs in mudflat, saltmarsh, and mangrove habitats in the highly productive zones of the Western Indo-Pacific Rim.

Ostreopsis cf. ovata Bloom in Currais, Brazil: Phylogeny, Toxin Profile and Contamination of Mussels and Marine Plastic Litter.

Ostreopsis cf. ovata is a toxic marine benthic dinoflagellate responsible for harmful blooms affecting ecosystem and human health, mostly in the Mediterranean Sea. In this study we report the occurrence of a summer O. cf. ovata bloom in Currais, a coastal archipelago located on the subtropical Brazilian coast (~25° S). This bloom was very similar to Mediterranean episodes in many aspects: (a) field-sampled and cultivated O. cf. ovata cells aligned phylogenetically (ITS and LSU regions) along with Mediterranean strains; (b) the bloom occurred at increasing temperature and irradiance, and decreasing wind speed; (c) cell densities reached up to 8.0 × 104 cell cm-2 on fiberglass screen and 5.6 × 105 cell g-1 fresh weight on seaweeds; (d) and toxin profiles were composed mostly of ovatoxin-a (58%) and ovatoxin-b (32%), up to 35.5 pg PLTX-eq. cell-1 in total. Mussels were contaminated during the bloom with unsafe toxin levels (up to 131 µg PLTX-eq. kg-1). Ostreopsis cells attached to different plastic litter, indicating an alternate route for toxin transfer to marine fauna via ingestion of biofilm-coated plastic debris.

Biofilm-forming capacity of two benthic microalgae, Navicula incerta and Navicula sp., on three substrates (Naviculales: Naviculaceae)

Benthic microalgae have the natural capacity to adhere to a diversity of fixed submerged substrates to form biofilms, which have important roles not only in natural ecosystems, but also in aquaculture systems. An experimental investigation was performed to assess the biofilm-forming capacity of two microalgae (Navicula incerta and Navicula sp.) on three different substrates (plastic net, fabric, and wood) under controlled temperature and light conditions. The substrates were arranged on curtains suspended from a wood stick, into plastic aquariums (45 L in capacity) filled with filtered marine water enriched with F/2 medium. The trial was carried out until the exponential growing phase of the microalgae was reached. After that, the incorporated biomass was gravimetrically calculated, and its biochemical composition was determined by standard methods. The greatest amount of incorporated dry matter was observed for Navicula sp. on fabric and the lowest was observed for wood. The highest number of cells associated with the biofilm was obtained for Navicula sp. on the plastic net (1.24 x 109 cells/m2), while the lowest was recorded for Navicula sp. on the wood (1.43 x 108 cells/m2). Significant differences in organic matter were found among the substrates, with the highest values for N. incerta on the fabric (3.22 g/m2) and the lowest for Navicula sp. on the wood (0.02 g/m2). The best biochemical profiles among the formed biofilms were observed for N. incerta on the plastic net and Navicula sp. on the fabric. The plastic net was considered the best substrate because of the stability of the biofilm and the easiness of harvesting the biomass.

Las microalgas bentónicas tienen la capacidad natural de adherirse a diversos sustratos fijos sumergidos para formar biopelículas, las cuales tienen roles importantes no solo en ecosistemas naturales sino también en sistemas de producción acuícolas. Se llevó a cabo una investigación experimental para evaluar la capacidad formadora de biopelículas de dos microalgas bentónicas (Navicula incerta y Navicula sp.) en tres diferentes sustratos (malla plástica, tela y madera), bajo condiciones controladas de temperatura y luz. Los sustratos fueron arreglados a manera de cortinas suspendidas de un tubo de PVC dentro de acuarios de plástico (45 L de capacidad) con agua marina enriquecida con el medio F/2. El experimento se llevó hasta que la fase de crecimiento exponencial de la microalga fue alcanzada. Posteriormente la biomasa incorporada fue calculada gravimétricamente, y su composición bioquímica fue determinada por métodos estándar. La mayor cantidad de materia seca se observó para N. incerta en el sustrato de tela y la menor se encontró en el de madera. El mayor número de células asociadas a la biopelícula fue registrado para Navicula sp. en malla plástica (1.24 x 109 cel/m2), mientras que el menor se encontró para Navicula sp. en madera (1.43 x 108 cels/m2). Diferencias significativas en cuanto a materia orgánica se encontraron entre los sustratos y las especies, con valores más altos para N. incerta en tela (3.22 g/m2) y más bajos para Navicula sp. en madera (0.02 ± 0.05 g/m2). Los mejores perfiles bioquímicos para las biopelículas correspondieron a las formadas por N. incerta sobre malla plástica y Navicula sp. sobre tela. La red de plástico se consideró el mejor sustrato debido a la estabilidad de la biopelícula y la facilidad para cosechar la biomasa.

Interactive Impacts of Silver and Phosphorus on Autotrophic Biofilm Elemental and Biochemical Quality for a Macroinvertebrate Consumer.

Autotrophic biofilms are complex and fundamental biological compartments of many aquatic ecosystems. In particular, these biofilms represent a major resource for many invertebrate consumers and the first ecological barrier against toxic metals. To date, very few studies have investigated the indirect effects of stressors on upper trophic levels through alterations of the quality of biofilms for their consumers. In a laboratory study, we investigated the single and combined effects of phosphorus (P) availability and silver, a re-emerging contaminant, on the elemental [carbon (C):nitrogen (N):P ratios] and biochemical (fatty acid profiles) compositions of a diatom-dominated biofilm initially collected in a shallow lake. We hypothesized that (1) P and silver, through the replacement of diatoms by more tolerant primary producer species, reduce the biochemical quality of biofilms for their consumers while (2) P enhances biofilm elemental quality and (3) silver contamination of biofilm has negative effects on consumers life history traits. The quality of biofilms for consumers was assessed for a common crustacean species, Gammarus fossarum, by measuring organisms' survival and growth rates during a 42-days feeding experiment. Results mainly showed that species replacement induced by both stressors affected biofilm fatty acid compositions, and that P immobilization permitted to achieve low C:P biofilms, whatever the level of silver contamination. Gammarids growth and survival rates were not significantly impacted by the ingestion of silver-contaminated resource. On the contrary, we found a significant positive relationship between the biofilm P-content and gammarids growth. This study underlines the large indirect consequences stressors could play on the quality of microbial biomass for consumers, and, in turn, on the whole food web.

Trophic transfer of metals in a seagrass food web: Bioaccumulation of essential and non-essential metals.

Metal concentrations are reported for a seagrass ecosystem receiving industrial inputs. δ13C and δ15N isotope ratios were used to establish trophic links. Copper concentrations (dry mass) ranged from <0.01 µg/g in fish species to 570 µg/g (µâ€¯= 49 ±â€¯SD = 90 µg/g) in the oyster Saccostrea glomerata. Zinc concentrations ranged from 0.6 µg/g in the seagrass Zostera capricorni to 10,800 µg/g in the mud oyster Ostrea angasi (µâ€¯= 434 ±â€¯1390 µg/g). Cadmium concentrations ranged from <0.01 µg/g in fish species to 268 µg/g in Ostrea angasi (µâ€¯= 6 ±â€¯25 µg/g). Lead concentrations ranged from <0.01 µg/g for most fish species to 20 µg/g in polychaetes (µâ€¯= 2 ±â€¯3 µg/g). Biomagnification of metals did not occur. Organisms that fed on particulate organic matter and benthic microalgae had higher metal concentrations than those that fed on detritus. Species physiology also played an important role in the bioaccumulation of metals.

Inhibition of Nitzschia ovalis biofilm settlement by a bacterial bioactive compound through alteration of EPS and epiphytic bacteria

Background: Marine ecosystems contain benthic microalgae and bacterial species that are capable of secreting extracellular polymeric substances (EPS), suggesting that settlement of these microorganisms can occur on submerged surfaces, a key part of the first stage of biofouling. Currently, anti-fouling treatments that help control this phenomenon involve the use of biocides or antifouling paints that contain heavy metals, which over a long period of exposure can spread to the environment. The bacterium Alteromonas sp. Ni1-LEM has an inhibitory effect on the adhesion of Nitzschia ovalis, an abundant diatom found on submerged surfaces. Results: We evaluated the effect of the bioactive compound secreted by this bacterium on the EPS of biofilms and associated epiphytic bacteria. Three methods of EPS extraction were evaluated to determine the most appropriate and efficient methodology based on the presence of soluble EPS and the total protein and carbohydrate concentrations. Microalgae were cultured with the bacterial compound to evaluate its effect on EPS secretion and variations in its protein and carbohydrate concentrations. An effect of the bacterial supernatant on EPS was observed by assessing biofilm formation and changes in the concentration of proteins and carbohydrates present in the biofilm. Conclusions: These results indicate that a possible mechanism for regulating biofouling could be through alteration of biofilm EPS and alteration of the epiphytic bacterial community associated with the microalga.

Effects of sandy vs muddy sediments on the vertical distribution of microphytobenthos in intertidal flats of the Fraser River Estuary, Canada.

Benthic algae or microphytobenthos (MPB) in intertidal flats play an important role in the sediment and overlying water ecosystems. We hypothesize that there are effects of sediment texture on the vertical distribution of MPB using chlorophyll a (chl a) as a proxy for MPB biomass and present results over a 2.5-year period. Four sites were sampled monthly: two sandy sites (A10 and A12) and two muddy sites (A0 and A14) on the intertidal flats of the Fraser River Estuary. At the two sandy sites, pigments were distributed down to 10 cm. High ratios of depth-integrated chl a to phaeopigments suggest that the chl a had been recently buried. In contrast, at the muddy sites, pigments were limited to the top 4 cm, with MBP in the top 1 cm contributing up to 60 % of the whole sediment core pigments. As a result, the depth-integrated chl a values were on average 2,044 mg m(-2) (160-4,200) at A10 and 882 mg m(-2) (183-2,569) at A12, the two sandy sites, and much higher than at the two muddy sites where averages of 84 mg m(-2) (41-174) and 235 mg m(-2) (77-854) were measured at A0 and A14, respectively. Despite these lower concentrations at the muddy sites than at the sandy sites, particulate organic carbon (POC) and nitrogen (PON) concentrations showed a homogenous vertical distribution at the two sandy sites. Such a homogeneous vertical distribution of chl a, POC, and PON suggests that vertical transport mechanisms were actively transporting organic material into and out of the sediment. These results suggest that MBP on sandy sediments play a very active role in providing food for herbivores and are interacting with the overlying water column in the sediment-water exchange processes during tidal cycles.

THE EFFECTS OF TEMPERATURE ON THE PHOTOSYNTHETIC PARAMETERS AND RECOVERY OF TWO TEMPERATE BENTHIC MICROALGAE, AMPHORA CF. COFFEAEFORMIS AND COCCONEIS CF. SUBLITTORALIS (BACILLARIOPHYCEAE)(1).

Temperature and irradiance are the most important factors affecting marine benthic microalgal photosynthetic rates in temperate intertidal areas. Two temperate benthic diatoms species, Amphora cf. coffeaeformis (C. Agardh) Kütz. and Cocconeis cf. sublittoralis Hendey, were investigated to determine how their photosynthesis responded to temperatures ranging from 5°C to 50°C after short-term exposure (1 h) to a range of irradiance levels (0, 500, and 1,100 µmol photons · m(-2 ) ·( ) s(-1) ). Significant differences were observed between the temperature responses of maximum relative electron transport rate (rETRmax), photoacclimation index (Ek ), photosynthetic efficiency (α), and effective quantum yield (ΔF/Fm' ) in both species. A. coffeaeformis had a greater tolerance to higher temperatures than C. sublittoralis, with nonphotochemical quenching (NPQ) activated at temperatures of 45°C and 50°C. C. sublittoralis, however, demonstrated a more rapid rate of recovery at ambient temperatures. Temperatures between 10°C and 20°C were determined to be optimal for photosynthesis for both species. High temperatures and irradiances caused a greater decrease in ΔF/Fm' values. These results suggest that the effects of temperature are species specific and that short-term exposure to adverse temperature slows the recovery process, which subsequently leads to photoinhibition.

Decoupling of nutrient and grazer impacts on a benthic estuarine diatom assemblage.

Strong interactions between top-down (consumptive) and bottom-up (resource supply) trophic factors occur in many aquatic communities, but these forces can act independently in some microphytobenthic communities. Within benthic estuarine diatom assemblages, the dynamics of these interactions and how they vary with abiotic environmental conditions are not well understood. We conducted a field experiment at two sites with varying habitat characteristics to investigate the interactive effects of grazers and nutrients on benthic estuarine diatoms. We crossed snail (Cerithidea californica) and nutrient (nitrogen and phosphorus) addition treatments in enclosures on a restored tidal sandflat and a reference tidal mudflat in Mugu Lagoon, southern California. We repeated the study in summer 2000 and spring 2001 to assess temporal variation in the interactions. Snails caused a large decrease in diatom relative abundance and biomass (estimated as surface area); nutrients increased diatom abundance but did not alter diatom biomass. Snails and nutrients both reduced average diatom length, although the nutrient effect was weaker and temporally variable, occurring in the reference mudflat in the spring. There were few interactions between snail and nutrient addition treatments, suggesting that links between top-down and bottom-up forces on the diatom community were weak. There were no consistent differences in diatom assemblage characteristics between the two study sites, despite marked differences in sediment grain size and other abiotic characteristics between the sites. The strong diatom response to herbivores and weaker responses to enrichment differed from the previous studies where cyanobacteria increased in response to nutrient enrichment, further dissolving the "black box" perception of microphytobenthic communities.

Human trampling as short-term disturbance on intertidal mudflats: effects on macrofauna biodiversity and population dynamics of bivalves.

The effect of physical disturbance in the form of trampling on the benthic environment of an intertidal mudflat was investigated. Intense trampling was created as unintended side-effect by benthic ecologists during field experiments in spring and summer 2005, when a mid-shore area of 25 × 25 m was visited twice per month by on average five researchers for a period of 8 months. At the putatively-impacted location (I) (25 × 25 m) and two nearby control locations (Cs) (25 × 25 m each), three sites (4 × 4 m) were randomly selected and at each site, three plots (50 × 50 cm) were sampled after 18 and 40 days from the end of the disturbance. Multivariate and univariate asymmetrical analyses tested for changes in the macrofaunal assemblage, biomass of microphytobenthos and various sediment properties (grain-size, water content, NH4 and NO3 concentrations in the pore water) between the two control locations (Cs) and the putatively-impacted location (I). There were no detectable changes in the sediment properties and microphytobenthos biomass, but variability at small scale was observed. Microphytobenthos and NH4 were correlated at I to the number of footprints, as estimated by the percentage cover of physical depressions. This indicated that trampling could have an impact at small scales, but more investigation is needed. Trampling, instead, clearly modified the abundance and population dynamics of the clam Macoma balthica (L.) and the cockle Cerastoderma edule (L.). There was a negative impact on adults of both species, probably because footsteps directly killed or buried the animals, provoking asphyxia. Conversely, trampling indirectly enhanced recruitment rate of M. balthica, while small-sized C. edule did not react to the trampling. It was likely that small animals could recover more quickly because trampling occurred during the growing season and there was a continuous supply of larvae and juveniles. In addition, trampling might have weakened negative adult-juvenile interactions between adult cockles and juvenile M. balthica, thus facilitating the recruitment. Our findings indicated that human trampling is a relevant source of disturbance for the conservation and management of mudflats. During the growing season recovery can be fast, but in the long-term it might lead towards the dominance of M. balthica to the cost of C. edule, thereby affecting ecosystem functioning.