Influence of nutrient enrichment on colonisation and photosynthetic parameters of hard substrate marine microphytobenthos.

This study aimed to assess the influence of nutrient enrichment on the development of microalgal biofilm on concrete and PVC cubes. Three mesocosms were utilized to create a nutrient gradient over a period of 28 days. Various parameters including biomass, photosynthetic activity, microtopography, and extracellular polymeric substances (EPS) were measured. Imaging PAM techniques were employed to obtain surface-wide data. Results revealed that nutrient availability had no significant impact on Chl a biomass and the maximum quantum efficiency of PSII (Fv/Fm). The photosynthetic capacity and efficiency were minimally affected by nutrient availability. Interestingly, the relationship between microphytobenthic (MPB) biomass and photosynthesis and surface rugosity exhibited distinct patterns. Negative reliefs showed a strong correlation with Fv/Fm, while no clear pattern emerged for biomass on rough concrete structures. Overall, our findings demonstrate that under conditions of heightened eutrophication, biofilm photosynthesis thrives in the fissures and crevasses of colonized structures regardless of nutrient levels. This investigation provides valuable insights into the interplay between nutrient availability and surface rugosity.

Aluminium-based galvanic anode impacts the photosynthesis of microphytobenthos and supports the bioaccumulation of metals released.

Very few studies have looked at the potential biological effects of degradation products of galvanic anodes particularly on primary producers which are central to food webs in marine ecosystems. The galvanic anode cathodic protection system (GACP) is widely used to protect submerged metallic structures from corrosion. Aluminium (Al) and zinc (Zn) are the main constituents of galvanic anodes and are therefore released in the marine environment by oxidation process to form ions or oxy-hydroxides. The main objective of our study was to evaluate the effects of the metals released from an aluminium-based galvanic anode on microphytobenthos performance in term of biofilm growing through the analysis of photosynthetic parameters, the determination of chlorophyll and extracellular polymeric substances (EPS). The bioaccumulation of Al and Zn were measured in the microphytobenthic compartment collected at the surface of polyvinyl chloride (PVC) plates exposed during 13 days to seawaters enriched in different concentrations of metals released from dissolution of one anode. Determination of bioconcentration factors confirmed that the microphytobenthos has incorporated Al. A significative effect was observed on the Chl a concentration for the higher tested concentration ([Al] = 210.1 ± 60.2 µg L - 1; [Zn] = 20.2 ± 1.4 µg L - 1). The seawater exposed to the anode affected the MPB productivity (ETRIImax) with consequences on acclimatation light (Ek), absorption cross section of PSII (σPII), Fv/Fm and NPQ. Regarding the EPS production, the anode degradation presented an impact on high and low molecular weight of both carbohydrates and protein fractions of microphytobenthos suggesting that EPS play an essential role in sequestering metal contaminants to maintain the integrity of the biological membranes and the functionality of the cellular organelles. The accumulation of Al released by GACP in microphytobenthos cells could lead to physiologic problems in photosynthetic organisms.

Colonisation of artificial structures by primary producers: competition and photosynthetic behaviour.

Colonisation of artificial structures by primary producers is an important determinant for eco-engineering projects. In this context, interactions between the colonisation by microphytobenthic biofilm and macroalgae were explored on 48 samples of marine infrastructures (MI) immersed for one year in the English Channel. Marine infrastructures samples with smooth and rough surface were compared to evaluate the influence of surface micro-scale rugosity. Microphytobenthos biomass (MPB), macroalgal diversity and photosynthetic parameters of both were assessed during colonisation. No significant differences were found as a function of the surface rugosity of MI samples, which was unexpected, but can be explained by biogenic rugosity provided by barnacles. Marine infrastructures were largely colonised by a red encrusting alga, Phymatolithon purpureum, which showed poor photosynthetic capacity compared to the microphytobenthos present next to it. Colonisation by monospecific encrusting algae tended to reduce the primary productivity of hard substrate.

Organic Phosphorus Scavenging Supports Efficient Growth of Diazotrophic Cyanobacteria Under Phosphate Depletion.

Considering the reported significant diazotrophic activities in open-ocean regions where primary production is strongly limited by phosphate, we explored the ability of diazotrophs to use other sources of phosphorus to alleviate the phosphate depletion. We tested the actual efficiency of the open-ocean, N2-fixer Crocosphaera watsonii to grow on organic phosphorus as the sole P source, and observed how the P source affects the cellular C, N, and P composition. We obtained equivalent growth efficiencies on AMP and DL-α-glycerophosphate as compared with identical cultures grown on phosphate, and survival of the population on phytic acid. Our results show that Crocosphaera cannot use all phosphomonoesters with the same efficiency, but it can grow without phosphate, provided that usable DOP and sufficient light energy are available. Also, results point out that organic phosphorus uptake is not proportional to alkaline phosphatase activity, demonstrating that the latter is not a suitable proxy to estimate DOP-based growth yields of organisms, whether in culture experiments or in the natural environment. The growth parameters obtained, as a function of the P source, will be critical to improve and calibrate mathematical models of diazotrophic growth and the distribution of nitrogen fixation in the global ocean.

A review of methods and indicators used to evaluate the ecological modifications generated by artificial structures on marine ecosystems.

The current development of human activities at sea (e.g. land reclamation, maritime activity and marine renewable energy) is leading to a significant increase in the number of infrastructures installed in marine settings. These artificial structures provide new hard-bottom habitats for many marine organisms and can thus modify the structure and functioning of coastal ecosystems. In order to better evaluate the nature of these modifications as well as the potential benefits and/or impacts generated, it becomes essential to develop assessment methods that can be applied to a wide variety of study sites from harbours to coastal offshore environments. In this context, our study aims to review the different methods and indicators available which are used to measure the modifications of biodiversity and ecological functioning generated by such structures. Among the methods reviewed, we highlight some that were developed specifically for artificial structures, and others intended for various primary uses but which have been successfully transposed to artificial structures. Nevertheless, we also point out the lack of reliable methods concerning some biological ecosystem components impacted by artificial structures. In this context, we require the adaptation or creation of brand-new indicators to achieve a better characterisation of the ecological impacts generated by these structures. Overall, this study highlights a very high number of existing methods, which provide stakeholders with useful tools to study the impacts of artificial structures, and identifies the need to develop integrative indicators to enhance the deployment of new artificial structures.

Colimitation assessment of phytoplankton growth using a resource use efficiency approach in the Bay of Seine (French-English Channel).

Eutrophication and dystrophy are two of the main problems affecting coastal ecosystems. In the Bay of Seine, phosphorus (P) inputs from the Seine estuary have been largely reduced in the last decade, in contrast to nitrogen (N), which leads to high N/P ratio inputs. To study the effect of dystrophy, an enrichment bioassay using water sampled from the Bay of Seine was repeated 19 times over a period of 18 months with six different enrichments. After a few days, chlorophyll a (chl a), alkaline phosphatase activity (APA), transparent exopolymeric particles (TEPs), cytometric size structure, and maximum quantum yield of photosystem II were measured. The data provide strong evidence for an N & P colimitation system in the vast majority of the incubations, as only the N + P and N + P + Si enrichments supported phytoplankton growth, and Si only appeared to play a secondary role in our incubations. A N/P ratio of 16 equal to the Redfield ratio was identified as the optimum for balanced growth, as chl a was the highest and TEP and APA production was the lowest at this ratio. To fit the requirements of the colimited system, a new resource use efficiency (RUENP) calculation was developed to account for N and P colimitation instead of only one nutrient, as is usually the case. This calculation allows better representation of RUE in dystrophic conditions, as found in many highly anthropized ecosystems. The relationships between RUENP and the size structure of the phytoplankton community were explored, and a significant positive correlation between RUENP and larger cells (>2 µm) and a negative correlation with smaller cells (<2 µm) were noted, showing a better use of nutrients by larger cells. This study highlights an increase of RUENP with the phytoplankton cell size in a colimited system.

Erratum to "Quantifying Cyanothece growth under DIC limitation" [Comput. Struct. Biotechnol. J. 19 (2021) 6456-6464].

[This corrects the article DOI: 10.1016/j.csbj.2021.11.036.].

Quantifying Cyanothece growth under DIC limitation.

The photoautotrophic, unicellular N2-fixer, Cyanothece, is a model organism that has been widely used to study photosynthesis regulation, the structure of photosystems, and the temporal segregation of carbon (C) and nitrogen (N) fixation in light and dark phases of the diel cycle. Here, we present a simple quantitative model and experimental data that together, suggest external dissolved inorganic carbon (DIC) concentration as a major limiting factor for Cyanothece growth, due to its high C-storage requirement. Using experimental data from a parallel laboratory study as a basis, we show that after the onset of the light period, DIC was rapidly consumed by photosynthesis, leading to a sharp drop in the rate of photosynthesis and C accumulation. In N2-fixing cultures, high rates of photosynthesis in the morning enabled rapid conversion of DIC to intracellular C storage, hastening DIC consumption to levels that limited further uptake. The N2-fixing condition allows only a small fraction of fixed C for cellular growth since a large fraction was reserved in storage to fuel night-time N2 fixation. Our model provides a framework for resolving DIC limitation in aquatic ecosystem simulations, where DIC as a growth-limiting factor has rarely been considered, and importantly emphasizes the effect of intracellular C allocation on growth rate that varies depending on the growth environment.

Influence of infrastructure material composition and microtopography on marine biofilm growth and photobiology.

The impact of concrete composition and roughness on the formation of microalgal biofilms and their photobiology were studied on marine infrastructures presenting four different compositions combined with two degrees of roughness (rough and smooth). The structures were first inoculated with a natural microphytobenthic biofilm and immersed in sterilised seawater with a controlled photoperiod for six days. Photosynthetic activity was assessed with an imaging PAM-(Pulse Amplitude Modulated) fluorometer and microtopography was monitored in parallel with a 3-D camera. The results indicated that roughness had an impact on the biofilm biomass, its physiological status and its photosynthetic efficiency and capacity. The assessment of surface roughness indicated that negative reliefs were preferably colonised by MPB (microphytobenthic) cells with better photosynthetic performances. Moreover, MPB biofilms showed better photoacclimation in these microhabitats than on the positive and smooth reliefs. This study confirms the importance of microhabitat for biofilm formation and their photobiology.

Electron & Biomass Dynamics of Cyanothece Under Interacting Nitrogen & Carbon Limitations.

Marine diazotrophs are a diverse group with key roles in biogeochemical fluxes linked to primary productivity. The unicellular, diazotrophic cyanobacterium Cyanothece is widely found in coastal, subtropical oceans. We analyze the consequences of diazotrophy on growth efficiency, compared to NO3 --supported growth in Cyanothece, to understand how cells cope with N2-fixation when they also have to face carbon limitation, which may transiently affect populations in coastal environments or during blooms of phytoplankton communities. When grown in obligate diazotrophy, cells face the double burden of a more ATP-demanding N-acquisition mode and additional metabolic losses imposed by the transient storage of reducing potential as carbohydrate, compared to a hypothetical N2 assimilation directly driven by photosynthetic electron transport. Further, this energetic burden imposed by N2-fixation could not be alleviated, despite the high irradiance level within the cultures, because photosynthesis was limited by the availability of dissolved inorganic carbon (DIC), and possibly by a constrained capacity for carbon storage. DIC limitation exacerbates the costs on growth imposed by nitrogen fixation. Therefore, the competitive efficiency of diazotrophs could be hindered in areas with insufficient renewal of dissolved gases and/or with intense phytoplankton biomass that both decrease available light energy and draw the DIC level down.

Coupling high frequency monitoring and bioassay experiments to investigate a harmful algal bloom in the Bay of Seine (French-English Channel).

Coastal ecosystems are increasingly threatened by eutrophication and dystrophy. In this context, the full pattern of a bloom dominated by the dinoflagellate, Lepidodinium chlorophorum, was investigated by a high frequency monitoring buoy equipped with sensors allowing nutrients and photosynthesis measurements. An increase of the N/P ratio affected phytoplankton physiology leading to bloom collapse with a slight oxygen depletion. In parallel, enrichment experiments were performed on the natural bloom population. After 5 days of incubation the community structure, using flow cytometry and several physiological parameters were analysed. The data reveal a potential N and P co-limitation and a decoupling between primary production and productivity in fully enriched conditions. Under unbalanced N/P inputs, high level of alkaline phosphatase activity and transparent exopolymeric particle production, which favour phytoplankton sedimentation, were observed. Nutrient inputs and their stoichiometry control phytoplankton growth, the community structure, physiological regulations, the fate of the bloom and consequences.

Temporal Patterns and Intra- and Inter-Cellular Variability in Carbon and Nitrogen Assimilation by the Unicellular Cyanobacterium Cyanothece sp. ATCC 51142.

Unicellular nitrogen fixing cyanobacteria (UCYN) are abundant members of phytoplankton communities in a wide range of marine environments, including those with rapidly changing nitrogen (N) concentrations. We hypothesized that differences in N availability (N2 vs. combined N) would cause UCYN to shift strategies of intracellular N and C allocation. We used transmission electron microscopy and nanoscale secondary ion mass spectrometry imaging to track assimilation and intracellular allocation of 13C-labeled CO2 and 15N-labeled N2 or NO3 at different periods across a diel cycle in Cyanothece sp. ATCC 51142. We present new ideas on interpreting these imaging data, including the influences of pre-incubation cellular C and N contents and turnover rates of inclusion bodies. Within cultures growing diazotrophically, distinct subpopulations were detected that fixed N2 at night or in the morning. Additional significant within-population heterogeneity was likely caused by differences in the relative amounts of N assimilated into cyanophycin from sources external and internal to the cells. Whether growing on N2 or NO3, cells prioritized cyanophycin synthesis when N assimilation rates were highest. N assimilation in cells growing on NO3 switched from cyanophycin synthesis to protein synthesis, suggesting that once a cyanophycin quota is met, it is bypassed in favor of protein synthesis. Growth on NO3 also revealed that at night, there is a very low level of CO2 assimilation into polysaccharides simultaneous with their catabolism for protein synthesis. This study revealed multiple, detailed mechanisms underlying C and N management in Cyanothece that facilitate its success in dynamic aquatic environments.

Evidence for better microphytobenthos dynamics in mixed sand/mud zones than in pure sand or mud intertidal flats (Seine estuary, Normandy, France).

Understanding the dynamics of microphytobenthos biomass and photosynthetic performances in intertidal ecosystems will help advance our understanding of how trophic networks function in order to optimize ecological management and restoration projects. The main objective of this study was to investigate microphytobenthic biomass and photosynthetic performances as a function of the sedimentary and environmental variabilities in the range of intertidal habitats in the downstream Seine estuary (Normandy, France). Our results highlight higher biomass associated with more stratified biofilms and better photosynthetic performances in areas characterized by a sand/mud mixture (40-60% of mud) compared to pure sand or pure mud environments. This type of sediment probably offers an efficient trade-off between the favorable characteristics of the two types of sediments (sand and mud) with respect to light penetration and nutrient accessibility. Moreover, the large quantities of exopolysaccharides produced in sand/mud mixtures emphasizes the functional role played by microphytobenthos in promoting sediment stability against erosion. This allows us to show that despite the strong increase in sand content of the downstream Seine estuary, intertidal flats are still productive since microphytobenthic biomass, photosynthetic performances and exopolysaccharides secretion are highest in sand-mud mixtures. This study also underlines the impact of ecosystem modifications due to human disturbance and climate change on the dynamics of key primary producers in estuaries.

Differential Influence of Life Cycle on Growth and Toxin Production of three Pseudo-nitzschia Species (Bacillariophyceae).

We used a multistrain approach to study the intra- and interspecific variability of the growth rates of three Pseudo-nitzschia species - P. australis, P. fraudulenta, and P. pungens - and of their domoic acid (DA) production. We carried out mating and batch experiments to investigate the respective effects of strain age and cell size, and thus the influence of their life cycle on the physiology of these species. The cell size - life cycle relationship was characteristic of each species. The influence of age and cell size on the intraspecific variability of growth rates suggests that these characteristics should be considered cautiously for the strains used in physiological studies on Pseudo-nitzschia species. The results from all three species do not support the hypothesis of a decrease in DA production with time since isolation from natural populations. In P. australis, the cellular DA content was rather a function of cell size. More particularly, cells at the gametangia stage of their life cycle contained up to six times more DA than smaller or larger cells incapable of sexual reproduction. These findings reveal a link between P. australis life cycle and cell toxicity. This suggest that life cycle dynamics in Pseudo-nitzschia natural populations may influence bloom toxicity.

Electron requirements for carbon incorporation along a diel light cycle in three marine diatom species.

Diatoms account for about 40% of primary production in highly productive ecosystems. The development of a new generation of fluorometers has made it possible to improve estimation of the electron transport rate from photosystem II, which, when coupled with the carbon incorporation rate enables estimation of the electrons required for carbon fixation. The aim of this study was to investigate the daily dynamics of these electron requirements as a function of the diel light cycle in three relevant diatom species and to apprehend if the method of estimating the electron transport rate can lead to different pictures of the dynamics. The results confirmed the species-dependent capacity for photoacclimation under increasing light levels. Despite daily variations in the photosynthetic parameters, the results of this study underline the low daily variability of the electron requirements estimated using functional absorption of the photosystem II compared to an estimation based on a specific absorption cross section of chlorophyll a. The stability of the electron requirements throughout the day would suggest it is potentially possible to estimate high-frequency primary production by using autonomous variable fluorescence measurements from ships-of-opportunity or moorings, without taking potential daily variation in this parameter into consideration, but this result has to be confirmed on natural phytoplankton assemblages. The results obtained in this study confirm the low electron requirements of diatoms to perform photosynthesis, and suggest a potential additional source of energy for carbon fixation, as recently described in the literature for this class.

Nutrient ratios influence variability in Pseudo-nitzschia species diversity and particulate domoic acid production in the Bay of Seine (France).

The population dynamics of different Pseudo-nitzschia species, along with particulate domoic acid (pDA) concentrations, were studied from May 2012 to December 2013 in the Bay of Seine (English Channel, Normandy). While Pseudo-nitzschia spp. blooms occurred during the two years of study, Pseudo-nitzschia species diversity and particulate domoic acid concentrations varied greatly. In 2012, three different species were identified during the spring bloom (P. australis, P. pungens and P. fraudulenta) with high pDA concentrations (∼1400ngl-1) resulting in shellfish harvesting closures. In contrast, the 2013 spring was characterised by a P. delicatissima bloom without any toxic event. Above all, the results show that high pDA concentrations coincided with the presence of P. australis and with potential silicate limitation (Si:N<1), while nitrate concentrations were still replete. The contrasting environmental conditions between 2012 and 2013 highlight different environmental controls that might favour the development of either P. delicatissima or P. australis. This study points to the key role of Pseudo-nitzschia diversity and cellular toxicity in the control of particulate domoic acid variations and highlights the fact that diversity and toxicity are influenced by nutrients, especially nutrient ratios.

Dynamics of phytoplankton productivity and exopolysaccharides (EPS and TEP) pools in the Seine Estuary (France, Normandy) over tidal cycles and over two contrasting seasons.

Exopolysaccharides (EPS) play an important role in the carbon flux and may be directly linked to phytoplankton and microphytobenthos production, most notably in estuarine systems. However the temporal and spatial dynamics of estuarine EPS are still not well understood, nor how primary productivity triggers this variability at these different scales. The aim of this study was to investigate the primary productivity of phytoplankton and EPS dynamics in the Seine estuary over a tidal cycle in three different haline zones over two contrasted seasons. The other objectives was to investigate the origin of pools of soluble carbohydrates (S-EPS) and transparent exopolymeric particles (TEP) in phytoplankton, microphytobenthos or other compartments. High frequency measurements of productivity were made in winter and summer 2015. Physical and chemical parameters, biomass and EPS were measured at hourly intervals in sub-surface waters and just above the water sediment-interface. Our results confirmed that high frequency measurements improve the accuracy of primary productivity estimations and associated carbon fluxes in estuaries. The photosynthetic parameters were shown to be strongly controlled by salinity and by the concentrations of suspended particle matter at the smallest temporal and at spatial scales. At these scales, our results showed an inverse relationship between EPS concentrations and biomass and productivity, and a positive relationship with sediment resuspension. Additionally, the distribution of EPS appears to be linked to hydrodynamics with the tide at daily scale and with the winter at seasonal scale. At spatial scale, the maximum turbidity zone played an important role in the distribution of TEP. Our results suggest that, in the Seine estuary, between 9% and 33% of the S-EPS pool in the water column can be attributed to phytoplankton excretion, while only 0.4%-1.6% (up to 6.14% in exceptional conditions) originates from the microphytobenthos compartments. Most EPS was attributed to remobilization of detrital carbon pools in the maximum turbidity zone and in the sediment or allochthonous origin.

Physiological adjustments and transcriptome reprogramming are involved in the acclimation to salinity gradients in diatoms.

Salinity regimes in estuaries and coastal areas vary with river discharge patterns, seawater evaporation, the morphology of the coastal waterways and the dynamics of marine water mixing. Therefore, microalgae have to respond to salinity variations at time scales ranging from daily to annual cycles. Microalgae may also have to adapt to physical alterations that induce the loss of connectivity between habitats and the enclosure of bodies of water. Here, we integrated physiological assays and measurements of morphological plasticity with a functional genomics approach to examine the regulatory changes that occur during the acclimation to salinity in the estuarine diatom Thalassiosira weissflogii. We found that cells exposed to different salinity regimes for a short or long period presented adjustments in their carbon fractions, silicon pools, pigment concentrations and/or photosynthetic parameters. Salinity-induced alterations in frustule symmetry were observed only in the long-term (LT) cultures. Whole transcriptome analyses revealed a down-regulation of nuclear and plastid encoded genes during the LT response and identified only a few regulated genes that were in common between the ST and LT responses. We propose that in diatoms, one strategy for acclimating to salinity gradients and maintaining optimal cellular fitness could be a reduction in the cost of transcription.

Dynamics of δ(15)N isotopic signatures of different intertidal macroalgal species: Assessment of bioindicators of N sources in coastal areas.

δ(15)N of annual (Ulva sp., Porphyra sp.) and perennial intertidal seaweed species (Chondrus crispus, Fucus sp.) collected on 17 sampling points along the French coast of the English Channel in 2012 and 2013 were assessed on their suitability as bioindicators of N pollution in coastal areas. A sine function applied for δ(15)N time series data showed for all the species the same seasonal trend with lowest δ(15)N values in April and highest in summer but with no significant interspecific differences of amplitude (α) and phase angle (ϕ). This model provides a useful tool for monitoring the inter-annual changes of N pollution. An interspecific variability of δ(15)N values was observed, probably due to their tolerance to emersion. An in vitro study for comparing the kinetic acquisition of the isotopic signal and N uptake mechanisms of each species underlined the influence of algal physiology on the δ(15)N interspecific variability.

Photosystem-II shutdown evolved with Nitrogen fixation in the unicellular diazotroph Crocosphaera watsonii.

Protection of nitrogenase from oxygen in unicellular Cyanobacteria is obtained by temporal separation of photosynthesis and diazotrophy through transcriptional and translational regulations of nitrogenase. But diazotrophs can face environmental situations in which N2 fixation occurs significantly in the light, and we believe that another control operates to make it possible. The night-time shutdown of PSII activity is a peculiar behaviour that discriminates Crocosphaera watsonii WH8501 from any other phototroph, whether prokaryote or eukaryote. This phenomenon is not only due to the plastoquinone pool redox status, and suggests that the sentinel D1 protein, expressed in periods of nitrogen fixation, is inactive. Results demonstrate a tight constraint of oxygen evolution in C. watsonii as additional protection of nitrogenase activity and suggest a possible recycling of cellular components.