Functional and structural responses of plankton communities toward consecutive experimental heatwaves in Mediterranean coastal waters.

The frequency of marine heatwaves (HWs) is projected to increase in the Mediterranean Sea over the next decades. An in situ mesocosm experiment was performed in a Mediterranean lagoon for 33 days. Three mesocosms were used as controls following the natural temperature of the lagoon. In three others, two HWs of + 5 °C compared to the controls were applied from experimental day (d) 1 to d5 (HW1) and from d11 to d15 (HW2). High-frequency data of oxygen, chlorophyll-a (chl-a), temperature, salinity and light from sensors immersed in all mesocosms were used to calculate gross primary production (GPP), respiration (R) and phytoplankton growth (µ) and loss (L) rates. Nutrients and phytoplankton community structure from pigments were also analyzed. HW1 significantly increased GPP, R, chl-a, µ and L by 7 to 38%. HW2 shifted the system toward heterotrophy by only enhancing R. Thus, the effects of the first HW resulted in the attenuation of those of a second HW on phytoplankton processes, but not on community respiration, which was strongly regulated by temperature. In addition, natural phytoplankton succession from diatoms to haptophytes was altered by both HWs as cyanobacteria and chlorophytes were favored at the expense of haptophytes. These results indicate that HWs have pronounced effects on Mediterranean plankton communities.

Metabolic responses of plankton to warming during different productive seasons in coastal Mediterranean waters revealed by in situ mesocosm experiments.

The response of coastal lagoon plankton communities to warming was studied during two in situ mesocosm experiments in spring and fall of 2018 in the Mediterranean. Phytoplankton biomass, gross primary production (GPP), community respiration (R), phytoplankton growth (µ), and loss (l) rates were estimated using high-frequency chlorophyll-a fluorescence and dissolved oxygen sensors, and daily sampling was used to evaluate the nutrient status and phytoplankton pigment functional groups. Warming strongly depressed the dominant phytoplankton functional groups, mainly the prymnesiophytes, diatoms (spring), and green flagellates (fall). It favored minor groups such as the dinoflagellates (spring) and diatoms (fall). In spring, warming depressed GPP and R by half; however, µ (+ 18%) and l (+ 37%) were enhanced. In contrast, both GPP and µ were enhanced by 21% and 28%, respectively, in fall, and no effects were observed for R and l. Warming strongly decreased phytoplankton biomass and oxygen production in spring, and enhanced them, to a lesser extent, in fall. This led to an overall loss of production over both seasons. This study improves understanding of the contrasting effects of warming during two productive seasons, which depend on plankton community composition and interactions between components and environmental conditions.

Co-occurrence networks reveal the central role of temperature in structuring the plankton community of the Thau Lagoon.

To identify the environmental factors that drive plankton community composition and structure in coastal waters, a shallow northwestern Mediterranean lagoon was monitored from winter to spring in two contrasting years. The campaign was based on high-frequency recordings of hydrological and meteorological parameters and weekly samplings of nutrients and the plankton community. The collected data allowed the construction of correlation networks, which revealed that water temperature was the most important factor governing community composition, structure and succession at different trophic levels, suggesting its ubiquitous food web control. Temperature favoured phytoplanktonic flagellates (Cryptophyceae, Chrysophyceae, and Chlorophyceae) and ciliates during winter and early spring. In contrast, it favoured Bacillariophyceae, dinoflagellates, phytoplankton < 6 µm and aloricate Choreotrichida during spring. The secondary factors were light, which influenced phytoplankton, and wind, which may regulate turbidity and the nutrient supply from land or sediment, thus affecting benthic species such as Nitzschia sp. and Uronema sp. or salinity-tolerant species such as Prorocentrum sp. The central role of temperature in structuring the co-occurrence network suggests that future global warming could deeply modify plankton communities in shallow coastal zones, affecting whole-food web functioning.

Marine Microbial Food Web Networks During Phytoplankton Bloom and Non-bloom Periods: Warming Favors Smaller Organism Interactions and Intensifies Trophic Cascade.

Microbial food web organisms are at the base of the functioning of pelagic ecosystems and support the whole marine food web. They are very reactive to environmental changes and their interactions are modified in response to different productive periods such as phytoplankton bloom and non-bloom as well as contrasted climatic years. To study ecological associations, identify potential interactions between microorganisms and study the structure of the microbial food web in coastal waters, a weekly monitoring was carried out in the Thau Lagoon on the French Mediterranean coast. The monitoring lasted from winter to late spring during two contrasting climatic years, a typical Mediterranean (2015) and a year with an extreme warm winter (2016). Correlation networks comprising 110 groups/taxa/species were constructed to characterize potential possible interactions between the microorganisms during bloom and non-bloom periods. Complex correlation networks during the bloom and dominated by negative intraguild correlations and positive correlations of phytoplankton with bacteria. Such pattern can be interpreted as a dominance of competition and mutualism. In contrast, correlation networks during the non-bloom period were less complex and mostly dominated by tintinnids associations with bacteria mostly referring to potential feeding on bacteria, which suggests a shift of biomass transfer from phytoplankton-dominated food webs during bloom to more bacterioplankton-based food webs during non-bloom. Inter-annual climatic conditions significantly modified the structure of microbial food webs. The warmer year favored relationships among smaller group/taxa/species at the expense of large phytoplankton and ciliates, possibly due to an intensification of the trophic cascade with a potential shift in energy circulation through microbial food web. Our study compares a typical Mediterranean spring with another mimicking the prospected intensification of global warming; if such consideration holds true, the dominance of future coastal marine ecosystems will be shifted from the highly productive herbivorous food web to the less productive microbial food web.

Water temperature drives phytoplankton blooms in coastal waters.

Phytoplankton blooms are an important, widespread phenomenon in open oceans, coastal waters and freshwaters, supporting food webs and essential ecosystem services. Blooms are even more important in exploited coastal waters for maintaining high resource production. However, the environmental factors driving blooms in shallow productive coastal waters are still unclear, making it difficult to assess how environmental fluctuations influence bloom phenology and productivity. To gain insights into bloom phenology, Chl a fluorescence and meteorological and hydrological parameters were monitored at high-frequency (15 min) and nutrient concentrations and phytoplankton abundance and diversity, were monitored weekly in a typical Mediterranean shallow coastal system (Thau Lagoon). This study was carried out from winter to late spring in two successive years with different climatic conditions: 2014/2015 was typical, but the winter of 2015/2016 was the warmest on record. Rising water temperature was the main driver of phytoplankton blooms. However, blooms were sometimes correlated with winds and sometimes correlated with salinity, suggesting nutrients were supplied by water transport via winds, saltier seawater intake, rain and water flow events. This finding indicates the joint role of these factors in determining the success of phytoplankton blooms. Furthermore, interannual variability showed that winter water temperature was higher in 2016 than in 2015, resulting in lower phytoplankton biomass accumulation in the following spring. Moreover, the phytoplankton abundances and diversity also changed: cyanobacteria (< 1 µm), picoeukaryotes (< 1 µm) and nanoeukaryotes (3-6 µm) increased to the detriment of larger phytoplankton such as diatoms. Water temperature is a key factor affecting phytoplankton bloom dynamics in shallow productive coastal waters and could become crucial with future global warming by modifying bloom phenology and changing phytoplankton community structure, in turn affecting the entire food web and ecosystem services.

Simulation Method Linking Dense Microalgal Culture Spectral Properties in the 400-750 nm Range to the Physiology of the Cells.

This work describes a method to model the optical properties over the (400-750 nm) spectral range of a dense microalgal culture using the chemical and physical properties of the algal cells. The method was based on a specific program called AlgaSim coupled with the adding-doubling method: at the individual cell scale, AlgaSim simulates the spectral properties of one model, three-layer spherical algal cell from its size and chemical composition. As a second step, the adding-doubling method makes it possible to retrieve the total transmittance of the algal medium from the optical properties of the individual algal cells. The method was tested by comparing the simulated total transmittance spectra for dense marine microalgal cultures of Isochrysis galbana (small flagellates) and Phaeodactylum tricornutum (diatoms) to spectra measured using an experimental spectrophotometric setup. Our study revealed that the total transmittance spectra simulated for the quasi-spherical cells of Isochrysis galbana were in good agreement with the measured spectra over the whole spectral range. For Phaeodactylum tricornutum, large differences between simulated and measured spectra were observed over the blue part of the transmittance spectra, probably due to non-spherical shape of the algal cells. Prediction of the algal cell density, mean size and pigment composition from the total transmittance spectra measured on algal samples was also investigated using the reversal of the method. Mean cell size was successfully predicted for both species. The cell density was also successfully predicted for spherical Isochrysis galbana, with a relative error below 7%, but not for elongated Phaeodactylum tricornutum with a relative error up to 26%. The pigments total quantity and composition, the carotenoids:chlorophyll ratio in particular, were also successfully predicted for Isochrysis galbana with a relative error below 8%. However, the pigment predictions and measurements for Phaeodactylum tricornutum showed large discrepancies, with a relative error up to 88%. These results give strong support for the development of a promising tool providing rapid and accurate estimations of biomass and physiological status of a dense microalgal culture based on only light transmittance properties.

Heterotrophic Bacteria Show Weak Competition for Nitrogen in Mediterranean Coastal Waters (Thau Lagoon) in Autumn.

The importance of heterotrophic bacteria relative to phytoplankton in the uptake of ammonium and nitrate was studied in Mediterranean coastal waters (Thau Lagoon) during autumn, when the Mediterranean Sea received the greatest allochthonous nutrient loads. Specific inhibitors and size-fractionation methods were used in combination with isotopic (15)N tracers. NO3 (-) and NH4 (+) uptake was dominated by phytoplankton (60 % on average) during the study period, which included a flood event. Despite lower biomass specific NH4 (+) and NO3 uptake rates, free-living heterotrophic bacteria contributed significantly (>30 %) to total microbial NH4 (+) and NO3 (-) uptake rates in low chlorophyll waters. Under these conditions, heterotrophic bacteria may be responsible for more than 50 % of primary production, using very little freshly produced phytoplankton exudates. In low chlorophyll coastal waters as reported during the present 3-month study, the heterotrophic bacteria seemed to depend to a greater extent on allochthonous N and C substrates than on autochthonous substrates derived from phytoplankton.

Bacterial carbon dependence on freshly produced phytoplankton exudates under different nutrient availability and grazing pressure conditions in coastal marine waters.

The effects of grazing pressure and inorganic nutrient availability on the direct carbon transfer from freshly produced phytoplankton exudates to heterotrophic bacteria biomass production were studied in Mediterranean coastal waters. The short-term incorporation of ¹³C (H¹³CO3) in phytoplankton and bacterial lipid biomarkers was measured as well as the total bacterial carbon production (BP), viral lysis and the microbial community structure under three experimental conditions: (1) High inorganic Nutrient and High Grazing (HN + HG), (2) High inorganic Nutrient and Low Grazing (HN + LG) and (3) under natural in situ conditions with Low inorganic Nutrient and High Grazing (LN + HG) during spring. Under phytoplankton bloom conditions (HN + LG), the bacterial use of freshly produced phytoplankton exudates as a source of carbon, estimated from ¹³C enrichment of bacterial lipids, contributed more than half of the total bacterial production. However, under conditions of high grazing pressure on phytoplankton with or without the addition of inorganic nutrients (HN + HG and LN + HG), the ¹³C enrichment of bacterial lipids was low compared with the high total bacterial production. BP therefore seems to depend mainly on freshly produced phytoplankton exudates during the early phase of phytoplankton bloom period. However, BP seems mainly relying on recycled carbon from viral lysis and predators under high grazing pressure.

Short-term responses of unicellular planktonic eukaryotes to increases in temperature and UVB radiation.

BACKGROUND: Small size eukaryotes play a fundamental role in the functioning of coastal ecosystems, however, the way in which these micro-organisms respond to combined effects of water temperature, UVB radiations (UVBR) and nutrient availability is still poorly investigated. RESULTS: We coupled molecular tools (18S rRNA gene sequencing and fingerprinting) with microscope-based identification and counting to experimentally investigate the short-term responses of small eukaryotes (<6 µm; from a coastal Mediterranean lagoon) to a warming treatment (+3°C) and UVB radiation increases (+20%) at two different nutrient levels. Interestingly, the increase in temperature resulted in higher pigmented eukaryotes abundances and in community structure changes clearly illustrated by molecular analyses. For most of the phylogenetic groups, some rearrangements occurred at the OTUs level even when their relative proportion (microscope counting) did not change significantly. Temperature explained almost 20% of the total variance of the small eukaryote community structure (while UVB explained only 8.4%). However, complex cumulative effects were detected. Some antagonistic or non additive effects were detected between temperature and nutrients, especially for Dinophyceae and Cryptophyceae. CONCLUSIONS: This multifactorial experiment highlights the potential impacts, over short time scales, of changing environmental factors on the structure of various functional groups like small primary producers, parasites and saprotrophs which, in response, can modify energy flow in the planktonic food webs.

Impact of ultraviolet-B radiation on planktonic fish larvae: alteration of the osmoregulatory function.

Coastal marine ecosystems are submitted to variations of several abiotic and biotic parameters, some of them related to global change. Among them the ultraviolet-B (UV-B) radiation (UVBR: 280-320 nm) may strongly impact planktonic fish larvae. The consequences of an increase of UVBR on the osmoregulatory function of Dicentrarchus labrax larvae have been investigated in this study. In young larvae of D. labrax, as in other teleosts, osmoregulation depends on tegumentary ion transporting cells, or ionocytes, mainly located on the skin of the trunk and of the yolk sac. As early D. labrax larvae passively drift in the top water column, ionocytes are exposed to solar radiation. The effect of UVBR on larval osmoregulation in seawater was evaluated through nanoosmometric measurements of the blood osmolality after exposure to different UV-B treatments. A loss of osmoregulatory capability occured in larvae after 2 days of low (50 µWcm(-2): 4 h L/20 h D) and medium (80 µWcm(-2): 4 h L/20 h D) UVBR exposure. Compared to control larvae kept in the darkness, a significant increase in blood osmolality, abnormal behavior and high mortalities were detected in larvae exposed to UVBR from 2 days on. At the cellular level, an important decrease in abundance of tegumentary ionocytes and mucous cells was observed after 2 days of exposure to UVBR. In the ionocytes, two major osmoeffectors were immunolocalized, the Na+/K(+)-ATPase and the Na+/K+/2Cl- cotransporter. Compared to controls, the fluorescent immunostaining was lower in UVBR-exposed larvae. We hypothesize that the impaired osmoregulation in UVBR-exposed larvae originates from the lower number of tegumentary ionocytes and mucous cells. This alteration of the osmoregulatory function could negatively impact the survival of young larvae at the surface water exposed to UVBR.