Effects of turbulence on diatoms of the genus Pseudo-nitzschia spp. and associated bacteria.

Turbulence is one of the least investigated environmental factors impacting the ecophysiology of phytoplankton, both at the community and individual species level. Here, we investigated, for the first time, the effect of a turbulence gradient (Reynolds number, from Reλ = 0 to Reλ = 360) on two species of the marine diatom Pseudo-nitzschia and their associated bacterial communities under laboratory conditions. Cell abundance, domoic acid (DA) production, chain formation, and Chl a content of P. fraudulenta and P. multiseries were higher for intermediate turbulence (Reλ = 160 or 240). DA was detectable only in P. multiseries samples. These observations were supported by transcriptomic analyses results, which suggested the turbulence related induction of the expression of the DA production locus, with a linkage to an increased photosynthetic activity of the total metatranscriptome. This study also highlighted a higher richness of the bacterial community associated with the nontoxic strain of P. fraudulenta in comparison to the toxic strain of P. multiseries. Bacillus was an important genus in P. multiseries cultures (relative abundance 15.5%) and its highest abundances coincided with the highest DA levels. However, associated bacterial communities of both Pseudo-nitzschia species did not show clear patterns relative to turbulence intensity.

Molecular phylogeny of the spiny-surfaced species of the dinoflagellate Prorocentrum with the description of P. thermophilum sp. nov. and P. criophilum sp. nov. (Prorocentrales, Dinophyceae).

Spiny-surfaced species of Prorocentrum form harmful algal blooms, and its taxonomic identity is obscure due to the size and shape variability. Molecular phylogenies reveal two major clades: one for P. cordatum with sequences mainly retrieved as P. minimum, and the other for P. shikokuense with sequences also retrieved as P. dentatum and P. donghaiense. Several closely related clades still need to be characterized. Here, we provide nuclear SSU and LSU rRNA genes, and nuclear ITS region (ITS1-5.8S gene-ITS2) sequences of the strain CCMP3122 isolated from Florida (initially named P. donghaiense) and strains Prorocentrum sp. RCC6871-2 from the Ross Sea, Antarctica. We describe Prorocentrum thermophilum sp. nov. based on the strain CCMP3122, a species also distributed in the open waters of the Gulf of Mexico, New Zealand, and the Arabian Gulf; and Prorocentrum criophilum sp. nov. based on the strain RCC6872, which is distributed in the Antarctic Ocean and Arctic Sea. Prorocentrum thermophilum is roundish (~14 µm long, ~12 µm wide), with an inconspicuous anterior spine-like prolongation under light microscopy, valves with tiny, short knobs (5-7 per µm2 ), and several (<7) large trichocyst pores (~0.3 µm) in the right valve, as well as smaller pores (~0.15 µm). Prorocentrum criophilum is round in valve view (~11 µm long, 10 µm wide) and asymmetrically roundish in lateral view, the periflagellar area was not discernible under light microscopy, valves with very tiny, short knobs (6-10 per µm2 ), and at least 12 large pores in the right valve. Other potentially undescribed species of spiny-surfaced Prorocentrum are discussed.

Larval growth of the polychaete Arenicola marina under different temperature and food conditions: consequences on bioenergetic models.

Arenicola marina, a marine benthic polychaete, is widespread on sandy beaches in Europe and considered as an ecosystem engineer despite commonly used as bait by fishermen. Data regarding the bioenergetics of the lugworm larval stages are still incomplete. Trochophore is initially lecithotroph and then becomes planktotroph while growing as metatrochophore on subtidal area, a quite stable daily temperature environment compared with the foreshore, where juveniles and adult live, with daily temperature fluctuating up to 15°C. These discrepancies in temperature ranges may influence the temperature corrections (TCs) that control metabolic rates during the life cycle of A. marina. We carried out laboratory experiments in microcosms by inducing artificial spawning of lugworms, and then undertaken in vitro fertilization to obtain embryos and, finally, to follow, the larval development up to 10 segments with chaetae for 50 days under three temperature conditions (13°C, 15°C and 17°C) and two food conditions ('fed' and 'non-fed'). The first feeding ('birth') of A. marina larvae was deciphered anatomically for a size between 450 and 500 µm and described at 17 days post-fertilization for larvae reared at 15°C and 17°C. Using a biphasic model with a von Bertalanffy growth before 'birth' and an exponential growth after 'birth', among the three temperature treatments, the 15°C condition exhibited the best larval performance. TC based on embryonic and larval metabolic rates gave an Arrhenius temperature of ~6661 K and a higher boundary temperature tolerance range of ~294.5 K. Both temperature values differ from those calculated from TC based mostly on juvenile and adult metabolic rates. We claim to use two sets of Arrhenius temperatures according to the life history stages of A. marina while using Dynamic Energy Budget model. This model was developed initially in order to manage the conservation of the lugworm species.

Do transparent exopolymeric particles (TEP) affect the toxicity of nanoplastics on Chaetoceros neogracile?

The potential presence of nanoplastics (NP) in aquatic environments represents a growing concern regarding their possible effects on aquatic organisms. The objective of this study was to assess the impact of polystyrene (PS) amino-modified particles (50  nm PSNH2) on the cellular and metabolic responses of the diatom Chaetoceros neogracile cultures at two essential phases of the growth cycle, i.e. exponential (division) and stationary (storage) phases. Both cultures were exposed for 4 days to low (0.05 µg mL-1) and high (5 µg mL-1) concentrations of PS-NH2. Exposure to NP impaired more drastically the major cellular and physiological parameters during exponential phase than during the stationary phase. Only an increase in ROS production was observed at both culture phases following NP exposures. In exponential phase cultures, large decreases in chlorophyll content, esterase activity, cellular growth and photosynthetic efficiency were recorded upon NP exposure, which could have consequences on the diatoms life cycle and higher food-web levels. The observed differential responses to NP exposure according to culture phase could reflect i) the higher concentration of Transparent Exopolymer Particles (TEP) at stationary phase leading to NP aggregation and thus, probably minimizing NP effects, and/or ii) the fact that dividing cells during exponential phase may be intrinsically more sensitive to stress. This work evidenced the importance of algae physiological state for assessing the NP impacts with interactions between NP and TEP being one key factor affecting the fate of NP in algal media and their impact to algal' cells.

Physico-chemical characterization and in vitro inflammatory and oxidative potency of atmospheric particles collected in Dakar city's (Senegal).

Exposure to atmospheric pollutants has been recognized as a major risk factor of respiratory and cardiovascular diseases. Fine particles (PM2.5) and a coarser fraction (PM>2.5) sampled at an urban site in Dakar (HLM), characterized by high road traffic emissions, were compared with particles sampled at a rural area, Toubab Dialaw located about 40 km from Dakar. The physicochemical characteristics of samples revealed that PMs differ for their physical (surface area) and chemical properties (in terms of CHN, metals, ions, paraffins, VOCs and PAHs) that were 65-75% higher in urban samples. Moreover the fine PMs contain higher amounts of anthropogenic related pollutants than the PM>2.5 one. These differences are sustained by the ratios reported for the analysed PAHs which suggest as predominant primary emission sources vehicle exhausts at urban site and biomass combustion at the rural site. The inflammatory response and the oxidative damages were evaluated in BEAS-2B cells by the quantification of 4 selected inflammatory cytokines (TNF-α, IL-1ß, IL-6, IL-8) and of total carbonylated proteins and the oxidative DNA adduct 8-OHdG after 8 or 24 h exposure. In accordance with the different sources and different physical and chemical properties, the inflammatory response and the oxidative damages were found higher in bronchial cells exposed to urban PMs. These data confirm the importance, also for West African countries, to evaluate the correlation between PM physico-chemical properties and potential biological impacts.

Evolution of the scattering properties of phytoplankton cells from flow cytometry measurements.

After the exponential growth phase, variability in the scattering efficiency of phytoplankton cells over their complete life cycle is not well characterised. Bulk measurements are impacted by senescent cells and detritrus. Thus the analysis of the evolution of the optical properties thanks to their morphological and/or intra-cellular variations remains poorly studied. Using the Cytosense flow cytometer (CytoBuoy b.v., NL), the temporal course of the forward and sideward efficiencies of two phytoplankton species (Thalassiosira pseudonana and Chlamydomonas concordia) were analyzed during a complete life-cycle. These two species differ considerably from a morphological point of view. Over the whole experiment, the forward and sideward efficiencies of Thalassiosira pseudonana were, on average, respectively 2.2 and 1.6 times higher than the efficiencies of Chlamydomonas concordia. Large intra-species variability of the efficiencies were observed over the life cycle of the considered species. It highlights the importance of considering the optical properties of phytoplankton cells as a function of the population growth stage of the considered species. Furthermore, flow cytometry measurements were combined with radiative transfer simulations and biogeochemical and optical measurements. Results showed that the real refractive index of the chloroplast is a key parameter driving the sideward signal and that a simplistic two-layered model (cytoplasm-chloroplast) seems particularly appropriate to represent the phytoplankton cells.

Characterization of manganese-bearing particles in the vicinities of a manganese alloy plant.

Numerous studies have associated air manganese (Mn) exposure with negative health effects, primarily neurotoxic disorders. Despite there is not a specific European regulation, institutions such as the World Health Organization (WHO) have proposed an annual average guideline value of 150 ng/m3. Bioaccessibility and toxicity mechanisms of Mn remain unclear, however it is generally agreed that adverse health effects are strongly linked to particle size and morphology, chemical composition and oxidation state. This study aims to deepen the understanding of the physico-chemical characteristics of PM10 and deposition samples collected in an urban area in the proximities of a ferromanganese alloy plant. Total Mn content was determined by ICP-MS after a microwave-assisted acid digestion. The size, morphology and chemical composition of individual particles were studied by SEM-EDX. XRD was used to identify the major crystalline phases. Most of the particles observed by SEM-EDX contain Mn. 60% of Mn-PM10 particles were spheres of small size and were attributed to condensation processes at the smelting unit. Mn-bearing particles present in deposition were characterized by irregular shapes and bigger sizes, most of them consisting of SiMn slags and Mn ores and alloys, and attributed to diffuse emissions from raw material and product handling and processing. Due to the differences in the characteristics of Mn-bearing particles found in the different matrices, further studies on the potential toxicity and health effects of these particles should be done, especially in relation with the small and spherical particles present in PM10, which are expected to be more problematic.

Fine and ultrafine atmospheric particulate matter at a multi-influenced urban site: Physicochemical characterization, mutagenicity and cytotoxicity.

Particulate Matter (PM) air pollution is one of the major concerns for environment and health. Understanding the heterogeneity and complexity of fine and ultrafine PM is a fundamental issue notably for the assessment of PM toxicological effects. The aim of this study was to evaluate mutagenicity and cytotoxicity of a multi-influenced urban site PM, with or without the ultrafine fraction. For this purpose, PM2.5-0.3 (PM with aerodynamic diameter ranging from 0.3 to 2.5 µm) and PM2.5 were collected in Dunkerque, a French coastal industrial city and were extensively characterized for their physico-chemical properties, including inorganic and organic species. In order to identify the possible sources of atmospheric pollution, specific criteria like Carbon Preference Index (CPI) and PAH characteristic ratios were investigated. Mutagenicity assays using Ames test with TA98, TA102 and YG1041 Salmonella strains with or without S9 activation were performed on native PM sample and PM organic extracts and water-soluble fractions. BEAS-2B cell viability and cell proliferation were evaluated measuring lactate dehydrogenase release and mitochondrial dehydrogenase activity after exposure to PM and their extracts. Several contributing sources were identified in PM: soil resuspension, marine emissions including sea-salt or shipping, road traffic and industrial activities, mainly related to steelmaking or petro-chemistry. Mutagenicity of PM was evidenced, especially for PM2.5, including ultrafine fraction, in relation to PAHs content and possibly nitro-aromatics compounds. PM induced cytotoxic effects at relatively high doses, while alteration of proliferation with low PM doses could be related to underlying mechanisms such as genotoxicity.

Characterisation and seasonal variations of particles in the atmosphere of rural, urban and industrial areas: Organic compounds.

Atmospheric aerosol samples (PM2.5-0.3, i.e., atmospheric particles ranging from 0.3 to 2.5µm) were collected during two periods: spring-summer 2008 and autumn-winter 2008-2009, using high volume samplers equipped with cascade impactors. Two sites located in the Northern France were compared in this study: a highly industrialised city (Dunkirk) and a rural site (Rubrouck). Physicochemical analysis of particulate matter (PM) was undertaken to propose parameters that could be used to distinguish the various sources and to exhibit seasonal variations but also to provide knowledge of chemical element composition for the interpretation of future toxicological studies. The study showed that PM2.5-0.3 concentration in the atmosphere of the rural area remains stable along the year and was significantly lower than in the urban or industrial ones, for which concentrations increase during winter. High concentrations of polycyclic aromatic hydrocarbons (PAHs), dioxins, furans and dioxin like polychlorinated biphenyls (DL-PCBs), generated by industrial activities, traffic and municipal wastes incineration were detected in the samples. Specific criteria like Carbon Preference Index (CPI) and Combustion PAHs/Total PAHs ratio (CPAHs/TPAHs) were used to identify the possible sources of atmospheric pollution. They revealed that paraffins are mainly emitted by biogenic sources in spring-summer whereas as in the case of PAHs, they have numerous anthropogenic emission sources in autumn-winter (mainly from traffic and domestic heating).

Determination of backscattering cross section of individual particles from cytometric measurements: a new methodology.

A methodology is developed to derive the backscattering cross section of individual particles as measured with the CytoSense (CytoBuoy b.v., NL). This in situ flow cytometer detects light scatter in forward and sideward directions and fluorescence in various spectral bands for a wide range of particles. First, the weighting functions are determined for the forward and sideward detectors to take into account their instrumental response as a function of the scattering angle. The CytoSense values are converted into forward and sideward scattering cross sections. The CytoSense estimates of uniform polystyrene microspheres from 1 to 90 µm are compared with Mie computations. The mean absolute relative differences ΔE are around 33.7% and 23.9% for forward and sideward scattering, respectively. Then, a theoretical relationship is developed to convert sideward scattering into backscattering cross section, from a synthetic database of 495,900 simulations including homogeneous and multi-layered spheres. The relationship follows a power law with a coefficient of determination of 0.95. To test the methodology, a laboratory experiment is carried out on a suspension of silica beads to compare backscattering cross section as measured by the WET Labs ECO-BB9 and derived from CytoSense. Relative differences are between 35% and 60%. They are of the same order of magnitude as the instrumental variability. Differences can be partly explained by the fact that the two instruments do not measure exactly the same parameter: the cross section of individual particles for the CytoSense and the bulk cross section for the ECO-BB9.

Harbour porpoises (Phocoena phocoena) stranded along the southern North Sea: an assessment through metallic contamination.

Throughout the last few years, the southern North Sea has witnessed an increase in the number of stranded marine mammals, particularly the harbour porpoise (Phocoena phocoena). This species is subject to several potential threats such as exposure to contaminants, changes in food supply, marine traffic and fishery by-catch. The aims of this study were to investigate potential associations between contaminants and health status and to analyze spatial and temporal trends of metal concentrations in harbour porpoises. Selected trace elements (As, Cd, Cr, Cu, Hg, Mn, Se, V and Zn) were measured in kidneys and livers of 105 harbour porpoises stranded along the southern North Sea (French and Belgian coasts from 2006 to 2013) and 27 stranded along the Bay of Biscay (French coast from 2009 to 2012). Porpoises that died from infectious disease displayed significant higher hepatic concentrations of Cd, Hg, Se and Zn compared to healthy porpoises that died from physical trauma. Adult porpoises displayed significant higher concentrations of Cd, Cr, Hg, Se and V in livers compared to juveniles. No spatial or temporal trends in metal concentrations were detected in our study. The results of the present study suggested that chemical contamination may represent one of many threats encountered by harbour porpoises, but it cannot explain alone the increase in the number of stranded individuals.

A scanning electron microscopy study of Argulus vittatus (Rafinesque-Schmaltz, 1814) (Crustacea: Branchiura) from Algerian coast.

A study of the Algerian Branchiura, Argulus vittatus (Rafinesque-Schmaltz, 1814) was conducted using the scanning electron microscopy (SEM). New morphological features are reported for the first time (mouth con, first maxilla, second maxilla, structures and ornamentation of thoracic segments, structure of semen papillae, etc.). The morphology of small and large female specimens was compared. Two new hosts, Pagellus erythrinus L. and Sparus aurata L., are reported for this species. Until now, six host species were reported for A. vittatus, and stenoxenic specificity for Sparid fishes was observed for Algerian specimens. The biogeographical distribution of this species seems to be restricted to the eastern coast of Algeria. Additional information about the host specificity, ecology, and life cycle of this parasitic species were given.

Chemical profile identification of fugitive and confined particle emissions from an integrated iron and steelmaking plant.

The aim of this study is to obtain the characteristic inorganic chemical profile of important particle sources identified in the integrated iron and steel process: sintering, blast furnace, steelmaking and desulfurization slag processing. A complete chemical and physical characterization program was developed: particle size distribution, chemical analysis, XRD, SEM-EDX and TGA/DTA. The sample collected from the sinter stack showed high levels of K and Cl(-), followed by Fe, NH4(+), Ca, Na and Pb. The profile of the dust samples taken from the sinter cake discharge zone was quite different, showing higher amounts of Fe, Ca and Al, and lower amounts of K, Cl(-), Na and Pb. Dust samples collected from the blast furnace (BF) and steelmaking cast house may be distinguished from each other based on the higher levels of Fe (hematite and magnetite) and lower levels of Ca, Zn and C (graphite) found in BF dust. High levels of Ca and Fe were found in samples taken from the desulfurization slag processing area. Such information can be useful for source apportionment studies at receptor sites that could be influenced by iron and steelmaking plant emissions.

An optimised protocol to prepare Phaeocystis globosa morphotypes for scanning electron microscopy observation.

A preparation protocol for scanning electron microscopy (SEM) was adapted and tested to the observation of Phaeocystis globosa morphotypes. For extra colonial cells, critical point drying (CPD) gave satisfactory results while for intra colonial cells, preservation with Lugol's iodine and/or glutaraldehyde followed by air-drying appeared as the most suitable method.

Impact of the Phaeocystis globosa spring bloom on the intertidal benthic compartment in the eastern English Channel: a synthesis.

From 1999 to 2005, studies carried out in the frame of regional and national French programs aimed to determine whether the Phaeocystis globosa bloom affected the intertidal benthic communities of the French coast of the eastern English Channel in terms of composition and/or functioning. Study sites were chosen to cover most of the typical shore types encountered on this coast (a rocky shore, an exposed sandy beach and a small estuary). Both the presence of active Phaeocystis cells and their degradation product (foam) did have a significant impact on the studied shores. The primary production and growth rates of the kelp Saccharina latissima decreased during the bloom because of a shortage of light and nutrient for the macroalgae. On sandy sediments, the benthic metabolism (community respiration and community primary production), as well as the nitrification rate, were enhanced during foam deposits, in relation with the presence of bacteria and active pelagic cells within the decaying colonies. In estuarine sediments, the most impressive impact was the formation of a crust at the sediment surface due to drying foam. This led to anoxic conditions in the surface sediment and resulted in a high mortality among the benthic community. Some organisms also tended to migrate upward and were then directly accessible to the higher trophic level represented by birds. Phaeocystis then created a shortcut in the estuarine trophic network. Most of these modifications lasted shortly and all the systems considered came back to their regular properties and activities a few weeks after the end of the bloom, except for the most impacted estuarine area.