Anastasia Tsiola

The Potential Impact of Saharan Dust and Polluted Aerosols on Microbial Populations in the East Mediterranean Sea, an Overview of a Mesocosm Experimental Approach

Recent estimates of nutrient budgets for the Eastern Mediterranean Sea (EMS) indicate that atmospheric aerosols play a significant role as suppliers of macro- and micro- nutrients to its Low Nutrient Low Chlorophyll water. Here we present the first mesocosm experimental study that examines the overall response of the oligotrophic EMS surface mixed layer (Cretan Sea, May 2012) to two different types of natural aerosol additions, “pure” Saharan dust (SD, 1.6 mg l-1) and mixed aerosols (A - polluted and desert origin, 1 mg l-1). We describe the rationale, the experimental set-up, the chemical characteristics of the ambient water and aerosols and the relative maximal biological impacts that resulted from the added aerosols. The two treatments, run in triplicates (3 m3 each), were compared to control-unamended runs. Leaching of approximately 2.1-2.8 and 2.2-3.7 nmol PO4 and 20-26 and 53-55 nmol NOx was measured per each milligram of SD and A, respectively, representing an addition of approximately 30% of the ambient phosphate concentrations. The nitrate/phosphate ratios added in the A treatment were twice than those added in the SD treatment. Both types of dry aerosols triggered a positive change (25-600% normalized per 1 mg l-1 addition) in most of the rate and state variables that were measured: bacterial abundance (BA), bacterial production (BP), Synechococcus (Syn) abundance, chlorophyll-a (chl-a), primary production (PP) and dinitrogen fixation (N2-fix), with relative changes among them following the sequence BP>PP≈N2-fix>chl-a≈BA≈Syn. Our results show that the ‘polluted’ aerosols triggered a relatively larger biological change compared to the SD amendments (per a similar amount of mass addition), especially regarding BP and PP. We speculate that despite the co-limitation of P and N in the EMS, the additional N released by the A treatment may have triggered the relatively larger response in most of the rate and state variables as compared to SD. An implication of our study is that a warmer atmosphere in the future may increase dust emissions and influence the intensity and length of the already well stratified water column in the EMS and hence the impact of the aerosols as a significant external source of new nutrients.

Shifts in Microbial Community Structure and Activity in the Ultra-Oligotrophic Eastern Mediterranean Sea Driven by the Deposition of Saharan Dust and European Aerosols

The atmospheric deposition of gases and particulates from the Sahara Desert and European landmass is an important source of nutrients for the Mediterranean Sea. In this study, we investigated how such atmospheric input might affect bacterial metabolic activities and community dynamics in the ultra-oligotrophic Eastern Mediterranean Sea. Thus a mesocosm simulation experiment was conducted using “pure” Saharan dust (SD) and mixed aerosols (A, polluted and desert origin). The cell specific bacterial production (BP) was stimulated soon after the addition of SD and A, with a higher degree of stimulation being observed in the activity of Alphaproteobacteria than in Gammaproteobacteria, and this lead to significant increases in community BP. Subsequently, a shift between these two dominating classes was observed (such that the proportion of Gammaproteobacteria increased while that of Alphaproteobacteria decreased), along with significant increases in bacterial abundance and chlorophyll a concentration. After a few days, although the abundance of bacteria was still significantly higher in the SD- or A-treated groups, differences in the active community composition between the treatment and control groups were reduced. The altered activity of the two dominating Proteobacteria classes observed, might reflect their different strategies in responding to external nutrient input: with Alphaproteobacteria being more responsive to the direct dust input, whereas Gammaproteobacteria seemed to benefit more from the increase in phytoplankton biomass. In addition, the input of A had a stronger immediate effect and longer lasting influence on changing the active bacterial community composition than did that of SD. Our findings show that episodic atmospheric deposition events might affect the microbial community with regards to their abundance, activity and composition over a short period of time, and thus regulate the function of the microbial community and carbon cycling in oligotrophic waters.

Response of the Eastern Mediterranean Microbial Ecosystem to Dust and Dust Affected by Acid Processing in the Atmosphere

Acid processes in the atmosphere, particularly those caused by anthropogenic acid gases, increase the amount of bioavailable P in dust and hence are predicted to increase microbial biomass and primary productivity when supplied to oceanic surface waters. This is likely to be particularly important in the Eastern Mediterranean Sea (EMS), which is P limited during the winter bloom and N&P co-limited for phytoplankton in summer. However, it is not clear how the acid processes acting on Saharan dust will affect the microbial biomass and primary productivity in the EMS. Here, we carried out bioassay manipulations on EMS surface water on which Saharan dust was added as dust (Z), acid treated dust (ZA), dust plus excess N (ZN) and acid treated dust with excess N (ZNA) during springtime (May 2012) and measured bacterioplankton biomass, metabolic and other relevant chemical and biological parameters. We show that acid treatment of Saharan dust increased the amount of bioavailable P supplied by a factor of ~40 compared to non-acidified dust (18.4 nmoles P mg-1 dust vs. 0.45 nmoles P mg-1 dust, respectively). The increase in chlorophyll, primary and bacterial productivity for treatments Z and ZA were controlled by the amount of N added with the dust while those for treatments ZN and ZNA (in which excessive N was added) were controlled by the amount of P added. These results confirm that the surface waters were N&P co-limited for phytoplankton during springtime. However, total chlorophyll and primary productivity in the acid treated dust additions (ZA and ZNA) were less than predicted from that calculated from the amount of the potentially limiting nutrient added. This biological inhibition was interpreted as being due to labile trace metals being added with the acidified dust. A probable cause for this biological inhibition was the addition of dissolved Al, which forms potentially toxic Al nanoparticles when added to seawater. Thus, the effect of anthropogenic acid processes in the atmosphere, while increasing the flux of bioavailable P from dust to the surface ocean, may also add toxic trace metals such as Al, which moderate the fertilizing effect of the added nutrients.

Saharan Dust Deposition Effects on the Microbial Food Web in the Eastern Mediterranean: A Study Based on a Mesocosm Experiment

The effect of episodicity of Saharan dust deposition on the pelagic microbial food web was studied in the oligotrophic Eastern Mediterranean by means of a mesocosm experiment in May 2014. Two different treatments in triplicates (addition of natural Saharan dust in a single-strong pulse or in three smaller consecutive doses of the same total quantity), and three unamended controls were employed; chemical and biological parameters were measured during a 10-day experiment. Temporal changes in primary (PP) and bacterial (BP) production, chlorophyll a (Chla) concentration and heterotrophic bacteria, Synechococcus and mesozooplankton abundance were studied. The results suggested that the auto- and hetero-trophic components of the food web (at least the prokaryotes) were enhanced by the dust addition (and by the nitrogen and phosphorus added through dust). Furthermore, a 1-day delay was observed for PP, BP and Chla increases when dust was added in three daily doses; however, the maximal values attained were similar in the two treatments. Although the effect was evident in the first osmotrophic level (phytoplankton and bacteria), it was lost further up the food web, masked under the impact of grazing exerted by predators such as heterotrophic flagellates, ciliates and dinoflagellates. This was partly proved by two dilution experiments. This study demonstrates the important role of atmospheric deposition and protist grazing when evaluating the effect on oligotrophic systems characterised by increased numbers of trophic levels.

Atmospheric deposition effects on plankton communities in the Eastern Mediterranean: A mesocosm experimental approach

The effects of atmospheric deposition on plankton community structure were examined during a mesocosm experiment using water from the Cretan Sea (Eastern Mediterranean), an area with a high frequency of atmospheric aerosol deposition events. The experiment was carried out under spring-summer conditions (May 2012). The main objective was to study the changes induced from a single deposition event, on the autotrophic and heterotrophic surface microbial populations, from viruses to zooplankton. To this end, the effects of Saharan dust addition were compared to the effects of mixed aerosol deposition on the plankton community over 9 days. The effects of the dust addition seemed to propagate throughout the food-web, with changes observed in nearly all of the measured parameters up to copepods. The dust input stimulated increased productivity, both bacterial and primary. Picoplankton, both autotrophic and heterotrophic capitalized on the changes in nutrient availability and microzooplankton abundance also increased due to increased availability of prey. Five days after the simulated deposition, copepods also responded, with an increase in egg production. The results suggest that nutrients were transported up the food web through autotrophs that were favored by the Nitrogen supplied through both treatments. Although the effects of individual events are generally short lived, increased deposition frequency and magnitude of events is expected in the area, due to predicted reduction in rainfall and increase in temperature, which can lead to more persistent changes in plankton community structure. Here we demonstrate how a single dust deposition event leads to enhancement of phytoplankton and microzooplankton and can eventually, through copepods, transport more nutrients up the food web in the Eastern Mediterranean Sea.

Evaluating the Impact of Atmospheric Depositions on Springtime Dinitrogen Fixation in the Cretan Sea (Eastern Mediterranean)—A Mesocosm Approach

Large amounts of dust and atmospheric aerosols, originating from surrounding desert areas (e.g., Sahara and Middle East) are deposited annually on the surface of the Eastern Mediterranean Sea. These depositions can provide high amounts of micro (such as Fe, Zn, Co) and macro nutrients (such as P and N) to supplement nutrient-poor surface waters- that typically limit primary productivity and also dinitrogen (N2) fixation in many marine environments. Here, we studied the impact of the atmospheric deposition of dust and aerosols on N2 fixation in the Cretan Sea (Eastern Mediterranean Sea). Mixed polluted aerosols (hereafter A) and Saharan dust (hereafter SD) were added to nine mesocosms (3-m3 each) containing surface mixed layer seawater (~10 m), and N2 fixation was evaluated for 6 days during May 2012 (springtime). The addition of SD triggered a rapid (30 h) and robust (2-4 fold) increase in N2 fixation rates that remained high for 6 days and contributed 3-8% of the primary productivity. The A addition also resulted in higher N2 fixation rates compared to the unamended control mesocosms, although the responses were less profound (1.5-2 fold) and accounted for only 2-4% of the primary productivity. The microbial community responded differently to the two additions. Heterotrophic bacterial N2 fixers dominated the diazotroph community in A and the control mesocosms, while the non-filamentous cyanobacterial group Trichodesmium prevailed in the SD treatment (68% of all the operational taxonomic units, verified by qPCR analyses). Our results indicate that the aerosol source, its route prior to deposition, and its specific chemical composition, can alter the diazotrophic diversity and activity in the Eastern Mediterranean Sea and may thus impact both the N and C dynamics in this impoverished environment.

The impact of silver nanoparticles on marine plankton dynamics: Dependence on coating, size and concentration

During this study, three microcosm experiments were carried out with natural coastal seawater, collected in the Eastern Mediterranean Sea, in order to assess the effect of silver nanoparticle (AgNP) exposure to natural plankton communities. The impact of coating (branched-polyethyleneimine: BPEI vs. poly-vinylpyrrolidone: PVP), size (40 vs. 60nm), concentration (200, 500, 2000, 5000 and 10,000ng Ag L-1) and silver form (dissolved Ag+ vs. AgNPs) were tested. The results of chlorophyll a concentration revealed that PVP AgNPs caused a higher toxicity than BPEI AgNPs, and this was possibly related to the measured higher dissolution rate. Additionally, toxicity of BPEI AgNPs was size-dependent, with 40 being more toxic than 60 nm AgNPs, which was nevertheless not seen clearly for PVP AgNPs. Interestingly, community composition altered in response to AgNP exposure: cyanobacterial abundance was negatively affected at concentrations ≥200ng Ag L-1, and dinoflagellate abundance and composition were altered at a 2000ng Ag L-1 concentration. Specifically, dinoflagellate (Gymnodinium, Prorocentrum and Gyrodinium) and diatom (Nitzschia, Navicula and Climacosphenia) genera either increased or decreased, highlighting taxa-specific effects, with some of them being able to tolerate, compensate or even benefit from AgNPs. Silver in either form (dissolved Ag+ or in NPs) caused almost identical results in the plankton community, further indicating that Ag+ release is the primary cause of AgNP toxicity. This study employed for the first time environmentally relevant AgNP concentrations (minimum 200ng Ag L-1) in natural seawater without pre-filtration steps and showed that community changes were driven by the exposure but were largely dependent on ambient physico-chemical characteristics and should be further investigated.

Phytoplankton Response to Saharan Dust Depositions in the Eastern Mediterranean Sea: A Mesocosm Study

The response of phytoplankton populations from surface ultra-oligotrophic waters of the Eastern Mediterranean Sea to Saharan dust additions was studied during a 10-day mesocosm experiment in May 2014. A set of triplicate mesocosms entitled ‘Single Addition’ treatment (SA) was amended with Saharan dust once, while another triplicate set entitled ‘Repetitive Addition’ treatment (RA) received the same amount of dust divided into three consecutive daily doses administered within the first three experimental days, both simulating patterns of dust deposition events taking place in the field. In both treatments, dust particles released small amounts of dissolved inorganic nitrogen and phosphorus which stimulated by two-fold both chlorophyll-a concentration and primary production for a time period of six days, as compared to a set of control mesocosms carried out without dust addition. Phytoplankton response was similar in both treatments, regardless of the dust addition pattern, and it evolved through two distinct phases in both cases. The first phase (i.e. 1 to 2 days after initial addition) was characterized by enhancement of picoplankton chlorophyll-normalized production rates as a result of elevated orthophosphate concentrations while the second phase (i.e. 3 to 4 days after initial dust addition), was characterized by elevated chlorophyll-normalized production rates corresponding to larger cells (> 5 μm) as a result of increased mineral nitrogen concentrations. The stimulated primary production of larger cells was not accompanied by a respective increase in carbon biomass suggesting important top-down control. The major phytoplankton taxa detected during the experiment were Synechococcus, Pelagophytes and Prymnesiophytes. Estimations of cellular pigment concentrations and carbon-to-chlorophyll ratios of identified groups and differences between prokaryotic and eukaryotic cells are discussed.

Low-dose addition of silver nanoparticles stresses marine plankton communities

The release of silver nanoparticles (AgNPs) is expected to rise in the near future, with possible negative effects on aquatic life and enhancement of microbial resistance against AgNPs. However, a realistic evaluation of the toxicity of AgNPs to the marine ecosystem is currently missing. Therefore, we designed a mesocosm experiment to assess the impact of AgNP exposure on natural microbial plankton community dynamics in a coastal marine site at environmentally relevant concentrations. We monitored changes in the composition of the planktonic community, from viruses to protists. Further, we analyzed the concentration and properties of AgNPs for the total time of exposure. We found that the addition of AgNPs even at a low dose affected the plankton communities. Specifically, the growth of Synechococcus was inhibited and bacterial community composition significantly changed. Additionally, the amount of a lysogeny-related gene increased and viral auxiliary metabolic genes that are involved in cyanobacterial photosynthesis decreased, revealing a damaged photosynthetic potential after AgNP exposure. Microbial plankton was significantly affected due to both increased dissolved silver ions and decreased AgNP size. Our results highlight that the release of AgNPs alters the functioning of the marine food web by hampering important viral and bacterial processes.