Seawater physics and chemistry along the Med-SHIP transects in the Mediterranean Sea in 2016.

The Mediterranean Sea has been sampled irregularly by research vessels in the past, mostly by national expeditions in regional waters. To monitor the hydrographic, biogeochemical and circulation changes in the Mediterranean Sea, a systematic repeat oceanographic survey programme called Med-SHIP was recommended by the Mediterranean Science Commission (CIESM) in 2011, as part of the Global Ocean Ship-based Hydrographic Investigations Program (GO-SHIP). Med-SHIP consists of zonal and meridional surveys with different frequencies, where comprehensive physical and biogeochemical properties are measured with the highest international standards. The first zonal survey was done in 2011 and repeated in 2018. In addition, a network of meridional (and other key) hydrographic sections were designed: the first cycle of these sections was completed in 2016, with three cruises funded by the EU project EUROFLEETS2. This paper presents the physical and chemical data of the meridional and key transects in the Western and Eastern Mediterranean Sea collected during those cruises.

Long-term patterns and drivers of microbial organic matter utilization in the northernmost basin of the Mediterranean Sea.

Marine heterotrophic prokaryotes degrade, transform, and utilize half of the organic matter (OM) produced by photosynthesis, either in dissolved or particulate form. Microbial metabolic rates are affected by a plethora of different factors, spanning from environmental variables to OM composition. To tease apart the environmental drivers underlying the observed organic matter utilization rates, we analysed a 21 year-long time series from the Gulf of Trieste (NE Adriatic Sea). Heterotrophic carbon production (HCP) time series analysis highlighted a long-term structure made up by three periods of coherent observations (1999-2007; 2008-2011; 2012-2019), shared also by OM concentration time series. Temporal patterns of HCP drivers, extracted with a random forest approach, demonstrated that a period of high salinity anomalies (2002-2008) was the main driver of this structure. The reduced river runoff and the consequent depletion of river-borne inorganic nutrients induced a long-term Chl a decline (2006-2009), followed by a steady increase until 2014. HCP driving features over the three periods substantially changed in their seasonal patterns, suggesting that the years following the draught period represented a transition between two long-term regimes. Overall, temperature and particulate organic carbon concentration were the main factors driving HCP rates. The emergence of these variables highlighted the strong control exerted by the temperature-substrate co-limitation on microbial growth. Further exploration revealed that HCP rates did not follow the Arrhenius' linear response to temperature between 2008 and 2011, demonstrating that microbial growth was substrate-limited following the draught event. By teasing apart the environmental drivers of microbial growth on a long-term perspective, we demonstrated that a substantial change happened in the biogeochemistry of one of the most productive areas of the Mediterranean Sea. As planktonic microbes are the foundation of marine ecosystems, understanding their past dynamics may help to explain present and future changes.

Uptake-release dynamics of the inorganic and organic carbon pool mediated by planktonic prokaryotes in the deep Mediterranean Sea.

Understanding the ecosystem functioning in the dark portion of the ocean is a challenge that microbial ecologists are still facing. Due to the large volume, the global deep Ocean plays a central role in the regulation of climate, possibly buffering the rise of atmospheric carbon dioxide if processes of CO2 fixation compensate for respiration. We investigated the rates of several prokaryotic activities (dissolved and particulate primary production, heterotrophic carbon production and respiration) in meso- and bathypelagic waters of the Mediterranean Sea, covering all sub-basins. Chemosynthesis was the main process for C uptake. The rates of organic C (OC) excretion (or viral-induced cell lysis) inferred from the dissolved primary production measurements were noteworthy, being comparable to particulate primary production, and possibly contributing to the formation of non-sinking particulate organic matter. Inorganic C fixation rates were significantly higher than those reported for other deep-sea systems, probably as a consequence of the persistently higher temperature of dark Mediterranean waters or to phylogenetically diverse communities involved in the process. Primary production was negatively correlated with dissolved organic carbon concentration and showed an inverse pattern to heterotrophic carbon production, indicating a niche partitioning between heterotrophs and autotrophs. In sum, the deep Mediterranean Sea harbors active autotrophic communities able to fix inorganic carbon faster than the heterotrophic carbon production rates.

Coping with seawater acidification and the emerging contaminant diclofenac at the larval stage: A tale from the clam Ruditapes philippinarum.

Seawater acidification could alter the susceptibility of marine organisms to emerging contaminants, such as pharmaceuticals. In this study, the combined effects of seawater acidification and the non-steroidal anti-inflammatory drug diclofenac on survival, growth and oxidative stress-related parameters (catalase activity and lipid peroxidation) in the larvae of the Manila clam Ruditapes philippinarum were investigated for the first time. An experimental flow-through system was set up to carry out a 96-h exposure of clam larvae. Two pH levels (pH 8.0, the control, and pH 7.8, the predicted pH by the end of this century) were tested with and without diclofenac (0.5 µg/L). After 4 days, mortality was dramatically higher under reduced pH, particularly in the presence of diclofenac (62% of the larvae dead). Shell morphology was negatively affected by both acidification and diclofenac from the first day of exposure. The percentage of abnormal larvae was always higher at pH 7.8 than in controls, peaking at 98% in the presence of diclofenac after 96 h. Instead, shell length, shell height or the ratio of these values were only negatively influenced by reduced pH throughout the whole experiment. After 96 h, catalase activity was significantly increased in all larvae kept at pH 7.8, whereas no significant difference in lipid peroxidation was found among the treatments. This study demonstrates a high susceptibility of R. philippinarum larvae to a slight reduction in seawater pH. Furthermore, the results obtained highlight that acidification enhances the sensitivity of clam larvae to environmentally relevant concentrations of diclofenac.

Organic aggregates formed by benthopleustophyte brown alga Acinetospora crinita (Acinetosporaceae, Ectocarpales).

This work presents the elemental, polysaccharide, and fatty acid compositions of benthic aggregates formed by the filamentous brown alga Acinetospora crinita, which are widely spread on the rocky bottoms of the Mediterranean Sea. The aggregates can be characterized as mineralized centers in which regeneration of nutrients and recycling of dissolved organic matter actively occur and favor the development of an abundant phytoplankton community. Analyses of the stable isotopes of C and N display their marine origin and could provide evidence of the processes that occur inside/outside of the aggregates. The monosaccharide compositions of Adriatic and Tyrrhenian mucilages produced by brown alga A. crinita were quite similar. In particular, the Adriatic sample compositions resembled the average composition of the Tyrrhenian high molecular weight exopolymers, and the observed differences could be ascribed to different degradation stages. The fatty acid patterns found for the aggregates were similar to those observed in the isolated A. crinita algae with variable contributions from embedded diatom species. The bacterial contribution to the fatty acid pool was quite low, most likely due to the known poor conditions for their heterotrophic growth.

Phytoplankton-bacterioplankton interactions and carbon fluxes through microbial communities in a microtidal lagoon.

The strength of the bacteria-phytoplankton coupling and the importance of the microbially mediated carbon fluxes have been investigated in a microtidal lagoon (Lagoon of Venice), with emphasis on the trophic variations, in a within-system perspective. The large trophic heterogeneity of the three stations considered corresponded to an elevated variability of phytoplankton biomass and production (from 0.1 up to 300 microg C L(-1) h(-1)), while bacteria standing stock and production (from 2 to 8 microg C L(-1) h(-1)) appeared, in comparison, to be much more constant. The relationships between bacteria and the phytoplankton community could not be related to the trophic state in a straightforward way; rather, some patterns common to the three stations could be evidenced. In particular, the two communities appeared to be clearly uncoupled, bacterial carbon demand (BCD) always exceeding dissolved primary production (DPP) and, mostly, also total primary production, independent of the station and the season considered. The occurrence of situations in which bacterial production was larger than primary production and the continuous prevalence of BCD over DPP implied that, quite independent of the trophic variability, sources of organic carbon other than phytoplankton production were necessary and available to sustain the bacterial metabolism in the Lagoon of Venice.

Methylmercury determination in marine sediment and organisms by Direct Mercury Analyser.

An analytical method for simple and rapid determination of methylmercury in sediment and organism samples is described. The proposed method employs the oxygen combustion-gold amalgamation using Direct Mercury Analyser (DMA-80) after complete removal of MeHg by organic extraction and back extraction to an aqueous medium. DMA-80 instrument is equally suitable for the analysis of solid and liquid materials and has a good detection limit. The analytical performance of this method was evaluated by analysis of certified reference materials (CRM-580, IAEA-405, DORM-2, DOLT-3, SRM-2976 and SRM-2977) assessing its quality in terms of accuracy, repeatability and quantification limit. Furthermore total mercury and methylmercury have been analysed in sediment and organism samples collected during the XXI Italian Antarctic Expedition in Terra Nova Bay (Ross Sea, Northern Victoria Land). The results obtained show the validity of the proposed method as ready-to-use analytical method to analyse real samples.

Chemical characterization of different typologies of mucilaginous aggregates in the Northern Adriatic Sea.

The chemical composition of mucilage aggregates found during summer 2000, 2001 and 2002 in the North Adriatic Sea depends on the nature of the organic matter during aggregation, on the environmental conditions of the site of formation and on the transformations during ageing. The mucilages were composed of organic matter, together with a significant inorganic fraction. Elemental analysis revealed 12.5-32.2% of organic carbon, 0-7.3% of inorganic carbon and 1.0-3.7% of nitrogen. The C(org)/N ratios of most aggregates were between 7.5 and 12.6, values close to those found in the suspended matter; higher ratios were found in large-size (>5 m) aggregates which are probably older. The content of carbohydrates and proteins determined in the aggregates, respectively, 15.4+/-8.9% and 7.9+/-4.8%, w/w, showed a prevalence of carbohydrates over proteins. Neutral carbohydrate analysis of purified polysaccharides from mucilage samples showed very similar signatures with high relative abundance of galactose and glucose. Humic, fulvic and humin substances extracted from the mucilages constitute an important fraction of the organic matter in the aggregates. The humin (a fraction insoluble in acidic and basic media) was present in all mucilage samples, indicating the refractory nature of a part of the organic matter in the mucilage. The iron and calcium could play a role during the aggregation process to form a complex with polysaccharides and humic fractions. The C(org)/N ratio 10+/-2 found in the humic acids extracted from the Adriatic aggregates disclosed a marine origin. The low phosphorus content and the high C(org)/P ratio found in the aggregates might depend from high bacteria activity or from the aggregation of organic fractions depleted of phosphorus. The principal inorganic species contained aluminium and silicon, part of which was of biogenic origin and was more significant in the offshore mucilage aggregates than in the coastal ones. The Si(biog)/C(org) ratio showed that diatoms were always present in the aggregates, although it cannot be established whether these are the producers or these develop within the aggregates.

Temporal dynamics of dissolved and particulate organic carbon in the northern Adriatic Sea in relation to the mucilage events.

The accumulation of dissolved and particulate organic matter may play an important role in mucilage formation in the northern Adriatic. Distributions of dissolved and particulate organic carbon were therefore investigated during the period June 1999-July 2002, when massive mucilage events occurred: in the summer of 2000 and, to a greater extent, of 2002. The seasonal variations in dissolved organic carbon (DOC) concentrations were significant, doubling in summer (up to 150 micromol L(-1)) with respect to winter. The particulate organic carbon (POC) variations were also very large, with a less pronounced seasonal pattern compared to DOC, because the POC changes are much more dependent both on river discharges and on phytoplankton blooms. The comparison of the concentrations between the period before (March-May) and after the onset of mucilage events (June-August) showed that DOC, but particularly POC, were higher in the period before the event of 2002, more markedly in the surface waters of low salinity. The POC increased, reaching mean concentrations of up to 36 micromol L(-1) in March 2002 before the outbreak of the massive mucilage formation in June. This suggests that POC may have a more important role in the mucilage formation than DOC. The highest seasonal variations of organic matter concentrations took place in the upper layer of lower salinity, stressing the importance of stratification and pycnoclines for accumulation and aggregation of the organic matter in the northern Adriatic. The POC contribution to the total organic carbon was low in the oligotrophic waters (DOC/POC ratio >15) and increased with the phytoplankton biomass in the productive waters (DOC/POC ratio <10). Particulate organic carbon predominated over the dissolved inside the mucilage aggregate (DOC/POC ratio <1), probably because aggregation processes, in which colloidal organic carbon is involved, are important. The organic carbon within the aggregates reached a concentration of 13.6 mmol L(-1) which was about 100 times more than in the surrounding waters or in the waters when the mucilages were absent. This indicates that distributions of organic carbon in the northern Adriatic can be extremely patchy during mucilage events.

Spectroscopic evidence of the marine origin of mucilages in the Northern Adriatic Sea.

Vibrational Fourier transform infrared (FTIR) and UV-Visible spectroscopies coupled to thermogravimetry were used to characterise the gelatinous aggregates as well as the humin and humic fractions extracted from the aggregates. Spectroscopic analysis made it possible to identify the main classes of compounds present in the samples. The FTIR spectra of the aggregates showed the presence of carbohydrates, saturated aliphatic compounds, aminic, esteric, amidic (proteins) and ketonic compounds, organic and inorganic phosphates, silica and, in some cases, carbonates. The FTIR spectra of humin and humic acids showed functional groups similar to those found in the spectra of the aggregates. The high aliphatic character and the absence of aromatic compounds observed in many aggregates sampled far from estuarine areas suggest the marine origin of mucilages. These results were also confirmed by the 270/407 nm (A2/A4) and 465/665 nm (E4/E6) absorbance ratio in humic acids.

Mucilaginous aggregates in the northern Adriatic in the period 1999-2002: typology and distribution.

The spatial and temporal distributions of different types of visible mucilaginous aggregates were investigated monthly by underwater video cameras in the northern Adriatic (NA) from June 1999 to July 2002. Small aggregates (flocs, macroflocs and stringers) were observed in all seasons with higher values in autumn and winter. Larger aggregates (ribbons, cobwebs, clouds, and false bottoms) formed only during late-spring and summer, particularly in 2000 and 2002, when the seasonal thermohaline stratification was marked. In fact, the process of mucilage formation took place mainly in the upper water column, above the main pycnocline. The spatial distribution shows that higher concentrations of small aggregates occur in the more productive coastal waters. The larger aggregates form or accumulate mainly in coastal waters in the southern part of the NA and in the central part of the Po River delta-Rovinj transect, where a gyre greatly influences the aggregates distribution. The fronts between low salinity coastal waters and high salinity waters of southern origin play an important role for accumulation and aggregation of the mucilage during spring and summer. Most of the aggregates accumulate in correspondence with strong pycnoclines with differences in density anomaly of 2 kg m(-3) or higher. False bottoms formed in correspondence with strong haloclines (DeltaS congruent with 2), while cobwebs and ribbons also occurred when the halocline was less marked (DeltaS<0.5). Meteorological conditions greatly influenced the aggregate formation. Calm weather and weak winds favour aggregation processes, while strong north-easterly winds, causing intense mixing, result in the dispersion of the mucilaginous aggregates, as occurred during the Bora event in July 2000.

Mucilage microcosms.

In the summers of 2000, 2001 and 2002, large amounts of sticky mucilaginous material aggregated to form masses of impressive dimensions over large areas of the Adriatic Sea, particularly in its northern part. Aggregates differing in size were sampled by SCUBA divers and submitted to chemical (nutrient and organic matter concentrations) and biological analysis (virus, bacteria and phytoplankton abundances and bacterial metabolism). Suspended and sinking mucilaginous aggregates were biota-rich environments where the abundance of planktonic organisms and the concentration of nutrients were orders of magnitude higher than in the surrounding seawater. The embedded phytoplankton was mostly composed of diatoms, but the dinoflagellate Gonyaulax fragilis, previously reported in association with the aggregates, was also present. A variety of processes occurred within the mucilaginous aggregates which resulted in the transformation of the organic matter composition and lability and contributed to a partial degradation of mucilage. For an efficient degradation of mucilage, several conditions are necessary: high bacterial abundance and activity and an efficient recycling of nutrients within the aggregates. Most of these conditions, appear to change depending on the type and age of the aggregate. During the first phase of aggregation (cobwebs and ribbons), bacterial activities addressed the degradation of organic matter, particularly that of the nitrogen fraction. The degradation products were rapidly taken up by bacteria, supporting an increase in their abundance and production. In aged mucilage (clouds), the degradation processes decreased and the bacterial metabolism suggested the presence of new organic labile compounds probably due to phytoplankton production. On the basis of our results, stringers, generally considered the first step of the aggregation process, seemed to be the result of a mechanical disruption of other types of aggregates.