Phytoplankton Size Structure and Diversity in the Transitional System of the Aquatina Lagoon (Southern Adriatic Sea, Mediterranean).

The Aquatina Lagoon (Southern Adriatic Sea, Mediterranean Ecoregion) is a transitional water ecosystem with great ecological and socio-economic interest. Anthropogenic activities around the lagoon (e.g., agriculture and tourism) and hydrology can affect the environmental quality and biodiversity of the lagoon. Herein, the dynamics and diversity of phytoplankton communities were studied before and after the opening of a new canal connecting the lagoon with the sea, by using different approaches based on an evaluation of the size and structure of the phytoplankton as well as the taxonomic analyses. The lagoon depicted time-related fluctuations in chemical-physical parameters. The phytoplankton trend was characterized by an increase in abundance and biomass in summer, when pico-sized autotrophs dominated. Generally, nano-sized phytoflagellates dominated the community, while micro-sized dinoflagellates and diatoms were less abundant. An increase in the phytoplankton taxa number was observed throughout the years. All the analyzed parameters were generally relatively homogeneous before the opening of the channel, while some quantitative differences among stations were observed in the second sampling period. Considering the statistical evidence, both environmental and biological parameters were affected by the "dilution" effect exerted by marine water inputs. This research supports the evidence that phytoplankton is a good indicator of the environmental status, and the obtained results contribute to the implementation of management strategies for the conservation of transitional water ecosystems.

Harmful algae and pressure-impact relationship: Noxious blooms and toxic microalgae occurrence from coastal waters of the Apulia region (Adriatic and Ionian Seas, Mediterranean).

The spatial distribution of harmful microalgal taxa along the coasts of the Apulia region (Mediterranean Sea) based on results of eight years (2012-2019) of routine monitoring program and a series of sporadic observations is presented. A total number of 69 potentially harmful taxa were found during the study period. Occurrence, abundance and richness of harmful taxa (toxic, potentially toxic and high biomass producers) varied along the Apulian coasts. The occurrence of harmful species was significantly higher where most of anthropogenic pressures overlap than only a few or no pressures existed. The physical alteration of coast is the most important pressure determining this pattern. Despite the variety and the abundances of the harmful microalgae, to our knowledge, no human health problems or risks have been ever recorded, nor were full-blown consequences on marine organisms such as fish kills during algal blooms. However, blooms coupled with water discoloration phenomena could become a big issue to tourism and recreational activities that have locally important socio-economic value.

Microalgae: A Promising Future.

Microalgae are photosynthetic unicellular microorganisms that represent an extremely important component of the aquatic ecosystem productivity, diversity, and functioning [...].

Microbiological and chemical characteristics of beaches along the Taranto Gulf (Ionian Sea, Southern Italy).

Coastal habitats provide important ecosystem services, such as the maintenance of ecological sustainability, water quality regulation, nutrient recycling, and sandy beaches which are important areas for recreation and tourism. The quality of seawater is generally measured by determining the concentrations of Escherichia coli and intestinal Enterococci, which might be affected by the persistent populations of these bacteria in sand. Sand might thus be a significant source of pathogen exposure to beachgoers. The quality of coastal recreational waters can also be affected by eutrophication, water discoloration, and harmful algal blooms, which pose additional human health risks. Here, we conducted a monitoring of the beaches quality along the Taranto Gulf by determining the concentrations of fecal indicator organisms, as well as other parameters that are not traditionally measured (physicochemical parameters, Pseudomonas aeruginosa, and harmful microalgae), in shallow seawater and sand sampled from three beaches. The concentrations of bacteria were determined using both standard microbiological methods and the IDEXX system. Our results demonstrate the utility of measuring a greater number of parameters in addition to those conventionally measured, as well as the importance of assessing the health risks posed by the sand matrix. Additional work is needed to develop rapid analytical techniques that could be used to monitor the microbiological parameters of solid matrices.

Filtration of the Microalga Amphidinium carterae by the Polychaetes Sabella spallanzanii and Branchiomma luctuosum: A New Tool for the Control of Harmful Algal Blooms?

Harmful algal blooms (HABs) are extreme biological events representing a major issue in marine, brackish, and freshwater systems worldwide. Their proliferation is certainly a problem from both ecological and socioeconomic contexts, as harmful algae can affect human health and activities, the marine ecosystem functioning, and the economy of coastal areas. Once HABs establish, valuable and environmentally friendly control actions are needed to reduce their negative impacts. In this study, the influence exerted by the filter-feeding activity of the two sabellid polychaetes Branchiomma luctuosum (Grube) and Sabella spallanzanii (Gmelin) on a harmful dinoflagellate was investigated. Clearance rates (C) and retention efficiencies were estimated by employing the microalga Amphidinium carterae Hulburt. The Cmax was 1.15 ± 0.204 L h-1 g-1 DW for B. luctuosum and 0.936 ± 0.151 L h-1 g-1 DW for S. spallanzanii. The retention efficiency was 72% for B. luctuosum and 68% for S. spallanzanii. Maximum retention was recorded after 30 min for both species. The obtained results contribute to the knowledge of the two polychaetes' filtration activity and to characterize the filtration process on harmful microalgae in light of the protection of water resources and human health. Both species, indeed, were extremely efficient in removing A. carterae from seawater, thus suggesting their employment as a new tool in mitigation technologies for the control of harmful algae in marine environments, as well as in the aquaculture facilities where HABs are one of the most critical threats.

Bioactive Potential of Two Marine Picocyanobacteria Belonging to Cyanobium and Synechococcus Genera.

Coccoid cyanobacteria produce a great variety of secondary metabolites, which may have useful properties, such as antibacterial, antiviral, anticoagulant or anticancer activities. These cyanobacterial metabolites have high ecological significance, and they could be considered responsible for the widespread occurrence of these microorganisms. Considering the great benefit derived from the identification of competent cyanobacteria for the extraction of bioactive compounds, two strains of picocyanobacteria (coccoid cyanobacteria < 3 µm) (Cyanobium sp. ITAC108 and Synechococcus sp. ITAC107) isolated from the Mediterranean sponge Petrosia ficiformis were analyzed. The biological effects of organic and aqueous extracts from these picocyanobacteria toward the nauplii of Artemia salina, sea urchin embryos and human cancer lines (HeLa cells) were evaluated. Methanolic and aqueous extracts from the two strains strongly inhibited larval development; on the contrary, in ethyl acetate and hexane extracts, the percentage of anomalous embryos was low. Moreover, all the extracts of the two strains inhibited HeLa cell proliferation, but methanol extracts exerted the highest activity. Gas chromatography-mass spectrometry analysis evidenced for the first time the presence of ß-N-methylamino-l-alanine and microcystin in these picocyanobacteria. The strong cytotoxic activity observed for aqueous and methanolic extracts of these two cyanobacteria laid the foundation for the production of bioactive compounds of pharmacological interest.

Two - stages bloom of Margalefidinium cf. polykrikoides in a Mediterranean shallow bay (Ionian Sea, Italy).

The emergence of a red tide resulting in yellow-brownish discoloration of waters in Porto Cesareo bay (Italy) during July-August 2018 is reported. The species responsible for the bloom was the dinoflagellate Margalefidinium cf. polykrikoides. Cell densities reached 9.1 × 106 cells L-1 during the initial outbreak. A second peak was observed about three weeks later reaching 6.7 × 105 cells L-1. Study of live specimens showed great variation in cell size and shape. Different cyst morphotypes were found in the water samples and in the sediment. For the first time, we followed several stages of the life cycle of M. cf. polykrikoides in natural samples. Fish die-offs in the bay were not observed, however this high-density bloom may have caused consequences on the ecosystem (amount of mucilage on the beach) and in turn, on tourism that is the main activity in the area during the summer season.

First bloom event of the small dinoflagellate Prorocentrum shikokuense in the Mediterranean Sea: cryptogenic or introduced?

A bloom of putatively non-indigenous species (NIS) Prorocentrum shikokuense was detected for the first time in the Mediterranean Sea at the Brindisi harbor (Southern Adriatic Sea) on September 2016, in the context of EU Marine Strategy Framework Directive monitoring in the ports. This species is usually observed in the East China Sea and Japanese and Korean waters. In the Brindisi harbor this dinoflagellate reached the concentration 105 cell/L and represented from 30 to 50% of the total phytoplankton population. Besides this event, Prorocentrum shikokuense has not been found blooming until today in Mediterranean waters. This study suggests the necessity to improve the monitoring surveys in areas that are known vulnerable systems to alien and invasive species, such as ports.

Plankton dynamics across the freshwater, transitional and marine research sites of the LTER-Italy Network. Patterns, fluctuations, drivers.

A first synoptic and trans-domain overview of plankton dynamics was conducted across the aquatic sites belonging to the Italian Long-Term Ecological Research Network (LTER-Italy). Based on published studies, checked and complemented with unpublished information, we investigated phytoplankton and zooplankton annual dynamics and long-term changes across domains: from the large subalpine lakes to mountain lakes and artificial lakes, from lagoons to marine coastal ecosystems. This study permitted identifying common and unique environmental drivers and ecological functional processes controlling seasonal and long-term temporal course. The most relevant patterns of plankton seasonal succession were revealed, showing that the driving factors were nutrient availability, stratification regime, and freshwater inflow. Phytoplankton and mesozooplankton displayed a wide interannual variability at most sites. Unidirectional or linear long-term trends were rarely detected but all sites were impacted across the years by at least one, but in many case several major stressor(s): nutrient inputs, meteo-climatic variability at the local and regional scale, and direct human activities at specific sites. Different climatic and anthropic forcings frequently co-occurred, whereby the responses of plankton communities were the result of this environmental complexity. Overall, the LTER investigations are providing an unparalleled framework of knowledge to evaluate changes in the aquatic pelagic systems and management options.

Hydrological conditions and phytoplankton community in the Lesina lagoon (southern Adriatic Sea, Mediterranean).

The Lesina lagoon (southern Adriatic Sea, Mediterranean) is a coastal lagoon located in a highly intensified farming and tourist area. A monthly sampling was carried out in 1998 in five stations, representative of different hydrological features, in order to analyse phytoplankton composition and its relation to environmental parameters. Our results showed high spatial variability of abiotic variables. Phytoplankton abundances and biomass trends showed marked seasonality, with annual peaks occurring in late winter-early spring and summer periods. Phytoplankton blooms were due to the diatom Thalassiosira pseudonana and the dinoflagellate Prorocentrum cordatum. Statistical analyses showed that salinity and nutrients were the main factors affecting phytoplankton abundance and biomass. Phytoplankton dynamics was associated both to seasonality and the hydrodynamic regime of the system. Moreover, chemical-physical data were analysed together with those collected in the same stations in 2007, to compare their dynamics under different hydrological regimes. The two periods corresponded to the closure and opening, respectively, of canals connecting the lagoon to the sea. In general, abiotic variables (salinity, dissolved oxygen, pH, nitrate, phosphate and silicate concentrations) were significantly affected by the hydrodynamic regime. These data could provide a useful basis to complement the knowledge gained through current monitoring within the framework of the European Directives, as well as to implement conservation and management strategies of these transitional waters.

'End to end' planktonic trophic web and its implications for the mussel farms in the Mar Piccolo of Taranto (Ionian Sea, Italy).

The Mar Piccolo is a semi-enclosed basin subject to different natural and anthropogenic stressors. In order to better understand plankton dynamics and preferential carbon pathways within the planktonic trophic web, an integrated approach was adopted for the first time by examining all trophic levels (virioplankton, the heterotrophic and phototrophic fractions of pico-, nano- and microplankton, as well as mesozooplankton). Plankton abundance and biomass were investigated during four surveys in the period 2013-2014. Beside unveiling the dynamics of different plankton groups in the Mar Piccolo, the study revealed that high portion of the plankton carbon (C) pool was constituted by small-sized (<2 µm) planktonic fractions. The prevalence of small-sized species within micro- and mesozooplankton communities was observed as well. The succession of planktonic communities was clearly driven by the seasonality, i.e. by the nutrient availability and physical features of the water column. Our hypothesis is that beside the 'bottom-up' control and the grazing pressure, inferred from the C pools of different plankton groups, the presence of mussel farms in the Mar Piccolo exerts a profound impact on plankton communities, not only due to the important sequestration of the plankton biomass but also by strongly influencing its structure.

Microbial assemblages for environmental quality assessment: Knowledge, gaps and usefulness in the European Marine Strategy Framework Directive.

The EU Marine Strategy Framework Directive 2008/56/EC (MSFD) defines a framework for Community actions in the field of marine environmental policy in order to achieve and/or maintain the Good Environmental Status (GES) of the European seas by 2020. Microbial assemblages (from viruses to microbial-sized metazoa) provide a major contribution to global biodiversity and play a crucial role in the functioning of marine ecosystems, but are largely ignored by the MSFD. Prokaryotes are only seen as "microbial pathogens," without defining their role in GES indicators. However, structural or functional prokaryotic variables (abundance, biodiversity and metabolism) can be easily incorporated into several MSFD descriptors (i.e. D1. biodiversity, D4. food webs, D5. eutrophication, D8. contaminants and D9. contaminants in seafood) with beneficial effects. This review provides a critical analysis of the current MSFD descriptors and illustrates the reliability and advantages of the potential incorporation of some prokaryotic variables within the set of indicators of marine environmental quality. Following a cost/benefit analysis against scientific and economic criteria, we conclude that marine microbial components, and particularly prokaryotes, are highly effective for detecting the effects of anthropogenic pressures on marine environments and for assessing changes in the environmental health status. Thus, we recommend the inclusion of these components in future implementations of the MSFD.

Phytoplankton dynamics with a special emphasis on harmful algal blooms in the Mar Piccolo of Taranto (Ionian Sea, Italy).

The response of phytoplankton assemblages to the closure of urban sewage outfalls (USOs) was examined for the Mar Piccolo of Taranto (Mediterranean Sea), a productive semi-enclosed coastal marine ecosystem devoted to shellfish farming. Phytoplankton dynamics were investigated in relation to environmental variables, with a particular emphasis on harmful algal blooms (HABs). Recent analyses evidenced a general reduction of the inorganic nutrient loads, except for nitrates and silicates. Also phytoplankton biomass (chlorophyll a) and abundances were characterized by a decrease of the values, except for the inner area of the basin (second inlet). The phytoplankton composition changed, with nano-sized species, indicators of oligotrophic conditions, becoming dominant over micro-sized species. If the closure of the USOs affected phytoplankton dynamics, however, it did not preserve the Mar Piccolo from HABs and anoxia crises. About 25 harmful species have been detected throughout the years, such as the potentially domoic acid producers Pseudo-nitzschia cf. galaxiae and P seudo-nitzschia cf. multistriata, identified for the first time in these waters. The presence of HABs represents a threat for human health and aquaculture. Urgent initiatives are needed to improve the communication with authorities responsible for environmental protection, economic development, and public health for a sustainable mussel culture in the Mar Piccolo.

The red seaweed Gracilaria gracilis as a multi products source.

In recent years seaweeds have increasingly attracted interest in the search for new drugs and have been shown to be a primary source of bioactive natural compounds and biomaterials. In the present investigation, the biochemical composition of the red seaweed Gracilaria gracilis, collected seasonally in the Lesina Lagoon (Southern Adriatic Sea, Lesina, Italy), was assayed by means of advanced analytical techniques, such as gas-chromatography coupled with mass spectrometry and spectrophotometric tests. In particular, analysis of lipids, fatty acids, sterols, proteins, phycobiliproteins and carbohydrates as well as phenolic content, antioxidant and radical scavenging activity were performed. In winter extracts of G. gracilis, a high content of R-phycoerythrin together with other valuable products such as arachidonic acid (PUFA ω-6), proteins and carbohydrates was observed. High antioxidant and radical scavenging activities were also detected in summer extracts of the seaweed together with a high content of total phenols. In conclusion, this study points out the possibility of using Gracilaria gracilis as a multi products source for biotechnological, nutraceutical and pharmaceutical applications even although more investigations are required for separating, purifying and characterizing these bioactive compounds.

Toxicity assessment of Amphidinium carterae, Coolia cfr. monotis and Ostreopsis cfr. ovata (Dinophyta) isolated from the northern Ionian Sea (Mediterranean Sea).

In many coastal areas the abundant proliferation of microalgae producing biotoxins determines the occurrence of Harmful Algal Blooms (HABs). Their presence in temperate waters is well documented and often associated with marine toxin-derived disease. The occurrence and toxicity of three harmful microalgae (Amphidinium carterae, Coolia cfr. monotis and Ostreopsis cfr. ovata) from the northern Ionian Sea (Mediterranean Sea) is hereby reported. The three dinoflagellates were sampled both on macroalgae and water and their morphology and occurrence were compared to those of other Mediterranean sites. The toxicity of the three cultured strains was tested by Artemia salina and hemolysis tests and their effects on the first stages of the sea urchin development was also evaluated. The contemporary presence of the three species inhibited the in vitro sea urchin embryonic development. But this action could be ascribed to the sole Ostreopsis as the addition of the single species to the sea urchins embryos evidenced no effects in presence of Amphidinium or Coolia cells, and an irregular segmentation in presence of Ostreopsis. In particular, this latter species exerted a cytotoxic effect in a dose-dependent manner, with a production of deformed embryos even at very low cell concentration (42 cells mL⁻¹). Nevertheless, when algal cell lysate was added, some effects on the sea urchin development was detected for each dinoflagellate, and also in this case Ostreopsis has proved to be the most toxic species. However, the lysate of Amphidinium and Ostreopsis strongly affects the A. salina nauplii vitality, while the hemolytic activity was very low for Amphidinium and Coolia lysate and very strong for Ostreopsis. Our results highlight the importance to monitoring the presence of these dinoflagellates whose effects may also be reflected on the early life stages of marine organisms, especially those species that are important from both an ecological and economic point of view, as the sea urchins are.

Cytotoxic and antimitotic activities in aqueous extracts of eight cyanobacterial strains isolated from the marine sponge Petrosia ficiformis.

Marine cyanobacteria are photosynthetic prokaryotes of significant ecological interest, living free or in association with invertebrates. They are also considered as excellent sources of antineoplastic, antibacterial, antiviral and antifungal compounds. In this work, aqueous extracts from eight cyanobacterial strains isolated from the Mediterranean sponge Petrosia ficiformis have been investigated for their bioactive properties. Bioassays with human erythrocytes, Artemia salina nauplii, and Paracentrotus lividus gametes and embryos were performed. Some aqueous extracts exhibited citolytic effect on human erythrocytes and toxic activity against A. salina nauplii. Furthermore antimitotic activity was evidenced during sea urchin embryos development and disorganization of blastomeres with altered cell-cell contact was also induced. Some of the isolated cyanobacterial strains, belonging to Leptolyngbya and Synechococcus genera with an high citotoxic activity, should be further investigated to better characterize their bioactive molecules. Our data confirm cyanobacteria as an interesting source of novel bioactive compounds with potential applications in pharmaceutics.

Impact of heavy metals and PCBs on marine picoplankton.

Synergistic/antagonistic effects of multiple contaminants in marine environments are almost completely unexplored. In the present study, we investigated the effects of heavy metals (Zn and Pb) and PCBs on picoplankton abundance, biomass, cell size distribution, and bacterial C production. Natural picoplankton assemblages were exposed to heavy metals (Zn or Pb), organic contaminants (PCBs, Aroclor 1260), and to a mixture of different contaminants. The results of the present study indicate that Zn addition stimulated heterotrophic growth, whereas Pb has a negative impact on heterotrophic picoplankton, particularly significant in the first 24 h. Heavy metals had no effects on the autotrophic component. The addition of Aroclor 1260 had a significant impact on abundance, biomass, and cell size of autotrophic and heterotrophic picoplankton, and reduced significantly bacterial secondary production. Three weeks after PCB treatment, heterotrophic bacteria displayed a clear resilience, both in terms of abundance and biomass, reaching values comparable to those of the controls, but not in terms of bacterial C production. Our results indicate that picoplankton can be sensitive indicators of impact determined by heavy metals and PCBs in coastal marine systems.

Monitoring of a coastal Mediterranean area: culturable bacteria, phytoplankton, environmental factors and their relationships in the Southern Adriatic Sea.

Culturable heterotrophic bacterial and phytoplanktonic densities were investigated at four sites in the Southern Adriatic Sea (Brindisi, S. Cataldo, Otranto and S. M. di Leuca) over an annual cycle. The main phytoplankton groups, the bacterial biodiversity, as well as the faecal contamination indicators were determined. Culturable bacterial numbers averaged 4.8 +/- 0.2 log CFU ml(-1) whereas phytoplankton numbers averaged 2.1 +/- 0.4 log cells ml(-1). Relationships between culturable bacteria, phytoplankton and the environmental factors were established. Bacterial and phytoplankton densities usually depended significantly on temperature, dissolved oxygen, phosphate and nitrite only in the S. Cataldo transect. In all the examined transects phytoplankton showed a bloom during the January-February period followed by a bacterial peak during the February-March period. Thus we can suppose that the phytoplankton winter bloom is responsible for the availabily of organic matter for bacterial populations in the following months in this oligotrophic ecosystem.