RNA/DNA ratios as estimate of metabolic and functional traits in diatom species from the northwestern Adriatic Sea

Different phytoplankton biomass estimations can provide information about abundance variation, but they are not able to describe the metabolic activity of species or groups within assemblages. Conversely, molecular traits are key for the metabolic dynamics in pelagic ecosystems. To investigate if the RNA/DNA and taxon-specific 18S ribosomal RNA (rRNA)/ribosomal DNA (rDNA) ratios could be used to assess and be indicators of metabolic activity in marine phytoplankton species, two Adriatic diatom species, Chaetoceros socialis and Skeletonema marinoi, were studied. Significant correlations between abundance, chlorophyll a, carbon content and proteins were found in individual and co-cultured growth experiments (from r(s) = 0.570 to r(s) = 0.986, P < 0.001). The biomass trend followed a logistic curve without providing additional information regarding diatom metabolic activity. In both experiments, the RNA/DNA and taxon-specific 18S rRNA/rDNA ratios of C. socialis and S. marinoi showed maximum values at the beginning of the growth phase, i.e as 23.2 +/- 1.5 and 15.3 +/- 0.8, and 16.2 +/- 1.6 and 30.1 +/- 5.4 after 2 and 6 days, respectively, in individual cultures, with a subsequent significant decrease in these values for both species in individual and co-culture experiments. Our results showed that these molecular rRNA/rDNA ratios expressed an activation of metabolism before the abundance increases, even in the presence of interspecific interaction between C. socialis and S. marinoi

Ecological implications beyond the ecotoxicity of plastic debris on marine phytoplankton assemblage structure and functioning

none7noPlastic pollution is a global issue posing a threat to marine biota with ecological implications on ecosystem functioning. Micro and nanoplastic impact on phytoplankton autotrophic species (e.g., cell growth inhibition, decrease in chlorophyll a and photosynthetic efficiency and hetero-aggregates formation) have been largely documented. However, the heterogeneity of data makes rather difficult a comparison based on size (i.e. micro vs nano). In addition, knowledge gaps on the ecological impact on phytoplankton assemblage structure and functioning are evident. A new virtual meta-analysis on cause-effect relationships of micro and nanoplastics on phytoplankton species revealed the significant effect posed by polymer type on reducing cell density for tested PVC, PS and PE plastics. Linked with autotrophic phytoplankton role in atmospheric CO2 fixation, a potential impact of plastics on marine carbon pump is discussed. The understanding of the effects of microplastics and nanoplastics on the phytoplankton functioning is fundamental to raise awareness on the overall impact on the first level of marine food web. Interactions between micro and nanoplastics and phytoplankton assemblages have been quite documented by in vitro examinations; but, further studies considering natural plankton assemblages and/or large mesocosm experiments should be performed to evaluate and try predicting ecological impacts on primary producers.openCasabianca Silvia, Bellingeri Arianna, Capellacci Samuela, Sbrana Alice, Russo Tommaso, Corsi Ilaria, Penna AntonellaCasabianca, Silvia; Bellingeri, Arianna; Capellacci, Samuela; Sbrana, Alice; Russo, Tommaso; Corsi, Ilaria; Penna, Antonell

Physical interactions between marine phytoplankton and PET plastics in seawater.

Plastics are the most abundant marine debris globally dispersed in the oceans and its production is rising with documented negative impacts in marine ecosystems. However, the chemical-physical and biological interactions occurring between plastic and planktonic communities of different types of microorganisms are poorly understood. In these respects, it is of paramount importance to understand, on a molecular level on the surface, what happens to plastic fragments when dispersed in the ocean and directly interacting with phytoplankton assemblages. This study presents a computer-aided analysis of electron paramagnetic resonance (EPR) spectra of selected spin probes able to enter the phyoplanktonic cell interface and interact with the plastic surface. Two different marine phytoplankton species were analyzed, such as the diatom Skeletonema marinoi and dinoflagellate Lingulodinium polyedrum, in absence and presence of polyethylene terephthalate (PET) fragments in synthetic seawater (ASPM), in order to insitu characterize the interactions occurring between the microalgal cells and plastic surfaces. The analysis was performed at increasing incubation times. The cellular growth and adhesion rates of microalgae in batch culture medium and on the plastic fragments were also evaluated. The data agreed with the EPR results, which showed a significant difference in terms of surface properties between the diatom and dinoflagellate species. Low-polar interactions of lipid aggregates with the plastic surface sites were mainly responsible for the cell-plastic adhesion by S. marinoi, which is exponentially growing on the plastic surface over the incubation time

Impact of polystyrene nanoparticles on marine diatom Skeletonema marinoi chain assemblages and consequences on their ecological role in marine ecosystems.

Marine diatoms have been identified among the most abundant taxa of microorganisms associated with plastic waste collected at sea. However, the impact of nano-sized plastic fragments (nanoplastics) at single cell and population level is almost unknown. We exposed the marine diatom Skeletonema marinoi to model polystyrene nanoparticles with carboxylic acid groups (PS-COOH NPs, 90 nm) for 15 days (1, 10, 50 mu g/mL). Growth, reactive oxygen species (ROS) production, and nano-bio-interactions were investigated. No effect on diatom growth was observed, however Dynamic light scattering (DLS) demonstrated the formation of large PS aggregates which were localized at the diatoms' fultoportula process (FPP), as shown by TEM images. Increase production of ROS and reduction in chain length were also observed upon PS NPs exposure (p < 0.005). The observed PS-diatom interaction could have serious consequences on diatoms ecological role on the biogeochemical cycle of carbon, by impairing the formation of fast-sinking aggregates responsible for atmospheric carbon fixation and sequestration in the ocean sea floor. S. marinoi exposure to PS NPs caused an increase of intracellular and extracellular oxidative stress, the reduction of diatom's chain length and the adhesion of PS NPs onto the algal surface

The sxt Gene and Paralytic Shellfish Poisoning Toxins as Markers for the Monitoring of Toxic Alexandrium Species Blooms

Paralytic shellfish poisoning (PSP) is a serious human illness caused by the ingestion of seafood contaminated with saxitoxin and its derivatives (STXs). These toxins are produced by some species of marine dinoflagellates within the genus Alexandrium. In the Mediterranean Sea, toxic Alexandrium spp. blooms, especially of A. minutum, are frequent and intense with negative impact to coastal ecosystem, aquaculture practices and other economic activities. We conducted a large scale study on the sxt gene and toxin distribution and content in toxic dinoflagellate A. minutum of the Mediterranean Sea using both quantitative PCR (qPCR) and HILIC-HRMS techniques. We developed a new qPCR assay for the estimation of the sxtA1 gene copy number in seawater samples during a bloom event in Syracuse Bay (Mediterranean Sea) with an analytical sensitivity of 2.0 × 10° sxtA1 gene copy number per reaction. The linear correlation between sxtA1 gene copy number and microalgal abundance and between the sxtA1 gene and STX content allowed us to rapidly determine the STX-producing cell concentrations of two Alexandrium species in environmental samples. In these samples, the amount of sxtA1 gene was in the range of 1.38 × 105 − 2.55 × 108 copies/L and the STX concentrations ranged from 41−201 nmol/L. This study described a potential PSP scenario in the Mediterranean Sea.Versión del editor5,228

Assessment of spatio-temporal variability of faecal pollution along coastal waters during and after rainfall events.

More than 80% of wastewaters are discharged into rivers or seas, with a negative impact on water quality along the coast due to the presence of potential pathogens of faecal origin. Escherichia coli and enterococci are important indicators to assess, monitor, and predict microbial water quality in natural ecosystems. During rainfall events, the amount of wastewater delivered to rivers and coastal systems is increased dramatically. This study implements measures capable of monitoring the pathways of wastewater discharge to rivers and the transport of faecal bacteria to the coastal area during and following extreme rainfall events. Spatio-temporal variability of faecal microorganisms and their relationship with environmental variables and sewage outflow in an area located in the western Adriatic coast (Fano, Italy) was monitored. The daily monitoring during the rainy events was carried out for two summer seasons, for a total of five sampling periods. These results highlight that faecal microbial contaminations were related to rainy events with a high flow of wastewater, with recovery times for the microbiological indicators varying between 24 and 72 h and influenced by a dynamic dispersion. The positive correlation between ammonium and faecal bacteria at the Arzilla River and the consequences in seawater can provide a theoretical basis for controlling ammonium levels in rivers as a proxy to monitor the potential risk of bathing waters pathogen pollution

DNA barcoding and phylogenetic relationships in marine toxic dinoflagellate genus Ostreopsis based on mithocondrial genes

The marine dinoflagellate genus Ostreopsis includes species producing potent toxic compounds, such as palytoxin and palytoxin analogs, which cause toxic blooms in Mediterranean and tropical or other temperate areas. Phylogenetic and phylogeographical analyses based on ribosomal data set revealed the existence of distinct species and, within them, clades related to geographical distribution. Due to a high morphological variability, species belonging to this genus are very difficult to identify and cryptic species could be present. Thus, developing a standardized DNA barcode approach for this marine dinoflagellate can allow confirming known species and uncover hidden variability with consequent description of new species. These information, besides leading to a better understanding of species diversity in natural environment, could assist in identification and detection of different species directly in field sample. This study represents the first attempt to assess the suitability of mitochondrial COI (cytochrome c oxidase 1) and cob (cytochrome b) as an identification tool for Ostreopsis species. For this purpose, we designed specific primers to amplify and sequence mitochondrial COI and cob genes from several Ostreopsis spp. isolates from worldwide areas. Phylogenetic analyses of single and concatenated mtDNA genes within genus Ostreopsis, as well as their intra and inter-specific divergences, were compared to those based on nuclear ribosomal genes LSU and 5.8S-ITS regions. Phylogenetic analyses of the genus Ostreopsis using mtDNA genes resulted uninformative, being not able to distinguish different species. Moreover, the so called 'barcode gap' that is interspecific variation exceeding intraspecific variation, was detected only in ribosomal genes. Thus, our original goal to apply the mtDNA barcode to recognize species within this toxic marine dinoflagellate was not accomplishe