Keeping track of phaeodactylum tricornutum (Bacillariophyta) culture contamination by potentiometric e-tongue

The large-scale cultivation of microalgae provides a wide spectrum of marketable bioproducts, profitably used in many fields, from the preparation of functional health products and feed supplement in aquaculture and animal husbandry to biofuels and green chemistry agents. The commercially successful algal biomass production requires effective strategies to maintain the process at desired productivity and stability levels. Hence, the development of effective early warning methods to timely indicate remedial actions and to undertake countermeasures is extremely important to avoid culture collapse and consequent economic losses. With the aim to develop an early warning method of algal contamination, the potentiometric E-tongue was applied to record the variations in the culture environments, over the whole growth process, of two unialgal cultures, Phaeodactylum tricornutum and a microalgal contaminant, along with those of their mixed culture. The E-tongue system ability to distinguish the cultures and to predict their growth stage, through the application of multivariate data analysis, was shown. A PLS regression method applied to the E-tongue output data allowed a good prediction of culture growth time, expressed as growth days, with R-2 values in a range from 0.913 to 0.960 and RMSEP of 1.97-2.38 days. Moreover, the SIMCA and PLS-DA techniques were useful for cultures contamination monitoring. The constructed PLS-DA model properly discriminated 67% of cultures through the analysis of their growth media, i.e., environments, thus proving the potential of the E-tongue system for a real time monitoring of contamination in microalgal intensive cultivation

Shedding Light on Diatom Photonics by means of Digital Holography

Diatoms are among the dominant phytoplankters in the worl's ocean, and their external silica investments, resembling artificial photonics crystal, are expected to play an active role in light manipulation. Digital holography allowed studying the interaction with light of Coscinodiscus wailesii cell wall reconstructing the light confinement inside the cell cytoplasm, condition that is hardly accessible via standard microscopy. The full characterization of the propagated beam, in terms of quantitative phase and intensity, removed a long-standing ambiguity about the origin of the light. The data were discussed in the light of living cell behavior in response to their environment

Capsular polysaccharides of cultured phototrophic biofilms

Phototrophic biofilm samples from an Italian wastewater treatment plant were studied in microcosm experiments under varying irradiances, temperatures and flow regimes to assess the effects of environmental variables and phototrophic biomass on capsular exopolysaccharides (CPS). The results, obtained from circular dichroism spectroscopy and High Performance Liquid Chromatography, suggest that CPS have a stable spatial conformation and a complex monosaccharide composition. The total amount present was positively correlated with the biomass of cyanobacteria and diatoms, and negatively with the biovolume of green algae. The proportion of uronic acids showed the same correlation with these taxon groups, indicating a potential role of cyanobacteria and diatoms in the removal of residual nutrients and noxious cations in wastewater treatment. While overall biofilm growth was limited by low irradiance, high temperature (30 degrees C) and low flow velocity (25 l h(-1)) yielded the highest phototrophic biomass, the largest amount of CPS produced, and the highest proportion of carboxylic acids present

Diversity and biomass accumulation in cultured phototrophic biofilms

In the present study, biomass development and changes in community composition of phototrophic biofilms grown under different controlled ambient conditions (light, temperature and flow) were examined. Source communities were taken from a wastewater treatment plant and used to inoculate growth surfaces in a semi-continuous-flow microcosm. We recorded biofilm growth curves in cultures over a period of 30 days across 12 experiments. Biovolume of phototrophs and community composition for taxonomic shifts were also obtained using light and electron microscopy. Species richness in the cultured biofilms was greatly reduced with respect to the natural samples, and diversity decreased even further during biofilm development. Diadesmis confervacea, Phormidium spp., Scenedesmus spp. and Synechocystis spp. were identified as key taxa in the microcosm. While a significant positive effect of irradiance on biofilm growth could be identified, impacts of temperature and flow rate on biofilm development and diversity were less evident. We discuss the hypothesis that biofilm development could have been subject to multistability, i.e. the existence of several possible stable biofilm configurations for the same set of environmental parameters; small variations in the species composition might have been sufficient to switch between these different configurations and thus have contributed to overwriting the original effects of temperature and flow velocity

Phosphorus and metal removal combined with lipid production by the green microalga Desmodesmus sp.: An integrated approach

This work focused on the potential of Desmodesmus sp. to be employed for wastewater 15 bioremediation and biodiesel production. The green microalga was grown in a culture medium with a phosphorus (P) content of 4.55 mg L-1 16 simulating an industrial effluent; it was also exposed to a bimetal solution of copper (Cu) and nickel (Ni) for 2 days. P removal was between 94 and 100%. After 2 days of exposure to metals, 94% of Cu and 85% of Ni were removed by Desmodesmus sp. Adsorption tests showed that the green microalga was able to remove up to 90% of Cu and 43% of Ni in less than 30 minutes. The presence of metals decreased the lipid yield, but biodiesel quality from the biomass obtained from metal exposed samples was higher than that grown without metals. This result revealed that this technology could offer a new alternative solution to environmental pollution and carbon-neutral fuel generation

Developing a microbial consortium for removing nutrients in dishwasher wastewater: towards a biofilter for its up-cycling

Microbial consortia are effective biofilters to treat wastewaters, allowing for resource recovery and water remediation. To re-use and save water in the domestic cycle, we assembled a suspended biofilm, a ‘biofilter’ to treat dishwasher wastewater. Bacterial monocultures of both photo- and hetero-trophs were assembled in an increasingly complex fashion to test their nutrient stripping capacity. This ‘biofilter’ is the core of an integrated system devoted to re-using and upcycling of reconditioned wastewater, partly in subsequent dishwasher cycles and partly into a vertical garden for plant food cultivation. The biofilter has been assembled based on a strain of the photosynthetic, filamentous cyanobacterium Trichormus variabilis, selected to produce an oxygen evolving scaffold, and three heterotrophic aerobic bacterial isolates coming from the dishwasher wastewater itself: Acinetobacter, Exiguobacterium and Pseudomonas spp. The consortium has been constructed starting with 16 isolates tested one-to-one with T. variabilis and then selecting the heterotrophic microbes up to a final one-to-three consortium, which included two dominant and a rare component of the wastewater community. This consortium thrives in the wastewater much better than T. variabilis alone, efficiently stripping N and P in short time, a pivotal step to the reuse and saving of water in household appliances

A7. SITUAZIONE NAZIONALE: CORPI IDRICI INTERESSATI DA CIANOBATTERI TOSSICI

I cianobatteri sono diffusi in moltissimi ambienti acquatici, dove possono produrre cianotossine con diversi profili tossicologici. Il presente rapporto riporta le linee guida per la gestione delle fioriture di cianobatteri nelle acque di balneazione, elaborate da un gruppo di esperti. La prima parte sintetizza le attuali conoscenze scientifiche su vari aspetti, fra cui la loro presenza nei laghi italiani, le caratteristiche chimiche e tossicologiche delle varie cianotossine, gli effetti osservati sulla salute dell\u2019uomo e la valutazione del rischio. La seconda parte definisce le linee guida per prevenire effetti dannosi per la salute dei bagnanti e gestire il rischio associato alle fioriture. Vengono fornite indicazioni per pianificare attivit\ue0 di monitoraggio ambientale e sorveglianza sanitaria nelle aree a maggiore criticit\ue0. Viene inoltre presentato un sistema di reportistica, ambientale e sanitario, anche allo scopo di uniformare le informazioni a livello nazionale. Il rapporto \ue8 completato dalle indicazioni tecniche rivolte alle strutture territoriali preposte.Cyanobacteria thrive in many aquatic environments, where they can produce cyanotoxins with different toxicological profiles. This report provides the guidelines for the management of cyanobacterial blooms in bathing water, put together by a group of experts. The first part summarizes the current scientific knowledge on various aspects, including their presence in the Italian lakes, chemical and toxicological characteristics of different cyanotoxins, the observed effects on human health and the risk assessment. The second part defines the guidelines to prevent harmful effects on the health of bathers and manage the risk associated with blooms. It provides recommendations for planning environmental monitoring activities and a health surveillance system in most critical areas. It also introduces an environmental and health reporting system, with the purpose to standardize the information at national level too. The report is supplemented by technical information aimed at territorial authorities in charge

DNA barcoding of Ceramiales (Rhodophyta) around the Maltese islands reveals hidden biodiversity in the central Mediterranean

The genetic biodiversity of Mediterranean macroalgae is generally understudied, a fact which is especially important in the case of morphologically cryptic taxa. As a result, it is often challenging to compare DNA barcodes from Mediterranean samples to references in online databases since, very often, no close hits are available. At a regional scale, of around 1124 records of algae in the Mediterranean Sea, only 114 species have been barcoded.peer-reviewe

Development of a PNA Probe for Fluorescence In Situ Hybridization Detection of Prorocentrum donghaiense

Prorocentrum donghaiense is a common but dominant harmful algal bloom (HAB) species, which is widely distributed along the China Sea coast. Development of methods for rapid and precise identification and quantification is prerequisite for early-stage warning and monitoring of blooms due to P. donghaiense. In this study, sequences representing the partial large subunit rDNA (D1–D2), small subunit rDNA and internal transcribed spacer region (ITS-1, 5.8S rDNA and ITS-2) of P. donghaiense were firstly obtained, and then seven candidate DNA probes were designed for performing fluorescence in situ hybridization (FISH) tests on P. donghaiense. Based on the fluorescent intensity of P. donghaiense cells labeled by the DNA probes, the probe DP0443A displayed the best hybridization performance. Therefore, a PNA probe (PP0443A) analogous to DP0443A was used in the further study. The cells labeled with the PNA probe displayed more intensive green fluorescence than that labeled with its DNA analog. The PNA probe was used to hybridize with thirteen microalgae belonging to five families, i.e., Dinophyceae, Prymnesiophyceae, Raphidophyceae, Chlorophyceae and Bacillariophyceae, and showed no visible cross-reaction. Finally, FISH with the probes PP0443A and DP0443A and light microscopy (LM) analysis aiming at enumerating P. donghaiense cells were performed on the field samples. Statistical comparisons of the cell densities (cells/L) of P. donghaiense in the natural samples determined by FISH and LM were performed using one-way ANOVA and Duncan's multiple comparisons of the means. The P. donghaiense cell densities determined by LM and the PNA probe are remarkably higher than (p<0.05) that determined by the DNA probe, while no significant difference is observed between LM and the PNA probe. All results suggest that the PNA probe is more sensitive that its DNA analog, and therefore is promising for the monitoring of harmful algal blooms of P. donghaiense in the future

FISH methods in phycology: Phototrophic biofilm and phytoplankton applications

Many photosynthetic microorganisms, living attached to immersed substrates or free in the water column, lack distinct morphological details, are small in size and often unculturable. Thus, whole-cell fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes has become a valuable and widely used technique to identify bacteria and protists within their natural communities. FISH methods not only allow direct, cultivation independent determination of community composition, but provide spatio-temporal quantification of microorganisms in the environment. Coupling of FISH techniques to Confocal Laser Scanning Microscopy (CLSM) has become essential for the assessment of diversity and structural integrity in three-dimensional complex biofilm samples. Combining FISH with microautoradiography (FISH-MAR) and microsensors also opens new perspectives in microbial ecology by providing new tools for revealing physiological properties of organisms with single-cell resolution. This paper briefly summarizes the application of FISH methods to phototrophic biofilm and phytoplankton research. The potential of DNA microarray technology in phycological research is highlighted, especially for the fast and accurate identification of HAB (Harmful Algal Bloom) species in marine phytoplankton. Some CLSM and FISH data from phototrophic biofilms from an Italian wastewater treatment plant are shown