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.

Confocal hyperspectral microscopic imager for the detection and classification of individual microalgae.

We propose a confocal hyperspectral microscopic imager (CHMI) that can measure both transmission and fluorescent spectra of individual microalgae, as well as obtain classical transmission images and corresponding fluorescent hyperspectral images with a high signal-to-noise ratio. Thus, the system can realize precise identification, classification, and location of microalgae in a free or symbiosis state. The CHMI works in a staring state, with two imaging modes, a confocal fluorescence hyperspectral imaging (CFHI) mode and a transmission hyperspectral imaging (THI) mode. The imaging modes share the main light path, and thus obtained fluorescence and transmission hyperspectral images have point-to-point correspondence. In the CFHI mode, a confocal technology to eliminate image blurring caused by interference of axial points is included. The CHMI has excellent performance with spectral and spatial resolutions of 3 nm and 2 µm, respectively (using a 10× microscope objective magnification). To demonstrate the capacity and versatility of the CHMI, we report on demonstration experiments on four species of microalgae in free form as well as three species of jellyfish with symbiotic microalgae. In the microalgae species classification experiments, transmission and fluorescence spectra collected by the CHMI were preprocessed using principal component analysis (PCA), and a support vector machine (SVM) model or deep learning was then used for classification. The accuracy of the SVM model and deep learning method to distinguish one species of individual microalgae from another was found to be 96.25% and 98.34%, respectively. Also, the ability of the CHMI to analyze the concentration, species, and distribution differences of symbiotic microalgae in symbionts is furthermore demonstrated.

Biodegradation of low-density polyethylene (LDPE) sheet by microalga, Uronema africanum Borge.

Plastic (polyethylene) pollution is a severe cause of deterioration of a healthy environment. For example, ingestion of plastics in the animal gut, clogging of water canals and retarded solid waste management. Many conventional methods of polyethylene degradation include UV photooxidation, thermal oxidation, incineration, chemical oxidation and landfill are being practiced. However, these methods are not feasible, costlier and not a complete solution for this global issue. Therefore, plausible, alternative solution for this issue is biodegradation. Microbes such as bacteria, fungi and algae are involved in polyethylene degradation in its natural habitat. Among them, algae were given very less importance. In our present study, a potential microalga, morphologically identified as Uronema africanum Borge, isolated from a waste plastic bag collected from a domestic waste dumping site in a freshwater lake. This microalga was further treated with the LDPE sheet in BBM culture medium. Based on the results obtained from light microscopy, dark field microscopy, GC-MS, FT-IR, SEM and AFM, it was concluded that the microalga has initiated degradation of LDPE sheet within 30 days of incubation. Concurrently, the configuration of corrosions, abrasions, grooves and ridges were found similar with the morphological features of the microalga. For example, the configuration of the radial disc-like attachment structure of the microalga was found corresponding to the abrasions on the surface of LDPE sheet at an average size of 20-30 µm in diameter. Whereas, the configuration of ridges and grooves were found similar with the filamentous nature of the microalga (10-15 µm width). This is a hitherto report on the biodegradation of LDPE sheet by the microalga Uronema africanum Borge.

Cyanobacteria and microalgae growing on monuments of UNESCO World Heritage site Champaner Pavagadh, India: biofilms and their exopolysaccharide composition.

The present study investigated the biofilm organisms growing on selected monuments of the Champaner Pavagadh complex (Gujarat, India), which is a UNESCO World Heritage Site. The cyanobacteria and microalgae were isolated from biofilms collected through non-destructive methods. The identification of these biological organisms was done using micro-morphological characters and confirmed by 16S rRNA gene sequencing. The exopolysaccharide of each of the isolated strains was extracted, hydrolysed and analysed by the HPTLC. Six isolated strains representing five cyanobacteria and one microalga belong to the genera Desmonostoc, Nostoc, Leptolyngbya, Chroococcidiopsis and Asterarcys. The relationships between substrates' specificity of these isolated biofilm organisms and those identified globally were evaluated using maximum parsimony analysis to generate a consensus phylogenetic tree. The five strains of cyanobacteria isolated were closely clustered with cyanobacteria belonging to a tropical region. At the generic level, no relationship between the species and substratum specificity was recorded. The exopolysaccharide analysis of the isolated strains revealed the presence of seven monosaccharides. While glucose was present in all the analysed species, the concentration of either fucose or arabinose was high. The current study presents a novel HPTLC-based method for determination of monosaccharides composition from the extracellular polymeric substances.

Large-scale screening of natural genetic resource in the hydrocarbon-producing microalga Botrycoccus braunii identified novel fast-growing strains.

Algal biofuel research aims to make a renewable, carbon-neutral biofuel by using oil-producing microalgae. The freshwater microalga Botryococcus braunii has received much attention due to its ability to accumulate large amounts of petroleum-like hydrocarbons but suffers from slow growth. We performed a large-scale screening of fast-growing strains with 180 strains isolated from 22 ponds located in a wide geographic range from the tropics to cool-temperate. A fast-growing strain, Showa, which recorded the highest productivities of algal hydrocarbons to date, was used as a benchmark. The initial screening was performed by monitoring optical densities in glass tubes and identified 9 wild strains with faster or equivalent growth rates to Showa. The biomass-based assessments showed that biomass and hydrocarbon productivities of these strains were 12-37% and 11-88% higher than that of Showa, respectively. One strain, OIT-678 established a new record of the fastest growth rate in the race B strains with a doubling time of 1.2 days. The OIT-678 had 36% higher biomass productivity, 34% higher hydrocarbon productivity, and 20% higher biomass density than Showa at the same cultivation conditions, suggesting the potential of the new strain to break the record for the highest productivities of hydrocarbons.

Molecular and Structural Characterizations of Lipases from Chlorella by Functional Genomics.

Microalgae have been poorly investigated for new-lipolytic enzymes of biotechnological interest. In silico study combining analysis of sequences homologies and bioinformatic tools allowed the identification and preliminary characterization of 14 putative lipases expressed by Chlorella vulagaris. These proteins have different molecular weights, subcellular localizations, low instability index range and at least 40% of sequence identity with other microalgal lipases. Sequence comparison indicated that the catalytic triad corresponded to residues Ser, Asp and His, with the nucleophilic residue Ser positioned within the consensus GXSXG pentapeptide. 3D models were generated using different approaches and templates and demonstrated that these putative enzymes share a similar core with common α/ß hydrolases fold belonging to family 3 lipases and class GX. Six lipases were predicted to have a transmembrane domain and a lysosomal acid lipase was identified. A similar mammalian enzyme plays an important role in breaking down cholesteryl esters and triglycerides and its deficiency causes serious digestive problems in human. More structural insight would provide important information on the enzyme characteristics.

The Effect of Abiotic Factors on Abundance and Photosynthetic Performance of Airborne Cyanobacteria and Microalgae Isolated from the Southern Baltic Sea Region.

Cyanobacteria and microalgae present in the aquatic or terrestrial environment may be emitted into the air and transported along with air masses over long distances. As a result of staying in the atmosphere, these organisms may develop a greater tolerance to stressful factors, but this topic is still relatively unknown. The main aim was to show an autecological characteristic of some airborne microalgae and cyanobacteria strains by a factorial laboratory experiment approach, including changes in irradiance, temperature, and salinity conditions. The additional purpose of this work was also to present part of the Culture Collection of Baltic Algae (CCBA) collection, which consists of airborne algae (AA) isolated from the atmospheric air of the southern Baltic Sea region. Altogether, 61 strains of airborne cyanobacteria and microalgae from the southern Baltic Sea region were isolated from May 2018 to August 2020. Selected microorganisms were tested in controlled laboratory conditions to identify their response to different irradiance (10-190 µmol photons m-2 s-1), temperature (13-23 °C), and salinity conditions (0-36 PSU). The highest numbers of cells (above 30 × 105 cell mL-1) were recorded for cyanobacterium Nostoc sp., and for diatoms Nitzschia sp., Amphora sp., and Halamphora sp. We found that for cyanobacterium Nostoc sp. as well as for green alga Coccomyxa sp. the maximum cell concentrations were recorded at the salinity of 0 PSU. Moreover, cyanobacteria Planktolyngbya contorta, Pseudanabaena catenata, Leptolyngbya foveolarum, Gloeocapsa sp., and Rivularia sp. were able to grow only at a salinity of 0 PSU. On the other hand, in the range of 16-24 PSU, the highest cell numbers of examined diatoms have been identified. Our research provided that deposited airborne microalgae and cyanobacteria showed full colonization potential. The present experiment suggests that the adaptive abilities of microorganisms, in particular those producing toxins, may contribute to the spread in the future. Thus, it may increase human exposure to their negative health effects. Any distinctive adaptations of the genera give them an additional competitive advantage and a greater chance for territorial expansion.

Potential of Exopolysaccharide from Porphyridium marinum to Contend with Bacterial Proliferation, Biofilm Formation, and Breast Cancer.

Exopolysaccharide (EPS) from marine microalgae are promising sources of a new generation of drugs. However, lot of them remain to be discovered and tested. In this study, EPS produced by Porphyridium marinum and its oligomers prepared by High Pressure Homogenizer have been tested for different biological activities, i.e., antibacterial, anti-fungal and antibiofilm activities on Candida albicans, as well as for their effects on the viability of murine breast cancer cells. Results have shown that all EPS samples present some biological activity. For antibacterial and antibiofilm activities, the native EPS exhibited a better efficiency with Minimum Inhibitory Concentration (MIC) from 62.5 µg/mL to 1000 µg/mL depending on the bacterial strain. For Candida albicans, the biofilm formation was reduced by about 90% by using only a 31.3 µg/mL concentration. Concerning breast cancer cells, lower molar masses fractions appeared to be more efficient, with a reduction of viability of up to 55%. Finally, analyses of polymers composition and viscosity measurements were conducted on all samples, in order to propose hypotheses involving the activities caused by the intrinsic properties of polymers.

The first characterization of airborne cyanobacteria and microalgae in the Adriatic Sea region.

The presence of airborne cyanobacteria and microalgae as well as their negative impacts on human health have been documented by many researchers worldwide. However, studies on cyanobacteria and microalgae are few compared with those on bacteria and viruses. Research is especially lacking on the presence and taxonomic composition of cyanobacteria and microalgae near economically important water bodies with much tourism, such as the Adriatic Sea region. Here, we present the first characterization of the airborne cyanobacteria and microalgae in this area. Sampling conducted between 11th and 15th June 2017 revealed a total of 15 taxa of airborne cyanobacteria and microalgae. Inhalation of many of the detected taxa, including Synechocystis sp., Synechococcus sp., Bracteacoccus sp., Chlorella sp., Chlorococcum sp., Stichococcus sp., and Amphora sp., poses potential threats to human health. Aside from two green algae, all identified organisms were capable of producing harmful metabolites, including toxins. Moreover, we documented the presence of the cyanobacterium Snowella sp. and the green alga Tetrastrum sp., taxa that had not been previously documented in the atmosphere by other researchers. Our study shows that the Adriatic Sea region seems to be a productive location for future research on airborne cyanobacteria and microalgae in the context of their impacts on human health, especially during the peak of tourism activity.

Revision of Coelastrella (Scenedesmaceae, Chlorophyta) and first register of this green coccoid microalga for continental Norway.

A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.

Use of organic exudates from two polar diatoms by bacterial isolates from the Arctic Ocean.

Global warming affects primary producers in the Arctic, with potential consequences for the bacterial community composition through the consumption of microalgae-derived dissolved organic matter (DOM). To determine the degree of specificity in the use of an exudate by bacterial taxa, we used simple microalgae-bacteria model systems. We isolated 92 bacterial strains from the sea ice bottom and the water column in spring-summer in the Baffin Bay (Arctic Ocean). The isolates were grouped into 42 species belonging to Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. Forty strains were tested for their capacity to grow on the exudate from two Arctic diatoms. Most of the strains tested (78%) were able to grow on the exudate from the pelagic diatom Chaetoceros neogracilis, and 33% were able to use the exudate from the sea ice diatom Fragilariopsis cylindrus. 17.5% of the strains were not able to grow with any exudate, while 27.5% of the strains were able to use both types of exudates. All strains belonging to Flavobacteriia (n = 10) were able to use the DOM provided by C. neogracilis, and this exudate sustained a growth capacity of up to 100 times higher than diluted Marine Broth medium, of two Pseudomonas sp. strains and one Sulfitobacter strain. The variable bioavailability of exudates to bacterial strains highlights the potential role of microalgae in shaping the bacterial community composition. This article is part of the theme issue 'The changing Arctic Ocean: consequences for biological communities, biogeochemical processes and ecosystem functioning'.

High-Throughput In Vitro Screening of Changed Algal Community Structure Using the PhotoBiobox.

In a previous study, the sequential optimization and regulation of environmental parameters using the PhotoBiobox were demonstrated with high-throughput screening tests. In this study, we estimated changes in the biovolume-based composition of a polyculture built in vitro and composed of three algal strains: Chlorella sp., Scenedesmus sp., and Parachlorella sp. We performed this work using the PhotoBiobox under different temperatures (10-36°C) and light intensities (50-700 µmol/m-2/s-1) in air and in 5% CO2. In 5% CO2, Chlorella sp. exhibited better adaptation to high temperatures than in air conditions. Pearson's correlation analysis showed that the composition of Parachlorella sp. was highly related to temperature whereas Chlorella sp. and Scenedesmus sp. showed negative correlations in both air and 5% CO2. Furthermore, light intensity slightly affected the composition of Scenedesmus sp., whereas no significant effect was observed in other species. Based on these results, it is speculated that temperature is an important factor in influencing changes in algal polyculture community structure (PCS). These results further confirm that the PhotoBiobox is a convenient and available tool for performance of lab-scale experiments on PCS changes. The application of the PhotoBiobox in PCS studies will provide new insight into polyculture-based ecology.

Assessing the response of micro-eukaryotic diversity to the Great Acceleration using lake sedimentary DNA.

Long-term time series have provided evidence that anthropogenic pressures can threaten lakes. Yet it remains unclear how and the extent to which lake biodiversity has changed during the Anthropocene, in particular for microbes. Here, we used DNA preserved in sediments to compare modern micro-eukaryotic communities with those from the end of the 19th century, i.e., before acceleration of the human imprint on ecosystems. Our results obtained for 48 lakes indicate drastic changes in the composition of microbial communities, coupled with a homogenization of their diversity between lakes. Remote high elevation lakes were globally less impacted than lowland lakes affected by local human activity. All functional groups (micro-algae, parasites, saprotrophs and consumers) underwent significant changes in diversity. However, we show that the effects of anthropogenic changes have benefited in particular phototrophic and mixotrophic species, which is consistent with the hypothesis of a global increase of primary productivity in lakes.

Photosynthetic Sponge-associated Eukaryotes in the Aegean Sea: A Culture-dependent Approach.

Symbioses between sponges and photosynthetic organisms are very diverse regarding the taxonomy and biogeography of both hosts and symbionts; to date, most research has focused on the exploration of bacterial diversity. The present study aims to characterize the culturable diversity of photosynthetic eukaryotes associated with sponges in the Aegean Sea, on which no information exists. Five microalgae strains were isolated from marine sponges; the strains were characterized by morphological features, and the 18S rRNA, 18S-28S Internal Transcribed Spacer, and ribulose-bisphosphate carboxylase large chain (rbcL) sequences. Our polyphasic approach showed that the strains belonged to the green-alga Acrochaete leptochaete, the diatom Nanofrustulum cf. shiloi, the rhodophyte Acrochaetium spongicola, and the chlorachniophyte Lotharella oceanica. A. leptochaete is reported for the first time in sponges, even though green algae are known to be associated with sponges. Nanofrustulum shiloi was found in association with the sponges Agelas oroides and Chondrilla nucula, whereas information existed only for its association with the species Aplysina aerophoba. Acrochaetium spongicola was found for the first time in association with sponges in the eastern Mediterranean. Moreover, we report herein for the first time a sponge-chlorarachniophycean association. Our research revealed new diversity of microalgae associated with sponges and added new records of sponge species, previously unknown for their association with microalgae.

Biological Characterization of Water in Damietta Branch of the Nile River, Egypt.

BACKGROUND AND OBJECTIVE: Damietta branch is one of the two main branches of the Nile River (Egypt), that often inhabited by many aquatic organisms, which affect and reflect its water characteristics. This study examine the relation between submerged macrophytes, their epiphytic microalgae and bacterial communities as well as the variations in their distribution and species composition with respect to season and location. MATERIALS AND METHODS: Macrophytes, epiphytes and water samples were collected from 5 sites distributed along Damietta branch. Macrophytes, epiphytes and bacterial indicators of pollution were identified using standard methods. RESULTS: Three submerged macrophytes (Myriophyllum spicatum, Ceratophyllum demersum L. and Potamogeton crispus) and 191 epiphytic algal taxa dominated by 87 Bacillariophyta and 62 Chlorophyta were recorded with significance seasonal and spatial variations. Myriophyllum spicatum was the most frequent macrophyte (p = 100%) and represent about 100, 100, 97.4 and 64.9% kg DW m-2 of the total collected macrophytes biomass during autumn, winter, spring and summer respectively. The relation between some epiphytic algal species and specific macrophytes was evident and the high organic pollution tolerant algal species like, Melosira granulata, Nitzschia palea, Synedra ulna, Oscillatoria limosa, Microcystis aeruginosa were recorded. Results of bacteriological analysis revealed a significance difference in total viable bacterial counts developed on either 22 or 37°C, total coliform, fecal coliform, fecal streptococci and Escherichia coli attributed to the seasons and sites. CONCLUSION: The results indicated different relations between macrophytes, epiphytes and bacteria, which is a useful biological tool for characterization of water quality in Damietta branch for different purposes.

Harvesting of marine microalgae using cationic cellulose nanocrystals.

Flocculation of marine microalgae is challenging because of the high ionic strength of the culture medium. We investigated cationic cellulose nanocrystals (CNCs) as biobased flocculants for the marine microalgae Nannochloropsis oculata, and compared its performance to chitosan. Cationic CNCs induced flocculation at a low dose of 11 mg L-1, while chitosan required a dose of 35 mg L-1. Our cationic CNCs possess a permanent positive charge, allowing flocculation over a wide pH-range (4 to 10). The CNC maximum flocculation efficiency was 90 %, while chitosan achieved > 95 %, attributed to small flocs remaining in suspension for CNCs. However, centrifugation for 1 min at 180 g or gravity filtration using a 30 µm nylon filter after CNC flocculation resulted in the removal of these small, stable flocs (∼150 µm diam) and > 95% harvesting efficiency. Cationic CNCs can thus serve as a sustainable alternative natural flocculant for harvesting both freshwater and marine microalgae.

Seasonal and latitudinal variations in sea ice algae deposition in the Northern Bering and Chukchi Seas determined by algal biomarkers.

An assessment of the production, distribution and fate of highly branched isoprenoid (HBI) biomarkers produced by sea ice and pelagic diatoms is necessary to interpret their detection and proportions in the northern Bering and Chukchi Seas. HBIs measured in surface sediments collected from 2012 to 2017 were used to determine the distribution and seasonality of the biomarkers relative to sea ice patterns. A northward gradient of increasing ice algae deposition was observed with localized occurrences of elevated IP25 (sympagic HBI) concentrations from 68-70°N and consistently strong sympagic signatures from 71-72.5°N. A declining sympagic signature was observed from 2012 to 2017 in the northeast Chukchi Sea, coincident with declining sea ice concentrations. HBI fluxes were investigated on the northeast Chukchi shelf with a moored sediment trap deployed from August 2015 to July 2016. Fluxes of sea ice exclusive diatoms (Nitzschia frigida and Melosira arctica) and HBI-producing taxa (Pleurosigma, Haslea and Rhizosolenia spp.) were measured to confirm HBI sources and ice associations. IP25 was detected year-round, increasing in March 2016 (10 ng m-2 d-1) and reaching a maximum in July 2016 (1331 ng m-2 d-1). Snowmelt triggered the release of sea ice algae into the water column in May 2016, while under-ice pelagic production contributed to the diatom export in June and July 2016. Sea ice diatom fluxes were strongly correlated with the IP25 flux, however associations between pelagic diatoms and HBI fluxes were inconclusive. Bioturbation likely facilitates sustained burial of sympagic organic matter on the shelf despite the occurrence of pelagic diatom blooms. These results suggest that sympagic diatoms may sustain the food web through winter on the northeast Chukchi shelf. The reduced relative proportions of sympagic HBIs in the northern Bering Sea are likely driven by sea ice persistence in the region.

A simple 18S rDNA approach for the identification of cultured eukaryotic microalgae with an emphasis on primers.

Any forms of valorization of microorganisms would require accurate identity recognition to ensure repeatability, reproducibility and quality assurance. This study aimed to evaluate the effectiveness of different primers for identifying cultured eukaryotic microalgae using a simple 18S rDNA approach. A total of 34 isolated microalgae and one culture collection were utilized in the search for an effective molecular identification method for microalgae. Ammonium formate was applied to marine microalgae prior to DNA extraction. The microalgal DNA was extracted using a commercial kit and subjected directly to PCR amplification using four different published 18S rDNA primers. The DNA sequences were analysed using Basic Local Alignment Search Tool (BLAST) and phylogenetic trees to determine the microalgae identity. The identity was further validated with conventional morphological taxonomic identification, and the relationship of microalgal morphology and genetic materials was also determined. The microalgal DNA was successfully amplified, including marine species without prior cleaning. In addition, the ss5 + ss3 primer pair was found to be an ideal primer set among the tested primers for identifying microalgae. Overall, molecular identification showed relative matching with morphological identification (82.86%). This study is important because it serves as a platform to develop a standardized eukaryotic microalgae identification method. In addition, this method could help to ease the eukaryotic microalgae identification process and enrich the current reference databases such as GenBank.

Kinetic characterization and modeling of a microalgae consortium isolated from landfill leachate under a high CO2 concentration in a bubble column photobioreactor

BACKGROUND: The determination of kinetic parameters and the development of mathematical models are of great interest to predict the growth of microalgae, the consumption of substrate and the design of photobioreactors focused on CO2 capture. However, most of the models in the literature have been developed for CO2 concentrations below 10%. RESULTS: A nonaxenic microalgal consortium was isolated from landfill leachate in order to study its kinetic behavior using a dynamic model. The model considered the CO2 mass transfer from the gas phase to the liquid phase and the effect of light intensity, assimilated nitrogen concentration, ammonium concentration and nitrate concentration. The proposed mathematical model was adjusted with 13 kinetic parameters and validated with a good fit obtained between experimental and simulated data. CONCLUSIONS: Good results were obtained, demonstrating the robustness of the proposed model. The assumption in the model of DIC inhibition in the ammonium and nitrate uptakes was correct, so this aspect should be considered when evaluating the kinetics with microalgae with high inlet CO2 concentrations.

Electrochemical immunosensor functionalized with nanobodies for the detection of the toxic microalgae Alexandrium minutum using glassy carbon electrode modified with gold nanoparticles.

In this work an electrochemical immunosensor for the toxic microalgae Alexandrium minutum (A. minutum AL9T) detection is described. A glassy carbon electrode (GCE) was modified by depositing gold nanoparticles followed by L-cysteine for obtaining a self-assembled monolayer. The SpyTagged nanobody C1, specific for the A. minutum toxic strain AL9T, was then covalently immobilized via SpyCatcher on the surface of the modified electrode and used for the selective capture of such microalgae strain. Electrochemical impedance spectroscopy (EIS) was used for the quantification of A. minutum cells present in water samples by measuring the charge-transfer resistance changes of the electrode with a hexacyanoferrate probe. Each electrode modification step was accompanied by cyclic voltammetry (CV) and scanning electron microscopy (SEM). The immunosensor provided highly reproducible data, was simple to fabricate at low cost, exhibited higher sensitivity than previously described alternative diagnostic methods and showed a broad linear range between 103 and 109 cells L-1 with detection limit of 3 × 103 cells L-1 of A. minutum AL9T. The immunosensor was successfully applied to quantify A. minutum AL9T in seawater and brackish water samples proving that it can be used for early detection of harmful microalgae without the necessity of pre-concentration or dialysis steps.