Costanza Baldisserotto

Coccolitophores, such as Gephyrocapsa huxleyi , can have multiple interesting biotechnological applications thanks to their ability to produce complex structures made of CaCO ₃ , called coccolith, and a biomass rich in valuable molecules. Unfortunately, they usually reach low cellular density in standard conditions, hindering actual industrial applications. To enhance growth, CaCO ₃ may be used instead of standard CaCl ₂ . Here, shell-derived CaCO ₃ was also tested to valorise oyster shells, which are not recyclable and can cause environmental harm if not disposed of correctly. G. huxleyi cultures were grown in f/2 medium prepared using standard artificial seawater, artificial seawater with pure CaCO ₃ in place of CaCl ₂ , and artificial seawater with the pulverised shells of Magallana gigas (Pacific oyster) instead of CaCl ₂ . Cultures were monitored for 14 days, and final dry biomass concentration, photosynthetic pigment and total protein concentrations were evaluated. Moreover, coccolith morphology was checked through scanning electron microscopy. Replacing CaCl ₂ with pure CaCO ₃ increased cell density roughly sixfold relative to the control; using pulverised oyster shells yielded the greatest effect, with cell densities about 26 times higher than CaCl ₂ . Cells grown in the medium added with pulverised oyster shells were smaller, exhibited higher pigment content per dry weight and a protein content comparable with the control. The bulk composition of the oyster shell revealed the presence of trace elements, like Mn, Fe and Cu, that could have enhanced the algal growth. Results indicate pulverised oyster shells can be a sustainable and effective calcium source to valorise waste and enhance G. huxleyi biomass production.

Microalgae are gaining increasing attention for wastewater (WW) depuration thanks to their ability to remove pollutants from WW. As environmental conditions change widely geographically and throughout the year, also reaching growth-limiting temperatures during the cold season, sites and seasons specific tests are needed to assess the actual implementation of microalgae phytoremediation. In this paper, two site-specific prototype-scale experiments were performed to test the ability of an autochthonous microalga to depurate urban WW efficiently during winter. Two setups were tested: one indoor and one outdoor. We evaluated dry biomass production, photosynthetic stress, and nitrogen (N) and phosphorus (P) removals from WW. In addition, Escherichia coli concentration was monitored on the effluent. Despite the limited growth in both conditions, N was largely removed from the medium, with the highest result recorded outdoors (almost 70%). No P removal was recorded, although P content in biomass increased both indoors and outdoors, meaning that multiple processes could occur at the same time. Moreover, a huge decrease in E. coli content was recorded in both conditions, suggesting potential for effluent disinfection.

The global population increase during the last century has significantly amplified freshwater demand, leading to higher wastewater (WW) production. European regulations necessitate treating WW before environmental. Microalgae have gained attention for wastewater treatment (WWT) due to their efficiency in remediating nutrients and pollutants, alongside producing valuable biomass. This study investigates the phycoremediation potential of a Chlorella-like strain isolated from urban WW in a 600L-scale system under winter conditions. Experiments in December 2021 and February 2022 tested the strain’s adaptability to varying environmental conditions, particularly temperatures (min-max temperature range: from −3.69 to 10.61 °C in December and −3.96 to 17.61 °C in February), and its ability to meet legal discharge limits. In December, low temperatures algal growth. Nitrates showed an RE of about 92%, while ammonia slightly decreased (RE, about 32%), and phosphorous remained unchanged. In February, mild temperatures increased algal density (33.3 × 10⁶ cell mL⁻¹) and, at the end of experiment, all nutrients were below legal limits with very high RE % (NH₄⁺, 91.43; PO₄³⁻ 97.32). Both trials showed an E. coli RE, % = 99%. The study highlights the potential of microalgae for WWT and the importance of considering seasonal variations when implementing these systems.

Microalgae are considered promising sustainable sources of natural bioactive compounds to be used in biotechnological sectors. In recent years, attention is increasingly given to the search of microalgae-derived compounds with antioxidant and anti-inflammatory properties for nutraceutical or pharmacological issues. In this context, attention is usually focused on the composition and bioactivity of algae or their extracts, while less interest is driven to their biological features, for example, those related to morphology and cultivation conditions. In addition, specific studies on the antioxidant and anti-inflammatory properties of microalgae mainly concern Chlorella or Spirulina. The present work was focused on the characterization of the Chlorophyta Neochloris oleoabundans under two combinations of cultivation modes: autotrophy and glucose-induced mixotrophy, each followed by starvation. Biomass for morphological and biochemical characterization, as well as for extract preparation, was harvested at the end of each cultivation phase. Analyses indicated a different content of the most important classes of bioactive compounds with antioxidant/anti-inflammatory properties (lipids, exo-polysaccharides, pigments, total phenolics, and proteins). In particular, the most promising condition able to prompt the production of antioxidant algal biomass with anti-inflammatory properties was the mixotrophic one. Under mixotrophy, beside an elevated algal biomass production, a strong photosynthetic metabolism with high appression of thylakoid membranes and characteristics of high photo-protection from oxidative damage was observed and linked to the overproduction of exo-polysaccharides and lipids rather than pigments. Overall, mixotrophy appears a good choice to produce natural bioactive extracts, potentially well tolerated by human metabolism and environmentally sustainable.

Within the ancient vascular plant lineage known as lycophytes, many Selaginella species contain only one giant chloroplast in the upper epidermal cells of the leaf. In deep-shade species, such as S. martensii, the chloroplast is cup-shaped and the thylakoid system differentiates into an upper lamellar region and a lower granal region (bizonoplast). In this report, we describe the ultrastructural changes occurring in the giant chloroplast hosted in the epidermal cells of S. martensii during the daily relocation of the organelle. The process occurs in up to ca. 40% of the microphylls without the plants being exposed to high-light flecks. The relocated chloroplast loses its cup shape: first, it flattens laterally toward the radial cell wall and then assumes a more globular shape. The loss of the conical cell shape, the side-by-side lateral positioning of vacuole and chloroplast, and the extensive rearrangement of the thylakoid system to only granal cooperate in limiting light absorption. While the cup-shaped chloroplast emphasizes the light-harvesting capacity in the morning, the relocated chloroplast is suggested to support the renewal of the thylakoid system during the afternoon, including the recovery of photosystem II (PSII) from photoinhibition. The giant chloroplast repositioning is part of a complex reversible reshaping of the whole epidermal cell.

The release of inadequately treated urban wastewater is the main cause of environmental pollution of aquatic ecosystems. Among efficient and environmentally friendly technologies to improve the remediation process, those based on microalgae represent an attractive alternative due to the potential of microalgae to remove nitrogen (N) and phosphorus (P) from wastewaters. In this work, microalgae were isolated from the centrate stream of an urban wastewater treatment plant and a native Chlorella-like species was selected for studies on nutrient removal from centrate streams. Comparative experiments were set up using 100% centrate and BG11 synthetic medium, modified with the same N and P as the effluent. Since microalgal growth in 100% effluent was inhibited, cultivation of microalgae was performed by mixing tap-freshwater with centrate at increasing percentages (50%, 60%, 70%, and 80%). While algal biomass and nutrient removal was little affected by the differently diluted effluent, morpho-physiological parameters (F_V/F_M ratio, carotenoids, chloroplast ultrastructure) showed that cell stress increased with increasing amounts of centrate. However, the production of an algal biomass enriched in carotenoids and P, together with N and P abatement in the effluent, supports promising microalgae applications that combine centrate remediation with the production of compounds of biotechnological interest; for example, for organic agriculture.

Wheat mutants with a reduced chlorophyll synthesis are affected by a defective control of the photosynthetic electron flow, but tend to recover a wild-type phenotype. The sensitivity of some mutants to light fluctuations suggested that cultivation outdoors could significantly impact productivity. Six mutant lines of Triticum durum or Triticum aestivum with their respective wild-type cultivars were cultivated with a regular seasonal cycle (October–May) in a semi-field experiment. Leaf chlorophyll content and fluorescence parameters were analysed at the early (November) and late (May) developmental stages, and checked for correlation with morphometric and grain-production parameters. The alleviation of the phenotype severity concerned primarily the recovery of the photosynthetic-membrane functionality, but not the leaf chlorophyll content. Photosystem II (PSII) was less photoprotected in the mutants, but a moderate PSII photoinhibition could help control the electron flow into the chain. The accumulation of interchain electron carriers was a primary acclimative response towards the naturally fluctuating environment, maximally exploited by the mature durum-wheat mutants. The mutation itself and/or the energy-consuming compensatory mechanisms markedly influenced the plant morphogenesis, leading especially to reduced tillering, which in turn resulted in lower grain production per plant. Consistently with the interrelation between early photosynthetic phenotype and grain-yield per plant, chlorophyll-fluorescence indexes related to the level of photoprotective thermal dissipation (pNPQ), photosystem II antenna size (ABS/RC), and pool of electron carriers (Sm) are proposed as good candidates for the in-field phenotyping of chlorophyll-deficient wheat.

Microalgae are proposed in several biotechnological fields because of their ability to produce biomass enriched in high-value compounds according to cultivation conditions. Regarding the health sector, an emerging area focuses on natural products exploitable against viruses. This work deals with the characterization of the green microalga Neochloris oleoabundans cultivated under autotrophic and mixotrophic conditions as a source of whole aqueous extracts, tested as antivirals against HCoV-229E (Coronaviridae family). Glucose was employed for mixotrophic cultures. Growth and maximum quantum yield of photosystem II were monitored for both cultivations. Algae extracts for antiviral tests were prepared using cultures harvested at the early stationary phase of growth. Biochemical and morphological analyses of algae indicated a different content of the most important classes of bioactive compounds with antiviral properties (lipids, exo-polysaccharides, and total phenolics, proteins and pigments). To clarify which phase of HCoV-229E infection on MRC-5 fibroblast cells was affected by N. oleoabundans extracts, four conditions were tested. Extracts gave excellent results, mainly against the first steps of virus infection. Notwithstanding the biochemical profile of algae/extracts deserves further investigation, the antiviral effect may have been mainly promoted by the combination of proteins/pigments/phenolics for the extract derived from autotrophic cultures and of proteins/acidic exo-polysaccharides/lipids in the case of mixotrophic ones.

Microalgae are studied as innovative sources of a wide range of highly valuable products, including proteins for the food/feed sectors. However, protein content varies depending on algal species, culture conditions and harvesting period. The Phylum Chlorophyta includes most of the described species of green algae. Due to their remarkable peculiarities, four Chlorophyta species belonging to two distinct classes were selected for the research: Chlorella vulgaris and Chlorella protothecoides as Trebouxiophyceae, and Neochloris oleoabundans and Scenedesmus acutus as Chlorophyceae. The algae were studied to obtain comparative results about their growth performance, and total protein content and profile under the same culture conditions. Since photosynthetic process directly influences biomass production, photosynthetic pigment, PSII maximum quantum yield and thylakoid protein content and profile were analysed. SDS-PAGE and 2D BN/SDS-PAGE were performed to expand information on the organization and assembly of the resolved thylakoid complexes of samples. Despite the algal species showed similar growth rates and photosynthetic efficiency, S. acutus showed the highest capability to accumulate proteins and photosynthetic pigments. Regarding the thylakoid protein profile, the two Trebouxiophyceae showed very similar pattern, whereas different amounts of LHCII occurred within the two Chlorophyceae. Finally, the separation of thylakoid protein complexes in 2D BN/SDS-PAGE revealed a more complex pattern in S. acutus as compared to the other species. Overall, it is suggested that a higher stability of the photosynthetic membranes can result in higher biomass and protein production. Altogether, results have highlighted the metabolic uniqueness of each strain, resulting in a non-obvious comparison with the other species.