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The bioremediation of digestate using microalgae presents a solution to the current eutrophication issue in Northwest Europe, where the use of digestate as soil fertiliser is limited, thus resulting in an excess of digestate. Ammonium is the main nutrient of interest in digestate for microalgal cultivation, and improving its availability and consequent uptake is crucial for optimal bioremediation. This work aimed to determine the influence of pH on ammonium availability in cultures of two green microalgae, additionally screened for their growth performances on three digestates produced from different feedstocks, demonstrating the importance of tailoring a microalgal strain and digestate for bioremediation purposes. Results showed that an acidic pH of 6–6.5 resulted in a better ammonium availability in the digestate media, translated into better growth yields for both S. obliquus (GR: 0.099 ± 0.001 day−1; DW: 0.23 ± 0.02 g L−1) and C. vulgaris (GR: 0.09 ± 0.001 day−1; DW: 0.49 ± 0.012 g L−1). This result was especially true when considering larger-scale applications where ammonium loss via evaporation should be avoided. The results also demonstrated that digestates from different feedstocks resulted in different growth yields and biomass composition, especially fatty acids, for which, a digestate produced from pig manure resulted in acid contents of 6.94 ± 0.033% DW and 4.91 ± 0.3% DW in S. obliquus and C. vulgaris, respectively. Finally, this work demonstrated that the acclimation of microalgae to novel nutrient sources should be carefully considered, as it could convey significant advantages in terms of biomass composition, especially fatty acids and carbohydrate, for which, this study also demonstrated the importance of harvesting time.

Carotenoids in cyanobacteria play an important role in protecting against and in repairing damage against low level UV-B radiation. Here we use transcriptomics and metabolomic HPLC pigment analysis to compare carotenoid pathway regulation in the filamentous cyanobacterium Chlorogloeopsis fritschii PCC 6912 exposed to white light and to white light supplemented with low level UV-B. Under UV-B changes in carotenoid transcription regulation were found associated with carotenogenesis (carotenoid synthesis), photoprotection and carotenoid cleavage. Transcriptional regulation was reflected in corresponding pigment signatures. All carotenogenesis pathway genes from geranylgeranyl-diphosphate to lycopene were upregulated. There were significant increases in expression of gene homologs (crtW, crtR, cruF, and cruG) associated with routes to ketolation to produce significant increases in echinenone and canthaxanthin concentrations. There were gene homologs for four β-carotene-ketolases (crtO and crtW) present but only one crtW was upregulated. Putative genes encoding enzymes (CruF, CrtR, and CruG) for the conversion of γ-carotene to myxol 2′-methylpentoside were upregulated. The hydroxylation pathway to nostaxanthin via zeaxanthin and caloxanthin (gene homologs for CrtR and CrtG) were not upregulated, reflected in the unchanged corresponding pigment concentrations in zeaxanthin, caloxanthin and nostaxanthin, Transcripts for the non-photochemical quenching related Orange-Carotenoid-Protein (OCP) and associated Fluoresence-Recovery-Protein (FRP) associated with photoprotection were upregulated, and one carotenoid binding Helical-Carotenoid-Protein (HCP) gene homolog was downregulated. Multiple copies of genes encoding putative apocarotenoid related carotenoid oxygenases responsible for carotenoid cleavage were identified, including an upregulated apo-β-carotenal-oxygenase gene homologous to a retinal producing enzyme. Our study provides holistic insight into the photoregulatory processes that modulate the synthesis, photoprotection and cleavage of carotenoids in cyanobacterial cells exposed to low level UV-B. This is important to understanding how regulation of metabolism responds to a changing environment and how metabolism can be modulated for biotechnological purposes.

Improving mass cultivation of cyanobacteria is a goal for industrial biotechnology. In this study, the mass cultivation of the thermophilic cyanobacterium Chlorogloeopsis fritschii was assessed for biomass production under light-emitting diode white light (LEDWL), far-red light (FRL), and combined white light and far-red light (WLFRL) adaptation. The induction of chl f was confirmed at 24 h after the transfer of culture from LEDWL to FRL. Using combined light (WLFRL), chl f, a, and d, maintained the same level of concentration in comparison to FRL conditions. However, phycocyanin and xanthophylls (echinone, caloxanthin, myxoxanthin, nostoxanthin) concentration increased 2.7−4.7 times compared to LEDWL conditions. The productivity of culture was double under WLFRL compared with LEDWL conditions. No significant changes in lipid, protein, and carbohydrate concentrations were found in the two different light conditions. The results are important for informing on optimum biomass cultivation of this species for biomass production and bioactive product development.

Several factors have the potential to influence microalgae growth. In the present study, nitrogen concentration and light intensity were evaluated in order to obtain high biomass production and high phycoerythrin accumulation from Porphyridium purpureum. The range of nitrogen concentrations evaluated in the culture medium was 0.075−0.450 g L−1 and light intensities ranged between 30 and 100 μmol m−2 s−1. Surprisingly, low nitrogen concentration and high light intensity resulted in high biomass yield and phycoerythrin accumulation. Thus, the best biomass productivity (0.386 g L−1 d−1) and biomass yield (5.403 g L−1) were achieved with NaNO3 at 0.075 g L−1 and 100 μmol m−2 s−1. In addition, phycoerythrin production was improved to obtain a concentration of 14.66 mg L−1 (2.71 mg g−1 of phycoerythrin over dry weight). The results of the present study indicate that it is possible to significantly improve biomass and pigment production in Porphyridium purpureum by limiting nitrogen concentration and light intensity.

Cyanobacteria have many defence strategies to overcome harmful ultraviolet (UV) stress including the production of secondary metabolites. Metabolomics can be used to investigate this altered metabolism via targeted and untargeted techniques. In this study we assessed the changes in the intra- and extracellular low molecular weight metabolite levels of Chlorogloeopsis fritschii (C. fritschii) during 48 h of photosynthetically active radiation (PAR) supplemented with UV-B (15 µmol m−2 s−1 of PAR plus 3 µmol m−2 s−1 of UV-B) and intracellular levels during 48 h of PAR only (15 µmol m−2 s−1) with sampling points at 0, 2, 6, 12, 24 and 48 h. Gas chromatography–mass spectrometry (GC–MS) was used as a metabolite profiling tool to investigate the global changes in metabolite levels. The UV-B time series experiment showed an overall significant reduction in intracellular metabolites involved with carbon and nitrogen metabolism such as the amino acids tyrosine and phenylalanine which have a role in secondary metabolite production. Significant accumulation of proline was observed with a potential role in stress mitigation as seen in other photosynthetic organisms. 12 commonly identified metabolites were measured in both UV-B exposed (PAR + UV-B) and PAR only experiments with differences in significance observed. Extracellular metabolites (PAR + UV-B) showed accumulation of sugars as seen in other cyanobacterial species as a stress response to UV-B. In conclusion, a snapshot of the metabolome of C. fritschii was measured. Little work has been undertaken on C. fritschii, a novel candidate for use in industrial biotechnology, with, to our knowledge, no previous literature on combined intra- and extracellular analysis during a UV-B treatment time-series. This study is important to build on experimental data already available for cyanobacteria and other photosynthetic organisms exposed to UV-B.

We evaluate the effects of nutrient limitation on cellular composition of polar lipid classes/species in Chlorella sp. using modern polar lipidomic profiling methods (liquid chromatography⁻tandem mass spectrometry; LC-MS/MS). Total polar lipid concentration was highest in nutrient-replete (HN) cultures with a significant reduction in monogalactosyldiacylglycerol (MGDG), phosphatidylglycerol (PG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE) class concentrations for nutrient-deplete (LN) cultures. Moreover, reductions in the abundance of MGDG relative to total polar lipids versus an increase in the relative abundance of digalactosyldiacylglycerol (DGDG) were recorded in LN cultures. In HN cultures, polar lipid species composition remained relatively constant throughout culture with high degrees of unsaturation associated with acyl moieties. Conversely, in LN cultures lipid species composition shifted towards greater saturation of acyl moieties. Multivariate analyses revealed that changes in the abundance of a number of species contributed to the dissimilarity between LN and HN cultures but with dominant effects from certain species, e.g., reduction in MGDG 34:7 (18:3/16:4). Results demonstrate that Chlorella sp. significantly alters its polar lipidome in response to nutrient limitation, and this is discussed in terms of physiological significance and polar lipids production for applied microalgal production systems.

Marine chemical ecology comprises the study of the production and interaction of bioactive molecules affecting organism behavior and function. Here we focus on bioactive compounds and interactions associated with phytoplankton, particularly bloom-forming diatoms, prymnesiophytes and dinoflagellates. Planktonic bioactive metabolites are structurally and functionally diverse and some may have multiple simultaneous functions including roles in chemical defense (antipredator, allelopathic and antibacterial compounds), and/or cell-to-cell signaling (e.g., polyunsaturated aldehydes (PUAs) of diatoms). Among inducible chemical defenses in response to grazing, there is high species-specific variability in the effects on grazers, ranging from severe physical incapacitation and/or death to no apparent physiological response, depending on predator susceptibility and detoxification capability. Most bioactive compounds are present in very low concentrations, in both the producing organism and the surrounding aqueous medium. Furthermore, bioactivity may be subject to synergistic interactions with other natural and anthropogenic environmental toxicants. Most, if not all phycotoxins are classic secondary metabolites, but many other bioactive metabolites are simple molecules derived from primary metabolism (e.g., PUAs in diatoms, dimethylsulfoniopropionate (DMSP) in prymnesiophytes). Producing cells do not seem to suffer physiological impact due to their synthesis. Functional genome sequence data and gene expression analysis will provide insights into regulatory and metabolic pathways in producer organisms, as well as identification of mechanisms of action in target organisms. Understanding chemical ecological responses to environmental triggers and chemically-mediated species interactions will help define crucial chemical and molecular processes that help maintain biodiversity and ecosystem functionality.

The nutrient removal efficiency of Chlorella vulgaris cultivated in domestic wastewater was investigated, along with the potential to use membrane filtration as a pre-treatment tool during the wastewater treatment process. Chlorella vulgaris was batch cultivated for 12 days in a bubble column system with two different wastewater treatments. Maximum uptake of 94.18% ammonium (NH4-N) and 97.69% ortho-phosphate (PO4-P) occurred in 0.2 μm membrane filtered primary wastewater. Membrane filtration enhanced the nutrient uptake performance of C. vulgaris by removing bacteria, protozoa, colloidal particles and suspended solids, thereby improving light availability for photosynthesis. The results of this study suggest that growing C. vulgaris in nutrient rich membrane filtered wastewater provides an option for domestic wastewater treatment to improve the quality of the final effluent.

The phytoplankton community assemblage in surface water of the English Channel (Station L4) was measured and compared from March 1999 to October 2002 using two different methods. Pigment-CHEMTAX was used to derive class apportioned chlorophyll a (Chl a) and cell counts obtained using microscopy were used to derive phytoplankton carbon (phyto-C) estimations. Phyto-C (10340 g C L1) showed a strong linear relationship with total Chl a (0.54.8 g Chl a L1) when the Chl a:phyto-C ratio was >0.04 (r2 = 0.80; average Chl a:phyto-C = 0.044) but the relationship was weaker when the Chl a:phyto-C ratio was <0.04 (r2 = 0.42; average Chl a:phyto-C = 0.013). Correlation between class biomass estimates for phyto-C and Chl a was strong for diatoms during winter (r2 = 0.80) but poor for other classes during both summer and winter. The Chl a:phyto-C ratio declined as irradiance increased with strongest correlation on clear sky days in 2001 when diatoms dominated (r2 = 0.80). The Chl a:phyto-C versus irradiance relationship agrees with that produced from an empirically based dynamic model of phytoplankton acclimation to light. Results are discussed in relation to differences in the techniques used, the species present, cellular pigment concentrations and surface mixed layer irradiance. [ABSTRACT FROM AUTHOR]

The microbial dynamics during a spring diatom bloom decline was monitored in the Northeast Atlantic during a 5-day Lagrangian study (8–12 April 2002). Phytoplankton abundance, composition and health status were related to viral and bacterial abundance, zooplankton abundance and grazing rates, as well as bacterial production. Phytoplankton reached maximum concentration on Day 3 (Chl a >5 μg L−1) and declined on Day 5 (Chl a ∼2 μg L−1) and was dominated (70% of Chl a) by diatoms. Bacterial production increased substantially to >20 μg C L−1 day−1 on Day 3 and concomitantly large viruses decreased in number by half to <10 × 103 mL−1. This was followed by a 5-fold increase in large viruses on Day 5, indicating infection and subsequent lysis on Days 3 and 5, respectively. Micro- and mesozooplankton grazing were not the principal cause for the decline of the bloom and pheophorbide-a showing little variation in concentration from Days 1–4 (∼100 ng L−1) although doubled on Day 5. The poor physiological status of the diatoms, indicated by the high chlorophyllide-a concentrations (50–480 ng L−1), likely promoted a series of closely interrelated events involving bacteria and viruses leading to the demise of the diatom bloom. [ABSTRACT FROM AUTHOR]

Using algal-derived mycosporine-like amino acids (MAAs) in sunscreen formulations is constrained by low cellular concentrations of MAAs and by the high costs associated with harvesting algal cells and extracting the MAAs. Here, we report an industrial scalable method using a membrane filtration approach to purify and concentrate aqueous extracts of MAAs. The method includes an additional biorefinery step enabling purification of phycocyanin, an established valuable natural product. Cultivated cells of the cyanobacterium Chlorogloeopsis fritschii (PCC 6912) were concentrated and homogenised to produce a feed for sequential processing through three membranes of decreasing pore size to obtain a retentate and permeate for each step. Microfiltration (0.2 µm) was used to remove cell debris. Ultrafiltration (10,000 Da) was used to remove large molecules and recover phycocyanin. Finally, nanofiltration (300–400 Da) was used to remove water and other small molecules. Permeate and retentate were analysed using UV-visible spectrophotometry and HPLC. The initial homogenised feed had a shinorine concentration of 5.6 ± 07 mg L−1. The final nanofiltered retentate resulted in a 3.3 times-purified concentrate (shinorine concentration of 18.71 ± 0.29 mg L−1). Significant process losses (35%) highlight scope for improvement. Results confirm the potential of membrane filtration to purify and concentrate aqueous solutions of MAAs with simultaneous separation of phycocyanin highlighting a biorefinery approach.

Ultraviolet A (UV-A) is the major component of UV radiation reaching the Earth’s surface, causing indirect damage to photosynthetic organisms via the production of reactive oxygen species (ROS). In comparison, UV-B causes both direct damage to biomolecules and indirect damage. UV-B is well studied in cyanobacterial research due to their long evolutionary history and adaptation to high levels of UV, with less work on the effects of UV-A. In this study, the response of key metabolites in Chlorogloeopsis fritschii (C. fritschii) during 48 h of photosynthetically active radiation (PAR, 15 µmol·m−2·s−1) supplemented with UV-A (11 µmol·m−2·s−1) was investigated using gas chromatography- mass spectrometry (GC-MS). Results showed an overall significant increase in metabolite levels up to 24 h of UV-A exposure. Compared with previously reported UV-B (PAR + UV-B) and PAR only results, UV-A showed more similarity compared to PAR only exposure as opposed to supplemented UV-B. The amino acids glutamate, phenylalanine and leucine showed differences in levels between UV (both supplemented UV-A and supplemented UV-B) and PAR only (non-supplemented PAR), hinting to their relevance in UV stress response. The fatty acids, palmitic and stearic acid, showed positive log2 fold-change (FC) in supplemented UV-A and PAR only experiments but negative log2 FC in UV-B, indicating the more harmful effect of UV-B on primary metabolism. Less research has been conducted on UV-A exposure and cyanobacteria, a potential environmental stimuli for the optimisation of metabolites for industrial biotechnology. This study will add to the literature and knowledge on UV-A stress response at the metabolite level in cyanobacteria, especially within the less well-known species C. fritschii.

We provide a direct comparison of the distribution and abundance of mycosporine-like amino acids (MAAs) in a diverse range of microalgal cultures (33 species across 13 classes) grown without supplementary ultraviolet radiation (UV). We compare the MAAs in cultures with those present in characterised natural phytoplankton populations from the English Channel. We detected 25 UV absorbing compounds including at least two with multiple absorption maxima. We used LC-MS to provide chemical characterisation of the six most commonly occurring MAAs, namely, palythene, palythine, mycosporine-glycine, palythenic acid, porphyra-334 and shinorine. MAAs were abundant (up to 7 pg MAA cell−1)in 10 species, with more minor and often unknown MAAs in a further 11 cultures. Shinorine was the most frequently occurring and abundant MAA (up to 6.5 pg cell−1) and was present in all but two of the MAA-containing species. The study provides further insight into the diversity and abundance of MAAs important from an ecological perspective and as potential source of natural alternatives to synthetic sunscreens.