Search Results (37)

Background/Objectives: Microalgae are recognized as prolific producers of bioactive metabolites with pharmaceutical potential. This study aimed to isolate and characterize cytotoxic constituents from selected cytotoxic microalgae, collected in Hue city, Vietnam. Methods: Microalgal samples were collected from freshwater bodies, morphologically identified, and maintained in laboratory culture. Thirteen strains were successfully isolated and cultivated in BG11, Z8, and BBM media to determine optimal growth conditions. Cytotoxic effects of extracts/compounds were determined using the sulforhodamine B assay on human lung cancer (SK-LU-1) and human liver cancer (HepG2) cell lines. The methanol extract was partitioned with n-hexane and CH₂Cl₂, followed by extensive chromatographic separation and HPLC purification to afford twelve compounds, including two new and ten known compounds. The structures were elucidated by HR-ESI-MS and NMR spectra, chemical methods, and comparing compounds in the literature. Results: From the phytoplankton samples collected across six freshwater bodies in Hue city, Vietnam, thirteen microalgal strains were successfully isolated and purified under laboratory conditions. These strains were morphologically and taxonomically identified to be Microcystis aeruginosa HU05, Microcystis viridis HU13, Anabaena circinalis HU08, Aphanizomenon flos-aquae HU02, Dolichospermum smithii HU04, Calothrix braunii HU14, Nostoc muscorum HU12, Nostoc punctiforme HU11, Raphidiopsis raciborskii HU03, Lyngbya spiralis HU15, Planktothrix stagnina HU16, Phormidium subtilis HU06, and Scenedesmus quadricauda HU07. All methanol extracts of those microalgae were evaluated for cytotoxic activity. The MeOH extracts of M. viridis (HU13) and D. smithii (HU04) exhibited significant cytotoxic effects, with IC₅₀ values of 6.19 ± 0.80 and 4.89 ± 0.76 µg/mL for M. viridis, and 9.51 ± 0.84 and 8.32 ± 0.94 µg/mL for D. smithii against SK-LU-1 and HepG2 cell lines, respectively. Furthermore, chemical studies of D. smithii HU04 led to the isolation of two new compounds, smithioside A (1) and smithioside B (2) and ten known ones, 3,4,5-trimethoxyphenyl-1-O-β-D-glucopyranoside (3), 4′-hydroxy-3′-methoxyphenol-β-D-[6-O-(4″-hydroxy-3″,5″-dimethoxylbenzoate)]-glucopyranoside (4), 4′-hydroxy-2′,6′-dimethoxyphenol 1-O-β-D-(6-O-syringoyl)glucopyranoside (5), mallophenol B (6), pisoninol II (7), guaiacylglycerol (8), (E)-asarone (9), deacetylsarmentamide B (10), (E)-2-hexenyl-β-D-glucopyranoside (11), and 5,6-dihydropyridin-2(1H)-one (12). The cytotoxic activity of all isolated compounds was also evaluated against SK-LU-1 and HepG2 cancer cell lines. Compound 12 showed the strongest activity, with IC₅₀ values of 9.13 ± 0.89 µM (SK-LU-1) and 7.64 ± 0.46 µM (HepG2). Compounds 5 and 6 exhibited moderate cytotoxic activity on both human cancer cell lines with IC₅₀ values ranging from 25.99 to 51.47 µM. Conclusions: These results highlight the potential of Dolichospermum smithii HU04 as a source of bioactive compounds, particularly in anticancer applications. These findings suggest that D. smithii HU04 extracts could be developed for therapeutic purposes targeting cancer. Full article

HCBs (Harmful Cyanobacterial Blooms) are increasing in freshwaters across the globe, particularly at lower latitudes. In Southern Europe, a decrease in annual precipitation and an increase in drought periods have enhanced the occurrence of HCBs, impacting both freshwater ecosystems and human health. This review gathers information on isolated cyanobacterial strains with the potential to form cyanobacterial blooms or to be toxic that have been reported over the past half-century in Portugal. Strains of Microcystis aeruginosa are the most represented ones, many of them microcystin producers. Toxic M. aeruginosa strains have been isolated from lakes (Mira, Barrinha de Mira, and Blue), river sections (Tâmega and Guadiana), and reservoirs (Torrão, Vilar, Montargil, Patudos, Caia, Monte da Barca, Corgas, and Magos). Many other strains from potentially toxic species are listed, namely from Aphanizomenon gracile, Aphanizomenon flos-aquae, Sphaerospermopsis aphanizomenoides, Cuspidothrix issatschenkoi, Dolichospermum flos-aquae, Dolichospermum circinalis, Chrysosporum bergii, Raphidiopsis raciborskii or Planktothrix agardhii. Many of the isolated strains were able to produce cyanotoxins such as microcystins, saxitoxins, cylindrospermopsin, or anatoxin. Most isolates belong to the Portuguese culture collections ESSACC (Estela Sousa e Silva Algal Culture Collection); LEGE-CC (Blue Biotechnology and Ecotoxicology Culture Collection); and ACOI (Coimbra Collection of Algae). Despite many strains already having associated molecular data corroborating a correct identification, a large number of strains are still lacking DNA-based information for phylogenetic affiliation. The present checklist is intended to facilitate access to information regarding strains of potentially toxic cyanobacterial species from Portugal in order to contribute to a better understanding of species-specific HCBs at both regional and global scales. Full article

Floodplain lakes play a significant role in maintaining biological diversity and providing a food base for aquatic organisms. In 2023–2024, for the first time, we studied phytoplankton of five floodplain lakes of the transboundary Irtysh River in Kazakhstan. A total of 149 species and forms of planktonic algae were recorded, with a low level of similarity between the lakes. The ratio of indicator species (predominance of eutraphents and meso-eutraphents), abundance (3301.6–168,961.1 thou. cells L⁻¹), biomass (2.41–83.67 mg L⁻¹) of phytoplankton communities, and composition of dominant phyla and species (Cyanobacteria: Microcystis pulverea, M. aeruginosa, Aphanizomenon flos-aquae; Chlorophyta: Volvox globator; Dinoflagellata: Ceratium hirundinella and others) testified to a high level of organic pollution of floodplain lakes. Chemical variables (nitrogen compound content, PI) supported this conclusion. Analysis of the RDA revealed that the biomass of Cyanobacteria was controlled by nitrate nitrogen, while phosphates controlled that of Chlorophyta. The applied integrated approach showed an improvement in the trophic status of lakes in a high-water year and can be useful in assessing the ecological state of aquatic ecosystems in other regions. Full article

Spirulina (Arthrospira spp.) and klamath (Aphanizomenon flos-aquae) are widely consumed cyanobacteria-based food supplements valued for their nutritional and health-promoting properties. However, these products are susceptible to contamination by cyanotoxins, which are potent toxins produced by co-occurring cyanobacteria that may pose health risks to consumers. In this study, we applied an integrated targeted and suspect screening approach to comprehensively assess the presence of cyanotoxins in commercial spirulina- and klamath-based food supplements. Targeted analysis was performed using UHPLC-QqQ under dynamic multiple reaction-monitoring conditions optimized for the determination of twelve cyclic peptide cyanotoxins. Suspect screening was conducted using high-resolution mass spectrometry (HRMS) with a Q-Orbitrap analyser, applying a specific workflow to detect additional related compounds lacking analytical standards. The method enabled the detection and identification of multiple cyanotoxins, including microcystins, nodularin, and anabaenopeptins. The combination of targeted and suspect workflows allowed for a broader coverage of potential related cyanotoxins. Several cyanotoxins were detected in a klamath-based supplement, with high concentrations of microcystin-RR, while additional variants were identified through suspect screening. These findings highlight the need for routine monitoring and stricter regulatory oversight of cyanobacteria-based supplements to ensure consumer safety. Full article

Gehu Lake in the lower reaches of the Taihu Lake Basin has experienced water quality degradation due to increasing human activities, pollutant discharge, and non-point source pollution, which requires ecosystem restoration. Currently, the community structure of aquatic organisms and their influencing environmental factors remain poorly understood. Thus, in this study, we conducted comprehensive fieldwork in June 2024 and analyzed the community structures of plankton (i.e., phytoplankton and zooplankton) and macroinvertebrates, and their influencing environmental factors in Gehu Lake and the inflowing river. The trophic level index (TLI) and biodiversity indices (Shannon–Wiener, Pielou, and Margalef) were utilized to assess water quality status. Pearson correlation analysis and redundancy analysis (RDA) were applied to identify key factors influencing plankton and macroinvertebrate community structures. The dominant phytoplankton species included Merismopedia tranquilla, Microcystis aeruginosa, Aphanizomenon flos-aquae, Aphanocapsa elachista, and Aulacoseira granulata. The dominant zooplankton species were mainly Brachionus diversicornis, Brachionus calyciflorus, and Asplanchna priodonta. The dominant macroinvertebrate species were Microchironomus tabarui and Chironomus flaviplumus. The findings suggest that Gehu Lake exhibited moderate pollution levels, while the diversity indices were significantly correlated with environmental factors. The Shannon–Wiener index of zooplankton displayed a markedly negative correlation with Chl-a (p < 0.05). The results from redundancy analysis showed that TP, TN, SD, COD_Mn, and Chl-a were key environmental factors shaping the aquatic community structure in the lake. Full article

This study presents a biorefinery approach for Aphanizomenon flos-aquae, demonstrating its potential as a dual source for phycocyanin and biogas. The antioxidant capacity of the extract was evaluated using the ABTS^•+ assay, while flow cytometry determined its cytotoxic effects on breast cancer (HCC1806) and brain glioma (U-118 MG) cell lines, comparing pure C-phycocyanin to the non-purified extract. The non-purified extract scavenged 77% of ABTS^•+ radicals at 2.4 mg/mL, compared to 22% for pure C-phycocyanin. In U-118 MG cells, pure C-phycocyanin accounted for 55.5% of the 29.9 ± 6.1% total mortality observed with the non-purified extract at 0.75 mg/mL. HCC1806 cytotoxicity (80.9 ± 5.1% at 1 mg/mL) was attributed to synergistic effects of other extract components. The spent biomass was valorized through anaerobic digestion for biogas production, enhancing process sustainability. At a 2:1 inoculum-to-substrate ratio, the anaerobic digestion of the spent biomass yielded 447 ± 18 mL CH₄/gVS, significantly higher than the 351 ± 19 mL CH₄/gVS from the initial biomass. LCA estimated the environmental impacts of the A. flos-aquae biorefinery for phycocyanin production, targeting the cosmetic, food, and nutraceutical sectors, and highlighting the benefits of spent biomass valorization to produce biogas within a circular economy framework. This integrated approach demonstrates the potential of A. flos-aquae for the sustainable production of high-value compounds and renewable energy. Full article

This study explores the in vitro effects of cyanotoxins from the methanolic extract of the cyanobacteria Aphanizomenon flos-aquae and Microcystis aeruginosa on human blood cells, with samples drawn from the Gruža reservoir in Serbia. These cyanobacteria, which made up 98.5% of the reservoir’s phytoplankton, reached densities of 4,656,450 cells mL⁻¹, with A. flos aquae (3,105,120 cells mL⁻¹) as the dominant species, followed by M. aeruginosa (1,480,130 cells mL⁻¹). A cyanotoxin analysis of biomass detected anatoxin-a (3.56 µg g⁻¹), cylindrospermopsin (6.86 µg g⁻¹), microcystin LR (0.87 µg g⁻¹), and microcystin RR (2.47 µg g⁻¹). This study assessed the genotoxic potential of the methanolic extract of the cyanobacterial biomass by evaluating the DNA damage and the Genetic Damage Index (GDI) in peripheral blood lymphocytes (PBLs) from healthy donors. The results showed a dose-dependent increase in the DNA damage, from 35.67 ± 4.93% at 10 µg mL⁻¹ to 95.67 ± 1.53% at 100 µg mL⁻¹, with a corresponding rise in the GDI from 0.61 ± 0.02 to 2.39 ± 0.07. The extract also caused the concentration-dependent hemolysis of red blood cells, with 5.63% hemolysis at the highest concentration (200 µg mL⁻¹). These findings underscore the significant genotoxic risks posed by cyanotoxins from biomass extracts of A. flos aquae and M. aeruginosa, particularly in water sources used for human consumption. Full article

The present study evaluated the ability of KlamExtra^®, an Aphanizomenon flos aquae (AFA) extract, to counteract metabolic dysfunctions due to a high fat diet (HFD) or to accelerate their reversion induced by switching an HFD to a normocaloric diet in mice with diet-induced obesity. A group of HFD mice was fed with an HFD supplemented with AFA (HFD-AFA) and another one was fed with regular chow (standard diet—STD) alone or supplemented with AFA (STD-AFA). AFA was able to significantly reduce body weight, hypertriglyceridemia, liver fat accumulation and adipocyte size in HFD mice. AFA also reduced hyperglycaemia, insulinaemia, HOMA-IR and ameliorated the glucose tolerance and the insulin response of obese mice. Furthermore, in obese mice AFA normalised the gene and the protein expression of factors involved in lipid metabolism (FAS, PPAR-γ, SREBP-1c and FAT-P mRNA), inflammation (TNF-α and IL-6 mRNA, NFkB and IL-10 proteins) and oxidative stress (ROS levels and SOD activity). Interestingly, AFA accelerated the STD-induced reversion of glucose dysmetabolism, hepatic and VAT inflammation and oxidative stress. In conclusion, AFA supplementation prevents HFD-induced dysmetabolism and accelerates the STD-dependent recovery of glucose dysmetabolism by positively modulating oxidative stress, inflammation and the expression of the genes linked to lipid metabolism. Full article

Cyanobacteria harmful algal blooms (cyanoHABs) can have impacts on human health, aquatic ecosystems, and the economy. Nutrient management is an important mitigation and even remediation strategy. In this work, the paralytic shellfish toxin (PST)-producing Aphanizomenon (Aphan.) flos-aquae (Linnaeus) Ralfs ex Bornet & Flahault (now identified as Aphan. sp.) single filament isolate NH-5 was grown in P-depleted media, N-depleted media, and complete BG-11 media. Growth and heterocyst and vegetative cells were monitored using dry weight and cell counts. Ultrasonication was used to separate heterocysts from vegetative cells. HPLC-FLD with post-column derivatization was used to determine the saxitoxin (STX) and neosaxitoxin (NEOSTX) concentration per cell. Aphan. sp. NH-5 biomass was lower in the P-depleted media than in the N-depleted media and the control, though higher heterocyst counts were detected in the N-depleted media. The heterocyst toxin concentration was significantly higher compared to the vegetative cells for the N-depleted media, control, and P-depleted media. However, no significant differences were found among all preparations with regard to the STX-to-NEOSTX ratio. We conclude that N limitation induced higher heterocyst numbers and that N fixation activity is a factor behind the increase in the STX and NEOSTX production of Aphan. sp. NH-5. Full article

Cyano-phycocyanin (C-PC) is a light-absorbing biliprotein found in cyanobacteria, commonly known as blue-green algae. Due to its antioxidative, anti-inflammatory, and anticancer properties, this protein is a promising substance in medicine and pharmaceuticals. However, cyanobacteria tend to bind heavy metals from the environment, making it necessary to ensure the safety of C-PC for the development of pharmaceutical products, with C-PC isolated from naturally collected cyanobacterial biomass. This study aimed to determine the content of the most toxic heavy metals, arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb) in C-PC isolated from different cyanobacterial biomasses collected in the Kaunas Lagoon during 2019–2022, and compare them with the content of heavy metals in C-PC isolated from cultivated Spirulina platensis (S. platensis). Cyanobacteria of Aphanizomenon flos-aquae (A. flos-aquae) dominated the biomass collected in 2019, while the genus Microcystis dominated the biomasses collected in the years 2020 and 2022. Heavy metals were determined using inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS analysis revealed higher levels of the most investigated heavy metals (Pb, Cd, and As) in C-PC isolated from the biomass with the dominant Microcystis spp. compared to C-PC isolated from the biomass with the predominant A. flos-aquae. Meanwhile, C-PC isolated from cultivated S. platensis exhibited lower concentrations of As and Pb than C-PC isolated from naturally collected cyanobacterial biomass. Full article

Sea urchins rely on chemical senses to localize suitable food resources, therefore representing model species for chemosensory studies. In the present study, we investigated the chemical sensitivity of the Mediterranean sea urchin Paracentrotus lividus to the blue-green alga Aphanizomenon flos-aquae, namely “Klamath”, and to a few amino acids chosen from the biochemical composition of the same algae. To this end, we used the “urchinogram” method, which estimates the movement rate of the sea urchins in response to chemicals. Our results showed that Klamath represents a strong chemical stimulus for P. lividus as it elicits an overall movement of spines, pedicellariae, and tube feet coupled, in some cases, to a coordinated locomotion of the animals. Sea urchins also displayed a sensitivity, even if to a lesser extent, to leucine, threonine, arginine, and proline, thus implying that the amino acids contained in Klamath may account, at least in part, for the stimulating effects exerted by the whole algae. Additionally, our results show that Klamath, as well as spirulina, another blue-green alga with high nutritional value, is very attractive for this sea urchin species. These findings gain further importance considering the potential profit of echinoderms for commercial consumers and their growing role in aquaculture. Klamath and spirulina combine high nutritional profiles with attractive and stimulating abilities and may be considered potential valuable feed supplements in sea urchin aquaculture. Full article

Obesity and related metabolic dysfunctions are associated with neurodegenerative diseases, such as Alzheimer’s disease. Aphanizomenon flos-aquae (AFA) is a cyanobacterium considered a suitable supplement for its nutritional profile and beneficial properties. The potential neuroprotective effect of an AFA extract, commercialized as KlamExtra^®, including the two AFA extracts Klamin^® and AphaMax^®, in High-Fat Diet (HFD)-fed mice was explored. Three groups of mice were provided with a standard diet (Lean), HFD or HFD supplemented with AFA extract (HFD + AFA) for 28 weeks. Metabolic parameters, brain insulin resistance, expression of apoptosis biomarkers, modulation of astrocytes and microglia activation markers, and Aβ deposition were analyzed and compared in the brains of different groups. AFA extract treatment attenuated HFD-induced neurodegeneration by reducing insulin resistance and loss of neurons. AFA supplementation improved the expression of synaptic proteins and reduced the HFD-induced astrocytes and microglia activation, and Aβ plaques accumulation. Together, these outcomes indicate that regular intake of AFA extract could benefit the metabolic and neuronal dysfunction caused by HFD, decreasing neuroinflammation and promoting Aβ plaques clearance. Full article

Global warming and eutrophication are the main factors driving the development of cyanobacterial dominance in aquatic ecosystems. We used a model linking water temperature, oxygen saturation, concentrations of PO₄³⁻, NO₃⁻, NH₄⁺, total dissolved iron (TDFe), and SO₄²⁻ to cyanobacteria to test the turnover patterns of cyanobacterial dominance of non-nitrogen-fixing (chroococcal species) and nitrogen-fixing (filamentous diazotrophic) species. Statistical analysis was performed using decision trees. The dominance patterns of the two morphologically and ecologically distinct cyanobacterial species were associated with different environmental factors. However, SO₄²⁻ was the most important factor that explained whether non-nitrogen-fixing or nitrogen-fixing species would dominate. Other important factors were water temperature, phosphate concentration, and oxygen saturation. The model for dominance of non-nitrogen-fixing species used SO₄²⁻, PO₄³⁻, and water temperature (upper layers), and SO₄²⁻, the ratio of PO₄³⁻/NH₄⁺, and oxygen saturation (bottom layers). In contrast, water temperature, SO₄²⁻, and NH₄⁺ in the upper layers and SO₄²⁻, NH₄⁺, and water temperature in the bottom layers were used for the dominance of nitrogen-fixing species. The dominance of Aphanizomenon flos-aquae was explained by different sets of variables, indicating the presence of different strains of this species. The other cyanobacteria species showed dominance patterns that could be explained by one set of variables. As cyanobacterial blooms proliferate due to climate change, it is important to know which factors, in addition to phosphorus and nitrogen, are crucial for the mass development of the various cyanobacterial species. Full article

Aphanizomenon flos-aquae can form extensive blooms from freshwater to the brackish environment and, being a diazotrophic species, contribute significantly to the nitrogen and carbon cycle. It occurs as single filaments or aggregates and could be used as an alternative nutrients source for bacteria and ciliates. Ciliates are a group of organisms playing a crucial role in the transfer of nitrogen from primary producers to higher trophic levels in aquatic food webs. The aim of the experiment was to study the effects of the cyanobacterium A. flos-aquae on the community of five ciliate species (Spirostomum minus, Euplotes aediculatus, Strobilidium sp., Vorticella sp. and Paramecium tetraurelia). The response of each species to the presence of a low/high cyanobacterial biomass and to the different morphological forms of A. flos-aquae (single filaments or aggregates) was demonstrated. The results of the experiment showed the variability of interactions between the cyanobacterium A. flos-aquae and ciliates and pointed out the possible benefits that A. flos-aquae provides to the ciliates (e.g., a substrate for the development of bacteria as food for ciliates or as a source of nitrogen and carbon). Full article