To investigate the modification of Pb2+ adsorption of the functional groups of Cladophora rupestris (C. rupestris) biomass by gamma radiation (60Coγ-ray), the interface structure, chemical properties, adsorption behaviors, and Pb2+ adsorption mechanisms of C. rupestris biomass were investigated after irradiation with varying doses of 60Coγ-ray. The results indicate that 60Coγ-ray significantly changed the surface characteristics and interfacial chemistry of the C. rupestris biomass.This led to fracturing and fragmentation that produced a larger specific surface area and more abundant pore structure, increasing the electronegativity in the C. rupestris biomass. The theoretical Pb2+ adsorption capacity increased significantly (2.6-2.9 times) after 60Coγ-ray irradiation. 60Coγ-ray caused preferential degradation of protein components in the dissolved organic matter of the C. rupestris biomass, and protein deamination increased the absorption sites of cations. In the C. rupestris biomass, 60Coγ-ray altered the elemental composition and functional groups, particularly the carbon- and oxygen-containing functional groups, to improve Pb2+ adsorption. In conclusion, 60Coγ-ray can activate the functional groups of C. rupestris biomass and improve their Pb2+ adsorption sites. This study provides new insight into modification of biomass materials for enhanced removal of heavy metals from waterbodies.

Abstract Cellulose from macroalgae is potential for the extraction of nanomaterials due to its availability and its great fraction in the biomass. Cladophora rupestris is a macroalgae and is a good material to which a cellulose may be extracted. The said macroalgae has no value-added products or uses yet. It has a lignocellulosic profile of: alpha-cellulose, 33.43 %; hemicellulose, 15.48%; holocellulose, 48.93; acid soluble lignin, 2.22%; acid insoluble lignin, 22.33%, extractives, 5.67% and total ash, 39.65%. Identifying the best method and conditions for the extraction of cellulose from the said material is a challenge that needs to be addressed. The process for the cellulose extraction considered variations on drying (oven, air, and sun drying), defatting (using solvents: methanol and hexane; and extraction time: 8 hrs and 4 days), solvent drying (oven and air drying), alkaline pre-treatment (0.1, 0.5, and 1M NaOH), fixed conditions on bleaching, bleached biomass oven drying. The biomass weight losses were monitored for some steps, attributing this on the removal of the lignin and extractives. The lignocellulosic profile of the crude cellulose extracted from the macroalgae using the best conditions was determined indicating an increased fraction of the cellulose components in the biomass and a transformation of the rigid biomass into a soft and paper-like texture. The functional groups present on the cellulose was determined. The extracted cellulose was used to produced cellulose nanofibril (CNF) with fiber diameter of the CNF ranges on 14.29 – 37.50 nm and crystallinity index of 92.48%. Attempts were done on extraction of cellulose nanocrystal (CNC) utilizing microwave-assisted acid hydrolysis (MAAH) and semi-batch acid hydrolysis (SBAH); and variation on the acid used (sulfuric acid, and oxalic acid), concentration of the acid, and the temperature conditions.

Cladophora rupestris is ubiquitous in many kinds of waterbodies, and C. rupestris biomass can serve as a carrier for adsorbing and transferring heavy metals. Batch experiments and characterization were performed. Results showed that the organic frameworks of C. rupestris (CROF) had a specific surface area of 2.58 m2/g and an external surface area of 2.06 m2/g. Many mesopores were present in CROF, mainly distributed in 2.5-7.5 nm. The zeta potentials were within the range of - 4.46 to - 13.98 mV in the tested pH of 2.0-9.0. CROF could effectively adsorb Pb2+ in large pH range. The maximum adsorption capacity (qmax) of Pb2+ on CROF was 15.02 mg/g, and 97% of Pb2+ was adsorbed onto CROF after 25 min. CROF had a preferential adsorption of Pb2+. The protein secondary structures and carbon skeletons of CROF all worked in adsorption. The main Pb2+ adsorption mechanisms were pore filling, electrostatic attraction, Pb-π interaction, and surface complexation. Therefore, it is valuable as a biosorbent for the removal of Pb2+ from waterbodies.

Near-infrared spectroscopy bioprobe estimation of metabolites’ responses to Pb2+ in Cladophora rupestris

The continental coasts and remote islands in the high-latitude Southern Hemisphere, including the subantarctic region, are characterized by many endemic species, high abundance of taxa, and intermediate levels of biodiversity. The macroalgal flora of these locations has received relatively little attention. Filamentous green algae are prolific in the intertidal of southern islands, but the taxonomy, distribution, and evolutionary history of these taxa are yet to be fully explored, mostly due to the difficulty of access to some of these locations. In this study, we examined specimens of the order Cladophorales from various locations in the high-latitude Southern Hemisphere including the subantarctic (the Auckland Islands, Bounty Islands, Campbell Island, Macquarie Island, and Kerguelen Islands), as well as mainland New Zealand, the Chatham Islands, Chile, and Tasmania. The analyses of the rDNA sequences of the samples revealed the existence of two new clades in a phylogeny of the Cladophoraceae. One of these clades is described as the novel genus Vandenhoekia gen. nov., which contains three species that are branched or unbranched. The amended genus Rama is reinstated to accommodate the other clade, and contains four species, including the Northern Hemisphere "Cladophora rupestris." In Rama both branched and unbranched morphologies are found. It is remarkable that gross morphology is not a predictor for generic affiliations in these algae. This study illustrates that much can still be learned about diversity in the Cladophorales and highlights the importance of new collections, especially in novel locations.

Phytochelatins (PCs) exist widely in plants and are closely related to plant resistance to heavy metals (HMs). Glutathione (GSH) and nonprotein thiols (NPTs) are the major components of PCs. This study investigates the role of the PCs of Cladophora rupestris in Pb 2+ accumulation and detoxification. The distribution of Pb 2+ in the PCs of C. rupestris was studied. FTIR and XPS are used to characterize the chelating power of Pb 2+ with PCs in C. rupestris. The curve fitting of the secondary protein structure is used to identify the functional groups with Pb 2+ . Results showed that the content of Pb 2+ in the PCs of C. rupestris increased with an increase in Pb 2+ stress. Pb content increased to 352 and 314 mg/kg in NPTs and GSH, respectively, when Pb stress concentration reached 7.5 mg/dm 3 . The Pb 2+ fraction of C. rupestris PCs reached a maximum of 10.8 and 9.3% in NPTs and GSH, respectively. The Pb 2+ uptake by GSH and NPTs was 40-48% and 52-60%, respectively. Pb 2+ bound with the PCs of C. rupestris, forming

This study aimed to analyze the transcriptome of C. rupestris under Pb2+ stress by using high-throughput sequencing technology, observe the changes of gene expression and metabolic pathway after three and five days under 1.0 and 5.0 mg/L of Pb2+ treatment, and analyze the differentially expressed genes (DEGs) and related functional genes after Pb2+ treatment. Metabolic pathways were revealed through Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Results show that DEGs increased significantly with the increase of Pb2+ concentration and stress time. A total of 32 genes were closely related to Pb2+ stress response. GO analysis identified two major transporter proteins, namely, ATP-binding transport protein-related (ABC transporters) and zinc finger CCHC domain containing protein (Zfp) in C. rupestris. Pthr19248, pthr19211, Zfp pthr23002, Zfp p48znf pthr12681, Zfp 294 pthr12389, and Zfp pthr23067 played important roles against Pb2+ toxicity and its absorption in C. rupestris. KEGG pathway analysis suggested that ABCA1, ATM, and ABCD3 were closely related to Pb2+ absorption. Pb2+ stress was mainly involved in metallothionein (MT), plant hormone signal transduction, ABC transporters, and glutathione (GSH) metabolism.

Algal biomass (AB) is prospective source of valuable compounds, however, Baltic Sea macroalgae have some challenges, because of their high microbial and chemical contamination. These problems can be solved, by using appropriate technologies for AG pre-treatment. The aim of this study was to evaluate the influence of two pre-treatments, solid-state fermentation with the Lactiplantibacillus plantarum LUHS135 and ultrasonication, on the antioxidant and antimicrobial characteristics of macro- (Cladophora rupestris, Cladophora glomerata, Furcellaria lumbricalis, Ulva intestinalis) and Spirulina (Arthrospira platensis) extracts. Also, combinations of extracts and LUHS135 were developed and their characteristics were evaluated. The total phenolic compound content was determined from the calibration curve and expressed in mg of gallic acid equivalents; antioxidant activity was measured by a Trolox equivalent antioxidant capacity assay using the DPPH• (1,1-diphenyl-2-picrylhydrazyl), ABTS•+ 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid), FRAP (Ferric Reducing Ability of Plasma) discoloration methods. Antimicrobial activity was measured by using agar well diffusion assay and in a liquid medium. The highest DPPH• and ABTS•+ was shown by C.rupestris and F.lumbricalis extract × LUHS135 combinations, the highest FRAP - by non-pretreated C.rupestris and F.lumbricalis extract × LUHS135 combinations. Ultrasonicated samples inhibited four out of seven tested pathogens. Finally, the tested pre-treatments showed good perspectives and can be recommended for AB valorization.

In this study, the combined forms of Pb in Cladophora rupestris (L.) (C. rupestris) were investigated via X-ray photoelectron spectroscopy (XPS) and nuclear magnetic resonance (NMR), different Pb concentrations (0, 0.5, and 5.0 mg/L), and C. rupestris subcells were explored. Results showed that combined forms of Pb mainly account for Pb-polysaccharides (Pb-OH of carbohydrates) in the cell wall, Pb-protein (Pb-N= and (C-N-)2Pb) in the organelle, and Pb-organic acid (Pb-sulfates, (CO)2-Pb and (COO)2-Pb) in the soluble fraction. Pb-S-containing group (Pb-C-S) could formed in subcelluar when C. rupestris was subjected to high Pb stress. Meanwhile, Pb2+ could penetrate the C. rupestris cells via the formed chelate between GSH/MT and -OH functional groups. Results could help understand the role of subcellular fraction in the algae remediation and detoxification to heavy metal.

The aim of this study was to evaluate the characteristics of macroalgae (Cladophora rupestris, Furcellaria lumbricalis, Ulva intestinalis) and microalgae (Arthrospira platensis (Sp1, Sp2), Chlorella vulgaris) extracts, including micro- and macroelement transition to extract, antioxidant, antimicrobial properties, the concentrations of chlorophyll (-a, -b), and the total carotenoid concentration (TCC). In macroalgae, the highest TCC and chlorophyll content were found in C. rupestris. In microalgae, the TCC was 10.1-times higher in C. vulgaris than in Sp1, Sp2; however, the chlorophyll contents in C. vulgaris samples were lower. A moderate negative correlation was found between the chlorophyll-a and TCC contents (r = −0.4644). In macroalgae extract samples, C. rupestris and F. lumbricalis showed the highest total phenolic compound content (TPCC). DPPH antioxidant activity and TPCC in microalgae was related to the TCC (r = 0.6191, r = 0.6439, respectively). Sp2 extracts inhibited Staphylococcus haemolyticus; C. rupestris, F. lumbricalis, U. intestinalis, and Sp2 extracts inhibited Bacillus subtilis; and U. intestinalis extracts inhibited Streptococcus mutans strains. This study showed that extraction is a suitable technology for toxic metal decontamination in algae; however, some of the desirable microelements are reduced during the extraction, and only the final products, could be applied in food, feed, and others.

Influence of fermentation on the characteristics of Baltic Sea macroalgae, including microbial profile and trace element content

This study investigated the effects of various Pb concentrations and different exposure times on the Cladophora rupestris, the subcellular distributions, and ultrastructures were investigated using air acetylene flame atomic absorption spectrometry, transmission electron microscopy (TEM), and energy dispersive X-ray (EDX). Results showed that Pb was mainly distributed in the cell wall, followed by organelle fractions under low concentration of Pb (≤5.0 mg/L), whereas its relative content in soluble fraction was higher than that in organelle when its concentration exceeded 5.0 mg/L. With the increase in exposure time, the proportion of Pb in the cell wall decreased by 13.14%, and the organelle and soluble fractions increased significantly. TEM and EDX revealed that C. rupestris could accumulate Pb intracellularly and extracellularly. The ultrastructure of C. rupestris changed markedly under 7.5 mg/L of Pb stress, the cells underwent plasmolysis, and some chloroplasts were swollen into a sphere containing starch granules or destroyed into fragmented structure. This study provided an evidence that C. rupestris uses cell wall sequestration and vacuolar compartmentalization to reduce the migration of Pb. Highlights Pb was mainly distributed in the cell wall, followed by organelle fractions under low concentration (≤5.0 mg/L), whereas its relative content in soluble fraction was higher than that in organelle when its concentration exceeded 5.0 mg/L. With the increase in exposure time, the organelle and soluble fractions increased significantly. TEM and EDX revealed that C. rupestris could accumulate Pb intracellularly and extracellularly. The ultrastructure of C. rupestris changed markedly under 7.5 mg/L of Pb stress.

Combined forms of Pb and its detoxification and absorption in Cladophora rupestris subcells

This study aimed to determine the role of sulfhydryl compounds in the subcells of C. rupestris under Pb stress. Different concentrations (0, 0.5, 1.0, 2.5, 5.0, 7.5, and 10mg/L) and different exposure days (1, 3, 5, and 7days) were designed to analyze the subcellular distribution of non-protein thiols (NPT), glutathione (GSH), and phytochelatins (PCs) in C. rupestris. NPT, GSH, and PCs increased significantly with increasing Pb stress in the cell wall and soluble fraction, especially NPT. NPT and GSH slowly increased, and PCs showed no significant difference in the organelle of C. rupestris at low concentrations (< 5.0mg/L). PCs slightly increased under 5.0mg/L of Pb stress. PCs/NPT gradually increased with Pb stress at a low Pb concentration. GSH detoxification response lagged behind those of NPT and PCs in response to time. PCs/NPT initially increased and then decreased with Pb stress duration. This study suggested that NPT, GSH, and PCs played an important role in the detoxification of the cell wall and the soluble fraction of C. rupestris under Pb stress. PCs were important in the organelle.

Simple SummaryThe use of seaweeds as ingredients of ruminant diets can be an alternative to conventional feedstuffs, but it is necessary to assess their nutritive value. The aim of this study was to analyze the chemical composition and in vitro rumen fermentation of eight brown, red and green seaweed species collected in Norway during both spring and autumn. The in vitro ruminal fermentation characteristics of 17 diets composed of oat hay:concentrate in a 1:1 ratio, with the concentrate containing no seaweed or including one of the 16 seaweed samples, was also studied. Species and season determined differences in chemical composition and in vitro fermentation of seaweeds. Most of the tested seaweeds can be included in the diet (up to 200 g/kg concentrate) without negative effects on in vitro ruminal fermentation.This study was designed to analyze the chemical composition and in vitro rumen fermentation of eight seaweed species (Brown: Alaria esculenta, Laminaria digitata, Pelvetia canaliculata, Saccharina latissima; Red: Mastocarpus stellatus, Palmaria palmata and Porphyra sp.; Green: Cladophora rupestris) collected in Norway during spring and autumn. Moreover, the in vitro ruminal fermentation of seventeen diets composed of 1:1 oat hay:concentrate, without (control diet) or including seaweeds was studied. The ash and N contents were greater (p < 0.001) in seaweeds collected during spring than in autumn, but autumn-seaweeds had greater total extractable polyphenols. Nitrogen in red and green seaweeds was greater than 2.20 and in brown seaweeds, it was lower than 1.92 g/kg DM. Degradability after 24 h of fermentation was greater in spring seaweeds than in autumn, with Palmaria palmata showing the greatest value and Pelvetia canaliculata the lowest. Seaweeds differed in their fermentation pattern, and autumn Alaria esculenta, Laminaria digitata, Saccharina latissima and Palmaria palmata were similar to high-starch feeds. The inclusion of seaweeds in the concentrate of a diet up to 200 g/kg concentrate produced only subtle effects on in vitro ruminal fermentation.

This study determined the subcellular distribution, chemical forms, and effects of metal homeostasis of excess Cd in Cladophora rupestris. Biosorption data were analyzed with Langmuir and Freundlich adsorption models and kinetic equations. Results showed that C. rupestris can accumulate Cd. Cd mainly localized in the cell wall and debris (42.8-68.2%) of C. rupestris, followed by the soluble fraction (22.1-38.4%) observed in C. rupestris. A large quantity of Cd ions existed as insoluble CdHPO4 complexed with organic acids, Cd(H2PO4)2, Cd-phosphate complexes (FHAC) (43.2-56.0%), and pectate and protein-integrated Cd (FNaCl) (30.8-43.2%). The adsorption data were well fitted by the Freundlich model (R2 = 0.933) and could be described by the pseudo-second-order reaction rate (R2 = 0.997) and Elovich (R2 = 0.972) equations. Related parameters indicated that Cd adsorption by C. rupestris is a heterogeneous diffusion. Cd promoted Ca and Zn uptake by C. rupestris. Cu, Fe, Mn, and Mg adsorption was promoted by low Cd concentrations and inhibited by high Cd concentrations. Results suggested that cell wall sequestration, vacuolar compartmentalization, and chemical morphological transformation are important mechanisms of Cd stress tolerance by C. rupestris. This study suggests that C. rupestris has bioremediation potential of Cd.

Amino acid profiles of nine seaweed species and their in situ degradability in dairy cows

Epifaunal community composition in five macroalgal species – What are the consequences if some algal species are lost?

Characteristics of unique HBr-hydrolyzed cellulose nanocrystals from freshwater green algae (Cladophora rupestris) and its reinforcement in starch-based film

Marine macroalgae represent a valuable natural resource for bioactive phytochemicals with promising applications in therapeutics, although they remain largely under-exploited. In this work, the potential of two marine green macroalgae (Cladophora rupestris and Codium fragile) as a source of bioactive phenolic compounds was explored, and antioxidant, mineralogenic, and osteogenic activities were evaluated. For each species, a crude hydroalcoholic extract (CE) was prepared by solid/liquid extraction and fractionated by liquid/liquid purification into an ethyl acetate fraction (EAF) enriched in phenolic compounds and an aqueous fraction (AF). Antioxidant activity, assessed through radical scavenging activity and reducing power assay, was increased in EAF fraction of both species and closely related to the phenolic content in each fraction. Mineralogenic activity, assessed through extracellular matrix mineralization of a fish bone-derived cell line, was induced by EAF fractions (up to 600 % for C. rupestris EAF). Quantitative analysis of operculum formation in zebrafish larvae stained with alizarin red S further confirmed the osteogenic potential of EAF fractions in vivo, with an increase of more than 1.5-fold for both C. fragile and C. rupestris fractions, similar to vitamin D (control). Our results demonstrated a positive correlation between phenolic fractions and biological activity, suggesting that phenolic compounds extracted from marine green macroalgae may represent promising molecules toward therapeutic applications in the field of bone biology.