Assessing the role of ulvan as immunonutrient in Solea senegalensis.

Immunonutrition is a promising and viable strategy for the development of prophylactic measures in aquaculture. Ulvan, a sulphated marine polysaccharide from green seaweeds, has many biological activities including the immunomodulatory ones. The aim of this study was to assess the short and long-term effects of an ulvan-rich extract obtained from U. ohnoi as immunonutrient in Senegalese sole juveniles. In this work, an ulvan-rich extract from Ulva ohnoi has been obtained by the hot water method and isolated by ethanol precipitation. The FTIR analysis revealed that the ulvan-rich extact had very similar characteristics to previously published ulvan spectra. The total sulfate and protein content was 24.85 ± 3.98 and 0.91 ± 0.04 %, respectively. In vitro assays performed in Senegalese sole (Solea senegalensis) macrophages showed that the ulvan obtained in this study did not compromise the cell viability at concentrations up to 1 mg ml-1 and expression levels of lyg, irf1, il6, il10, c7, tf and txn were significantly upregulated in a concentration dependent-manner. Finally, S. senegalensis juveniles were fed basal diets and diets supplemented with the ulvan-rich extract at ratios 1 and 2 % for 30 days and then, challenged with Photobacterium damselae subsp. piscicida (Phdp). Thereafter, ulvan was withdrawn from the diet and all juveniles were fed the basal diet for 30 days. At 30 days post withdrawal (dpw), juveniles were challenged with Phdp. The expression profiles of a set of genes related to the immune system in spleen were evaluated as well as the lysozyme, peroxidase and bactericidal activity in plasma. Dietary effects of 1 % ulvan resulted in a boost of the immune response and increased disease resistance at short-term whereas juveniles fed diets supplemented with 2 % ulvan showed a significant decrease in the bactericidal activity and lack of protection against Phdp. At long-term (30 days after the withdrawal of ulvan), an improved response was observed in juveniles previously fed 1 % ulvan.

Prospective Antiviral Effect of Ulva lactuca Aqueous Extract against COVID-19 Infection.

Marine algal extracts exhibit a potent inhibitory effect against several enveloped and non-enveloped viruses. The infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has several adverse effects, including an increased mortality rate. The anti-COVID-19 agents are still limited; this issue requires exploring novel, effective anti-SARS-CoV-2 therapeutic approaches. This study investigated the antiviral activity of an aqueous extract of Ulva lactuca, which was collected from the Gulf of Suez, Egypt. The aqueous extract of Ulva lactuca was characterized by high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and Energy Dispersive X-ray (EDX) analyses. According to the HPLC analysis, the extract comprises several sugars, mostly rhamnose (32.88%). The FTIR spectra showed numerous bands related to the functional groups. EDX analysis confirmed the presence of different elements, such as oxygen (O), carbon (C), sulfur (S), magnesium (Mg), potassium (K), calcium (Ca), and sodium (Na), with different concentrations. The aqueous extract of U. lactuca (0.0312 mg/mL) exhibited potent anti-SARS-CoV-2 activity via virucidal activity, inhibition of viral replication, and interference with viral adsorption (% inhibitions of 64%, 33.3%, and 31.1%, respectively). Consequently, ulvan could be a promising compound for preclinical study in the drug development process to combat SARS-CoV-2.

Biochemical Characterization of a Novel Thermostable Ulvan Lyase from Tamlana fucoidanivorans CW2-9.

Ulvan is a complex sulfated polysaccharide extracted from Ulva, and ulvan lyases can degrade ulvan through a ß-elimination mechanism to obtain oligosaccharides. In this study, a new ulvan lyase, EPL15085, which belongs to the polysaccharide lyase (PL) 28 family from Tamlana fucoidanivorans CW2-9, was characterized in detail. The optimal pH and salinity are 9.0 and 0.4 M NaCl, respectively. The Km and Vmax of recombinant EPL15085 toward ulvan are 0.80 mg·mL-1 and 11.22 µmol·min -1 mg-1·mL-1, respectively. Unexpectedly, it is very resistant to high temperatures. After treatment at 100 °C, EPL15085 maintained its ability to degrade ulvan. Molecular dynamics simulation analysis and site-directed mutagenesis analysis indicated that the strong rigidity of the disulfide bond between Cys74-Cys102 in the N-terminus is related to its thermostability. In addition, oligosaccharides with disaccharides and tetrasaccharides were the end products of EPL15085. Based on molecular docking and site-directed mutagenesis analysis, Tyr177 and Leu134 are considered to be the crucial residues for enzyme activity. In conclusion, our study identified a new PL28 family of ulvan lyases, EPL15085, with excellent heat resistance that can expand the database of ulvan lyases and provide the possibility to make full use of ulvan.

Immunostimulating effects of ulvan type polysaccharide isolated from Korean Ulva pertusa in cyclophosphamide-induced immunosuppressed BALB/c mice.

This study aimed to determine the immunostimulatory activities of ulvan type polysaccharides isolated from Ulva pertusa. First, U. pertusa polysaccharide (UPP) mainly consists of rhamnose, glucuronic acid, iduronic acid, and xylose, which are typical ulvan type monosaccharides. UPP induced phosphorylation of the mitogen-activated protein kinase and nuclear factor-kappa B pathways in macrophages, subsequently triggering cytokine release and phagocytosis. The effects were closely associated with pattern recognition receptors such as dectin-1, mannose receptor, CD11b, CD14, and Toll-like receptors 2 and 4. Moreover, prophylactic administration of UPP was found to protect against body weight loss and lymphatic organ damage in cyclophosphamide-induced immunosuppressed mice. In addition, UPP demonstrated significant stimulatory effects on various immunocytes, such as T cells, B cells, macrophages, and natural killer cells derived from the spleen. These effects were closely related to the mitogen-activated protein kinase and nuclear factor-kappa B pathways, and significant secretion of immunostimulatory cytokines such as IL-6, -12, and TNF-α was noted in both blood and spleen samples. Impairment of the short-chain fatty acid balance in the cecum was prevented by UPP administration in a dose-dependent manner. Consequently, these results suggest that the UPP isolated from U. pertusa contributes to immune system activation.

Biochemical characterisation of a PL24 ulvan lyase from seaweed-associated Vibrio sp. FNV38.

Ulvan is a green macroalgal cell wall polysaccharide that has tremendous potential for valorisation due to its unique composition of sulphated rhamnose, glucuronic acid, iduronic acid and xylose. Several potential applications such as production of biofuels, bioplastics and other value-added products necessitate the breakdown of the polysaccharide to oligomers or monomers. Research on ulvan saccharifying enzymes has been continually increasing over the last decade, with the increasing focus on valorisation of seaweed biomass for a biobased economy. Lyases are the first of several enzymes that are involved in saccharifying the polysaccharide and several ulvan lyases have been structurally and biochemically characterised to enable their effective use in the valorisation processes. This study investigates the whole genome of Vibrio sp. FNV38, an ulvan metabolising organism and biochemical characteristics of a PL24 ulvan lyase that it possesses. The genome of Vibrio sp. FNV38 has a diverse CAZy profile with several genes involved in the metabolism of ulvan, cellulose, agar, and alginate. The enzyme exhibits optimal activity at pH 8.5 in 100 mM Tris-HCl buffer and 30 °C. However, its thermal stability is poor with significant loss of activity after 2 h of incubation at temperatures above 25 °C. Breakdown product analysis reveals that the enzyme depolymerised the polysaccharide predominantly to disaccharides and tetrasaccharides. Supplementary Information: The online version contains supplementary material available at 10.1007/s10811-023-03136-3.

Genome sequence of Vibrio sp. strain 10N, an ulvan-degrading bacterium isolated from coastal seawater collected at Ise Bay, Japan.

Genome sequence of an ulvan-degrading bacterium, Vibrio sp. strain 10N, is presented. The genome is 5,358,550 bp with a G + C content of 46.5%. A total of 4,712 coding sequences, including two novel ulvan lyase genes encoding a BNR4 and a glycoside hydrolase (GH88) motif, are known to be involved in the degradation of green algae.

Potentials of Ulva spp.-derived sulfated polysaccharides as gelling agents with promising therapeutic effects.

Ulvan, a sulfated polysaccharide extracted from Ulva spp., has garnered significant attention in the food and pharmaceutical industries due to its potential health benefits. These include immunomodulation, antiviral, anti-inflammatory, anti-hyperlipidemic, and anti-cancer effects. Nonetheless, practical applications in these fields remain limited due to an incomplete understanding of its gelation mechanisms. Additionally, the underlying mechanisms of its gelation have not been completely understood and thoroughly reviewed. The primary objective is to provide current insights into ulvan's gelling mechanisms and potential health impacts. This review also delves into the existing applications of ulvan polysaccharides. By unraveling these aspects, the information provided in this work is expected to deepen our understanding of ulvan's gelation mechanisms and its prospective role in enhancing health, holding promise for advancements in the fields of food science and disease prevention. This work's theoretical insights contribute significantly to a deeper understanding of these aspects, which holds paramount importance in unleashing the full potential of ulvan and elevating its scientific significance.

Characterization and Biological Activities of the Ulvan Polysaccharide-Rich Fraction Obtained from Ulva rigida and Ulva pseudorotundata and Their Potential for Pharmaceutical Application.

Seaweed from the genus Ulva (Ulvales, Chlorophyta) has a worldwide distribution and represents a potential biomass source for biotechnological applications. In the present study, we investigated the ulvan polysaccharide-rich fraction (UPRF) isolated from two Ulva species (U. rigida and U. pseudorotundata), naturally occurring on the Spanish Mediterranean coast. Chemical characterization of UPRFs was performed in order to explore the polysaccharides' composition. Biological assessments of UPRFs were compared by antioxidant activity and in vitro toxicity tests in the human cell lines: HCT-116 (colon cancer), G-361 (malignant melanoma), U-937 (leukemia), and HaCaT cells (immortalized keratinocytes). Chemical analysis revealed that both UPRFs presented rhamnose as the major relative sugar constituent, followed by glucose in U. rigida and xylose in U. pseudorotundata. Both also presented glucuronic acid, galactose, ribose, and mannose as the remaining monosaccharides. Similar antioxidant activity was obtained, where we observed increased activity in response to increased polysaccharide concentrations. Both UPRFs presented moderate toxicity against HCT-116 cell lines and a selectivity index ≥ 3, suggesting a good potential for use in pharmaceutical products.

Ulvans are not equal - Linkage and substitution patterns in ulvan polysaccharides differ with Ulva morphology.

Ulva are hardy green seaweeds that contain the sulfated polysaccharide ulvan and grow in two distinct morphologies: foliose and tubular. The authors hypothesise that ulvan from tubular species are more structurally complex than ulvans from foliose species. Herein, using standardised methods, the glycosyl linkage positions and sulfate ester substitutions of constituent monosaccharides of ulvan isolated from foliose (U. lacinulata and U. stenophylloides) and tubular (U. prolifera and U. ralfsii) species of Ulva were investigated. Comparison of native ulvans with 80 and 100 °C desulfated counterparts indicated that 4-linked rhamnose is predominantly 3-O-sulfated in all four ulvans. Ulvans from the foliose species predominantly contained →3,4)-Rhap-(1→, →4)-GlcAp-(1→ and →4)-IdoAp-(1→, collectively accounting for 67 to 81 mol% of the total linkages. In contrast, these same linkages in ulvans from the tubular species only collectively accounted for 29 to 36 mol%. Instead, ulvan from tubular species contained a combination of →2,3,4)-Rhap-(1→, terminal Rhap-(1→, →4)-GlcAp-(1→, →4)-Xylp-(1→, and/or →4)-Galp-(1→ in high proportions; some of the latter three residues were also likely O-2 sulfated. The results presented here suggest that ulvan from foliose species are predominantly unbranched polysaccharides composed of repeat disaccharides while ulvans from tubular species contain a greater diversity of branch and sulfate substitution locations.

Anti-Bacterial and Immunostimulatory Properties of Ulvan-Loaded Chitosan Nanoparticles for Use in Aquaculture.

Alternative prophylactic strategies to limit farm animal infection are needed in order to avoid the use of antibiotics. Anti-bacterial and immunostimulatory properties of bioactive compounds are of great interest in aquaculture. Marine derived polysaccharides, such as chitosan and ulvan, together with nanotechnology, have become the focus of attention in the scientific community due to their wide range of biological properties. In this work, chitosan and ulvan-loaded chitosan nanoparticles (referred as CS-TPP NPs and CS-UL-TPP NPs, respectively), obtained by the ionotropic gelation method, had round shape, and the mean sizes were 137.00 ± 5.44 and 325.50 ± 4.95 nm, respectively. No study about the anti-bacterial activity of both types of NPs against Photobacterium damselae subsp. piscicida, an important fish pathogen, has been reported so far. Furthermore, the potential immunostimulatory effects of CS-UL-TPP NPs after oral administration in fish have not yet been evaluated. The percentage of bacterial inhibition against P. damselae subsp. piscicida was determined through in vitro assays, and it was significantly higher in CS-UL-TPP NPs than in CS-TPP NPs at concentrations below 0.03 mg mL-1. The effects on the immune system of CS-TPP and CS-UL-TPP NPs were evaluated in Solea senegalensis juveniles at 30 days after oral administration. Lysozyme activity as well as gene expression levels of il1b, il6, hamp1, tf and c3 was significantly higher in CS-UL-TPP NP-treated groups than in the controls, and no significant differences were observed in CS-TPP NP-treated groups. Thus, ulvan extracted from the macroalgae Ulva ohnoi could improve anti-bacterial and immunostimulant properties of CS-TPP NPs thereby making them suitable to be used as vaccine adjuvant or as immunostimulant.

Functionalization of seaweed bloom-derived Ulvan using response surface methodology with application in flocculation of oil-in-water pollution.

Utilizing macroalgal waste biomass for pollution management is a highly efficient method for addressing the environmental difficulties associated with its disposal. To accomplish this, we have attempted to synthesize a graft copolymer by combining ulvan, a sulfated polysaccharide isolated from seaweed, with acrylates. A one-pot synthesis method using UV-initiated graft polymerization with V-50 as the photoinitiator resulted in the production of a distinctive, high-performance, and eco-friendly flocculant, Ulvan-g-Poly (acrylamide-co-acrylic acid) referred as P(U_AAm_AAc). The synthesis was optimized using the CCD-RSM approach, employing molecular weight and inherent viscosity as indicators to optimize the parameters. The structural and physio-chemical properties of the synthesized P(U_AAm_AAc) were characterized utilizing XRD, ATR-FTIR, ζ-potential, and H1 NMR spectroscopy. The flocculation performance of P(U_AAm_AAc) was further examined for the removal of oils from samples with high neem oil in urea solution and low crude oil in seawater. By employing a coagulant-flocculant combination of poly-aluminium chloride (PAC) and P(U_AAm_AAc), it was noted that more than 94% of oil was effectively eliminated in both samples. Optimization of the dosage of P(U_AAm_AAc) resulted in enhanced turbidity reduction and improved dewatering efficiency of the filter cake generated following flocculation. An evaluation of performance was conducted using the commercial flocculant APAM, where synthesized P(U_AAm_AAc) demonstrated similar results. In conclusion, the findings of this research highlight the potential of P(U_AAm_AAc) as a sustainable alternative to commercial flocculants with multifaceted solution to coastal waste management, paving the way for a cleaner and healthier marine ecosystem to mitigate oil emulsion pollution.

Characterization of a novel sulfated-rhamnoglucuronan isolated from Korean seaweed Ulva pertusa and its efficacy for treatment of inflammatory bowel disease in mice.

This study aimed to isolate Ulva pertusa polysaccharide (UPP), which elicits anti-inflammatory bowel disease (IBD) effects, from the Korea seaweed U. pertusa and identify its structure. Firstly, UPP was isolated from U. pertusa using hydrothermal extraction and ethanol precipitation. UPP is a novel polysaccharide that exhibits unique structural features such as 3-sulfated rhamnose, glucuronic acid, iduronic acid, and 3-sulfated xylose, which are repeated in 1,4-glycosidic bonds. Prophylactic oral administration of UPP in mice with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) suppressed the levels of inflammatory cytokines and MAPK- and NF-κB-related factors in the serum and colon tissue. Tight junction (TJ)-related factors such as occludin, claudin-1, and mucin were effectively augmented by UPP in the colon tissue. In addition, UPP administration prevented the DSS treatment-led cecal short chain fatty acid imbalance, and this effect was most evident for propionic acid. In conclusion, UPP isolated from the Korean U. pertusa demonstrates potent anti-IBD activity. Characterization of this ulvan revealed its unique structure. Moreover, its efficacy may be associated with its anti-inflammatory effects and regulation of gut microbiota and TJ proteins. Thus, this study provides new insights into the biological effects of UPP in IBD.

Ulvan and Ulva oligosaccharides: a systematic review of structure, preparation, biological activities and applications.

Ulva is one of the main green algae causing green tide disasters. Ulvan is the primarily component polysaccharide of the cell wall of Ulva and its complex structure and monosaccharide composition resulted in various biological activities. However, the high-value and effective utilization of extracted ulvan have been obstructed by limitations ranging from large molecular weight and low solubility to poor bioavailability. Ulva oligosaccharide obtained by degrading ulvan can not only ideally retain the various biological activities of ulvan very well but also effectively solve the problems of low solubility and poor bioavailability. The preparation and biological activity studies of ulvan and Ulva oligosaccharides have become a hot spot in the field of marine biological resources development research. At present, the comprehensive reviews of ulvan and Ulva oligosaccharides are still scarce. What are overviewed in this paper are the chemical composition, structure, extraction, and purification of ulvan and Ulva oligosaccharides, where research progress on the biological activities of ulvan and Ulva oligosaccharides is summarized and prospected. A theoretical and practical basis has been provided for further research on ulvan and Ulva oligosaccharides, as well as the high-value development and effective utilization of marine algae resources.

Durabilidade da resistência induzida por ulvana e efeito da concentração de inóculo no controle da antracnose do feijão / Persistence of ulvan-induced resistance and effect of inoculum concentration in the control of bean anthracnose

O presente trabalho teve por objetivo avaliar a duração da resistência induzida, a eficiência do número e intervalo de aplicações de ulvana, bem como a influência de diferentes concentrações de inóculo no controle e na severidade da antracnose do feijão. Para tanto, foram realizados três experimentos independentes. Plantas de feijão (P. vulgaris, cv. Uirapuru) foram cultivadas em condições de casa-de-vegetação e, no primeiro experimento, foram pulverizadas com ulvana em uma única aplicação aos: nove dias antes da inoculação (dai) V3(i); seis dai V3(m); três dai V3(f); ou duas aplicações consecutivas realizadas aos nove e seis dai V3(i)/(m); nove e três dai V3(i)/(f) ou seis e três dai V3(m)/(f) e inoculadas em um mesmo momento. No segundo e terceiro experimentos, plantas de feijão foram tratadas duas vezes (seis e três dai) e inoculadas em diferentes momentos ou com diferentes concentrações de inóculo, respectivamente. A pulverização de ulvana reduziu a severidade da antracnose em cerca de 50%. Duas aplicações de ulvana foram mais eficientes em induzir respostas de defesa em feijão contra a antracnose que somente uma. A maior redução (96%) da doença ocorreu com pulverizações sucessivas em V3(m)/(f). O efeito de duas aplicações do polissacarídeo persistiu até nove dias após o tratamento. A redução na severidade da antracnose foi maior quando foram utilizadas concentrações intermediárias de inóculo (105 e 106 conídios por mL).

The present work aimed to evaluate the persistence of the induced resistance, the efficiency of the number and the interval of ulvan applications and the influence of different inoculum concentrations in the control of bean anthracnose. Three independent experiments were carried out. Bean plants (P. vulgaris, cv. Uirapuru) were cultivated under green house conditions and, in the first experiment, they were treated with ulvan once at nine days before inoculation (dbi) V3(i); six dbi V3(m) or three dbi V3(f) or they were treated twice at nine and six dbi V3(i)/(m); nine and three V3(i)/(f) or six and three (dbi) V3(m)/(f), the they were inoculated at the same time. In the second and third experiments, bean plants were treated twice (six and three dbi) then inoculated in different moments or with different inoculums concentrations, respectively. The application of ulvan reduced the anthracnose severity by 50%. Two ulvan applications were more efficient in inducing defense responses in bean plants than a single application. The highest disease reduction (96%) was achieved by spraying ulvan twice at V3(m)/(f). The effect of two applications persists until nine days after the treatment. The reduction on the anthracnose severity was higher when intermediate (105 e 106 conidia per mL) inoculum concentrations were used.