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Carbohydrates are known to act as analog receptors for bacteria and therefore are promising alternatives for the control and prevention of bacterial infections. The present study evaluated the chemical structure of modified soybean meal polysaccharides and their capacity to adhere enterobacteria (Salmonella Typhimurium) and to interfere with the bacteria adhesion to the known analogue receptors, using in vitro assays. For this, soybean meal suspensions were subjected to a thermochemical extraction process and structural analyses showed that the fraction with higher adhesion and adhesion-inhibition potential, SAP, was constituted by two types of polysaccharides: a partially depolymerized pectin, of high molar mass, composed of xylogalacturonan and rhamnogalacturonan regions (SAP1, 545.5 kDa), and a (1 → 4)-linked-β-D-galactan of low molar mass (SAP2, 8.7 kDa). The results showed a high affinity of Salmonella for galactans, while high molar mass pectins showed no adhesion capacity. The chemical compositions of the fractions suggested that galactose could be responsible for the recognition process in the adhesion process. Other factors, such as structure and degree of polymerization of the polymers, may also be influencing the adhesion process. Modified soybean meal polysaccharides appear to be a promising alternative agent to antibiotics for the control and prevention of foodborne diseases.

Aqueous and KCl-soluble polysaccharides were extracted from Laurencia dendroidea (Rhodomelaceae, Ceramiales) and their chemical profile was accessed by anion-exchange chromatography, chemical and spectroscopic analyses. The homogeneous agaran DHS-4 (181.3 × 103 g. mol−1, 21.3% of NaSO3) presents A units mostly 2-sulfated (18.9 mol%), nonsubstituted (15.3 mol%) and 6-O-methylated (10.1 mol%), while B units are l -sugars composed predominantly by galactose 6-sulfate precursor units (19.2 mol%) and 3,6-anhydrogalactose (13.8 mol%), besides non-precursor galactose 6-sulfate units bearing d -xylose substituents on C-3 (8.1 mol%). The crude KCl-soluble DHS agaran (20.5% of NaSO3) inhibited proteolysis and hemolysis induced by Lachesis muta and Bothrops jararaca venoms. DHS was able to inhibit up to 75% the L. muta venom hemorrhagic effect and to reduce the lethality displayed by B. jararaca venom, increasing the mice survival time up to 3 times. Therefore, this agaran has high potential to be used as an additional tool to treat snakebite envenomation.

Natural elicitors from macroalgae may affect plant secondary metabolites. Ulvan is a sulfated heteropolysaccharide extracted from green seaweed, acting as both a plant biotic protecting agent, and a plant elicitor, leading to the synthesis of signal molecules. In this work, the aqueous extract of Ulva intestinalis L., mainly composed of ulvan, was used as foliar-spraying treatment and its eliciting effect was investigated in basil (Ocimum basilicum L.) and parsley (Petroselinum crispum L.). Antioxidant metabolites (polyphenols and carotenoids), volatile compounds (both in headspace emissions and hydrodistilled essential oils), and hormones (jasmonic acid, salicylic acid, salicylic acid 2-O-β-D-glucoside, abscisic acid, and azelaic acid) were quantified. The foliar-spraying treatment with U. intestinalis extract increased salicylic acid and its β-glucoside in parsley, in basil, it induced the accumulation of jasmonic and abscisic acids, indicating the presence of a priming effect. In basil, the elicitation caused a change of the essential oil (EO) chemotype from methyl eugenol/eugenol to epi-α-cadinol and increased sesquiterpenes. In parsley EO it caused a significant accumulation of 1,3,8-p-menthatriene, responsible of the typical “parsley-like” smell. In both species, the phenylpropanoids decreased in headspace and EO compositions, while the salicylic acid concentration increased, this could indicate a primarily defensive response to U. intestinalis extract. Due to the evidenced significant biological activity, U. intestinalis extract used as an elicitor may represent a suitable tool to obtain higher amounts of metabolites for optimizing plant flavor metabolites.

The current global and economic scenario has driven the search for alternative source of natural colorants produced by microalgae. Astaxanthin (3,3′-dihydroxy-β,β′-carotene-4,4′-dione) is a secondary carotenoid widely used in food, pharmaceutical, and aquaculture industries with antioxidant effects. The green microalga Haematococcus pluvialis is the richest source worldwide of natural astaxanthin, produced with lower cost than the synthetic molecules currently used. This chapter reviews some laboratory parameters related to H. pluvialis cell enhancement, astaxanthin production, and cost. The review begins with an introduction about astaxanthin-producing H. pluvialis cells followed by upstream process conditions (e.g., microalgal media effects on biomass productivity and carotenogenesis). Subsequently, downstream process parameters are evaluated (e.g., astaxanthin extraction methods and efficiency), and laboratory production cost and future prospects of this topic are presented. Economic challenges and laboratory conditions of astaxanthin production by H. pluvialis have been reviewed and discussed in view of a cost-effective astaxanthin facility.

The development of new strategies in microalgal studies represents an outstanding opportunity to mitigate environmental problems coupled with biomass production at a reduced cost. Here we present a combined bioprocess for the treatment of rice vinasse using immobilized cyanobacteria Synechococcus pevalekii in alginate beads followed by the use of the treated vinasse as a culture medium for Dunaliella salina biomass production. Cyanobacterial-alginate beads showed a chlorophyll a production of 0.68 × 10–3 mg bead−1 and a total carotenoid production of 0.64 × 10–3 mg bead−1. The first step showed a decrease in nitrate (91%), total solids (29%), and ions. Addition of treated vinasse into D. salina cultivation resulted in a significant increase in cell replication of about 175% (optimized cultivation). The use of natural seawater drastically reduced the medium cost to US$4.75 per m3 and the addition of treated vinasse has the potential to reduce it even more (up to 69%). This study not only provides an insight on the use of cyanobacteria for rice vinasse treatment but also demonstrates a promising lower-cost medium for marine microalgal biomass production with biotechnological purposes.

Different molecules have been tested as analog receptors due to their capacity to bind bacteria and prevent cell adhesion. By using in vitro assays, the present study characterized the aqueous and alkaline extracts from microalgae Pavlova lutheri and Pavlova gyrans and evaluated the capacity of these extracts to adhere to enterobacteria (Salmonella Typhimurium). The aqueous and alkaline extracts of both species were fractionated via freeze-thawing, giving rise to soluble and insoluble (precipitate) fractions in cold water. The obtained fractions were studied using thermogravimetric, methylation analyses, and using 1D and 2D NMR techniques. The cold-water-soluble fractions obtained from the aqueous extracts were mainly composed of highly branched (1&rarr, 3),(1&rarr, 6)-&beta, glucans, whereas the cold-water-precipitate fractions were constituted by (1&rarr, 3)-&beta, glucans. The alkaline extract fractions showed similar compositions with a high protein content, and the presence of glycosides (sulfoquinovosylglycerol (SQG), digalactosylglycerol (DGG)), and free fatty acids. The linear (1&rarr, glucans and the alkaline extract fractions showed an adhesion capacity toward Salmonella. The chemical composition of the active fractions suggested that the presence of three-linked &beta, glucose units, as well as microalgal proteins and glycosides, could be important in the adhesion process. Therefore, these microalgal species possess a high potential to serve as a source of anti-adhesive compounds.

Crude extracts using hexane, dichloromethane, methanol, 80% methanol or water of two agarophytes (Gracilaria caudata and Gracilaria domingensis) and their phase partitions were evaluated on early growth of lettuce. Hexane, methanol, 80% methanol and aqueous extracts of G. caudata and dichloromethane and aqueous extracts of G. domingensis were biostimulants. Palmitic acid, the major compound of non-polar extracts and phase partition, showed a significant stimulant activity in the concentrations tested (ranging from 0.49 mM to 1.95 mM), increasing 83% of lettuce root length, comparing to control, at the highest concentration. The promoting effect of the aqueous extracts is probably related to the presence of agaran, a typical hydrocolloid polysaccharide from red algae, which is mostly an exclusive constituent of these extracts. The agarans of G. domingensis and G. caudata promoted an increase in lettuce root length of 60% and 40%, respectively (both at 1 mg·mL−1 of extract). Differences in the effects promoted by these two agarans are probably related to the different content of sulfate groups (higher for polysaccharide from G. domingensis). These results suggest that structural features of agaran-type polysaccharides can elicit distinct responses. To the best of our knowledge, this is the first report on the identification of compounds with biostimulating potential from Gracilaria.

The plant biostimulant activity of the green microalga Desmodesmus subspicatus was investigated, focusing on its potential application to organic crops. Different concentrations of D. subspicatus biomass suspensions were applied to tomato seedlings (Solanum lycopersicum) via a foliar spray. The treatments promoted root growth and foliar area, which increased when the suspension was applied at concentrations of 1 g L−1 and 0.4 g L−1, respectively. To elucidate the major constituents responsible for this potential activity, the biomass of D. subspicatus was submitted to aqueous extraction and dialysis (1 kDa cut-off), giving rise to the non-dialyzable fraction E (12.3%). Size-exclusion chromatography performed on fraction E using a Bio-Gel P-2 column yielded six subfractions. Gas chromatography-mass spectrometry (GC–MS) and nuclear magnetic resonance (NMR) analyses of the E subfractions revealed the presence of glycosides as major constituents (approx. 81%), including one acidic glycoside, [6-sulfo-α- D -quinovopyranosyl-(1→1)-glycerol], and two neutral galactosyl glycosides, [α- D -galactopyranosyl-(1→6)-β- D -galactopyranosyl-(1→1)-glycerol] and [β- D -galactopyranosyl-(1→1)-glycerol]. Ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) analyses of fraction E resulted in the identification of the phytohormone zeatin (35 μg g−1). To evaluate the biostimulant potential of fraction E, different concentrations (0.5, 1.0, 1.5, and 2.0 g L−1) were tested for their effects on tomato seed germination, hypocotyl volume, and length. All tested concentrations induced increased hypocotyl lengths and volumes compared with standard treatment (water), at rates equivalent to those in response to a commercial product derived from Ascophyllum nodosum brown seaweed. These results indicated that the biomass suspension and aqueous extract of D. subspicatus showed plant growth biostimulant activity. Glycosides, together with zeatin, may play important roles in biostimulant activity.

Snakebite is a serious occupational hazard affecting mainly rural populations of tropical and subtropical developing countries. Lachesis muta (Bushmaster) bites are extremely serious but are rarely reported in the literature. Bushmaster envenomings are characterized by intense local pain, edema, neurotoxicity, hypotension, local hemorrhage, and dramatic systemic alterations. Antivenom treatment has regularly been used for more than a century; however, it fails to neutralize local tissue damage and hemorrhage, leading to morbidity or disabilities in victims. Thus, the production and clinical use of antivenom must be improved. The present work characterizes, for the first time, a sulfated polysaccharide from the red seaweed, Laurencia aldingensis, including its neutralizing effect on some toxic activities of L. muta venom. Chemical and spectroscopic analyses showed that L. aldingensis produces sulfated agarans with the A-units partially C-2 sulfated or 6-O-methoxylated presetting the B-units in the cyclized (3,6-anhydro-α-L-galactose) or in the non-cyclized form (α-L-galactose). The latter is significantly substituted by sulfate groups on C-6. In vitro and in vivo assays showed that this sulfated agaran inhibited hemolysis, coagulation, proteolysis, edema, and hemorrhage of L. muta venom. Neutralization of hemorrhagic activity was also observed when the agaran was administered by different routes and after or before the venom injection. Furthermore, the agaran blocked the edema caused by a phospholipase A2 isolated from the L. muta venom. Experimental evidence therefore indicates that the sulfated agaran of L. aldingensis has potential to aid antivenom therapy of accidents caused by L. muta venom and may help to develop more effective antivenom treatments of snake bites in general.

Graphical abstract, Highlights • Comparison among well-known marine microalgae culture media was performed. • Conway medium increased cellular growth of Dunaliella salina. • Johnson medium improved accumulation of photosynthetic pigments. • Microalgal biomass showed a cost variation of US$4.64–301.61 per kg in lab scale. • Monitoring of pigment profile proved to be essential in microalgal parameters., Microalgal cultivation system depends on optimal parameters obtained from laboratory conditions to overcome high-value compounds production and decrease cost. In this study, the laboratory-scale cultivation of Dunaliella salina was performed to explore the cost and biochemical effects on biomass cultivated using F/2, Conway, and Johnson media. D. salina cultivation was monitored by cell counting, dry biomass measurements and biochemical analyses. Photosynthetic pigment profiles were identified and quantified through high-performance liquid chromatography (HPLC) whereas economic evaluation was based on media cost. Cellular growth parameters were significantly improved by Conway medium, whereas maximum accumulation of pigments, especially violaxanthin (0.855 mg g−1) and chlorophyll a (14.255 mg g−1) were observed when Johnson medium was used. Conway-biomass showed 43 % of total lipid content. This value represents 1.5 and 1.4 times higher than lipid content found in Johnson- and F/2-biomass, respectively. Furthermore, media cost presented a variation of US$1.17–49.62 for each 103 L of media mainly due to NaNO3, KNO3, and cyanocobalamin supplementation. Thus, biomass production showed a cost variation of US$4.64–301.61 per kg on a dry weight basis. These outcomes emphasize the suitability of laboratory parameters and cost comparison among biomass produced by different media.

The present study investigated in vitro the effects of sulphated heterorhamnan (Go3), iota-/nu-carrageenans (G3d and EHW-I) and arabinogalactan (ARAGAL) polysaccharides on macrophage activation and inhibition of intracellular amastigotes of Leishmania (L.) amazonensis . All the sulphated polysaccharides (Go3, G3d and EHW-I) promoted increased nitric oxide production varying from 71 to 110%. The leishmanicidal activity of all compounds was compared to the inhibition effect of Meglumine Antimoniate at 300 μg/mL (∼79%), used as positive control. Inhibition of Leishmania ( L.) amazonensis growth was 55% with 5 μg/mL of Go3, 50% and 98% to G3d and EHW-I, respectively at 10 μg/mL, and 88% with 10 μg/mL of ARAGAL. The superoxide anion scavenging activity for the sulphated polysaccharides varied from approximately 30–55% at 10 μg/mL. In conclusion, the results of the present study indicate the promising potential of these polysaccharides for the development of new alternative therapeutic agents against leishmaniasis.

Sulfated heterorhamnans produced by Gayralia oxysperma were utilized for the preparation of two homogeneous and highly sulfated Smith-degraded products (Mwof 109 and 251 kDa), which were constituted principally by 3-linked α- l -rhamnosyl units 2- or 4-sulfate and 2-linked α- l -rhamnosyl units 4- or 3,4-sulfate, in different percentages. The homogeneous products and the crude extracts containing the sulfated heterorhamnans showed cytotoxic effect against U87MG cells. These sulfated polysaccharides induced an increase in the number of cells in G1 phase with concomitant increase of the mRNA levels of p53 and p21. The presence of 2-linked disulfated rhamnose residues together with the molecular weight could be important factors to be correlated with the inhibitory effect on human glioblastoma cells.

The aim of this study was to conduct an economic analysis and evaluate the physiological features and growth performance of the biomass produced from Haematococcus pluvialis at vegetative stage using CHU, BG-11 and WC media under batch culture. Higher cell density was achieved by using CHU and BG-11 media by the 16th day of cultivation, whereas CHU medium promoted the highest pigment content during the microalgae cultivation period. Biochemical analysis showed an increase of 26.3% on protein production when cells were cultivated on BG-11 medium compared to WC medium cultivation. Economic analysis showed that the NPK components contributed to the major participation cost in the media composition. Media cost had shown a variation of US$2.61–18.36 each 103 L of produced medium with a biomass production cost of US$6.20 (WC medium), US$21.73 (CHU medium), and US$40.36 (BG-11 medium)/kg in a dry basis.

The anti-HMPV (human metapneumovirus) activity was determined for sulfated dl -hybrid galactans obtained from the red seaweed Cryptonemia seminervis and their depolymerized products obtained by reductive partial hydrolysis. Structural studies carried out in three homogeneous depolymerized fractions DS-1, DS-2e and DS-3 (Mw of 51.6–63.8 kDa) showed that these galactans present different chemical characteristics, as monosaccharide composition, content of sulfate groups (14.1–29.9%) and agaran:carrageenan molar ratio diads, 2.7:1 for DS-1 and DS-2e and 1:1 for DS-3. The sulfate groups are located principally on C-2 of β- d -galactopyranose and 4,6-O-(1′-carboxyethylidene)-β- d -galactopyranose residues and on C-6 of α-galactose residues. Sulfated dl -galactans and their depolymerized products exhibited antiviral activity at a very early stage of the viral infection cycle. All fractions, except DS-2e inhibited HMPV replication by binding to the viral particle. Besides depolymerized galactans DS-2e and DS-3 inhibited the recognition of cell receptor by HMPV and penetration to the host cell, respectively.

Photodynamic inactivation (PDI) is an efficient approach for the elimination of a series of microorganisms; however, PDI involving phytopathogenic filamentous fungi is scarce in the literature. In the present study, we have demonstrated the photoinactivating properties of five cationic meso-(1-methyl-4-pyridinio)porphyrins on conidia of the phytopathogen Colletotrichum graminicola. For this purpose, photophysical properties (photostability and (1)O2 singlet production) of the porphyrins under study were first evaluated. PDI assays were then performed with a fluence of 30, 60, 90 and 120 J cm(-2) and varying the porphyrin concentration from 1 to 25 μmol L(-1). Considering the lowest concentration that enabled the best photoinactivation, with the respective lowest effective irradiation time, the meso-(1-methyl-4-pyridinio)porphyrins herein studied could be ranked as follows: triple-charged 4 (1 μmol L(-1) with a fluence of 30 J cm(-2))double-charged-trans2 (1 μmol L(-1) with 60 J cm(-2))tetra-charged 5 (15 μmol L(-1) with 90 J cm(-2))mono-charged 1 (25 μmol L(-1) with 120 J cm(-2)). Double-charged-cis-porphyrin 3 inactivated C. graminicola conidia in the absence of light. Evaluation of the porphyrin binding to the conidia and fluorescence microscopic analysis were also performed, which were in agreement with the PDI results. In conclusion, the cationic porphyrins herein studied were considered efficient photosensitizers to inactivate C. graminicola conidia. The amount and position of positive charges are related to the compounds' amphiphilicity and therefore to their photodynamic activity.

A homogeneous agaran fraction from Palisada flagellifera (Laurencia complex, Rhodomelaceae, Ceramiales) was obtained by aqueous room-temperature extraction, followed by ion-exchange chromatography. This galactan presents a highly complex structure with at least 18 different types of derivatives. The A units were found mostly pyruvylated, 2-sulfated (∼34%), and 6-methylated (∼34%), with the latter partially 2- and 2,4-sulfated. Minor amounts of β-D-galactopyranosyl units 2-, 6- and 2,6-sulfated, 6-glycosylated, and non-substituted are also present. The B-units are L-sugars composed predominantly of their cyclized derivatives, 3,6-anhydrogalactose and 3,6-anhydro-2-O-methylgalactose (∼56%). The former are linked to β-D-galactosyl (6-methyl) (6-glycosylated) units, as well as to 4,6-O-(1-carboxyethylidene)-β-D-galactose 2-sulfate in the proportion of 3:1.8, respectively. A significant amount (∼18%) of the α-L-galactopyranosyl units are linked to pyruvylated β-D-galactose 2-sulfate residues. An important part of the B-units (20%) is represented by α-L-galactose 6-sulfate substituted on C-3 by xylosyl, galactosyl and/or 2,3-di-O-methylgalactose units or sulfate groups that preclude their cyclization to 3,6-anhydrogalactosyl derivative. The precursor units are present in relatively low percentages. Kinetic studies suggest that in P. flagellifera agaran the cyclizable units are linked to 6-O-methyl-β-D-galactosyl and/or β-D-galactosyl units (6-glycosylated). The structural complexity of this polysaccharide is increased by the presence of 2- and 3,6-sulfated α-L-galactoses, with the latter additionally 2-O-methylated. Therefore, the major subfraction obtained from the cold extract contains structurally complex sulfated, methylated, and pyruvylated agaran.

Seaweed compounds can play important roles in either plant growth promoting or plant-pathogen interactions. The aim of this study was to evaluate the potential of crude extracts and sulfated polysaccharides isolated from the green macroalga Ulva fasciata in the control of bean anthracnose caused by Colletotrichum lindemuthianum as well the influence in the plant growth. Sulfated polysaccharides, called ulvan, were extracted with hot water, precipitated in ethanol and identified by chemical and spectroscopic methods (13C NMR). The contents of sulfate, uronic acid, protein and monosaccharides were determined. In order to obtain the crude extracts, the dried alga was extracted with methanol in Soxhlet apparatus or with ethanol at room temperature. The effect of ulvan (0.1; 1; and 10 mg ml−1) and crude extracts were tested in vitro on conidial germination and mycelial growth of C. lindemuthianum and also on the seed germination and seedling length of bean. Under greenhouse conditions, bean plants (Phaseolus vulgaris L.) were sprayed twice with ulvan or extracts and inoculated two days after the second application. The anthracnose severity was evaluated 7 days and aerial plant dry weight 14 days after inoculation. The soluble methanolic extract inhibited the mycelial growth of C. lindemuthianum in vitro, but did not reduce the disease severity under greenhouse conditions. Foliar spray of methanolic extracts enhanced the dry weight of bean plants by 20%. In contrast, ulvan increased in vitro the mycelium growth and the conidia germination of the fungus, but in greenhouse, the spray of 10 mg ml−1 ulvan reduced the anthracnose severity by 38% without affecting plant growth. The results indicate that ulvan is probably able to induce resistance to bean anthracnose.

We studied the chemical structure and yield of low-molecular-mass carbohydrates (LMMC) and soluble polysaccharides produced by green and red morphs of Gracilaria domingensis. Red plants had higher amounts of the LMMC floridoside than green morphs (324.4±8.1 and 220.1±6.0 μmol g-1 dry biomass, respectively). The yield of polysaccharides was similar between morphs (46.3±5.8% for the red and 41.4±8.3% for the green) as was the monosaccharide composition. The same agaran (6-O-methylated and sulphated) was isolated from both morphs, as determined by 13C NMR spectroscopy analysis. The floridean starch content (mg g-1 dry biomass) for the green morphs (116.2±1.7) was lower than that of the red morphs (143.9±1.8). This study showed that green and red morphs of G. domingensis are similar in their polysaccharide composition, but differ in the short-term storage compounds, starch and floridoside. These differences may be related to light and shade photosynthetic strategies, which lead to different storage accumulation.

In Brazil, snakebites are a public health problem and accidents caused by Lachesis muta have the highest mortality index. Envenomation by L. muta is characterized by systemic (hypotension, bleeding and renal failure) and local effects (necrosis, pain and edema). The treatment to reverse the evolution of all the toxic effects is performed by injection of antivenom. However, such therapy does not effectively neutralize tissue damage or any other local effect, since in most cases victims delay seeking appropriate medical care. In this way, alternative therapies are in demand, and molecules from natural sources have been exhaustively tested. In this paper, we analyzed the inhibitory effect of a sulfated galactan obtained from the red seaweed Palisada flagellifera against some toxic activities of L. muta venom. Incubation of sulfated galactan with venom resulted in inhibition of hemolysis, coagulation, proteolysis, edema and hemorrhage. Neutralization of hemorrhage was also observed when the galactan was administered after or before the venom injection, thus mimicking a real in vivo situation. Moreover, the galactan blocked the edema caused by a phospholipase A2 isolated from the same venom. Therefore, the galactan from P. flagellifera may represent a promising tool to treat envenomation by L. muta as a coadjuvant for the conventional antivenom.

The present work aimed to evaluate the defense responses induced by chemically sulfated ulvans in Arabidopsis thaliana plants against the phytopathogenic fungi Alternaria brassicicola and Colletotrichum higginsianum. Derivatives with growing sulfate content (from 20.9 to 36.6%) were prepared with SO3-pyridine complex in formamide. NMR and FTIR spectroscopic analyses confirmed the increase of sulfate groups after the chemical sulfation process. The native sulfated polysaccharide (18.9% of sulfate) and its chemically sulfated derivatives similarly reduced the severity of both pathogenic fungi infections. Collectively, our results suggest that ulvans induce resistance against both fungal pathogens independently of its sulfation degree.

The phycocolloid iota-carrageenan (ι-CAR) was extracted from the Brazilian seaweed Meristiella gelidium . At a concentration of 5 mg ml −1 it was mixed with solutions of common corn starch (CS), high amylose starch (MS), waxy corn starch (WS) and YS starch (YS), to give systems at a final concentration of 25 g l −1 , which were subjected to rheological measurements. Each mixture gave rise to a different behaviour under dynamic rheological conditions showing a distinct degree of interaction between ι-CAR and the starch. The presence of ι-CAR in ι-CAR-WS, ι-CAR-MS and ι-CAR-CS systems resulted in synergy manifested as enhancements in thermal stability (absence of hysteresis), resulting in thermoreversible gels softer and more stable between 45 and 85 °C. In the ι-CAR-YS system, the synergistic effect occurred as an improvement in solid character, increasing the elastic nature of the gel, suggesting that the structural characteristic of YS, more regular structure with amylopectin having a low molar mass, can contribute to enhanced intermolecular binding.

Two homogeneous sulfated polysaccharides obtained from the red seaweeds Gymnogongrus griffithsiae and Cryptonemia crenulata, the kappa/iota/nu carrageenan G3d and the dl-galactan hybrid C2S-3, were assayed for their antiviral properties against the four serotypes of dengue virus (DENV) in different host cell types. Both seaweed derivatives were selective inhibitors of DENV-2 multiplication in Vero cells with inhibitory concentration 50% (IC50) values around 1 microg/ml and selectivity indices > 1000. The compounds had a lower antiviral effect against DENV-3 (IC50 values in the range 13.9-14.2 microg/ml), an even lower effect against DENV-4 (IC50 values in the range 29.3 to > 50 microg/ml) and were totally inactive against DENV-1. With respect to the host cell, the polysulfates were inhibitors of DENV-2 and DENV-3 in the human hepatoma HepG2 and foreskin PH cells, with similar antiviral effectiveness as in Vero cells, but were totally inactive in mosquito C6/36 HT cells. Mechanistic studies demonstrated that G3d and C2S-3 were active DENV-2 inhibitors only when added together with the virus or early after infection, and both initial processes of virus adsorption and internalization are the main targets of these compounds. Therefore, the variations in antiviral activity of the polysaccharides depending on the viral serotype and the host cell may be ascribed to differences in the virus-cell interaction leading to virus entry.

A homogeneous iota/nu-hybrid carrageenan (71% iota- and 21% nu-) isolated from Eucheuma denticulatum was used as a model compound to study the cyclization reaction of α- d -galactose 2,6-disulfate units to 3,6-anhydro-α- d -galactose 2-sulfate. The rate of cyclization, at 70 °C, of this carrageenan is about 50 times faster than that of a porphyran (non-sulfated β- d -galactose linked to α- l -galactose 6-sulfate) and 210 times faster when compared with a lambda-carrageenan (2-sulfated β- d -units linked to α- d -galactose 2,6-disulfate). The use of this model compound confirms the previous hypothesis of the accelerating effect of the β- d -4-sulfate group as well as suggests the influence of the 2-sulfate of the α- d -galactose units on the 4 C 1 → 1 C 4 chair forms interchange. The easy of cyclization indicates that to produce commercial iota-carrageenans milder alkaline treatments could be used, avoiding degradation and increasing the gel strength.

The sulfated agaran isolated by water extraction from the red seaweed, Acanthophora spicifera (Rhodomelaceae, Ceramiales), is made up of A-units highly substituted with sulfate groups on C-2 (28-30%), sulfates on C-2 and 4,6-O-(1'-carboxyethylidene) groups (9-15%), and only the C-2 sulfate groups (5-8%) with small amounts of C-6 sulfate, 6-O-methyl, and nonsubstituted residues. B-units are formed mainly by 3,6-anhydro-alpha-L-galactose (15-16%) and its precursor, alpha-L-galactose 6-sulfate (10-17%), together with lesser amounts of 3,6-anhydro-alpha-L-galactose 2-sulfate, alpha-L-galactose 2,6-disulfate, alpha-L-galactose 2,3,6-tri-sulfate, alpha-L-galactose 2,6-disulfate 3-xylose, 2-O-methyl-alpha-L-galactose, and unsubstituted alpha-L-galactose. Small, but significant quantities of beta-D-xylose were found in all the fractions, together with small amounts to traces of D-glucose. Some of the fractions have high antiviral activity. Attempts to correlate structure and antiviral activity in agarans are presented.

Xylans from five seaweeds belonging to the order Nemaliales ( Galaxaura marginata , Galaxaura obtusata , Tricleocarpa cylindrica , Tricleocarpa fragilis, and Scinaia halliae ) and one of the order Palmariales ( Palmaria palmata ) collected on the Brazilian coasts were extracted with hot water and purified from acid xylomannans and/or xylogalactans through Cetavlon precipitation of the acid polysaccharides. The β- d -(1→4), β- d -(1→3) ‘mixed linkage’ structures were determined using methylation analysis and 1D and 2D NMR spectroscopy. The presence of large sequences of β-(1→4)-linked units suggests transient aggregates of ribbon- or helical-ordered structures that would explain the low optical rotations.

Brown algae are often used as heavy metal biomonitors and biosorbents because they can accumulate high concentrations of metals. Cation-exchange performed by cell wall polysaccharides is pointed out as the main chemical mechanism for the metal sequestration. Here, we biochemically investigated if the brown alga Padina gymnospora living in a heavy metal contaminated area would modify their polysaccharidic content. We exposed non-living biomass to Cd and Pb and studied the metals adsorption and localization. We found that raw dried polysaccharides, sulfate groups, uronic acids, fucose, mannose, and galactose were significantly higher in contaminated algae compared with the control ones. Metal concentrations adsorbed by non-living biomass were rising comparatively to the tested concentrations. Electron microscopy showed numerous granules in the cell walls and X-ray microanalysis revealed Cd as the main element. We concluded that P. gymnospora overproduces cell wall polysaccharides when exposed to high metal concentrations as a defense mechanism.

The DL-galactan hybrid C2S-3, isolated from the red seaweed Cryptonemia crenulata (Halymeniaceae, Halymeniales), is a potent and selective inhibitor of the multiplication of diverse strains of DENV-2 in Vero cells with higher effectiveness than the reference polysaccharide heparin. The presence of the compound either only at virus adsorption or at virus internalization exerted a significant and dose-dependent inhibition in DENV-2 plaque number. The compound failed to inactivate DENV-2 directly by incubation of virus before cell infection as well as to induce a refractory state by cell pretreatment. Thus, the inhibitory effect was exclusively exerted through a blockade in virus multiplication during the infectious process. When the entry of DENV-2 particles into the cell is bypassed, as occurs in virus RNA transfection, the polysaccharide C2S-3 failed to block the completion of the multiplication cycle. Furthermore, the antiviral properties of C2S-3 are not correlated with anticoagulant activity. Fil: Talarico, Laura Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Duarte, María E. R.. Universidade Federal do Paraná; Brasil Fil: Zibetti, Rosiane G. M.. Universidade Federal do Paraná; Brasil. Instituto de Ensino Superior Pequeno Príncipe; Brasil Fil: Noseda, Miguel D.. Universidade Federal do Paraná; Brasil Fil: Damonte, Elsa Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Sulfated xylomannans were isolated from two species of genus Chondrophycus by aqueous extraction followed by KCl fractionation. Structural determination of the native, desulfated and Smith-degraded KCl-precipitated polysaccharides carried out by composition and methylation analysis and NMR spectroscopy (1D and 2D experiments) showed the following general structure: Download : Download full-size image These xylomannans present different degrees of branching (15–25%) by β- d -Xylp (70–80%) and β- d -Manp-2-S (20–30%) and molecular weights (33–222 kDa). This is the first report of the presence of a sulfated xylomannan in species of order Ceramiales.

Chemical and spectroscopic methods showed that the major KCl-precipitated galactans from Meristiella gelidium (Solieriaceae) are iota/kappa/nu-hybrid carrageenans with the former one in higher proportion. These carrageenans showed, by HPSEC-MALLS analysis, unimodal symmetrical peaks with MW of 425.6–956.7 kDa. The effectiveness of the crude extracts from M. gelidium against HSV-2 was higher than the corresponding extract from G. griffithsiae, previously determined. However, when considering the homogeneous carrageenans, the fractions obtained from both seaweeds showed the same level of activity. The extracts and carrageenan derived from M. gelidium were more effective inhibitors of DENV-2 if compared with G. griffithsiae samples and reference polysaccharides. The most active fraction obtained from M. gelidium showed a selectivity index against HSV-2 of 25,000, a value high enough to consider this carrageenan as a promising agent to be evaluated for the treatment of genital HSV-2 infections. Fil: De S.F. Tischer, Paula Cristina. Universidade Federal do Paraná; Brasil Fil: Talarico, Laura Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de Virología; Argentina Fil: Noseda, Miguel D.. Universidade Federal do Paraná; Brasil Fil: Pita B. Guimarães, Silvia Maria. Instituto de Botânica; Brasil Fil: Damonte, Elsa Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de Virología; Argentina Fil: Duarte, Maria Eugênia R.. Universidade Federal do Paraná; Brasil

The sulfated, methylated galactan isolated from the red seaweed Bostrychia montagnei, showed an unusually narrow structural dispersion. This agaran has the defining linear backbone of alternating 3-linked beta-D-galactopyranosyl units and 4-linked alpha-L-galactopyranosyl and 3,6-anhydrogalactopyranosyl residues. The D-units have C-6 methylation, C-6 single stubs of xylopyranosyl and minor to trace amounts of (possible) C-6 linked single stubs of galactopyranosyl. These units are mainly sulfated on C-4 with lesser sulfation at C-6 and minor at C-2. The L-residues are mainly methylated on C-2 of the 3,6-anhydrogalactopyranosyl and sulfated on C-3 of the L-galactopyranosyl; minor amounts of 2,3- and 3,6-disulfated and 2-O-methyl or 2-O-glycosyl 3-sulfated L-galactopyranosyl were also found.

The alginate from fronds of the brown seaweed Laminaria brasiliensis contains residues of (1 → 4)-linked β-d-mannopyranosyl (uronic acid) and α-l-gulopyranosil, (uronic acid) in a molar ratio 1.2:1. Strong acid hydrolysis gave rise to oligosaccharide blocks of β-d-ManpA (yield 6%; DP 19–26) and of α-l-GulpA (yield 17%; DP 16–20). These contained 8%; and < 2%; of the other uronic acid respectively, indicating preferential hydrolysis of glycosidic bonds in the region containing mixed uronic acids.