Investigating non-fungal eukaryotic diversity in snow in the Antarctic Peninsula region using DNA metabarcoding.

Snow is a unique microhabitat, despite being a harsh environment, multiple life forms have adapted to survive in it. While algae, bacteria and fungi are dominant microorganisms in Antarctic snow, little is known about other organisms that may be present in this habitat. We used metabarcoding to investigate DNA sequence diversity of non-fungal eukaryotes present in snow obtained from six different sites across the Maritime Antarctica. A total of 20 taxa were assigned to obtained sequences, representing five Kingdoms (Chromista, Protozoa, Viridiplantae and Metazoa) and four phyla (Ciliophora, Cercozoa, Chlorophyta and Cnidaria). The highest diversity indices were detected in Trinity Peninsula followed by Robert Island, Arctowski Peninsula, Deception Island, King George Island and Snow Island. The most abundant assignments were to Trebouxiophyceae, followed by Chlamydomonas nivalis and Chlamidomonadales. No taxa were detected at all sites. Three potentially new records for Antarctica were detected: two Ciliophora (Aspidisca magna and Stokesia sp.) and the green algae Trebouxia potteri. Our data suggested that similarities found between the sites may be more related with snow physicochemical properties rather than geographic proximity or latitude. This study provides new insights into the diversity and distribution of eukaryotic organisms in Antarctic snow.

Identification and Characterization of a Novel Microalgal Strain from the Antofagasta Coast Tetraselmis marina AC16-MESO (Chlorophyta) for Biotechnological Applications.

The wide rocky coastline of the Antofagasta hosts an intertidal ecosystem in which the species that inhabit it are routinely exposed to a wide range of physical and chemical conditions and have therefore evolved to tolerate extremes. In the search for new species of potential biotechnological interest with adaptations to a wide range of environmental conditions, the isolation and characterization of microalgae from these ecosystems is of great interest. Here, a new microalgal strain, Tetraselmis marina AC16-MESO, is described, which was isolated from a biofilm collected on the intertidal rocks of the Antofagasta coast (23°36'57.2″ S, 70°23'33.8″ W). In addition to the morphological characterization, 18S and ITS sequence as well as ITS-2 secondary structure analysis revealed an identity of 99.76% and 100% with the species Tetraselmis marina, respectively. The analyses of the culture characteristics and biochemical content showed similarities with other strains that are frequently used in aquaculture, such as the species Tetraselmis suecica. In addition, it is tolerant of a wide range of salinities, thus allowing its culture in water of varying quality. On the other hand, added to these characteristics, the results of the improvement of the lipid content in stressful situations of salinity observed in this study, together with other antecedents such as the potential in bioremediation already published for this strain by the same research group, present a clear example of its biotechnological plasticity. It is noteworthy that this strain, due to its characteristics, allows easy collection of its biomass by decantation and, therefore, a more cost-efficient harvesting than for other microalgal strains. Therefore, this new strain of Tetraselmis marina, first report of this species in Chile, and its morphologically, molecularly and biochemically description, presents promising characteristics for its use in biotechnology and as feed for aquaculture.

Urea as a source of nitrogen and carbon leads to increased photosynthesis rates in Chlamydomonas reinhardtii under mixotrophy.

Microalgae is a potential source of bioproducts, including feedstock to biofuels. Urea has been pointed as potential N source for microalgae growth. Considering that urea metabolism releases HCO3- to the medium, we tested the hypothesis that this carbon source could improve photosynthesis and consequently growth rates of Chlamydomonas reinhardtii. In this sense, the metabolic responses of C. reinhardtii grown with ammonium and urea as nitrogen sources under mixotrophic and autotrophic conditions were investigated. Overall, the mixotrophy led to increased cell growth as well as to a higher accumulation of lipids independent of N source, followed by a decrease in photosynthesis over the growth phases. In mixotrophy, urea stimulates growth in terms of cell number and dry weight. Furthermore, higher photosynthesis was verified in late logarithmic phase compared to ammonium. Under autotrophy conditions, although cell number and biomass were reduced, there was higher production of starch independent of N source. Nonetheless, urea-based autotrophic treatments stimulated biomass production compared to ammonium-based treatment. Under mixotrophy higher input of carbon into the cell from acetate and urea optimized photosynthesis and consequently promoted cell growth. Together, these results suggest urea as alternative source of carbon, improving photosynthesis and cell growth in C. reinhardtii.

Antimicrobial activity of some macroalgae of the Veracruzano Reef System (SAV), Mexico.

The study of macroalgae antimicrobial agents is limited to Mexico and scarce in the Veracruzano Reef System (SAV). It is necessary to devote efforts towards this field of applied phycology. The aim was to evaluate the antimicrobial activity of some phyla of Rhodophyta, Chlorophyta and Ochrophyta from SAV. Methanolic extracts from 23 marine macroalgae species (7 Chlorophyta, 4 Phaeophyta and 12 Rhodophyta) from the Veracruzano Reef System (SAV) (Mexico) were evaluated for antimicrobial activity. Antibacterial and antifungal activity were assessed by agar diffusion and agar dilution methods. The differences between mean values obtained for experimental groups was done by analysis of variance (ANOVA multifactorial model), p-values of 0.001 or less were considered statistically significant. Two new records are recognized for SAV (Laurencia gracilis and Sebdenia flabellata) and Compsothamnion thuioides for the Gulf of Mexico coasts. 16 species showed antibacterial activity, of which Caulerpa sertularioides, Ulva lactuca and Laurencia obtuse had significant activity on Gram-positive bacteria. 43.7% belong to the phyla Chlorophyta (7 species), 50% Rhodophyta (8 species) and 6.25% Ochrophyta (1 species). This indicates that the extracts of the algae of the Rhodophyta and Chlorophyta are the ones that showed the greatest activity. Regarding the yeasts, 16.6% of the total algae collected were active in the different yeast strains. 43.7% belongs to Chlorophyta species and for Rhodophyta were 60%. The macroalgae with the highest antifungal activity were: Cymopolia barbata, Ulva lactuca and Laurencia gracilis. The macroalgae of the Veracruzano Reef System present antimicrobial activity. This study is the first investigation of macroalgae's bioactive components from SAV, where they could be sources for future medical applications.

Green algae (Viridiplantae) in sediments from three lakes on Vega Island, Antarctica, assessed using DNA metabarcoding.

BACKGROUND: Vega Island is located off the eastern tip of the Antarctic Peninsula (Maritime Antarctica), in the Weddell Sea. In this study, we used metabarcoding to investigate green algal DNA sequence diversity present in sediments from three lakes on Vega Island (Esmeralda, Copépodo, and Pan Negro Lakes). METHODS AND RESULTS: Total DNA was extracted and the internal transcribed spacer 2 region of the nuclear ribosomal DNA was used as a DNA barcode for molecular identification. Green algae were represented by sequences representing 78 taxa belonging to Phylum Chlorophyta, of which 32% have not previously been recorded from Antarctica. Sediment from Pan Negro Lake generated the highest number of DNA reads (11,205), followed by Esmeralda (9085) and Copépodo (1595) Lakes. Esmeralda Lake was the richest in terms of number of taxa (59), with Copépodo and Pan Negro Lakes having 30 taxa each. Bray-Curtis dissimilarity among lakes was high (~ 0.80). The Order Chlamydomonadales (Chlorophyceae) gave the highest contribution in terms of numbers of taxa and DNA reads in all lakes. The most abundant taxon was Chlorococcum microstigmatum. CONCLUSIONS: The study confirms the utility of DNA metabarcoding in assessing potential green algal diversity in Antarctic lakes, generating new Antarctic records.

Fairy ring disease affects epiphytic algal assemblages associated with the moss Sanionia uncinata (Hedw.) Loeske (Bryophyta) on King George Island, Antarctica.

Since the nineteenth century, a ring-forming disease attacking Antarctic mosses has been reported. However, to date, only the effects on the mosses themselves are known. In this study, we used DNA metabarcoding to investigate the effects on the moss epiphytic algal community at different stages of disease progression. As the disease progressed, algal species richness decreased, although overall abundance was not significantly affected. Prasiolales appeared unaffected, whereas Ulotrichales were more sensitive. Trebouxiales dominated the advanced disease stage, suggesting a possible benefit from the disease, either through the elimination of competition or creation of new niches. Infection is responsible for moss death, leading to habitat loss for other organisms, but pathogenic effects on algae cannot be ruled out. Our data indicate that the disease not only impacts mosses but also other groups, potentially resulting in loss of Antarctic biodiversity. This study provides the first report of the disease effects on epiphytic algal communities of Antarctic bryophytes.

Physiological and metabolic responses to hypersalinity reveal interpopulation tolerance in the green macroalga Ulva compressa with different pollution histories.

There is scarce investigation addressing interpopulation tolerance responses to address the influence of a history of chronic stress exposure, as that occurring in polluted environments, in photoautotrophs. We evaluated ecophysiological (photosynthetic activity) and metabolic (oxidative stress and damage) responses of two populations of green macroalga Ulva compressa from polluted (Ventanas) and non-polluted (Cachagua) localions of central Chile, and exposed to controlled hypersalinity conditions of 32 (control), 42, 62 and 82 psu (practical salinity units) for 6 h, 48 h and 6 d. Both primary production (ETRmax) and photosynthetic efficiency (αETR) were generally higher in the population from Cachagua compared to Ventanas at all times and salinities. Moreover, at most experimental times and salinities the population from Ventanas had greater levels of H2O2 and lipid peroxidation that individuals from Cachagua. Total ascorbate was higher in the population of Cachagua than Ventanas at 42 and 82 psu after 6 and 48 h, respectively, while at 6 d concentrations were similar between both populations at all salinities. Total glutathione was greater in both populations after 6 h at all salinities, but at 48 h its concentrations were higher only in the population from Cachagua, a trend that was maintained at 6 d under 82 psu only. Reduced and oxidized ascorbate (ASC and DHA, respectively) and glutathione (GSH and GSSG, respectively) demonstrated similar patterns between U. compressa populations, with an increase oxidation with greater salinities but efficient recycling to maintain sufficient batch of ASC and GSH. When assessing the expression of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR), while the population of Ventanas displayed a general trend of upregulation with increasing salinities along the experiments, U. compressa from Cachagua revealed patterns of downregulation. Results demonstrated that although both populations were still viable after the applied hypersalinities during all experimental times, biological performance was usually more affected in the population from the Ventanas than Cachagua, likely due to a depressed baseline metabolism after a long history of exposition to environmental pollution.

Alternative fertilizer-based growth media support high lipid contents without growth impairment in Scenedesmus obliquus BR003.

Nitrogen (N) sources have been target in microalgae cultivation studies, considering their nutritional impact on growth and high costs. Here, we have evaluated the growth of Scenedesmus obliquus BR003, applying alternative low-cost culture media containing ammonium and urea, or combinations of both N sources. The culture media were applied for indoor and outdoor cultivation, followed by growth analyses and metabolic characterization. The alternative culture media B4 and L4 supported higher biomass production (1.4 g L-1) compared to BG11 (nitrate-based medium). In addition, the lipid percentage was higher for B4 (ammonium-based culture medium), reaching up to 25% DW. High contents of carbohydrates (60%) and proteins (40%) were also obtained in media with ammonium and urea, respectively. Considering the lower costs of alternative fertilizer-based media, using ammonium and/or urea as N sources, and the high lipid content observed, we suggest these media as viable for large-scale production of S. obliquus.

Antioxidant activity and total phenolic content in Caulerpa filiformis (Chlorophyta) from Sechura Bay and Paracas Bay, Peru / Actividad antioxidante y contenido fenólico total en Caulerpa filiformis (Chlorophyta) de Bahía de Sechura y Bahía de Paracas, Perú

Abstract In Peru, Caulerpa filiformis is a marine algae listed as an invasive species. For years, its distribution has been considered to be in the north coast (Isla Lobos de Afuera and Piura) until a recent report of its distribution in the central coast (Ancash, Lima, and Ica). The present investigation aims to determine the main groups of secondary metabolites, total phenol content, and antioxidant activity of the methanolic extract of C. filiformis from Sechura Bay (Piura) and Paracas Bay (Ica). The main chemical groups were determined through phytochemical screening, the content of phenols by the Folin-Ciocalteu method, and antioxidant activity by the ABTS method (2,2-azinobis-[3-ethylbenzothiazoline-6-sulfonic acid]) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH). The phytochemical screening of the methanolic extract of C. filiformis from Sechura Bay and Paracas Bay revealed the presence of carbohydrates, polyphenols, tannins, flavonoids, lipids, alkaloids, steroids, and triterpenes for both extracts. The total phenol content of the extract of C. filiformis from Sechura Bay (39.31 ± 0.39 mg of AGE/g extract) was significantly higher (p < 0.05) than that from Paracas Bay (18.78 ± 0.31 mg of AGE/g extract). In the ABTS and DPPH assays, the antioxidant capacity of the Sechura C. filiformis extract (IC50 = 3.49 ± 0.01 and 2.18 ± 0.02 mg/mL) was significantly higher (p < 0.05) than that of the Paracas C. filiformis extract (IC50 = 6.41 ± 0.02 and 2.42 ± 0.04 mg /mL). These findings suggest that the methanolic extract of C. filiformis is a source of secondary metabolites with an antioxidant potential.

Resumen En Perú, Caulerpa filiformis es una macroalga catalogada como especie invasora. Durante años, su distribución fue considerada en la costa norte (Isla Lobos de Afuera y Piura) hasta un informe reciente de su distribución en la costa central (Ancash, Lima e Ica). El objetivo de esta investigación es determinar los principales grupos de metabolitos secundarios, contenido total de fenol y actividad antioxidante del extracto metanólico de C. filiformis de Bahía de Sechura (Piura) y Bahía de Paracas (Ica). Los principales grupos químicos se determinaron mediante análisis fitoquímico, el contenido de fenoles mediante el método Folin-Ciocalteu y la actividad antioxidante mediante el método ABTS (ácido 2,2-azinobis- [3-etilbenzotiazolina-6-sulfónico]) y 2, 2′-difenil-1-picrylhydrazyl (DPPH). El examen fitoquímico del extracto metanólico de C. filiformis de ambas bahías revelaron la presencia de carbohidratos, polifenoles, taninos, flavonoides, lípidos, alcaloides, esteroides y triterpenos. El contenido total de fenol del extracto de C. filiformis de Bahía de Sechura (39.31 ± 0.39 mg de extracto de AGE / g) fue significativamente mayor (p <0.05) que el de Bahía de Paracas (18.78 ± 0.31 mg de extracto de AGE / g). En los ensayos ABTS y DPPH, la capacidad antioxidante del extracto de Sechura (IC50 = 3.49 ± 0.01 y 2.18 ± 0.02 mg / mL) fue significativamente mayor (p <0.05) que la del extracto de Paracas C. filiformis (IC50 = 6.41 ± 0.02 y 2.42 ± 0.04 mg / mL). Estos hallazgos sugieren que el extracto metanólico de C. filiformis es una fuente de metabolitos secundarios con potencial antioxidante.

In vitro inactivation of thrombin generation by polysulfated fractions isolated from the tropical coenocytic green seaweed Caulerpa racemosa (Caulerpaceae, Bryopsidales) / Inativação in vitro da geração de trombina por frações polissulfatadas isoladas damacroalga verde cenocítica tropical Caulerpa racemosa (Caulerpaceae; Bryopsidales)

Pharmacological efficacy of Caulerpa racemosa (Chlorophyta) sulfated polysaccharidic (SPs) fractions (F IIII) on models of coagulation and inflammation has been demonstrated, but not their effects on thrombin generation (TG). This study examined fractions for composition and physical-chemical characteristics and in vitro inactivation of TG by F I and F II in 60-fold diluted human plasma using continuous method. Papain-extraction yield of 0.7% revealed F IIII by DEAE-cellulose chromatography, with differences among the relative proportions of sulfate (17.37-24.00%), total sugars (30.03-48.34%) and absence of proteins. Charge density patterns and molecular sizes > 100 kDa of the fractions were verified by both agarose/polyacrylamide analyses, respectively. These electrophoreses combined with toluidine blue/Stains-All also indicated nonSPs. Anticoagulant effects of 4.76 (F I), 12.00 (F II) and 2.32 (F II) IU mg-1 by activated partial thromboplastin time test were recorded against heparin (193 IU mg-1), without changes in prothrombin time. Diluted plasma treated with F I and F II reduced concentration-dependent and sulfation pattern TG by both intrinsic and extrinsic pathways, with 50% inactivation by intrinsic pathway of F II even at 4.1 µg. Heparin abolished TG at least 4-fold lower. Therefore, C. racemosa produces SPs with TG inhibition.(AU)

Eficácia farmacológica de frações (F I→III) polissacarídicas sulfatadas (PSs) da Chlorophyta Caulerpa racemosa sobre modelos de coagulação e inflamação tem sido demonstrada, exceto seus efeitos sobre geração de trombina (GT). Examinaram-se frações quanto à composição, características físico-químicas e inativação in vitro de GT por F I e F II, em plasma humano diluído 60 vezes usando método contínuo. Rendimento de extração-papaína (0,7%) revelou, por cromatografia de DEAE-celulose, F I→III com diferenças entre as proporções relativas de sulfato (17,37-24,00%), açúcares totais (30,03-48,34%) e ausência de proteínas. Foram verificados, por ambas as análises agarose/poliacrilamida, graus de densidade de carga e tamanhos moleculares > 100 kDa das frações, respectivamente. Também essas eletroforeses, combinadas com azul de toluidina/Stains-All, indicaram polissacarídeos não sulfatados. Foram registrados, pelo teste do tempo de tromboplastina parcial ativada, efeitos anticoagulantes de 4,76 (F I), 12,00 (F II) e 2,32 (F II) UI mg-1 contra heparina (193 UI mg-1), porém não modificando tempo de protrombina. Plasma diluído tratado com F I e F II reduziu GT por ambas as vias intrinsíca/extrínsica, dependente de concentração e grau de sulfatação, com F II em 4,1 μg apresentando eficácia de 50% pela via intrínsica. Heparina, quatro vezes menos, aboliu GT. Portanto, C. racemosa produz PSs com inibição de GT.(AU)

In vitro inactivation of thrombin generation by polysulfated fractions isolated from the tropical coenocytic green seaweed Caulerpa racemosa (Caulerpaceae, Bryopsidales) / Inativação in vitro da geração de trombina por frações polissulfatadas isoladas da macroalga verde cenocítica tropical Caulerpa racemosa (Caulerpaceae; Bryopsidales)

Pharmacological efficacy of Caulerpa racemosa (Chlorophyta) sulfated polysaccharidic (SPs) fractions (F I→III) on models of coagulation and inflammation has been demonstrated, but not their effects on thrombin generation (TG). This study examined fractions for composition and physical-chemical characteristics and in vitro inactivation of TG by F I and F II in 60-fold diluted human plasma using continuous method. Papain-extraction yield of 0.7% revealed F I→III by DEAE-cellulose chromatography, with differences among the relative proportions of sulfate (17.37-24.00%), total sugars (30.03-48.34%) and absence of proteins. Charge density patterns and molecular sizes > 100 kDa of the fractions were verified by both agarose/polyacrylamide analyses, respectively. These electrophoreses combined with toluidine blue/Stains-All also indicated nonSPs. Anticoagulant effects of 4.76 (F I), 12.00 (F II) and 2.32 (F II) IU mg-1 by activated partial thromboplastin time test were recorded against heparin (193 IU mg-1), without changes in prothrombin time. Diluted plasma treated with F I and F II reduced concentration-dependent and sulfation pattern TG by both intrinsic and extrinsic pathways, with 50% inactivation by intrinsic pathway of F II even at 4.1 µg. Heparin abolished TG at least 4-fold lower. Therefore, C. racemosa produces SPs with TG inhibition.

Eficácia farmacológica de frações (F I→III) polissacarídicas sulfatadas (PSs) da Chlorophyta Caulerpa racemosa sobre modelos de coagulação e inflamação tem sido demonstrada, exceto seus efeitos sobre geração de trombina (GT). Examinaram-se frações quanto à composição, características físico-químicas e inativação in vitro de GT por F I e F II, em plasma humano diluído 60 vezes usando método contínuo. Rendimento de extração-papaína (0,7%) revelou, por cromatografia de DEAE -celulose, F I→III com diferenças entre as proporções relativas de sulfato (17,37-24,00%), açúcares totais (30,03-48,34%) e ausência de proteínas. Foram verificados, por ambas as análises agarose/poliacrilamida, graus de densidade de carga e tamanhos moleculares > 100 kDa das frações, respectivamente. Também essas eletroforeses, combinadas com azul de toluidina/Stains-All, indicaram polissacarídeos não sulfatados. Foram registrados, pelo teste do tempo de tromboplastina parcial ativada, efeitos anticoagulantes de 4,76 (F I), 12,00 (F II) e 2,32 (F II) UI mg-1 contra heparina (193 UI mg- 1), porém não modificando tempo de protrombina. Plasma diluído tratado com F I e F II reduziu GT por ambas as vias intrinsíca/extrínsica, dependente de concentração e grau de sulfatação, com F II em 4,1 µg apresentando eficácia de 50% pela via intrínsica. Heparina, quatro vezes menos, aboliu GT. Portanto, C. racemosa produz PSs com inibição de GT.

Bulk hydrogen stable isotope composition of seaweeds: Clear separation between Ulvophyceae and other classes.

Little is known about the bulk hydrogen stable isotope composition (δ2 H) of seaweeds. This study investigated the bulk δ2 H in several different seaweed species collected from three different beaches in Brazil, Australia, and Argentina. Here, we show that Ulvophyceae (a group of green algae) had lower δ2 H values (between -94‰ and -130‰) than red algae (Florideophyceae), brown algae (Phaeophyceae), and species from the class Bryopsidophyceae (another group of green algae). Overall the latter three groups of seaweeds had δ2 H values between -50‰ and -90‰. These findings were similar at the three different geographic locations. Observed differences in δ2 H values were probably related to differences in hydrogen (H) metabolism among algal groups, also observed in the δ2 H values of their lipids. The marked difference between the δ2 H values of Ulvophyecae and those of the other groups could be useful to trace the food source of food webs in coastal rocky shores, to assess the impacts of green tides on coastal ecosystems, and to help clarify aspects of their phylogeny. However, reference materials for seaweed δ2 H are required before the full potential of using the δ2 H of seaweeds for ecological studies can be exploited.

Structural features and assessment of zymosan-induced arthritis in rat temporomandibular joint model using sulfated polysaccharide / Características estruturais e ensaio de artrite induzida por zimosam em modelo de articulação temporomandibular de ratos usando polissacarídeo sulfatado

The green seaweed Caulerpa cupressoides var. lycopodium contains three SPs fractions (Cc-SP1, Cc-SP2 and Cc-SP3). Cc-SP1andCc-SP2 had anticoagulant (in vitro), pro- and antithrombotic, antinociceptive and/or anti-inflammatory (in vivo) effects. This study analyzed structural features and the antinociceptive and anti-inflammatory effects of Cc-SP1 on zymosan-induced acute arthritis of the rat temporomandibular joint (TMJ). Cc-SP1 was investigated by infrared technique. Male Wistar rats (200-240 g) received subcutaneously (s.c.) Cc-SP1 1h prior to intra-articular (i.art.) injection of zymosan (2 mg joint-1) or saline (0.9%) into the left TMJ. Mechanical hypernociception was measured by the electronic Von Frey method in the basal and 4h after zymosan injection. Animals were euthanized 6h after zymosan injection and the TMJ cavity was removed for total leukocyte counts from the synovial fluid and myeloperoxidase (MPO) activity assessment. Cc-SP1 (1, 3 or 9 mg kg-1) containing sulfate ester, galactose-6-sulfate, uronic acid and glycosidic linkages reduced zymosan-induced hypernociception (78.12, 81.13 and 87.43%, respectively, p 0.01), and inhibited the total leukocyte influx (85, 88.14 and 89.95%, respectively, p 0.01), being confirmed by MPO activity (p 0.05). Therefore, Cc-SP1 reveals a pharmacological tool for treating inflammatory arthropathies.(AU)

A alga marinha verde Caulerpa cupressoides var. lycopodium contém três frações de PSs (Cc-PS1; Cc-PS2 e Cc-PS3). Cc-PS1 e Cc-PS2 apresentaram efeitos anticoagulante (in vitro), pró- e antitrombótico, antinociceptivo e/ou anti-inflammatório (in vivo). Analisaram-se as características estruturais e os efeitos antinociceptivo e anti-inflamatório de Cc-PS1 sobre artrite aguda induzida por zimosam na articulação temporomandibular (ATM) de ratos. Foi investigada a Cc-PS1 por técnica de infravermelho. Ratos machos Wistar (200-240 g) receberam subcutaneamente (s.c.) Cc-PS1 1h antes de injeção intra-articular (i.art.) de zimosam (2 mg articulação-1) ou salina (0,9%) na ATM esquerda. A hipernocicepção mecânica foi mensurada por método Von Frey elétrico em zero e 4h após injeção de zimosam. Os animais foram entanasiados após 6h de injeção de zimosam e a cavidade da ATM foi removida para contagem de leucócitos totais do fluído sinovial e ensaio da atividade de mieloperoxidase (MPO). A Cc-PS1 (1; 3 ou 9 mg kg-1) contendo éster sulfato, galactose-6-sulfato, ácido urônico e ligações glicosídicas reduziu a hipernocicepção induzida por zimosam (78,12; 81,13 e 87,4%, respectivamente; p 0,01), além de inibir o influxo de leucócitos totais (85; 88,14 e 89,95%, respectivamente; p 0,01), sendo, ainda, confirmado pela atividade de MPO (p 0,05). Portanto, a Cc-PS1 revela como uma ferramenta farmacológica para tratar de artropatias inflamatórias.(AU)

Structural features and assessment of zymosan-induced arthritis in rat temporomandibular joint model using sulfated polysaccharide / Características estruturais e ensaio de artrite induzida por zimosam em modelo de articulação temporomandibular de ratos usando polissacarídeo sulfatado

The green seaweed Caulerpa cupressoides var. lycopodium contains three SPs fractions (Cc-SP1, Cc-SP2 and Cc-SP3). Cc-SP1 and Cc-SP2 had anticoagulant (in vitro), pro- and antithrombotic, antinociceptive and/or anti-inflammatory (in vivo ) effects. This study analyzed structural features and the antinociceptive and anti-inflammatory effects of Cc-SP1 on zymosan-induced acute arthritis of the rat temporomandibular joint (TMJ). Cc -SP1 was investigated by infrared technique. Male Wistar rats (200-240 g) received subcutaneously (s.c.) Cc-SP1 1h prior to intra-articular (i.art.) injection of zymosan (2 mg joint -1) or saline (0.9%) into the left TMJ. Mechanical hypernociception was measured by the electronic Von Frey method in the basal and 4h after zymosan injection. Animals were euthanized 6h after zymosan injection and the TMJ cavity was removed for total leukocyte counts from the synovial fluid and myeloperoxidase (MPO) activity assessment. Cc-SP1 (1, 3 or 9 mg kg-1) containing sulfate ester, galactose-6-sulfate, uronic acid and glycosidic linkages reduced zymosan-induced hypernociception (78.12, 81.13 and 87.43%, respectively, p < 0.01), and inhibited the total leukocyte influx (85, 88.14 and 89.95%, respectively, p < 0.01), being confirmed by MPO activity (p < 0.05). Therefore, Cc-SP1 reveals a pharmacological tool for treating inflammatory arthropathies.

A alga marinha verde Caulerpa cupressoides var. lycopodium contém três frações de PSs (Cc-PS1; Cc-PS2 e Cc-PS3). Cc-PS1 e Cc-PS2 apresentaram efeitos anticoagulante (in vitro), pró- e antitrombótico, antinociceptivo e/ou anti-inflammatório (in vivo). Analisaram-se as características estruturais e os efeitos antinociceptivo e anti-inflamatório de Cc-PS1 sobre artrite aguda induzida por zimosam na articulação temporomandibular (ATM) de ratos. Foi investigada a Cc-PS1 por técnica de infravermelho. Ratos machos Wistar (200-240 g) receberam subcutaneamente (s.c.) Cc-PS1 1h antes de injeção intra-articular (i.art.) de zimosam (2 mg articulação-1) ou salina (0,9%) na ATM esquerda. A hipernocicepção mecânica foi mensurada por método Von Frey elétrico em zero e 4h após injeção de zimosam. Os animais foram entanasiados após 6h de injeção de zimosam e a cavidade da ATM foi removida para contagem de leucócitos totais do fluído sinovial e ensaio da atividade de mieloperoxidase (MPO). A Cc-PS1 (1; 3 ou 9 mg kg-1) contendo éster sulfato, galactose-6-sulfato, ácido urônico e ligações glicosídicas reduziu a hipernocicepção induzida por zimosam (78,12; 81,13 e 87,4%, respectivamente; p < 0,01), além de inibir o influxo de leucócitos totais (85; 88,14 e 89,95%, respectivamente; p < 0,01), sendo, ainda, confirmado pela atividade de MPO (p < 0,05). Portanto, a Cc-PS1 revela como uma ferramenta farmacológica para tratar de artropatias inflamatórias.

Algas marinas bentónicas de la Isla Gorgona, costa pacífica colombiana / Benthic marine algae of Gorgona island, Colombian Pacific coast

The knowledge of the biodiversity of Gorgona, a continental island in Colombia, is very limited in the case of algae. We present an updated list of the benthic marine algae of Gorgona Island, associated with different marine environments and type of substrates, such as coral reefs, rocks, and soft bottoms. Field samplings were taken between October 2010 and June 2011. We estimated algal cover, species composition and algal diversity for each reef environment using 0.25 m² quadrats. Ecological and environmental aspects are also included. A total of 43 species of algae for the three types of habitats are documented: 24 Rhodophyta, 12 Clorophyta, six Ochrophyta and one species of Cyanobacteria, and 55% of the algal composition corresponded to red algae. Rev. Biol. Trop. 62 (Suppl. 1): 27-41. Epub 2014 February 01.

Se presenta el listado de las algas marinas bénticas del Parque Nacional Natural Isla Gorgona, el inventario corresponde a la flora algal asociada a los diferentes ambientes marinos de la isla, a saber: arrecifes coralinos y fondos blandos. El monitoreo de las comunidades de algas se llevó a cabo entre octubre de 2010 y mayo de 2011, se realizó una estimación de abundancia y cobertura para lo cual se establecieron transeptos paralelos a la línea de costa y se evaluó el porcentaje de cobertura en cuadrantes de 0.25m². Se reportó un total de 43 especies para los dos ambientes estudiados (coralino y fondos blandos), 24 Rhodophyta, 12 Clorophyta, 6 Ochrophyta y una Cyanobacteria. El grupo más representativo fue el perteneciente a la clase Rhodophyceae, que correspondió al 55.8% de la flora inventariada. Los resultados de nuevos registros en la isla ratifican su importancia fitogeográfica, por cuanto la composición florística insular contribuye significativamente a la biodiversidad de la costa pacífica colombiana, destacándose la presencia de arrecifes coralinos como los principales ambientes marinos para el desarrollo de la flora algal de la región.

Comparative study of sulfated polysaccharides from Caulerpa spp. (Chlorophyceae). Biotechnological tool for species identification? / Estudo comparativo dos polissacarídeos sulfatados de clorofíceas Caulerpa spp. (Chlorophyceae). Ferramenta biotecnológica na identificação de espécies?

Studies on macromolecules isolated from marine algae suggested sulfated polysaccharides (SPs) as possible molecular markers for species. We evaluated isolated and fractionated SPs from the green marine algae Caulerpa cupressoides, C. prolifera and C. racemosa collected at Pacheco Beach, as possible taxonomic molecular indicators. Total SPs were extracted with papain in 100 mM sodium acetate buffer (pH 5.0) containing cysteine and EDTA (both 5 mM), followed by ion-exchange chromatography on DEAE-cellulose using a NaCl gradient. The obtained fractions were analyzed by 0.5% agarose gel electrophoresis. Anticoagulant assays employing normal human plasma and standard heparin (193 IU mg-1) by the activated partial thromboplastin time (APTT) test were also performed as comparison parameters. Low yields, and similar chromatographic profiles were found among species' SPs, but electrophoresis revealed distinct SPs resolution patterns. The changes in APTT of SP fractions were dependent on charge density as showed by electrophoresis profiles. Activities were 17.37 (C. cupressoides), 22.17 (C. racemosa) and 25.64 (C. prolifera) IU mg-1, respectively, similar to a previous study using the first and second species. The results suggest that comparative studies of SPs isolated from seaweeds may be an important tool for the identification of Caulerpaceae.

A utilização de macromoléculas isoladas de organismos marinhos sugere correlacionar características em estudos taxonômicos e a investigação comparativa de polissacarídeos sulfatados (PSs) de algas despertam seu interesse como marcadores moleculares. Objetivou-se avaliar PSs isolados e fracionados das algas marinhas verdes Caulerpa cupressoides, C. prolifera e C. racemosa, coletadas na Praia do Pacheco, Estado do Ceará, como possíveis indicadores moleculares taxonômicos. Os PSs totais foram extraídos com papaína em tampão acetato de sódio 100 mM (pH 5,0) contendo cisteína e EDTA (ambos 5 mM), seguido por cromatografia de troca iônica em coluna de DEAE-celulose utilizando um gradiente de NaCl. As frações obtidas foram analisadas por eletroforese em gel de agarose a 0,5%. Ensaios anticoagulantes, utilizando o teste do tempo de tromboplastina parcial ativada (TTPA) com plasma humano normal e heparina padrão (193 UI mg-1), também foram realizados como parâmetros de comparação. Verificaram-se baixos rendimentos e semelhantes perfis cromatográficos entre os PSs das espécies, porém revelando, por eletroforese, diferenças moleculares marcantes. As alterações no TTPA das frações de PS foram dependentes da densidade de cargas negativas mostradas nos perfis eletroforéticos, cujas atividades foram 17,37 (C. cupressoides), 22,17 (C. racemosa) e 25,64 (C. prolifera) UI mg-1, respectivamente, e tal propriedade justificou um estudo já realizado utilizando a primeira e segunda espécies. Os resultados sugerem que estudos comparativos de PSs isolados de algas marinhas possam vir a ser uma ferramenta importante na identificação de Caulerpaceae.

Extraction and anticoagulant activity of sulfated polysaccharides fromCaulerpa cupressoides var. lycopodium (Vahl) C. Agardh (Chlorophyceae) / Extração e atividade anticoagulante dos polissacarídeos sulfatados da clorofícea Caulerpa cupressoides var. lycopodium (Vahl) C. Agardh

The reportedly low standard quality of heparin (HEP) for use in cardiac surgerieshas led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) fromseaweeds have been regarded as promising substitutes for HEP. The aim of this study was tosequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed byfractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate theanticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) usingnormal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions werechemically characterized by chemical composition and agarose gel electrophoresis. The yield was4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl,respectively, were observed on chromatography profiles; however, differences in charge densitiespatterns and degree of resolution among them were revealed by electrophoresis. SPs were capableof modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C.cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.(AU)

O baixo padrão dequalidade outrora declarado da heparina (HEP) para o uso em cirurgias cardíacas tem levadopreocupação nos mercados nacional e internacional. Os polissacarídeos sulfatados (PSs) de algasmarinhas têm sido considerados como promissores substitutos para HEP. Objetivou-se a extrairsequencialmente PSs totais (PSTs) da clorofícea Caulerpa cupressoides com papaína em tampãoacetato de sódio 100 mM (pH 5,0) contendo cisteína 5 mM e EDTA 5 mM, fracionar porcromatografia de troca iônica (DEAE-celulose) e avaliar o potencial anticoagulante das frações dePS por meio do tempo de tromboplastina parcial ativada (TTPA), utilizando plasma humanonormal e comparando-se à HEP padrão (193 IU mg-1). As frações obtidas foram caracterizadasquimicamente em composição química e por eletroforese em gel de agarose. O rendimento dePSTs foi 4,61% e os perfis cromatográficos, em DEAE-celulose, indicaram a separação de trêsfrações de PS (F I; F II e F III) eluídas nas concentrações 0,50; 0,75 e 1,00 M de NaCl,respectivamente, revelando, por eletroforese, diferenças em termos de densidade de cargas e graude resolução. Os PSs foram capazes de modificar o TTPA somente nas frações eluídas com 0,75M de NaCl, cujas atividades foram 23,37 e 25,76 IU mg-1, respectivamente, quando a densidadede cargas foi pré-requisito para atividade. Portanto, C. cupressoides é uma fonte de PSs com baixospotenciais anticoagulantes comparados à HEP.(AU)

Extraction and anticoagulant activity of sulfated polysaccharides from Caulerpa cupressoides var lycopodium (Vahl) C Agardh (Chlorophyceae) / Extração e atividade anticoagulante dos polissacarídeos sulfatados da clorofícea Caulerpa cupressoides var lycopodium (Vahl) C Agardh

The reportedly low standard quality of heparin (HEP) for use in cardiac surgeries has led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) from seaweeds have been regarded as promising substitutes for HEP. The aim of this study was to sequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100 mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed by fractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate the anticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) using normal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions were chemically characterized by chemical composition and agarose gel electrophoresis. The yield was 4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl, respectively, were observed on chromatography profiles; however, differences in charge densities patterns and degree of resolution among them were revealed by electrophoresis. SPs were capable of modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and 25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C. cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.

O baixo padrão de qualidade outrora declarado da heparina (HEP) para o uso em cirurgias cardíacas tem levado preocupação nos mercados nacional e internacional. Os polissacarídeos sulfatados (PSs) de algas marinhas têm sido considerados como promissores substitutos para HEP. Objetivou-se a extrair sequencialmente PSs totais (PSTs) da clorofícea Caulerpa cupressoides com papaína em tampão acetato de sódio 100 mM (pH 5,0) contendo cisteína 5 mM e EDTA 5 mM, fracionar por cromatografia de troca iônica (DEAE-celulose) e avaliar o potencial anticoagulante das frações de PS por meio do tempo de tromboplastina parcial ativada (TTPA), utilizando plasma humano normal e comparando-se à HEP padrão (193 IU mg-1). As frações obtidas foram caracterizadas quimicamente em composição química e por eletroforese em gel de agarose. O rendimento de PSTs foi 4,61% e os perfis cromatográficos, em DEAE-celulose, indicaram a separação de três frações de PS (F I; F II e F III) eluídas nas concentrações 0,50; 0,75 e 1,00 M de NaCl, respectivamente, revelando, por eletroforese, diferenças em termos de densidade de cargas e grau de resolução. Os PSs foram capazes de modificar o TTPA somente nas frações eluídas com 0,75 M de NaCl, cujas atividades foram 23,37 e 25,76 IU mg-1, respectivamente, quando a densidade de cargas foi pré-requisito para atividade. Portanto, C. cupressoides é uma fonte de PSs com baixos potenciais anticoagulantes comparados à HEP.

Chemical composition of cultivated seaweed Ulva clathrata (Roth) C. Agardh.

Samples of cultivated Ulva clathrata were collected from a medium scale system (MSS, 1.5×1.5m tank), or from a large scale system (LSS, 0.8ha earthen pond). MSS samples were dried directly while the LSS sample was washed in freshwater and pressed before drying. Crude protein content ranged 20-26%, essential amino acids accounting for 32-36% of crude protein. The main analysed monosaccharides were rhamnose (36-40%), uronic acids (27-29%), xylose (10-13%) and glucose (10-16%). Some notable variations between MSS and LSS samples were observed for total dietary fibre (26% vs 41%), saturated fatty acids (31% vs 51%), PUFAS (33% vs 13%), carotenoids (358 vs 169mgkg-1dw) and for Ca (9 vs 19gkg-1), Fe (0.6 vs 4.2gkg-1), Cu (44 vs 14mgkg-1), Zn (93 vs 17mgkg-1) and As (2 vs 9mgkg-1). The chemical composition of U. clathrata indicates that it has a good potential for its use in human and animal food.

Structure and dynamics of phytoplankton in an Amazon lake, Brazil

Natural lake systems represent important reservoirs for residential water supply, fish production, recreational activities and enjoyment of their natural beauty. Nevertheless, human impacts may affect their health status resulting in degradation and loss of biodiversity. The aim of the present study was to obtain data on the health status of a natural lake located in an indigenous reservation in the Brazilian Amazon, using the phytoplankton community changes along the rainy (June) and dry (November) seasons of 2006. We collected water (temperature, pH, Secchi depth and conductivity) and phytoplankton samples from the subsurface, middle of the water column, and approximately 30cm above the bottom, over 24-hour sampling periods, from a central station in the lake. Samples taken from biotic and abiotic variables were correlated using canonical correspondence analysis (CCA). Results showed that the lake exhibited high temperatures in both seasons, and showed thermal stratification only during the rainy season. Dissolved oxygen exhibited a clinograde pattern in the rainy season and high oxygen in the hypolimnion in the dry season. In the rainy season, the water near the bottom was acidic, turbid and had a greater concentration of phosphorus. Dissolved oxygen, conductivity, pH, nitrite, total phosphorus and total dissolved phosphorus exhibited diel variations in the rainy season, whereas water temperature, dissolved oxygen, total nitrogen and total dissolved phosphorus exhibited significant differences between hours of the day in the dry season. The phytoplankton was represented by 39 taxa, and Chlorophyta showed the greatest species richness, totaling 25 taxa. Among Chlorophyta, desmids were the most diverse, accounting 52%. Bacillariophyta (nine species) was the second most diverse group. Cyanophyta was represented by three species, including Merismopedia tenuissima, the most abundant taxon. Despite the occurrence of taxa that indicate organic pollution, their biomass and frequency indicate that the system is not currently threatened. Lake Caracaranã is an oligotrophic system, with low algal density and isolated blooming episodes due to its shallow depth. Rev. Biol. Trop. 58 (4): 1421-1436. Epub 2010 December 01.

Los sistemas de lagos naturales constituyen reservorios importantes para el abastecimiento de agua residencial, la producción de peces, actividades recreativas y el disfrute de su belleza natural. Sin embargo, el impacto humano puede afectar su estado de salud como resultado de la degradación y la pérdida de biodiversidad. El objetivo del estudio fue determinar el estado de salud de un lago natural en una reserva indígena de la Amazonia brasileña, usando los cambios de la comunidad fitoplanctónica a lo largo de la época lluviosa (junio) y seca (de noviembre) en 2006. Se recogieron muestras de agua (temperatura, pH, conductividad y profundidad de Secchi) y fitoplancton del subsuelo, columna media del agua y a 30cm por encima del fondo, cada 24horas, en una estación central en el lago. Las variables bióticas y abióticas se correlacionaron mediante análisis de correspondencia canónica (CCA). Los resultados mostraron que el lago exhibió altas temperaturas en ambas temporadas, una estratificación térmica solamente durante la época lluviosa así como un patrón vertical de oxígeno disuelto, mientras que en la estación seca se observó una alta cantidad de oxígeno en el hipolimnion. En la época lluviosa, el agua cercana al fondo era ácida, turbia y tenía una mayor concentración de fósforo. Oxígeno disuelto, conductividad, pH, nitritos, fósforo total y disuelto mostraron variaciones diarias en la época lluviosa, mientras que la temperatura del agua, oxígeno disuelto, nitrógeno total y fósforo disuelto mostraron una diferencia significativa en las horas del día durante la estación seca. El fitoplancton estuvo representado por 39 táxones, y Chlorophyta mostró la mayor riqueza de especies, un total de 25 táxones. En Chlorophyta, los desmidios fueron los más diversos, lo que representa el 52%. Bacillariophyta (nueve especies) fue el segundo grupo más diverso. En Cyanophyta se encontraron tres especies, incluyendo Merismopedia tenuissima, el taxon más abundante. A pesar de la ocurrencia de taxones que indican contaminación orgánica, la biomasa y frecuencia sugieren que el sistema actualmente no está en peligro. Lago Caracaranã es un sistema oligotrófico, con baja densidad de algas y aislados episodios de afloramiento debido a su poca profundidad.