Heavy Metal Bioaccumulation in Enteromorpha intestinalis, (L) Nees, a Macrophytic Algae: The Example of Kadin Creek (Western Anatolia)

ABSTRACT This study aims to specify the amount of ten heavy metals (Al, Cr, Mn, Ni, Cu, Zn, As, Cd, Hg, Pb) in water, sediment and Enteromorpha intestinalis samples gathered from Kadin Creek in seasonal periods in 2011 and 2012 and to reveal bioaccumulation capability of this macrophytic algae. In this regard, heavy metal concentrations in the gathered samples were determined first with mineralization and then via inductively coupled plasma mass spectroscopy (ICP-MS) method. Findings suggested that algae could be used as an indicator in freshwater ecosystems in terms of studied heavy metals. With regard to bioconcentration factor values, in terms of water, algae accumulated aluminum the most and cadmium the least (Al > Cu > As > Zn > Cr > Mn > Ni > Pb > Hg > Cd), in terms of sediment, it accumulated mercury the most and aluminum the least (Hg > Cd > As > Zn > Pb > Mn > Ni > Cr > Cu > Al). The highest rate of heavy metal concentration in algae was observed in summer when contaminants in water and dynamics of plant development increased. It was also observed that the mean mercury and cadmium concentrations exceeded the maximum residue limit values.

Influence of bioactive sulfated polysaccharide-protein complexes on hepatocarcinogenesis, angiogenesis and immunomodulatory activities

OBJECTIVE@#To explore the in vivo anticancer, anti-angiogenesis and immunomodulatory efficacies of the bioactive polysaccharide isolated from cold aqueous extract of Jania rubens (JCEM) and Pterocladia capillacea (PCEM) as well as hot aqueous extract of Enteromorpha intestinalis (EHEM) against hepatocellular carcinoma rat model (HCC) and to study their chemical composition.@*METHODS@#The sugars and amino acids composition of the bioactive polysaccharides of JCEM, PCEM and EHEM were determined using gas liquid chromatography and amino acid analyzer, respectively. These polysaccharide extracts (20 mg/kg b.wt. for 5 weeks) were assessed on hepatocarcinogenesis in rats and α-fetoprotein (AFP), carcinoembryonic antigen (CEA), glypican-3 (GPC-3), hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) and Ig G levels were evaluated.@*RESULTS@#The GLC analysis of JCEM, PCEM and EHEM polysaccharide revealed the presence of 10, 9 and 10 sugars, in addition the amino acid analyzer enable identification of 16, 15 and 15 amino acids, respectively. These polysaccharide extracts of JCEM, PCEM and EHEM produced significant decrease in serum AFP, CEA, GPC-3, HGF and VEGF compared with untreated HCC group. JCEM, PCEM and EHEM had an immunostimulatory responses by increasing the IgG levels as compared by naïve value (1.23, 1.53 and 1.17 folds), respectively. The bioactive polysaccharides in HCC induced rats improved the humoral immune response. The photomicrographs of liver tissue sections of the groups of HCC treated with polysaccharide extracts of Jania rubens and Enteromorpha intestinalis showed intact histological structure. Moreover, fractions HE1, HE4, HE7 obtained from polysaccharide of EHEM showed moderate cytotoxic activity against HepG2 in vitro with IC 73.1, 42.6, 76.2 μg/mL. However, fractions of PCEM and JCEM show no or weak cytotoxicity against HepG2 in vitro where the cytotoxic activity of their crude polysaccharide extract proved synergetic effect.@*CONCLUSIONS@#The pronounced antitumor activity of sulfated polysaccharide-protein complexes of JCEM and EHEM is due to direct cytotoxic activity, anti-hepatocarcinogenesis, and anti-angiogenesis. In addition, JCEM, PCEM and EHEM had an immunostimulatory response and improved the humoral immune response in HCC induced rats.

Influence of bioactive sulfated polysaccharide-protein complexes on hepatocarcinogenesis, angiogenesis and immunomodulatory activities

Objective To explore the in vivo anticancer, anti-angiogenesis and immunomodulatory efficacies of the bioactive polysaccharide isolated from cold aqueous extract of Jania rubens (JCEM) and Pterocladia capillacea (PCEM) as well as hot aqueous extract of Enteromorpha intestinalis (EHEM) against hepatocellular carcinoma rat model (HCC) and to study their chemical composition. Methods The sugars and amino acids composition of the bioactive polysaccharides of JCEM, PCEM and EHEM were determined using gas liquid chromatography and amino acid analyzer, respectively. These polysaccharide extracts (20 mg/kg b.wt. for 5 weeks) were assessed on hepatocarcinogenesis in rats and α-fetoprotein (AFP), carcinoembryonic antigen (CEA), glypican-3 (GPC-3), hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) and Ig G levels were evaluated. Results The GLC analysis of JCEM, PCEM and EHEM polysaccharide revealed the presence of 10, 9 and 10 sugars, in addition the amino acid analyzer enable identification of 16, 15 and 15 amino acids, respectively. These polysaccharide extracts of JCEM, PCEM and EHEM produced significant decrease in serum AFP, CEA, GPC-3, HGF and VEGF compared with untreated HCC group. JCEM, PCEM and EHEM had an immunostimulatory responses by increasing the IgG levels as compared by naïve value (1.23, 1.53 and 1.17 folds), respectively. The bioactive polysaccharides in HCC induced rats improved the humoral immune response. The photomicrographs of liver tissue sections of the groups of HCC treated with polysaccharide extracts of Jania rubens and Enteromorpha intestinalis showed intact histological structure. Moreover, fractions HE1, HE4, HE7 obtained from polysaccharide of EHEM showed moderate cytotoxic activity against HepG2 in vitro with IC

Enteromorpha intestinalis Derived Seaweed Liquid Fertilizers as Prospective Biostimulant for Glycine max

ABSTRACT In the present study, the potential of seaweed liquid fertilizer (SLF) of marine algae Enteromorpha intestinalis was evaluated for its effect on seed germination, yield, biochemical parameters and pigment characteristics of Glycine maxE. intestinalis was collected form Mandapam coast of Gulf of Mannar, Tamil Nadu, and the dried seaweeds were used for the preparation of SLF. G. max seeds were germinated with four different concentrations (20, 40, 60, and 100%) of SLF; its growth and yield parameters were evaluated and compared with chemical fertilizer and control. The morphological and bio-chemical parameters such as seed germination (100%), root (6.6cm) and shoot length (5.4 cm), carbohydrates (0.098 mg/g), protein (0.56 mg/g), pigment (0.444 mg/g chl a; 1.073 mg/g chl b; 3.70 mg/g carotenoids) of the plant was found maximum at a concentration of 60% SLF. The phenol content (3.25 mg/g) was maximum in 40% SLF. The GC-MS analysis of SLF revealed the presence of notable benzoic compounds involved in plant growth promotion. Results showed thatE. intestinalis derived SLF was potential biostimulant forG. max. Thus, marine algae based fertilizer could be an effective and alternate to the chemical fertilizers emphasizing the need for systematic evaluation programme for SLF on various crops.

Comparative assessment of bacterial diversity associated with co-occurring eukaryotic hosts of Palk Bay origin.

Epibacterial communities of co-occurring eukaryotic hosts of Palk Bay origin (five seaweed species (Gracilaria sp, Padina sp, Enteromorpha sp, Sargassum sp, and Turbinaria sp) and one seagrass [Cymodaceae sp]) were analyzed for diversity and compared using 16S rRNA based Denaturant Gradient Gel Electrophoresis analysis. Diversity index revealed that Turbinaria sp hosts highest bacterial diversity while it was least in Gracilaria sp. The DGGE band profile showed that the epibacterial community differed considerably among the studied species. Statistical assessment using cluster analysis and Non-metric multidimensional scale analysis also authenticated the observed variability. Despite huge overlap, the composition of bacterial community structure differed significantly among the three closely related species namely Sargassum, Turbinaria and Padina. In addition, Enteromorpha and Sargassum, one being chlorophyta and the other phaeophyta showed about 80% similarity in bacterial composition. This differs from the general notion that epibacterial community composition will vary widely depending on the host phyla. The results extended the phenomenon of host specific epibacterial community irrespective of phylogeny and similarity in geographical location.

In vitro antimicrobial activities of methanolic extract from marine alga Enteromorpha intestinalis

To extract the bioactive compound from Enteromorpha intestinalis (E. intestinalis) and determine its in vitro antimicrobial activity. Methods: E. intestinalis was extracted by methanol and subjected to antimicrobial screening. The antimicrobial activity was studied by using disc diffusion and broth dilution method. The effect of the extract on the growth profile of the bacterial was also examined via time-kill assay. Microscopy observations using SEM was done to determine the major alterations in the microstructure of methicillin-resistant Staphylococcus aureus (MRSA). Results: The results showed methanolic extract of E. intestinalis exhibited a favourable antimicrobial activity against tested bacteria with produced inhibition zone ranging from 8.0-19.0 mm. However, all the tested fungi and yeast were resistant to the extract treatment. Time kill assay suggested that methanolic extract of E. intestinalis had completely inhibited MRSA growth and also exhibited prolonged antibacterial activity. The main abnormalities noted from the microscopic observations were the structural deterioration in the normal morphology and complete collapsed of the bacteria cells after 36 h of treatment. Conclusions: The significant antibacterial activity shown by crude extract suggested its potential against MRSA infection. The extract may have potential to develop as antibacterial agent in pharmaceutical use.

In vitro antimicrobial activities of methanolic extract from marine alga Enteromorpha intestinalis

OBJECTIVE: To extract the bioactive compound from Enteromorpha intestinalis (E. intestinalis) and determine its in vitro antimicrobial activity. METHODS: E. intestinalis was extracted by methanol and subjected to antimicrobial screening. The antimicrobial activity was studied by using disc diffusion and broth dilution method. The effect of the extract on the growth profile of the bacterial was also examined via time-kill assay. Microscopy observations using SEM was done to determine the major alterations in the microstructure of methicillin-resistant Staphylococcus aureus (MRSA). RESULTS: The results showed methanolic extract of E. intestinalis exhibited a favourable antimicrobial activity against tested bacteria with produced inhibition zone ranging from 8.0 to 19.0 mm. However, all the tested fungi and yeast were resistant to the extract treatment. Time kill assay suggested that methanolic extract of E. intestinalis had completely inhibited MRSA growth and also exhibited prolonged antibacterial activity. The main abnormalities noted from the microscopic observations were the structural deterioration in the normal morphology and complete collapsed of the bacteria cells after 36 h of treatment. CONCLUSIONS: The significant antibacterial activity shown by crude extract suggested its potential against MRSA infection. The extract may have potential to develop as antibacterial agent in pharmaceutical use.

Optimization of Microwave-assisted Extraction of Polyphenols from Enteromorpha prolifra by Orthogonal Test / 中草药·英文版

Objective To optimize microwave-assisted extraction of polyphenols from Enteromorpha prolifra.Methods Based on single-factor tests,an efficient microwave-assisted extraction(MAE)technique was developed to extract bioactive polyphenols from E.prolifra through orthogonal L16(4)5 test.Results The highest yield(0.923±0.013)mg/g was obtained when microwave power,solvent to raw material ratio,irradiation time,ethanol concentration,and extraction cycles were 500 W,25 mL/g,25 min,40%,and 3,respectively,which was higher than that of Soxhlet extraction with methanol for 6 h,ultrasound-assisted extraction with 40% ethanol for 1 h twice and heat reflux extraction with 40%ethanol for 2 h twice.Conclusion This finding indicates that MAE is a superior technique for the extraction of polyphenols due to less impurity,higher time efficiency and yield.

Flora béntica y reproducción de las algas Batophora spp. (Chlorophyta: Dasycladaceae) de una laguna costera contaminada (Bahía de Chetumal, México) / Benthic flora and reproduction of Batophora spp. algae (Chlorophyta: Dasycladaceae) in a polluted coastal lagoon (Chetumal Bay, Mexico)

The benthic flora, and the vegetative and reproductive characters of the algae Batophora oerstedii and B. occidentalis (Chlorophyta) were recorded from five sites of Chetumal Bay, Quintana Roo, Mexico. A sewage gradient has been reported along those sites. Plants were sampled in May and October 1999, which corresponded to dry and rainy seasons, respectively. Forty taxa were found, 11 are new records for the Chetumal Bay, and 6 are new records for the Mexican Caribbean. Enteromorpha species were present in sites known as rich in organic matter (both from anthropogenic and natural sources). Batophora spp. is the dominant algae in all Chetumal Bay. However, it was absent next to sewage outfalls. The morphological characters of B. oerstedii and B. occidentalis did not change significantly along the sites reported as polluted. The length and width of gametophores, as well as the diameter of the gametangia were clearly different for both species. Different reproductive strategies may help B. oerstedii and B. occidentalis to closely coexist in the Chetumal Bay.

Los componentes de la flora béntica, y mediciones de partes vegetativas y reproductivas de Batophora oerstedii y B. occidentalis (Chlorophyta) se registraron en cinco sitios de la costa Oeste de la Bahía de Chetumal, donde se ha registrado la existencia de un gradiente de contaminación orgánica. La colecta de flora se realizó en mayo y Octubre de 1999, meses incluidos en las épocas climáticas e secas y lluvias, respectivamente. Se registraron 40 taxa de vegetación béntica, de los cuales 11 representan nuevos registros para la Bahía de Chetumal, y 6 para el Caribe Mexicano. Especies de Enteromorpha estuvieron presentes en ambientes ricos en materia orgánica, tanto de origen urbano, como natural. En los desagües de la Bahía de Chetumal existen especies indicadoras de contaminación, pero la ausencia de Batophora spp., dominante en esta laguna costera. Los caracteres morfológicos de B. occidentalis y B. oerstedii a lo largo de los cinco sitios de muestreo no reflejaron la presencia de contaminantes. Ambas especies presentaron diferencias significativas en su largo y ancho de gametóforos, y en el diámetro de gametangios Las diferencias en estrategias reproductivas probablemente ayudan a que B. oerstedii y B. occidentalis puedan coexistir cercanamente en la Bahía de Chetumal.

Study on the antioxidant activity of extracts from Enteromorpha prolifera / 中国海洋药物

Objective To investigate the antioxidant activity of Enteromorpha prolifera by ABTS radical scavenging assay in vitro and provide scientific basis for exploitation of Enteromorpha prolifera.Methods The antioxidant activities of extracts by different solvents from Enteromorpha prolifera and water extracts from different Enteromorpha prolifera samples were investigated by ABTS radical scavenging assay.Moreover,the antioxidant ability of the water extract from Enteromorpha prolifera was evaluated by using caffeic acid and ascorbic acid as positive control standards.Results The water extract from Enteromorpha prolifera provided the highest radical scavenging activity,and the antioxidant capability of 1mL of water extract from Enteromorpha prolifera was equivalent to 1mL of 2.55 10~(-3) mg?mL~(-1) caffeic acid or 1mL of 2.89?10~(-3)mg?mL~(-1) ascorbic acid.However,the extraction methods,dry sample or not and different kinds of Enteromorpha prolifera samples had certain influence on the antioxidant activity of water extract from Enteromorpha prolifera. Conclusion There are abundant antioxidant compounds present in the aqueous extract of Enteromorpha prolifera,and it can be exploited as latent oxidation inhibitor.

Isolation, purification and characterization of lectin from Enteromorpha intestinalis / 中国海洋药物

Lectin from green marine algae Enteromorpha intestinalis (EIL) was isolated by extraction with PBS, followed by ammonium sulfate fraction and a combination of ion-exchange chromatography on DEAE-52 fiber and gel filtration on SephadexG-200. Lectin showed single band on both PAGE and isoelectrofocusing electrophoresis. Its isoelectric point was 8.9 and molecular weight was 17783 tested on SephadexG-200. EIL displayed some agglutination activities with Chlamydomonas and erythrocytes of rabbit, common carp, crucian carp and human (A, B, AB, O), but the maximum agglutination was with crucian carp. The hemagglutinating activity for rabbit erythrocyte could not be inhibited by D-fructose, D-galactose, glucose, sucrose, mannose, lactose, bovine-thyroglobulin, egg albumin and gamma-globulin. The hemagglutinating activity of lectin would be showed at pH 4.0~10.2, and more potent at pH 6.9~8.0. The hemagglutinating activity was not affected at 90℃ for 1h, so it could be said that the lectin had high heat resistance.