Biosynthesis of silver nanoparticles using novel Bacillus sp. SBT8

Biosynthesis of silver nanoparticles (AgNPs) using microorganisms is an important application of nanobiotechnology and green chemistry because of interest by pharmaceutical and food manufacturers. In this study, biosynthesis of AgNPs by a novel Bacillus strain isolated from a soil sample from Sakarya district in Turkey was investigated. Biosynthesis was performed using cell-free supernatant of the bacterium following 24?h growth. Effects of varying AgNO₃ concentration (1–10?mM), pH (5–10), and temperature (30–40°C) on the synthesis of AgNPs were determined. Formation of AgNPs was monitored by UV–VIS spectroscopy. Field emission scanning electron microscopy was used to compare morphologies among the various culture conditions. The peaks created by surface plasmon resonance (SPR) of metals were obtained only at 4 and 6?mM AgNO₃ concentrations and the maximum concentration for the biosynthesis was observed at 6?mM. The highest yield was achieved at pH 10 and larger nanoparticles were obtained at this pH. The optimum temperatures for biosynthesis were 33 and 37°C. Fourier transform infrared spectroscopy analysis and transmission electron microcopy images confirmed that the proteins served as capping. Energy-dispersive spectroscopy analysis validated the formation of AgNPs. AgNPs exhibited antibacterial activity toward Gram-positive and Gram-negative pathogens.     

Novel marine Nocardiopsis dassonvillei-DS013 mediated silver nanoparticles characterization and its bactericidal potential against clinical isolates

The sediment marine samples were obtained from several places along the coastline of the Tuticorin shoreline, Tamil Nadu, India were separated for the presence of bioactive compound producing actinobacteria. The actinobacterial strain was subjected to 16Sr RNA sequence cluster analysis and identified as Nocardiopsis dassonvillei- DS013 NCBI accession number: KM098151. Bacterial mediated synthesis of nanoparticles gaining research attention owing its wide applications in nonmedical biotechnology. In the current study, a single step eco-friendly silver nanoparticles (AgNPs) were synthesized from novel actinobacteria Nocardiopsis dassonvillei- DS013 has been attempted. The actinobacterial mediated silver nanoparticles were characterized by TEM, UV–Visible, XRD, FT-IR spectroscopy. The initial detection of AgNPs was identified using UV–Vis spectrum and confirmed by the appearance of absorbance peak at 408 nm. A Fourier transform infrared spectroscopy (FT-IR) result reveals the presence of protein component in the culture supernatant may act as protecting agents. The XRD pattern indicated that the typical peaks reveal the presence of nanoparticles. The TEM morphology confirms the formation of circular and non uniform distributions of AgNPs with the size range from 30 to 80 nm. The antibacterial activity of both isolated actinobacterial (IA) and silver nanoparticles mediated actinobacterial (SNA) of Nocardiopsis dassonvillei- DS013 were done by well diffusion method against selected clinical isolates of bacteria, namely Escherichia coli, Enterococcus sp., Pseudomonas sp., Klebsiella sp., Proteus sp., Shigella sp., Bacillus subtilis, and Streptococcus sp. When compared to isolated actinobacteria, the SNA shows the better antibacterial activity against clinical isolates.     

Eco-friendly synthesis of silver nanoparticles by Bacillus subtilis and their antibacterial activity

Bacillus subtilis was used for biogenic of silver nanoparticles. Characterization of the prepared silver nanoparticles was done by UV–Vis spectroscopy, Transmission Electron Microscopy (TEM), and Fourier Transform Infrared Spectroscopy (FT-IR). The particle size of the prepared nanoparticles ranges from 3 to 20 nm with spherical or roughly spherical forms. The antimicrobial efficacy of the produced nanoparticles was investigated against five strains of multidrug resistant microorganisms including: Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Klebsiella. pneumoniae, Escherichia coli and Candida albicans tested as yeast. During this study, the minimum inhibitory concentrations (MICs) and the minimum lethal concentrations (MLCs) of synthesized silver nanoparticles were detected using selected strains of the genus Bacillus by a broth dilution method. The rate of MIC of the prepared silver nano-particles versus the investigated clinical isolates exhibit a massive anti-microbial efficacy; (230 µgml⁻¹) for MRSA; 180 for Staphylococcus epidermidis, 200 for Escherichia coli and 100 µgml⁻¹ for Candida albicans. On the other hand, the lowest anti-microbial efficacy (300 µgml⁻¹) was appeared for Klebsiella pneumonia. The obtained results demonstrated the effectiveness of the biogenic nanoparticles and the possibility of using them as a new method in combating infectious diseases.     

Sterols of the senegalese brown alga Padina vickersiae

Contrary to all the brown algae already studied for sterol content the Senegalese Padina vickersiae contains a great amount of cholesterol, chiefly in the later stages of growth. The seasonal variation of the sterols is studied and compared to other Phaeophyceae belonging to the genus Cystoseira.     

Phyconanotechnology: synthesis of silver nanoparticles using brown marine algae Cystophora moniliformis and their characterisation

Biosynthesis of metallic nanoparticles is a relatively new developing area of nanotechnology which has economic and environmentally friendly advantages over conventional chemical and physical methods of synthesis. In this paper, we report for the first time, on the synthesis of silver nanoparticles (AgNPs) using the Australasian brown marine algae Cystophora moniliformis. An extract of this alga was used as a reducing and stabilising agent. Temperature-dependent variation of the size of the AgNPs was observed. Agglomeration of the nanoparticles was observed at high temperatures. The average size of the AgNPs formed at temperatures?<?65°C was 75 nm, whereas they were >2 μm at higher temperatures. The X-ray diffraction (XRD) pattern revealed face-centered cubic structure of the formed Ag nanoparticles.     

Biosynthesis of silver nanoparticles using Malva parviflora and their antifungal activity

Cheeseweed mallow (Malva parviflora L.) was used to biosynthesize silver nanoparticles. The biosynthesized silver nanoparticles were classified by UV–vis Spectroscopy and Fourier-Transform Infrared Spectroscopy (FT-IR). The shape and size distribution were visualized by Transmission Electron Microscopy (TEM), Field Emission Scanning Electron Microscopy (FE-SEM), and Zeta potential analysis. The chemical composition of M. parviflora leaf extract was identified by Gas Chromatography and Mass Spectroscopy (GC/MS). Finally, in vitro antifungal assay was done to assess the potential of biosynthesized silver nanoparticles and crude leaf extract of M. parviflora for inhibiting the mycelial growth of phytopathogenic fungi. The UV–vis analysis manifests the formation of silver nanoparticles. FTIR analysis established that chemicals of the leaf extract stabilized the biosynthesized silver nanoparticles by binding with the free silver ions. The TEM, FE-SEM and zeta potential analyzer confirmed that the biosynthesized silver nanoparticles were mostly spherical with an average diameter of 50.6 nm. The biosynthesized silver nanoparticles and leaf extract of M. parviflora effectively mitigate the mycelial growth of Helminthosporium rostratum, Fusarium solani, Fusarium oxysporum, and Alternaria alternata. The maximum reduction in mycelial growth by biosynthesized nanoparticles was observed against H. rostratum (88.6%). Whereas, the leaf extract of M. parviflora was most effective against F. solani (65.3%). Thus, the biosynthesis of nanoparticle assisted by M. parviflora is a feasible and eco-friendly method for the synthesis of silver nanoparticles. Further the silver nanoparticles and leaf extract of M. parviflora could be explored for the development of the fungicide.     

Biosynthesis and characterization of silver nanoparticles produced by Pleurotus ostreatus and their anticandidal and anticancer activities

The biosynthesis of nanoparticles has received increasing interest because of the growing need to develop safe, cost-effective and environmentally friendly technologies for the synthesis of nano-materials. In this study, silver nanoparticles (AgNPs) were synthesized using a reduction of aqueous Ag⁺ ions with culture supernatant from Pleurotus ostreatus. The bioreduction of AgNPs was monitored by ultra violet-visible spectroscopy and the obtained AgNPs were characterized by transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy techniques. TEM studies showed the size of the AgNPs to be in the range of 4–15 nm. The formation of AgNPs might be an enzyme-mediated extracellular reaction process. Furthermore, the antifungal effect of AgNPs against Candida albicans as compared with commercially antifungal drugs was examined. The effect of AgNPs on dimorphic transition of C. albicans was tested. The anticancer properties of AgNPs against cells (MCF-7) were also evaluated. AgNPs caused a significant decrease in cell viability of an MCF-7 cell line (breast carcinoma). Exposure of MCF-7 cells with AgNPs resulted in a dose-dependent increase in cell growth inhibition varying from 5 to 78 % at concentrations in the range of 10–640 μg ml^?1. The present study demonstrated that AgNPs have potent antifungal, antidimorphic, and anticancer activities. The current research opens a new avenue for the green synthesis of nano-materials.     

Biosynthesis of silver nanoparticles using Penicillium verrucosum and analysis of their antifungal activity

The present study describes the biosynthesis of silver nanoparticles, using the fungus Penicillium verrucosum. The silver nanoparticles were synthesised by reacting silver nitrate (AgNO₃) with the cell free filtrates of the fungal culture, and were then characterized by UV–visible spectroscopy, transmission electron microscopy, scanning electron microscopy, energy-dispersive, and X-ray diffraction analysis to further evaluate their successful biosynthesis, optical and morphological features (size and shape), and crystallinity. The bioactivity of the synthesized nanoparticles against two phytopathogenic fungi i.e: Fusarium chlamydosporum and Aspergillus flavus was evaluated using nanomaterial seeding media. These biogenic silver nanoparticles were polydisperse in nature, with a size of 10–12 nm. With regard to the antifungal activity, 150 ppm of the nanoparticles suppressed the growth of F. chlamydosporum and A. flavus by about 50%. To the best of our knowledge, this is the first report on the use of P. verrucosum to synthesise silver nanoparticles. The present study demonstrates a novel, simple, and eco-friendly process for the generation of biofunctionally useful biogenic nanoparticles.     

Effects of solar radiation and solar radiation deprived of UV-B and total UV on photosynthetic oxygen production and pulse amplitude modulated fluorescence in the brown alga Padina pavonia

Abstract: The effects of solar radiation on photosynthetic oxygen production and pulse amplitude modulated (PAM) fluorescence were measured in the marine brown macroalga Padina pavonia harvested from different depths from the Greek coast near Korinth. In fluence rate-response curves the light compensation point for photosynthetic oxygen production increased and the saturation level decreased with increasing exposure time to solar radiation. Cutting off the UV-B wavelength range (280–315 nm) from solar radiation reduced the inhibition of photosynthesis, and the organisms were less affected when all of the UV radiation was filtered out. Algae collected from 7 m depth were much more prone to photoinhibition than those harvested from rock pools exposed to unfiltered solar radiation. During continuous exposure to solar radiation, rock pool algae showed photoinhibition after longer periods of time than specimens from 7 m or from dark adapted habitats. When subjected to unfiltered solar radiation the ratio of the variable fluorescence to the maximal fluorescence     (F_v = F_m− F_o) rapidly declined with increasing exposure time. However, again algae from 7 m depth were more prone to photoinhibition than rock pool algae. The differences between the two ecological strains were less obvious when UV-B or total UV was removed from solar radiation. Only in the latter case a complete recovery was observed after 2 h while, when exposed to unifiltered sunlight, only the rock pool algae recovered completely within that time.     

Extracellular biosynthesis,characterization, optimization of silver nanoparticles (AgNPs) using Bacillus mojavensis BTCB15 and its antimicrobial activity against multidrug resistant pathogens

Abstract

Biosynthesis of metal nanoparticles is an area of interest among researchers because of its eco-friendly approach. Current study focuses at biosynthesis of silver nanoparticles (AgNPs) and optimization of physico-chemical conditions to obtain mono-dispersed and stable AgNPs having antimicrobial activity. Initially Bacillus mojavensis BTCB15 produced silver nanoparticles (AgNPs) of 105?nm. Silver nanoparticles (AgNPs) were characterized by particle size analyzer, UV-Vis Spectroscopy, Fourier transforms infrared spectroscopy (FTIR), Atomic force microscopy (AFM), and X-ray diffraction (XRD). Whereas, under optimal conditions of temperature 55?°C, pH 8, addition of surfactant Tween 20, and metal ion K₂SO₄, about 104% size reduction was achieved with average size of 2.3nm. Molecular characterization revealed 98% sequence homology with Bacillus mojavensis. AgNPs exhibited antibacterial activity at concentrations ranging from 0.5 to 2.5?µg/µl against Escherichia coli BTCB03, Klebsiella pneumonia BTCB04, Acinetobacter sp. BTCB05, and Pseudomonas aeruginosa BTCB01 but none against Staphylococcus aureus BTCB02. Highest antibacterial activity was observed at 0.27?µg/µl and lowest at 0.05?µg/µl of AgNPs indicated by zone of inhibition. Conclusively, under optimum conditions, Bacillus mojavensis BTCB15 was able to produce AgNPs of 2.3?nm size and had antibacterial activity against multi drug resistant pathogens.     

Fruit extract capped colloidal silver nanoparticles and their application in reduction of methylene blue dye

Green synthesis of silver nanoparticles (AgNPs) has become a promising environmentally benign synthetic route in nanoscience and nanotechnology during recent years. In the present work, we have developed an environment-friendly and low-cost method for synthesis of silver nanoparticles from silver nitrate using aqueous fruit extract of Dillenia indica. The as-synthesized nanoparticles were characterized by UV-Vis spectrophotometer, transmission electron microscopy (TEM) and X-ray diffraction (XRD). FTIR study was performed to know the interaction of bio-molecules present in the fruit extract with AgNPs. The catalytic application of the as-synthesized AgNPs was demonstrated against degradation of methylene blue (MB) in aqueous system. The absorption spectra of colloidal suspension of AgNPs showed characteristic surface plasmon resonance (SPR) band centred at a wavelength of 416?nm. TEM image showed that the AgNPs were almost spherical in shape having an average diameter of 10.78?±?.48?nm. XRD pattern and selected area electron diffraction (SAED) pattern with bright spots signify the crystalline nature of nanoparticles. The fruit extract-capped AgNPs was highly stable and have showed the effective catalytic activity in reduction of MB dye.     

Biosynthesis of silver nanoparticles by natural precursor from clove and their antimicrobial activity

Silver nanoparticles (AgNPs) have attracted the attention of researchers because of their unique properties and applications in various fields, such as medicine, catalysis, textile engineering, and pollution treatment. The green synthesis of AgNPs has many advantages, such as less time requirement, highly stable AgNPs, better control over crystal growth, morphology, ease for scale up, and economic viability. Syzygium aromaticum (clove) was used for the extracellular biosynthesis of AgNPs. Eugenols are the active biomolecules present in clove, responsible for the bioreduction of AgNO₃ (Ag⁺) leading to the formation and capping of AgNPs (Ag⁰). One molecule of eugenol releases two electrons and these two electrons will be taken by 2 Ag⁺ ions and these will get reduced to 2 Ag⁰. The synthesis of AgNPs was confirmed by the appearance of brown colour. The synthesized AgNPs were characterised by various techniques, such as UV-VIS spectroscopy, transmission electron microscopy, X-ray diffraction and Fourier transformed infrared spectroscopy. The synthesised AgNPs have λ _max of 440 nm. It was evaluated that the AgNPs were biphasic in nature (cubic + hexagonal) with an average size of 50.0 nm. The synthesized AgNPs showed significant antimicrobial activity against Bacillus cereus NCDC 240 as they are nano-sized and have high surface area to volume ratio. AgNPs inhibit the growth of bacteria by various ways, such as by disrupting the cell membrane of bacteria, uncoupling the oxidative phosphorylation, inhibiting the DNA replication, forming free radicals and affecting the cellular signalling of bacteria leading to cell death.     

钩状木霉生物合成纳米银及其杀菌性能

【目的】以钩状木霉为微生物材料合成纳米银粒子,并对其杀菌性能进行测定。【方法】将钩状木霉与2 mmol/L的Ag NO3溶液混合暗培养合成纳米银,采用UV-vis、XRD和TEM等方法对纳米银进行表征;利用原子吸收光谱仪和热重分析仪测定并计算银离子的转化率和纳米银的产率;以大肠杆菌和枯草芽孢杆菌为受试菌株检测纳米银的杀菌性能。【结果】钩状木霉与硝酸银混合的培养液颜色为红褐色,UV-vis图谱显示在420 nm左右出现了强的吸收峰;XRD图谱出现了4个特征性衍射峰,分别对应纳米银的4个晶面;TEM照片可以看出纳米银多数为球形,具有单分散性;粒度分布仪显示纳米银具有很窄的粒径分布,在1-13 nm之间,平均粒径为6.69 nm;根据原子光谱吸收仪测定的结果得到银的转化率为84.41%,根据热重分析结果得到纳米银的产率为67.12%;纳米银对大肠杆菌的MBC为10 mg/L,MIC为7 mg/L;对枯草芽孢杆菌的MBC为5 mg/L,MIC为4 mg/L。【结论】钩状木霉与Ag NO3溶液混合培养可以合成纳米银。合成的纳米银大小均匀,粒径小且分布很窄,具有面心立方结构,是纯净的,产率约为67.12%;纳米银对枯草芽孢杆菌的致死效果好于对大肠杆菌的致死效果。     

Biogenic synthesis of silver nanoparticles: Antibacterial and cytotoxic potential

In green chemistry, the application of a biogenic material as a mediator in nanoparticles formation is an innovative nanotechnology. Our current investigation aimed at testing the cytotoxic potential and antimicrobial ability of silver nanoparticles (AgNPs) that were prepared using Calligonum comosum roots and Azadirachta indica leaf extracts as stabilizing and reducing agents. An agar well diffusion technique was employed to detect synthesized AgNPs antibacterial ability on Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus bacterial strains. Furthermore, their cytotoxic capability against LoVo, MDA-MB231 and HepG2 ca cells was investigated. For phyto-chemical detection in the biogenic AgNPs the Fourier-transform infrared spectroscopy (FT-IR) was considered. Zeta sizer, TEM (Transmission Electron Microscope) and FE-SEM (Field Emission Scanning Electron Microscope) were used to detect biogenic AgNPs’ size and morphology. The current results showed the capability of tested plant extract for conversion of Ag ions to AgNPs with a mean size ranging between 90.8 ± 0.8 and 183.2 ± 0.7 nm in diameter. Furthermore, prepared AgNPs exhibited apoptotic potential against HepG2, LoVo, and MDA-MB 231cell with IC₅₀ ranging between 10.9 and 21.4 μg/ml and antibacterial ability in the range of 16.0 ± 0.1 to 22.0 ± 1.8 mm diameter. Activation of caspases in AgNPs treated cells could be the main indicator for their positive effect causing apoptosis. The current investigation suggested that the green production of AgNPs could be a suitable substitute to large-scale production of AgNPs, since stable and active nanoparticles could be obtained.     

A cytotoxic oxysterol from the marine alga Padina pavonica (L.) Thivy

The dichloromethane extract of Padina pavonica showed a cytotoxic activity against KB cells. An oxysterol, hydroperoxy-24 vinyl-24 cholesterol, 1, was identified as being responsible for this activity. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization of silver nanoparticles synthesized by using marine isolate Streptomyces albidoflavus

Silver nanoparticles production by the green chemistry approach was investigated using an isolated marine actinomycetes strain. The isolated strain was identified as Streptomyces albidoflavus based on chemotaxonomic and ribotyping properties. The strain revealed production of silver nanoparticles both extracellular and intracellularly. Surface Plasmon Resonance analysis with the function of time revealed that particle synthesis by this strain is reaction time dependent. The produced particles were spherical shaped and monodispersive in nature and showed a single surface plasmon resonance peak at 410 nm. Size distribution histograms indicated production of 10-40- nm-size nanoparticles with a mean size of 14.5 nm. FT-IR spectra of nanopartilces showed N-H, C-H, and C-N stretching vibrations, denoting the presence of amino acid/ peptide compounds on the surface of silver nanoparticles produced by S. albidoflavus. Synthesized nanoparticles revealed a mean negative zeta potential and electrophoretic mobility of -8.5 mV and -0.000066 cm2/Vs, respectively. The nanoparticles produced were proteinaceous compounds as capping agents with -8.5 mV zeta potential and revealed antimicrobial activity against both Gram-negative and -positive bacterial strains. Owing to their small size, these particles have greater impact on industrial application spectra.     

Biosynthesis of silver nanoparticles by the endophytic fungus Epicoccum nigrum and their activity against pathogenic fungi

There is an enormous interest in developing safe, cost-effective and environmentally friendly technologies for nano-materials synthesis. In the present study, extracellular biosynthesis of silver nanoparticles was achieved by Epicoccum nigrum, an endophytic fungus isolated from the cambium of Phellodendron amurense. The reduction of the silver ions was monitored by UV–visible spectrophotometry, and the characterization of the Ag NPs was carried out by X-ray diffraction and transmission electron microscopy. The synthesized Ag NPs were exceptionally stable. It was found that an alkaline pH favored the formation of Ag NPs and elevated temperature accelerated the reduction process. Furthermore, the antifungal activity of the Ag NPs was assessed using a microdilution method. The biosynthesized Ag NPs showed considerable activity against the pathogenic fungi. The current research opens a new path for the green synthesis of Ag NPs and the process is easy to scale up for biomedical applications.     

Biosynthesis,characterization and antimicrobial activity of silver nanoparticles by a halotolerant Bacillus endophyticus SCU-L

Abstract

We have conducted a thorough study on extracellular biosynthesis of silver nanoparticles (AgNPs) by a halotolerant bacterium Bacillus endophyticus SCU-L, which was identified by 16S rRNA gene sequencing analysis. This strain was selected during an ongoing research programme aimed at finding a novel biological method for green nanosynthetic routes using the extremophiles in unexplored hypersaline habitats. The biosynthesized AgNPs were characterized and analyzed with UV–vis spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, atomic force microscopy and X-ray diffraction. Further, the AgNPs were found to be spherical in shape with an average particle size of about 5.1?nm, and it was stable in aqueous solution for three months period of storage at room temperature under dark condition. Also, the synthesized AgNPs significantly presented antimicrobial activity against Candida albicans, Escherichia coli, Salmonella typhi and Staphylococcus aureus. The above results suggested that the present work may provide a valuable reference and theoretical basis for further exploration on microbial biosynthesis of AgNPs by halotolerant bacteria.     

Biosynthesis,characterization and antimicrobial activity of silver nanoparticles by Streptomyces sp. SS2

In the present study the microbial biosynthesis of silver nanoparticles (AgNPs) by secondary metabolites of Streptomyces sp. SS2 in an eco-friendly approach has been reported. The Streptomyces sp. SS2 was isolated from the soil sediment of Similipal Biosphere Reserve. The identification of this strain was determined by phenotypical characteristics (morphological and biochemical) and molecular characterization method using 16 s rDNA sequencing. The morphological study was also done by high-resolution scanning electron microscopy. The preliminary characterization of biosynthesized silver nanoparticle was carried out using UV–Vis spectrum analysis, which showed an absorption peak at 420 nm corresponding to plasmon absorption of silver. The average size and charge (zeta potential) of the particles were found to be 67.95 ± 18.52 nm and ?17.7 ± 5.30 mV, respectively. The functional groups were identified by FTIR studies and their morphology (round and spherical shape) was determined by scanning electron microscopy. The synthesized AgNPs exhibited excellent antibacterial activity against Escherichia coli (MTCC 1089), Bacillus subtilis (MTCC 7164), Staphylococcus epidermis (MTCC 3615), Vibrio cholerae (MTCC 3904) and Staphylococcus aureus (MTCC 1144). These biotechnological approaches of synthesis of nanoparticles can direct a new path in biomaterial sciences and enrich biomedical applications.     

Localization of specific monosaccharides in cells of the brown alga Padina gymnospora and the relation to heavy-metal accumulation

The brown alga Padina gymnospora has been studied due to their ecological significance and biochemical characteristics, including its high capability of heavy-metal accumulation. It has been suggested that the fucans are among the main polysaccharides related to metal binding and precipitation in cell walls. The main purpose of this work was to determine the localization of specific monosaccharides in P. gymnospora cells. In this way, the lectins Ulex europaeus agglutinin and Canavalia ensiformis concanavalin A with specificity to alpha-L-fucose and to terminal residues of alpha-D-glucosyl and alpha-D-mannosyl, respectively, were applied in young individuals. These revealed a preferential distribution of alpha-L-fucose at cell walls near the external surface in cortical cells and near the plasmalemma in cortical and medullar cells. The distribution of alpha-L-fucose in cell walls indicates the distribution of sulfated polysaccharides (sulfated fucans) that colocalize with the heavy-metal granules (Zn and Cd) described in previous works. Therefore, our results suggest that alpha-L-fucose participates in the nucleation and immobilization of heavy metals in P. gymnospora cell walls. An intense labeling of U. europaeus agglutinin and a weak labeling of concanavalin A was also observed in physodes. X-ray microanalysis revealed the presence of zinc, sulfur, and calcium in physodes of algae collected in a heavy-metal-contaminated area. Besides the affinity between polyphenolic compounds and heavy metals, it is suggested that the mechanism of metal binding by physodes could be related to the presence of sulfated fucans.