Production and particle characterization of the frustules of Cyclotella cryptica in comparison with siliceous earth

Diatoms are photoautotrophic micro-organisms that use inorganic carbon sources and light in photosynthesis. Diatom frustules were characterized in terms of particle techniques and compared with siliceous earth, i.e. depositions of diatoms that have wide technical applications. To obtain enough biomass for frustule characterization Cyclotella cryptica has been cultivated in a 15 l photobioreactor under controlled conditions. Native diatom frustules are characterized by a 1.5-fold lower density and 80-fold higher specific surface than siliceous earth. Therefore, native diatom frustules provides a material with novel properties which might be interesting for special technical application.     

硅藻纳米技术

硅藻是一类微小的单细胞藻类,具有由无定形氧化硅组成的坚硬细胞壁(硅壳).硅壳具有精致的形态和结构,且随硅藻种类和生长条件不同而千变万化.目前估算的硅藻种类超过200 000种,其独特的纳米结构对光子结构、化学生物传感器、新纳米材料和器件的开发具有启发意义.同时硅藻形态形成学和分子生物学的研究,可以推动硅质材料的仿生合成...     

Beyond micromachining: the potential of diatoms.

Diatoms are microscopic, single-celled algae that possess rigid cell walls (frustules) composed of amorphous silica. Depending on the species of diatom and the growth conditions, these frustules can display a wide range of different morphologies. It is possible to design and produce specific frustule morphologies that have potential applications in nanotechnology.     

Comportamento Di Un Carattere Teratologico Comparso in Navicula Minima Grun

Abstract

Behaviour of a teratological character present in «Navicula minima» Grun. — A teratological form with curved cells like the kidneys has been isolated from a culture of Navicula minima Grun, to follow the behaviour during the biological cycle. Alternately varied cells in the morphology of the frustules appeared in the descendants, deriving from a modification more or less evident in the initial teratological character. The main cause of this phenomenon can be found in the normalization process of the frustules that has been put in relation to a probable plasticity of the cellular Silicon walls. The cells, even though more or less deformed, have given origin to auxozygotes which, having terminated their growth, have formed perfectly normal frustules.     

The ratio of diatom frustules to chrysophycean statospores: A useful paleolimnological index

Stratigraphic analyses of the postglacial sediments of Little Round Lake and the recent sediments of two additional southern Ontario lakes were used to compare the relative abundance of fossil diatom frustules to chrysophycean statospores. Relatively few statospores were recorded during eutrophic phases in the lakes' histories such as during periods of anthropogenic activity in the lakes' catch me nt, when diatom frustules were abundant. The increased relative abundance of statospores during the early postglacial in Little Round Lake was attributed to both oligotrophy and a short growing season, conditions that currently exist in many high arctic lakes. From these data it is seen that the ratio of fossil diatom frustules to chrysophycean statospores may provide a useful paleolimnological index of trophic status in temperate lakes. The advantages of this index include its simplicity, the fact that frustules and statospores can be enumerated from the same microscope slides, and that the ratio is independent of sedimentation rates.     

EVIDENCE OF AN ORGANIC MATRIX FROM DIATOM BIOSILICA1

Most biominerals appear to be composites of organic material and mineral. Whether biosilica is such a composite is unresolved because of a lack of evidence for such organic components. We present evidence that organic material exists within diatom biosilica and can be extracted using HF/NH₄F solutions from frustules isolated from Cyclotella meneghiniana Kütz and diatomaceous earth. To eliminate organic casing on the silicified frustules as a source of organic materials, the casing was removed by oxidation of frustules with NaOCl before extraction. The removal of the casing was confirmed in that oxidized frustules no longer displayed the ability to be stained with ruthenium red and fluorescamine. Frustules examined with EDXA showed an emission peak from sulfur before treatment but no peak following treatment, indicating that oxidation removed organic sulfur. The organic material obtained from extracts of fresh frustules contained both soluble and insoluble components. Only soluble material was evident in extracts from diatomaceous earth. The soluble material appears to contain glycoproteins with relatively high levels of serine and glycine. The soluble proteins from fresh frustules also appear to be phosphorylated. Indirect evidence is presented that suggests the soluble proteins may contain regions of primary structure enriched in anionic amino acids. The soluble extracts differ from general cell contents when the two fractions are compared, suggesting that frustules contain specialized organic material. The identification of silica-specific organic material suggests that mineralization in diatoms may be in part matrix-mediated.     

ULTRASTRUCTURE OF CHAETOCEROS MUELLERI (BACILLARIOPHYCEAE): AUXOSPORE,RESTING SPORE AND VEGETATIVE CELL MORPHOLOGY1

The structure of the frustule of auxospores, resting spores and vegetative cells of Chaetoceros muelleri Lemm. are described with LM and SEM. Vegetative frustules are relatively small and lightly silicified, are not united into filaments, and appear unornamented under LM and SEM. The setae are circular to subcircular in transverse section with spines and puncta arranged in a spiral pattern. The resting spore and auxospore frustules are more silicified than the vegetative frustules and appear unornamented under LM and SEM. The auxospores of C. muelleri were previously unknown.     

Density gradient separation of microbenthos from estuarine sediments

Summary The main drawback in investigations on microbenthos in estuarine regions are formed by the presence of detritus, clay minerals and aggregates of these components. Moreover, counts of benthic diatoms in these areas are also made difficult by the presence of empty frustules. Separation of empty frustules from living benthic diatoms is necessary, because identification of specimens must be carried out with empty frustules. Several techniques have been developed to isolate specific groups of organisms from the sediments.For benthic epipelic diatoms the lens-tissue technique was developed; the technique failed for estuarine sediments. Harvest percentages ranged from about 15 to 75%. A more adequate method was obtained by using density gradient centrifugation with the polymer silica Ludox. With a newly developed technique, which will be published in detail elsewhere, it appeared possible to separate about 87% of the total amount of benthic epipelic as well as epipsammic diatoms from empty diatom frustules, clay minerals and sand particles. The diatom fraction still contained a great deal of detritus which, however, does not disturb the microscopical identification of specimens because it can be oxidized.Some meiobenthic groups (copepods, nematodes) can be isolated quantitatively from sediment samples with the density technique. Such fractions contained only a small amount of detritus.     

The Diatom Staurosirella pinnata for Photoactive Material Production

A native isolate of the colonial benthic diatom Staurosirella pinnata was cultivated for biosilica production. The silicified cell walls (frustules) were used as a source of homogeneous and structurally predictable porous biosilica for dye trapping and random laser applications. This was coupled with the extraction of lipids from biomass showing potential to fabricate photoactive composite materials sustainably. The strain was selected for its ease of growth in culture and harvesting. Biosilica and lipids were obtained at the end of growth in indoor photobioreactors. Frustules were structurally characterized microscopically and their chemistry analyzed with Fourier Transform Infrared Spectroscopy. Frustule capacity of binding laser dyes was evaluated on a set of frustules/Rhodamine B (Rho B) solutions and with respect to silicon dioxide and diatomite by Fluorescence Spectroscopy demonstrating a high affinity for the organic dye. The effect of dye trapping property in conveying Rho B emission to frustules, with enhancement of scattering events, was analyzed on Rho B doped polyacrylamide gels filled or not with frustules. Amplified spontaneous emission was recorded at increasing pump power indicating the onset of a random laser effect in frustule filled gels at lower power threshold compared to unfilled matrices.     

Luminescence properties of a nanoporous freshwater diatom

Freshwater diatom frustules show special optical properties. In this paper we observed luminescence properties of the freshwater diatom Cyclotella meneghiniana. To confirm the morphological properties we present scanning electron microscopy (SEM) images. X‐ray diffraction (XRD) studies were carried out to visualize the structural properties of the frustules, confirming that silica present in diatom frustules crystallizes in an α‐quartz structure. Study of the optical properties of the silica frustules of diatoms using ultra‐violet‐visible (UV‐vis) spectroscopy and photoluminescence spectroscopy confirmed that the diatom C. meneghiniana shows luminescence in the blue region of the electromagnetic spectrum when irradiated with UV light. This property of diatoms can be exploited to obtain many applications in day‐to‐day life. Also, using time‐resolved photoluminescence spectroscopy (TRPL) it was confirmed that this species of diatom shows bi‐exponential decay. Copyright © 2011 John Wiley & Sons, Ltd.     

New tools for labeling silica in living diatoms

Silicon biomineralization is a widespread mechanism found in several kingdoms that concerns both unicellular and multicellular organisms. As a result of genomic and molecular tools, diatoms have emerged as a good model for biomineralization studies and have provided most of the current knowledge on this process. However, the number of techniques available to study its dynamics at the cellular level is still rather limited. Here, new probes were developed specifically to label the pre-existing or the newly synthesized silica frustule of several diatoms species. It is shown that the LysoTracker Yellow HCK-123, which can be used to visualize silica frustules with common filter sets, presents an enhanced signal-to-noise ratio and allows details of the frustules to be imaged without of the use of ionophores. It is also demonstrated that methoxysilane derivatives can be coupled to fluorescein-5-isothiocyanate (FITC) to preferentially label the silica components of living cells. The coupling of labeling procedures might help to address the challenging question of the process of frustule exocytosis.     

MORPHOLOGICAL VARIATION,SPECIES CONCEPTS,AND CLASSIFICATION OF AN UNUSUAL FOSSIL CENTRIC DIATOM (BACILLARIOPHYTA) FROM WESTERN NORTH AMERICA1

The classification system of diatoms (Bacillariophyta) is based almost entirely on the organization and structure of their silica cell walls. We have discovered Miocene freshwater fossils that contain valves of two different orders within the same organism. Within frustules of the genus Ellerbeckia Crawford, a genus of the order Melosirales, are contained frustules of the genus Actinocyclus Ehrenberg, a member of the Coscinodiscales. Based on the abundance of each morphotype, they appear to be able to reproduce themselves. The ability of a single diatom taxon to express this degree of morphological variability is unprecedented. These results suggest species concepts in diatoms may be in need of review and challenge the basis of the classification system for this widely distributed and ecologically important group of organisms.     

A test on different aspects of diatom processing techniques

A factorial experiment was carried out in order to assess the effects of four treatment steps in diatom processing techniques (the quantity of hydrogen peroxide added to the sample, the time allowed to the digestion reaction, the use of centrifugation and the method used to settle the frustules onto the coverslip) on different aspects of the quality of the resulting microscopic slide: the concentration of organic and inorganic matter, the density, randomness and homogeneity of the frustule distribution in the slide, and the abundance of broken frustules and girdle views. The results show no significant effects of any of the experimental treatments on the proportion of broken frustules in the samples. No significant effects were related to the duration of the oxidation reaction in any of the aspects evaluated in the quality of the slides. A sample:hydrogen peroxide ratio of 1:1 for a 6-h oxidizing reaction can be used to optimize the protocol. Despite being more time consuming, the use of successive decantations instead of centrifuging is more appropriate to achieve better results. Slide quality can also be improved by leaving the coverslips to dry at room temperature in order to improve the distribution of frustules.     

New insights into the complex architecture of siliceous copepod teeth

Copepods belong to the dominant marine zooplankton taxa and play an important role in particle and energy fluxes of the marine water column. Their mandibular gnathobases possess tooth-like structures, so-called teeth. In species feeding on large proportions of diatoms these teeth often contain silica, which is very probably the result of a coevolution with the siliceous diatom frustules. Detailed knowledge of the morphology and composition of the siliceous teeth is essential for understanding their functioning and their significance in the context of feeding interactions between copepods and diatoms. Based on analyses of the gnathobases of the Antarctic copepod Rhincalanus gigas, the present study clearly shows, for the first time, that the silica in the siliceous teeth features large proportions of crystalline silica that is consistent with the mineral α-cristobalite and is doped with aluminium. The siliceous structures have internal chitinous fibre networks, which are assumed to serve as scaffolds during the silicification process. The compact siliceous teeth of R. gigas are accompanied by structures with large proportions of the elastic protein resilin, likely reducing the mechanical damage of the teeth when the copepods feed on diatoms with very stable frustules. The results indicate that the coevolution with diatom frustules has resulted in gnathobases exhibiting highly sophisticated composite structures.     

POLYMORPHISM IN MASTOGLOIA1

Cells in a -wild population of a species belonging to the diatom germs Mastogloia may form frustules of 2 strikingly different morphologic types during their vegetative life cycle. Total conversion from coarsely structured morphologic form to a more finely structured form takes place during a single division; hence no inter gradations between the 2 forms are evident. Intact frustules with 1 valve of each type leave no reasonable doubt of the ability to make the change.     

Ultra‐sensitive immunoassay biosensors using hybrid plasmonic‐biosilica nanostructured materials

We experimentally demonstrate an ultra‐sensitive immunoassay biosensor using diatom biosilica with self‐assembled plasmonic nanoparticles. As the nature‐created photonic crystal structures, diatoms have been adopted to enhance surface plasmon resonances of metal nanoparticles on the surfaces of diatom frustules and to increase the sensitivity of surface‐enhanced Raman scattering (SERS). In this study, a sandwich SERS immunoassay is developed based on the hybrid plasmonic‐biosilica nanostructured materials that are functionalized with goat anti‐mouse IgG. Our experimental results show that diatom frustules improve the detection limit of mouse IgG to 10 pg/mL, which is ?100× better than conventional colloidal SERS sensors on flat glass.

Ultra‐sensitive immunoassay biosensor using diatom biosilica with self‐assembled plasmonic nanoparticles.     

The Probable Mechanism for Silicon Capture by Diatom Algae: Assimilation of Polycarbonic Acids with Diatoms—Is Endocytosis a Key Stage in Building of Siliceous Frustules?

Many organisms including unicellular (diatoms, radiolaria, and chrysophytes), higher plants (rice and horsetail) and animals (sponges) use silica as a main part of skeletons. The bioavailable form of silicon is silicic acid and the mechanism of silicic acid penetration into living cells is still an enigma. Macropinocytosis was assumed as a key stage of the silicon capture by diatoms but assimilation of monomeric silicic acid by this way requires enormous amounts of water to be passed through the cell. We hypothesized that silicon can be captured by diatoms via endocytosis in the form of partially condensed silicic acid (oligosilicates) whose formation on the diatom surface was supposed. Oligosilicates are negatively charged nanoparticles and similar to coils of poly(acrylic acid) (PAA). We have synthesized fluorescent tagged PAA as well as several neutral and positively charged polymers. Cultivation of the diatom Ulnaria ferefusiformis in the presence of these polymers showed that only PAA is able to penetrate into siliceous frustules. The presence of PAA in the frustules was confirmed with chromatography and PAA causes various aberrations of the valve morphology. Growth of U. ferefusiformis and two other diatoms in the presence of tri- and tetracarbonic fluorescent tagged acids points to the ability of diatoms to recognize substances that bear four acidic groups and to include them into siliceous frustules. Thus, partial condensation of silicic acid is a plausible first stage of silicon assimilation.     

Processing samples of benthic marine diatoms from Mediterranean oligotrophic areas

The processing of benthic diatoms is tedious and involves several potentially damaging steps for cells. Although the preservation of siliceous frustules is of paramount importance in the implementation of biotic indices, only few studies quantified treatment-induced cell losses. We assumed that commonly used treatments may lead to mechanical (centrifugation, sedimentation, boiling, sonication and mounting in Naphrax) and chemical (cold H₂O₂ digestion) damages on diatoms. We analysed the potential adverse effects of these treatments and the cleaning efficiency of H₂O₂ and incineration in order to find out the most suitable technique to process lightly silicified Mediterranean populations. Results showed that successive resuspensions of material after each concentration treatment (sedimentation and centrifugation) and low speed centrifugation did not alter the physical integrity of frustules. In contrast, boiling and sonication exhibited adverse effects especially on the preservation of large frustules and Naphrax mounting proved to be the most damaging step whatever the size of diatoms. For cleaning treatments, incineration provided the most satisfactory results and acted on a non-selective way as opposed to hydrogen peroxide which led to either a large number of non-cleaned frustules or dissolved valves. Our recommendations for processing samples of lightly silicified Mediterranean benthic diatoms include the use of low-speed centrifugations, dehydration at room temperature, incineration and dry mounting.     

Benthic algal mats of some lakes of Inexpressible Island (northern Victoria Land,Antarctica)

 Benthic algal mats from four lakes located on Inexpressible Island (northern Victoria Land, Antarctica) were studied, analysing their structure and floristic composition. Thirty taxa of algae were identified: 11 Cyanophyta, 15 Bacillariophyta and 4 Chlorophyta. The lake mats contained from 15 (lake C) to 26 species (lake D). The number of diatom frustules in the mats varied from 771×10³ (lake C) to 9428×10³ frustules g^-1 (lake B). In terms of floristic composition and macromorphology, these mats were observed to be very similar to moat mats described from Lake Gondwana (northern Victoria Land) and the Dry Valleys lakes of southern Victoria Land. Received: 9 July 1995/Accepted: 12 February 1996     

Silicon nanotechnologies of pigmented heterokonts

Many pigmented heterokonts are able to synthesize elements of their cell walls (the frustules) of dense biogenic silica. These include diatom algae, which occupy a significant place in the biosphere. The siliceous frustules of diatoms have species-specific patterns of surface structures between 10 and a few hundred nanometers. The present review considers possible mechanisms of uptake of silicic acid from the aquatic environment, its transport across the plasmalemma, and intracellular transport and deposition of silica inside the specialized Silica Deposition Vesicle (SDV) where elements of the new frustule are formed. It is proposed that a complex of silicic acid with positively charged proteins silaffins and polypropylamines remains a homogeneous solution during the intracellular transport to SDV, where biogenic silica precipitates. The high density of the deposited biogenic silica may be due to removal of water from the SDV by aquaporins followed by syneresis--a process during which pore water is expelled from the network of the contracting gel. The pattern of aquaporins in the silicalemma, the membrane embracing the SDV, can determine the pattern of species-specific siliceous nanostructures.