Barnacle settlement and the adhesion of protein and diatom microfouling to xerogel films with varying surface energy and water wettability

Previous work has shown that organosilica-based xerogels have the potential to control biofouling. In this study, modifications of chemistry were investigated with respect to their resistance to marine slimes and to settlement of barnacle cyprids. Adhesion force measurements of bovine serum albumin (BSA)-coated atomic force microscopy (AFM) tips to xerogel surfaces prepared from aminopropylsilyl-, fluorocarbonsilyl-, and hydrocarbonsilyl-containing precursors, indicated that adhesion was significantly less on the xerogel surfaces in comparison to a poly(dimethylsiloxane) elastomer (PDMSE) standard. The strength of adhesion of BSA on the xerogels was highest on surfaces with the highest and the lowest critical surface tensions, γ_C and surface energies, γ_S, and duplicated the ‘Baier curve’. The attachment to and removal of cells of the diatom Navicula perminuta from a similar series of xerogel surfaces were examined. Initial attachment of cells was comparable on all of the xerogel surfaces, but the percentage removal of attached cells by hydrodynamic shear stress increased with γ_C and increased wettability as measured by the static water contact angle, θ_Ws, of the xerogel surfaces. The percentage removal of cells of Navicula was linearly correlated with both properties (R ² = 0.74 for percentage removal as a function of θ_Ws and R ² = 0.69 for percentage removal as a function of γ_C). Several of the aminopropylsilyl-containing xerogels showed significantly greater removal of Navicula compared to a PDMSE standard. Cypris larvae of the barnacle B. amphitrite showed preferred settlement on hydrophilic/higher energy surfaces. Settlement was linearly correlated with θ_Ws (R ² = 0.84) and γ_C (R ² = 0.84). Hydrophilic xerogels should prove useful as coatings for boats in regions where fouling is dominated by microfouling (protein and diatom slimes).     

Surface chemistry induces mitochondria-mediated apoptosis of breast cancer cells via PTEN/PI3K/AKT signaling pathway

Tumor cell can be significantly influenced by various chemical groups of the extracellular matrix proteins. However, the underlying molecular mechanisms involved in the interaction between cancer cells and functional groups in the extracellular matrix remain unknown. Using chemically modified surfaces with biological functional groups (CH₃, NH₂, OH), it was found that hydrophobic surfaces modified with CH₃ and NH₂ suppressed cell proliferation and induced the number of apoptotic cells. Mitochondrial dysfunction, cytochrome c release, Bax upregulation, cleaved caspase-3 and PARP, and Bcl-2 downregulation indicated that hydrophobic surfaces with CH₃ and NH₂ triggered the activation of intrinsic apoptotic signaling pathway. Cells on the CH₃- and NH₂-modified hydrophobic surfaces showed downregulated expression and activation of integrin β1, with a subsequent decrease of focal adhesion kinase (FAK) activity. The RhoA/ROCK/PTEN signaling was then activated to inhibit the phosphorylation of PI3K and AKT, which are essential for cell proliferation. However, pretreatment of MDA-MB-231 cells with SF1670, a PTEN inhibitor, abolished the hydrophobic surface-induced activation of the intrinsic pathway. Taken together, the present results indicate that CH₃- and NH₂-modified hydrophobic surfaces induce mitochondria-mediated apoptosis by suppressing the PTEN/PI3K/AKT pathway, but not OH surfaces. These findings are helpful to understand the interaction between extracellular matrix and cancer cells, which might provide new insights into the mechanism potential intervention strategies for tumor prognosis.     

Testing attachment point theory: diatom attachment on microtextured polyimide biomimics

Abstract

This paper explores diatom attachment to a range of laser etched polyimide surfaces to directly test ‘attachment point theory’. Static bioassays were conducted on microtextured polyimide surfaces using four diatom species, Fallacia carpentariae, Nitzschia cf. paleacea, Amphora sp. and Navicula jeffreyi with cell sizes ranging from 1 – 14 μm. The microtextured polyimides were modelled from natural fouling resistant bivalve surfaces and had wavelengths above, below and at the same scale as the diatom cell sizes. Diatoms attached in significantly higher numbers to treatments where the numbers of attachment points was highest. The lowest diatom attachment occurred where cells were slightly larger than the microtexture wavelength, resulting in only two theoretical points of attachment. The results support attachment point theory and highlight the need to address larval/cell size in relation to the number of attachment points on a surface. Further studies examining a range of microtexture scales are needed to apply attachment point theory to a suite of fouling organisms and to develop structured surfaces to control the attachment and development of fouling communities.     

Diatoms as water quality indicators in the upper reaches of the Great Fish River,Eastern Cape,South Africa

This study focused on the spring-fed upper reaches of the economically important Great Fish River with the aim of determining if diatoms could be used for biomonitoring in semi-arid conditions in southern Africa. Five sites were monitored monthly from 2010 to 2012. Of the 269 diatom taxa belonging to 51 genera identified, the dominant taxa were mostly those considered to be pollution-tolerant: Amphora pediculus, Craticula buderi, Fragilaria biceps, Nitzschia frustulum, Nitzschia paleacea, Planothidium lanceolatum and Rhopalodia gibba. A number of diatom-based numerical indices were used to infer water quality, including the generic diatom index, the specific pollution sensitivity index, the biological diatom index, and percentage pollution-tolerant valves, which forms part of the UK trophic diatom index. All index scores showed the Great Fish River to be impacted, and showed significant correlations of diatom species abundance with pH, NO₃-N, electrical conductivity, NH4-N and CaCO_3. Analysis revealed EC and NO3-N as the main environmental drivers affecting diatom community composition, followed by pH and PO4-P. The percentage of diatom deformities at all sites was high, at 3.5%. Diatom indices showed the river to be impacted by decades of agricultural activity, which was confirmed by chemical water analysis. Thus diatom indices can be used for biomonitoring in semi-arid areas.     

EFFECTS OF LIGHT AND SUBSTRATE ON THE BENTHIC DIATOMS IN AN OLIGOTROPHIC LAKE: A COMPARISON BETWEEN NATURAL AND ARTIFICIAL SUBSTRATES1

Benthic diatoms form a particularly important community in oligotrophic lakes, but factors influencing their distribution are not well known. This study reports the depth distribution of living motile and total diatoms (living plus dead diatoms) on both natural (from sand to fine organic mud) and artificial substrates in an oligotrophic lake. On artificial substrates, motile diatom densities peaked in abundance (24–30 cells · mm^?2) between 0.6 and 1.9 m depth; on natural sediment surfaces, motile diatoms were generally more numerous and peaked in abundance (925 cells · mm^?2) at 1.3 m depth. Total diatom densities on artificial substrates were highest (1260 valves · mm^?2) at 0.6 m depth, with very low values below 3 m depth; on natural sediment surfaces, total diatom abundances were generally much higher (21600 valves · mm^?2) at 3 m depth and declined gradually with depth. Significant relationships were found between light and diatom densities on the artificial substrate. Ordination analysis indicated that substrate type significantly correlated with the variation of diatom composition on artificial and natural substrates. Our results suggest that in oligotrophic lakes, light influences benthic diatom abundance, whereas substrate type has more influence on benthic diatom composition.     

Influence of the sulfur species reactivity on biofilm conformation during pyrite colonization by Acidithiobacillus thiooxidans

Massive pyrite (FeS₂) electrodes were potentiostatically modified by means of variable oxidation pulse to induce formation of diverse surface sulfur species (S _n ^2?, S⁰). The evolution of reactivity of the resulting surfaces considers transition from passive (e.g., Fe_1?x S₂) to active sulfur species (e.g., Fe_1?x S_2?y , S⁰). Selected modified pyrite surfaces were incubated with cells of sulfur-oxidizing Acidithiobacillus thiooxidans for 24 h in a specific culture medium (pH 2). Abiotic control experiments were also performed to compare chemical and biological oxidation. After incubation, the attached cells density and their exopolysaccharides were analyzed by confocal laser scanning microscopy (CLMS) and atomic force microscopy (AFM) on bio-oxidized surfaces; additionally, S _n ^2?/S⁰ speciation was carried out on bio-oxidized and abiotic pyrite surfaces using Raman spectroscopy. Our results indicate an important correlation between the evolution of S _n ^2?/S⁰ surface species ratio and biofilm formation. Hence, pyrite surfaces with mainly passive-sulfur species were less colonized by A. thiooxidans as compared to surfaces with active sulfur species. These results provide knowledge that may contribute to establishing interfacial conditions that enhance or delay metal sulfide (MS) dissolution, as a function of the biofilm formed by sulfur-oxidizing bacteria.     

Effect of surfaces on amyloid fibril formation

Using atomic force microscopy (AFM) we investigated the interaction of amyloid beta (Aβ) (1–42) peptide with chemically modified surfaces in order to better understand the mechanism of amyloid toxicity, which involves interaction of amyloid with cell membrane surfaces. We compared the structure and density of Aβ fibrils on positively and negatively charged as well as hydrophobic chemically-modified surfaces at physiologically relevant conditions. We report that due to the complex distribution of charge and hydrophobicity amyloid oligomers bind to all types of surfaces investigated (CH₃, COOH, and NH₂) although the charge and hydrophobicity of surfaces affected the structure and size of amyloid deposits as well as surface coverage. Hydrophobic surfaces promote formation of spherical amorphous clusters, while charged surfaces promote protofibril formation. We used the nonlinear Poisson-Boltzmann equation (PBE) approach to analyze the electrostatic interactions of amyloid monomers and oligomers with modified surfaces to complement our AFM data.     

Isolation and characterization of oxygen-evolving thylakoid membranes and Photosystem II particles from a marine diatom Chaetoceros gracilis

Thylakoid membranes retaining high oxygen-evolving activity (about 250 μmol O₂/mg Chl/h) were prepared from a marine centric diatom, Chaetoceros gracilis, after disruption of the cells by freeze-thawing. We also succeeded in purification of Photosystem II (PSII) particles by differential centrifugation of the thylakoid membranes after treatment with 1% Triton X-100. The diatom PSII particles showed an oxygen-evolving activity of 850 and 1045 μmol O₂/mg Chl/h in the absence and presence of CaCl₂, respectively. The PSII particles contained fucoxanthin chlorophyll a/c-binding proteins in addition to main intrinsic proteins of CP47, CP43, D2, D1, cytochrome b559, and the antenna size was estimated to be 229 Chl a per 2 molecules of pheophytin. Five extrinsic proteins were stoichiometrically released from the diatom PSII particles by alkaline Tris-treatment. Among these five extrinsic proteins, four proteins were red algal-type extrinsic proteins, namely, PsbO, PsbQ', PsbV and PsbU, whereas the other one was a novel, hypothetical protein. This is the first report on isolation and characterization of diatom PSII particles that are highly active in oxygen evolution and retain the full set of extrinsic proteins including an unknown protein.     

The diatom flora of the Salton Sea, California

We report on diatom species of the Salton Sea, a highly saline (43 g l^–1) inland lake in California. We identified and photographed all diatom taxa encountered in the phytoplankton and benthos of the Salton Sea and its immediate tributaries. Ninety-four taxa were distinguished based on their morphological features using light- and electron microscopy. In the Salton Sea, there are four general categories of diatom assemblages related to their habitats: (1) A planktonic assemblage composed of Chaetoceros muelleri var. subsalsum, Cyclotella choctawhatcheeana, Cyclotella sp., Cylindrotheca closterium, Pleurosigma ambrosianum, Thalassionema sp.; (2) a benthic assemblage with diatoms that live on the bottom (e.g. genera Caloneis, Diploneis, Entomoneis, Gyrosigma, Plagiotropis, Pleurosigma, Surirella and Tryblionella), or in algal mats (Proschkinia bulnheimii, several species of Navicula and Seminavis gracilenta); (3) an epiphytic community attached to the macroscopic green algae which grow on the rocks and other hard surfaces near shore (e.g. Achnanthes brevipes, Licmophora ehrenbergii, Tabularia parva); and (4) a freshwater assemblage composed of species that get washed in by the rivers and other inflows discharging into the Sea (e.g. Achnanthidium minutissimum, Cocconeis pediculus, Cyclotella atomus, C. scaldensis, Nitzschia elegantula, T. weissflogii). The most striking feature of the phytoplankton is the abundance of species formerly known only from marine environments; this is not surprising given the high salinity and the peculiar history of the lake.     

Photosystem I P700 chlorophyll a apoprotein A1 as PCR marker to identify diatoms and their associated lineage

The morphological characteristics of diatoms are useful for studying their taxonomy. However, the distinction between closely related diatom taxa can be very difficult, especially when the morphological characters are modified by environmental constraints. In the present study, 13 fresh water diatoms were identified morphologically and cultured under axenic conditions. To check this, PCR primers specific for multilocus genes were designed to amplify and screen 13 fresh water diatom monocultures. Multilocus PCR primers (DRR3, scfcpA, Lhcf11, SIT1, SIT3, SIT4, LOC101218388, COI-5P, rbcL, rbcL-3P, LSU D2/D3, UPA, psaA, and 18S rRNA) were tested. It was found that psaA gene, a plant pigment chlorophyll-based PCR marker, amplified in all the diatoms. Out of 13 diatom amplicons, only two fresh water diatoms DNA were sequenced. This included Cyclotella meneghiniana and Sellaphora pupula. The Sanger sequencing results thus established that morphologically identified diatom, Sellaphora pupula, exhibited close phylogeny to Sellaphora whereas fresh water Cyclotella meneghiniana has close lineage to marine diatom Thallosiosira.     

在种与属两个级别评价太子河硅藻群落与环境因子的关系

2009年8—9月,对辽宁省太子河流域67个点位进行采样调查,以硅藻群落为研究对象,比较硅藻属级水平与种级水平相对多度、物种丰富度以及生物多样性指数间的相关性,并比较硅藻属级与种级属性与环境因子的相关性。实验结果表明,太子河流域硅藻属级水平的相对多度、丰富度和多样性指数与硅藻种级水平都极显著相关。Pearson相关性分析表明,硅藻属级水平的丰富度与环境显著相关的因子与种级水平丰富度与环境显著相关的因子相一致。Mantel-Test相关性分析表明,硅藻属级水平的相对多度与环境因子的相关性弱于种级水平与环境因子相关性。典范对应分析结果显示,影响硅藻属级和种级群落结构分布的主要环境因子均为悬浮物。基于硅藻生物评价指数的流域健康评价结果表明,应用硅藻属级和种级属性对太子河流域进行健康评价,其评价结果相一致。     

Distribution of Epilithic Diatoms in Response to Environmental Conditions in an Urban Tropical Stream, Central Kenya

Use of diatoms in monitoring water quality is well acknowledged in developed countries, but only recently has the assessment started gaining importance in developing countries. Diatoms can be obtained from natural and artificial substrates. Appreciating the differences and similarities of diatom assemblages on both substrates may contribute to a better understanding and standardization particularly during monitoring of water quality. During this study we assessed diatom assemblages, biodiversity and trophic indices in relation to water quality along the Nairobi River. Fifteen sites were sampled in September 2000 during the dry season. Diatoms were collected from natural substrates (stones, pebbles) and artificial substrates (100% acrylic wool). On artificial and natural substrates, a total of 190 and 151 taxa were found, respectively, the majority of these taxa (80%) have cosmopolitan distribution and are also widespread throughout tropical African. Species composition changed downstream, five taxa dominated upper and mid stream sites whereas lower stream sites were dominated by one or two taxa. Species richness, diversity, dominance and evenness were positively correlated with NO₃, O₂ and altitude but decreased markedly downstream with a simultaneous increase in total dissolved solids, alkalinity, chemical oxygen demand and PO₄. Ordination and classification (CANOCO and TWINSPAN) showed that diatom assemblages in the Nairobi River responded strongly to water quality changes with respect to concentrations of NO₃, NO₂, total dissolved solids and temperature. Taxa common at less impacted upstream sites included Gomphonema gracilis, Anomoeoneis brachysira and Fragilaria biceps; while common taxa at midstream sites with agricultural catchments were Gomphonema parvulum, Navicula cryptocephala, N. schroeteri, N. bryophila, N. halophila, Nitzschia linearis var. linearis and Cymbella silesica. Achnanthes minutissima var. saprophila, Gomphonema angustum, Navicula subminuscula, N. arvensis, Nitzschia palea and N. umbonata were most common at urban sites, which were polluted by residential and industrial effluents. Trophic diatom indices suggested that water quality was poor at most sites in the Nairobi River. Most sites along the river had low Generic Diatom Index values, GDI (<12) and high Trophic Diatom Index values, TDI 73–78 (median = 76) and 75–84 (median = 77) for artificial and natural substrates, respectively. This study showed that diatoms' response on natural and artificial substrates were similar and reflected environmental conditions correctly.     

Influence of light quality and gassing on the vertical migration of diatoms inhabiting the Wadden Sea

Diatoms inhabiting the Wadden Sea show a rhythmic migration pattern, which is superimposed by the tidal rhythm. In addition to light intensity, light quality has a pronounced influence on the upward-directed migration, thus giving some information on the nature of the relevant photoreceptors. Maximum diatom migration occurred when sediment surfaces were illuminated with blue light. The cell densities in blue light exceeded those of white light control experiments 1.8-fold. Furthermore, we registered a minor peak in the red light region, which reached approximately 60% of the white light controls. Cryptochrome and/or phototropin may thus be involved and act as photoreceptors for the vertical migration pattern. Flushing sediment surfaces of freshly mixed Wadden Sea sediments with air, O₂, CO₂ or N₂ did not show a significant influence of O₂ on the upward migration. The disappearance of diatoms from sediment surfaces which were flushed with CO₂ is most probably caused by the acidification of the sediment bed.Communicated by E. Rachor     

Responses and structural recovery of periphytic diatom communities after short-term disturbance in some rivers (Hanoi, Vietnam)

Field transfer experiments of periphytic diatom assemblages developed on artificial substrates were set up to assess the responses of those communities to environmental disturbances. The glass slides were positioned for colonization at the relatively unpolluted site (Red, in the Red River) and at the heavily polluted site (TL, in the To Lich River) in the beginning of the experiment. After a period of 2?weeks, the colonized glass slides were concomitantly transferred from the unpolluted Red site to the heavily polluted TL site and to the moderate polluted site (NT₂, in the Nhue River) and, conversely, from the TL site to the Red site, and then to the NT₂ site. The responses and the adapting capacity of periphytic diatom communities to environmental changes were assessed through the cell density, diversity index, species richness, taxonomic composition, and diatom indices after 2 and 4?weeks of transfer periods. For all transfers except for the transfer from the Red to the TL site in which the growth inhibition of diatom cells was found, the diatom density significantly increased until the end of the experiment. Thus, the diatom communities have expressed their pollution tolerance or sensitivities by changing their composition to adapt themselves to environmental changes. Characteristic species of the Red site (Gyrosigma scalproides, Navicula recens) were replaced by Nitzschia palea, Nitzschia umbonata, Aulacoseira granulate typical species of the NT₂ site, in the biofilm transferred from the Red site to the NT₂ site. The relative abundances of typical diatom species of the Red site proliferated in the biofilm transferred from the TL site to the Red site. The replacement of periphytic diatom communities appeared after the transfer from the second week at the different sites. The slow shift of the species towards the typical species at the TL site could result from the organized structure of diatoms within biofilm before the transfer from the Red site to the TL site. The shifts in values of the Index of Specific Polluosensitivity and Diatom Assemblage Index to organic pollution throughout the experiment indicated the clear sensitivity of these indices to water quality changes.     

Mono‐ and digalactosyldiacylglycerol composition of the marennine‐producing diatom,Haslea ostrearia: Comparison to a selection of pennate and centric diatoms

Diatoms are one of the largest groups of primary producers in the oceans, yet despite their environmental importance little is known about their plastidial lipid biochemistry. It has been previously reported that Skeletonema species contain primarily C₁₆/C₁₆ and C₂₀/C₁₆ forms of mono‐ and digalactosyldiacylglycerol (MGDG and DGDG, respectively). Likewise, it was also reported that Phaeodactylum tricornutum contains primarily C₁₆/C₁₆ and C₂₀/C₂₀ forms of MGDG and DGDG. We seek to relate their studies to other diatoms, both in the centrics and pennates, with particular focus on the marennine‐producing pennate diatom, Haslea ostrearia. To this end, the composition and positional distribution of fatty acids of MGDG and DGDG were examined using positive‐ion electrospray ionization/mass spectrometry (ESI/MS). Two centric diatoms, Skeletonema marinoi and Thalassiosira weissflogii, and the pennate diatom, P. tricornutum, contained primarily C₂₀/C₁₆ (sn‐1/sn‐2) and C₁₈/C₁₆ forms of MGDG and DGDG. The other pennate diatoms, H. ostrearia and Navicula perminuta, contained primarily C₁₈/C₁₆ or C₁₈/C₁₈ forms of MGDG and DGDG, indicating a previously unrecognized fatty acid diversity in diatom MGDG and DGDG.     

Rare earth element uptake by the marine diatom

Abstract

Rare earth distribution coefficients, DT = (moles cm^-3, cells)/(moles cm^-3, solution), obtained using seawater (S = 36.4, t = 25°C, pH ～ 8.2, pCO₂ ～ 345 μatm) and the marine diatom, Skeletonema costatum, exhibited a strong tendency toward the order Ce > Gd > Yb. Observations of rapid initial uptake, with subsequent gradual uptake over time, are suggestive of initial adsorption onto cell surfaces followed by slow transport to interior cell sites. The average volume concentration factors (DT) obtained in our study are: DTCe = (3.33 ± 0.9) × 10⁵; DTGd = (2.41 ± 0.7) × 10⁵; DTYb = (1.64 ± 0.3) × 10⁵. Distribution coefficient results, expressed as a competition between solution and solid-state complexation terms, indicate that rare earth element complexation, both in solution and on surfaces, strongly increases with atomic number. Relatively small differences in rare earth element distribution coefficients (DT) with atomic number are the result of small differences between large solution and solid-state complexation terms.     

Effects of nutrient (N,P, C) enrichment upon the littoral diatom community of an oligotrophic high-mountain lake

The effects of nutrient additions upon the epilithic diatom communities and the algal standing crop were investigated in the oligotrophic, softwater Lake Piccolo Naret, situated in the Swiss alps. Nutrient-diffusing flower pot substrates were filled with either N (0.15 mol NaNO₃), P (0.015 mol Na₂HPO₄) or C (0.15 mol NaHCO₃) or combinations of them. Twenty-five pots representing eight treatments were placed into the lake in July 1991 and sampled after 42 days of exposure.On the surfaces of all pots containing P we measured higher algal biomasses as on the control pots. The chlorophyll-a maximum of 12.9 µg cm^{– 2} was obtained on NPC pots (0.47 µg cm^{– 2} on control pots). On pots with P, NP or NPC supply high amounts of green algae were detected, also reflected in an increased chl-b/chl-c ratio related to the controls which showed algal communities dominated by diatoms.The diatom communities on the control pots as well as on the pots with N, P and NP had a structure similar to the epilithic community in Lake Piccolo Naret (dominance identity > 58 %). However, the community structures of the diatoms from pots with C addition (C, NC, PC and NPC) differed considerably. This is discussed in view of the cell densities of dominant diatom species. For further comparisons the results of two additional high-mountain lakes are used. By means of a cluster analysis it could be shown that epilithic diatom communities were considerably influenced by C addition, while N and P supply were of minor importance.     

Taxonomic characterization of the microorganisms associated with the cultivable diatom Synedra acus from Lake Baikal

Determination of the taxonomic position of microorganisms associated with diatoms was carried out using microscopic, microbiological, and phylogenetic methods. Examination of the cultures of the fresh-water diatom Synedra acus grown in Lake Baikal water by epifluorescence and electron microscopy revealed that bacterial cells colonized the mucilage surrounding the cells of living microalgae and their cell surfaces and penetrate into the frustules of dead diatoms. A total of 13 strains of heterotrophic bacteria were isolated in pure cultures and described. Based on the results of analysis of their morphological, physiological, and biochemical properties, as well as on data obtained by 16S rRNA gene analysis, the following strains were identified to the species level: Sphingomonas sp., Variovorax paradoxus, Pseudomonas fluorescens, Microbacterium trichothecenolyticum, Rhodococcus sp., Caulobacter vibrioides, and Brevundimonas vesicularis.