Lectins probe molecular films in biofouling:characterization of early films on non-living and living surfaces

Michael, T.; Smith, C. W.

doi:10.3354/meps119229

Cited by 41 publications

(21 citation statements)

References 24 publications

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“…Natural marine biofilm systems on inert and living surfaces were also the subject of lectin analyses (Michael & Smith 1995). It was found that the marine films showed a spatial and chemical heterogeneity of glycoconjugates similar to the results of the present study.…”

Section: Discussionsupporting

confidence: 86%

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In situ cell and glycoconjugate distribution in river snow studied by confocal laser scanning microscopy

Neu¹

2000

Aquat. Microb. Ecol.

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Aggregates from the river Elbe (Germany) were collected and examined by confocal laser scanning microscopy. The river snow was directly transferred into coverslip chambers and observed without any embedding and fixation procedure. River snow samples were examined in the reflection and fluorescence mode to record mineral content and autofluorescence signals. Spatial distribution of microb~al cells within the aggregates was shown with general nucleic acid specific stains (SYTO 9, SYTO 64). The polymeric matnx of the river snow was demonstrated by using a panel of fluorescently labelled lectins. Staining with FITC and TRITC labelled lectins and simultaneous dual channel unaging showed the distribution of different types of glycoconjugates. The binding pattern observed included bacterial cell surface labelling and cloud-type labelling of extracellular polymeric substances (EPS). In addition, dual staining combined with triple-channel recording was employed to clearly separate the signal in the red channel originating from TRITC-lectin or SYTO 64 nucleic acid stain from the far red autofluorescence originating from chlorophyll-containing organisms. With this approach the fully hydrated, living, 3-dimensional architecture of aquatic aggregates was investigated. Application of lectins demonstrated the heterogeneity of the EPS matrix in between the cellular constituents of the river snow community. The chemical heterogeneity of river snow may be significant for sorption and transport of nutrients and contaminants in lotlc aquatic environments. This in situ technique may be also useful to examine interfacial microbial consortia in other habitats. Finally, it is suggested to employ in situ lectin staining to probe for the glycoconjugate distribution at the polymer level similar to in situ hybridisation with rRNA targeted oligonucleotides at the cellular level.

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Section: Discussionsupporting

confidence: 86%

“…Control experiments clearly showed the inhibiting effect of methyl-a-D-mannopyranoside on the binding of Canavalid ensiformis FITC-lectin to river snow. This is in agreement with another report where the identical lectin and carbohydrate combination was employed in control experiments with environmental biofilms (Michael & Smith 1995).…”

Section: Discussionsupporting

confidence: 82%

In situ cell and glycoconjugate distribution in river snow studied by confocal laser scanning microscopy

Neu¹

2000

Aquat. Microb. Ecol.

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“…Likewise, glycoconjugate-blinded lectins, in which the binding sites of the lectins are blocked with target sugars, are recommended by Hartmann et al (7) as control experiments for nonspecific binding of lectins to biofilms. Michael and Smith (17) found that emission from ConA-fluorescein isothiocyanate (ConA-FITC) was undetectable following coincubation of biofilms with ConA-FITC and the sugar competitor ␣-methyl mannoside.…”

Section: Resultsmentioning

confidence: 99%

“…Excretion of adhesive polymers during attachment of bacterial cells to surfaces has been described using a panel of fluorescent lectins (9,14,20). The formation of biofilms on living and nonliving surfaces has been investigated with lectins (17). Lectins in conjunction with confocal laser scanning microscopy (CLSM) have been valuable tools in the study of the threedimensional structure of biofilms (12,15) or of the composition of extracellular polymeric substances (EPS) involved in accumulation of chlorinated organic compounds (24).…”

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confidence: 99%

Evaluation of Fluorescently Labeled Lectins for Noninvasive Localization of Extracellular Polymeric Substances in Sphingomonas Biofilms

Johnsen¹,

Hausner

Schnell

et al. 2000

Appl Environ Microbiol

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Lectins are a group of diverse proteins which bind to specific configurations of sugar residues. The binding of lectins to sugar residues present in polysaccharides resembles the specific binding of antibodies to antigens (4). Lectins have been widely used to characterize surfaces of eucaryotic cells and polysaccharides. Recently, fluorescently labeled lectins have been applied in the study of biofilm formation and biofilm composition. Excretion of adhesive polymers during attachment of bacterial cells to surfaces has been described using a panel of fluorescent lectins (9,14,20). The formation of biofilms on living and nonliving surfaces has been investigated with lectins (17). Lectins in conjunction with confocal laser scanning microscopy (CLSM) have been valuable tools in the study of the threedimensional structure of biofilms (12,15) or of the composition of extracellular polymeric substances (EPS) involved in accumulation of chlorinated organic compounds (24). Strains of Sphingomonas spp. are known for their interesting catabolic capabilities to degrade a wide variety of environmentally hazardous compounds, including polycyclic aromatics (25), dioxine compounds (6), and chlorinated phenols (3). Theoretically, lectins may be used to study the interaction between Sphingomonas cells and environmental surfaces during biofilm formation or to investigate the interaction of EPS with organic compounds. The common approach has been to deduce the structure or composition of biofilm EPS on the basis of the specific binding of lectins to different sugar residues. In this study, we evaluate the use of lectins for the characterization of Sphingomonas biofilms by investigating the binding of five fluorescent lectins with known specificities to Sphingomonas biofilms and to industrially produced Sphingomonas exopolysaccharides (sphingans) with known molecular structures. MATERIALS AND METHODSBacterial strains and growth conditions. Sphingomonas paucimobilis EPA505 was obtained from J. Mueller (19), and Sphingomonas sp. strain LH128 and Sphingomonas sp. strain LB126 were received from L. Bastiaens (1). All strains were stored in 43% glycerol at Ϫ80°C. The bacteria were grown at room temperature in phosphate minimal medium supplemented with glucose as the sole carbon source (PMMG) containing (in grams/liter) the following: glucose, 2; Na 2 HPO 4 ⅐ 2H 2 O, 0.875; KH 2 PO 4 , 0.1; (NH 4 ) 2 SO 4 , 0.25; MgCl 2 ⅐ 6H 2 O, 0.05; CaCl 2 ⅐ 2H 2 O, 0.015; NaNO 3 , 0.018. The medium was amended with 5 ml of a trace element solution consisting of (in milligrams/liter) the following: Na-EDTA, 800; FeCl 2 , 300; MnCl 2 ⅐ 4H 2 O, 10; CoCl 2 ⅐ 6H 2 O, 4; CuSO 4 , 1; Na 2 MoO 4 ⅐ 2H 2 O, 3; ZnCl 2 , 2; LiCl, 0.5; SnCl 2 ⅐ 2H 2 O, 0.5; H 3 BO 3 , 1; KBr, 2; KI, 2; BaCl 2 , 0.5. Phosphate and glucose were autoclaved separately.Cultivation of biofilms on microscope slides. Single-species biofilms were grown on Cel-Line HTC printed microscope slides with six wells on each slide (Cel-Line Associates, Inc., Newfield, N.J.). The slides were sterili...

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“…Not only the EPS yield and occurrence of cell rupture, but also the chemical composition of EPS depends on the extraction method, which is the first and most important step in a chemical characterization of EPS (12,30). FLBA in combination with LSM was suggested to be a powerful tool for in situ, three-dimensional visualization and characterization of EPS in pure culture, as well as environmental fully hydrated biofilms (34,36,37). Quite recently, it was shown that by combining FLBA with fluorescence in situ hybridization (FISH) it was possible to segregate EPS microdomains within bacterial colonies (31).…”

Section: Discussionmentioning

confidence: 99%

Characterization of Glycoconjugates of Extracellular Polymeric Substances in Tufa-Associated Biofilms by Using Fluorescence Lectin-Binding Analysis

Zippel

Neu

2011

Appl Environ Microbiol

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Freshwater tufa deposits are the result of calcification associated with biofilms dominated by cyanobacteria. Recent investigations highlighted the fact that the formation of microbial calcium carbonates is mainly dependent on the saturation index, which is determined by pH, the ion activity of Ca 2؉ and CO 3 2؊ , and the occurrence of extracellular polymeric substances (EPS) produced by microorganisms. EPS, which contain carboxyl and/or hydroxyl groups, can strongly bind cations. This may result in inhibition of CaCO 3 precipitation. In contrast, the formation of templates for crystal nucleation was reported by many previous investigations. The purposes of this study were (i) to characterize the in situ distribution of EPS glycoconjugates in tufa-associated biofilms of two German hard-water creeks by employing fluorescence lectin-binding analysis (FLBA), (ii) to verify the specific lectin-binding pattern by competitive-inhibition assays, and (iii) to assess whether carbonates are associated with structural EPS domains. Three major in situ EPS domains (cyanobacterial, network-like, and cloud-like structures) were detected by FLBA in combination with laser scanning microscopy (LSM). Based on lectin specificity, the EPS glycoconjugates produced by cyanobacteria contained mainly fucose, amino sugars (N-acetyl-glucosamine and N-acetyl-galactosamine), and sialic acid. Tufa deposits were irregularly covered by network-like EPS structures, which may originate from cyanobacterial EPS secretions. Cloud-like EPS glycoconjugates were dominated by sialic acid, amino sugars, and galactose. In some cases calcium carbonate crystals were associated with cyanobacterial EPS glycoconjugates. The detection of amino sugars and calcium carbonate in close association with decaying sheath material indicated that microbially mediated processes might be important for calcium carbonate precipitation in freshwater tufa systems.

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Lectins probe molecular films in biofouling:characterization of early films on non-living and living surfaces

Cited by 41 publications

References 24 publications

In situ cell and glycoconjugate distribution in river snow studied by confocal laser scanning microscopy

In situ cell and glycoconjugate distribution in river snow studied by confocal laser scanning microscopy

Evaluation of Fluorescently Labeled Lectins for Noninvasive Localization of Extracellular Polymeric Substances in Sphingomonas Biofilms

Characterization of Glycoconjugates of Extracellular Polymeric Substances in Tufa-Associated Biofilms by Using Fluorescence Lectin-Binding Analysis

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