Specificity of Lectins Labeled with Colloidal Gold to the Exopolymeric Matrix Carbohydrates of the Sulfate-Reducing Bacteria Biofilm Formed on Steel

2020 ◽  
Vol 82 (5) ◽  
pp. 11-20
Author(s):  
D.R. Abdulina ◽  
◽  
L.M. Purish ◽  
G.O. Iutynska ◽  
◽  
...  

The studies of the carbohydrate composition of the sulfate-reducing bacteria (SRB) biofilms formed on the steel surface, which are a factor of microbial corrosion, are significant. Since exopolymers synthesized by bacteria could activate corrosive processes. The aim of the study was to investigate the specificity of commercial lectins, labeled with colloidal gold to carbohydrates in the biofilm exopolymeric matrix produced by the corrosive-relevant SRB strains from man-caused ecotopes. Methods. Microbiological methods (obtaining of the SRB biofilms during cultivation in liquid Postgate B media under microaerophilic conditions), biochemical methods (lectin-binding analysis of 10 commercial lectins, labeled with colloidal gold), transmission electron microscopy using JEM-1400 JEOL. Results. It was shown using transmission electron microscopy that the binding of lectins with carbohydrates in the biofilm of the studied SRB strains occurred directly in the exopolymerіс matrix, as well as on the surfaces of bacterial cells, as seen by the presence of colloidal gold particles. For detection of the neutral carbohydrates (D-glucose and D-mannose) in the biofilm of almost all studied bacterial strains PSA lectin was the most specific. This lectin binding in biofilms of Desulfotomaculum sp. К1/3 and Desulfovibrio sp. 10 strains was higher in 90.8% and 94.4%, respectively, then for ConA lectin. The presence of fucose in the SRB biofilms was detected using LABA lectin, that showed specificity to the biofilm EPS of all the studied strains. LBA lectin was the most specific to N-аcetyl-D-galactosamine for determination of amino sugars in the biofilm. The amount of this lectin binding in D. vulgaris DSM644 biofilm was 30.3, 10.1 and 9.3 times higher than SBA, SNA and PNA lectins, respectively. STA, LVA and WGA lectins were used to detect the N-acetyl-Dglucosamine and sialic acid in the biofilm. WGA lectin showed specificity to N-acetyl-D-glucosamine in the biofilm of all the studied SRB; maximum number of bounded colloidal gold particles (175 particles/μm2) was found in the Desulfotomaculum sp. TC3 biofilm. STA lectin was interacted most actively with N-acetyl-D-glucosamine in Desulfotomaculum sp. TC3 and Desulfomicrobium sp. TC4 biofilms. The number of bounded colloidal gold particles was in 9.2 and 7.4 times higher, respectively, than using LVA lectin. The lowest binding of colloidal gold particles was observed for LVA lectin. Conclusions. It was identified the individual specificity of the 10 commercial lectins to the carbohydrates of biofilm matrix on the steel surface, produced by SRB. It was estimated that lectins with identical carbohydrates specificity had variation in binding to the biofilm carbohydrates of different SRB strains. Establishing of the lectin range selected for each culture lead to the reduction of the scope of studies and labor time in the researching of the peculiarities of exopolymeric matrix composition of biofilms formed by corrosiverelevant SRB.

2011 ◽  
Vol 199-200 ◽  
pp. 102-105
Author(s):  
Xiao Dong Zhao ◽  
Jie Yang ◽  
Xi Qiu Fan

A kind of sulfate-reducing bacteria was isolated from the actual marine environment, cultured and enriched for phylogenetic analysis by molecular biology methods, and observed under fluorescent microscopy and transmission electron microscopy to determine the species and morphology. Taking the bacteria as the main object, the influenced corrosion behavior of steels in marine environment was studied in follow-up experiments.


Coatings ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 625
Author(s):  
Lijuan Chen ◽  
Bo Wei ◽  
Xianghong Xu

The influence of sulfate-reducing bacteria (SRB) on the corrosion behaviors of X80 pipeline steel was investigated in a soil environment by electrochemical techniques and surface analysis. It was found that SRB grew well in the acidic soil environment and further attached to the coupon surface, resulting in microbiologically influenced corrosion (MIC) of the steel. The corrosion process of X80 steel was significantly affected by the SRB biofilm on the steel surface. Steel corrosion was inhibited by the highly bioactive SRB biofilm at the early stage of the experiment, while SRB can accelerate the corrosion of steel at the later stage of the experiment. The steel surface suffered severe pitting corrosion in the SRB-containing soil solution.


2009 ◽  
Vol 5 ◽  
pp. 13-23 ◽  
Author(s):  
C. Guzmán ◽  
Gloria Del Angel ◽  
Ricardo Gómez ◽  
F. Galindo ◽  
R. Zanella ◽  
...  

Au/TiO2 and Au/TiO2-CeO2 catalysts were prepared by the sol-gel method and carbon monoxide, hydrogen chemisorption and TEM spectroscopy have been exploited to determine the size of gold particles. The gold nanoparticles (8.1 to 2.1 nm) were deposited by using the deposition-precipitation method. The XRD characterization shows the presence of anatase as the TiO2 crystalline phase; while by XPS spectroscopy, the presence of Au°, Au2O3, Ce3+ and Ce4+ species co-existing in the Au/TiO2-CeO2 catalysts is shown. The characterizations by TPD-CO as well as by TPD-H2 (temperature programmed desorption) showed that on catalysts containing cerium, the gold particle size can be determined with great accuracy by using these chemisorption methods. The gold particle size calculated from either the CO or H2 thermodesorption values is in good agreement with that obtained by High Resolution Transmission Electron Microscopy (HRTEM) and Scanning Transmission Electron Microscopy (STEM) analyses. It was proposed that the TPD-CO and/or TPD-H2 techniques could be helpful for the characterization of the gold particles by TEM; especially when the high contrast in the pictures of the supports containing CeO2 prevents the particle size from being determined.


1990 ◽  
Vol 36 (6) ◽  
pp. 400-408 ◽  
Author(s):  
Judy D. Wall ◽  
Barbara J. Rapp-Giles ◽  
Merton F. Brown ◽  
Jerry A. White

Oxygen tolerance of the strictly anaerobic sulfate-reducing bacteria is well documented and poorly understood. This capacity for surviving brief exposures to oxygen must be a major factor in the diversity of environmental niches observed for these bacteria. We observed that viable cells of Desulfovibrio desulfuricans (ATCC 27774) could be found in colonies on the surface of solidified medium exposed to air for periods as long as 1 month. During exposure to air, the originally black colonies became greyish white, presumably as a result of the air oxidation of the metal sulfide deposits. A black, brittle deposit formed at the bottom of the colony and, simultaneously, the colony descended into a dimple that developed into a well in the agar. Eventually the colony reached the bottom of the Petri dish. These changes did not take place when the colonies were maintained in an anaerobic chamber. The morphological changes took place with all strains tested: three strains of D. desulfuricans and one strain of Desulfovibrio gigas and Desulfovibrio multispirans. Continued sulfate reduction appeared to be essential. Cyclic sulfate (thiosulfate or sulfite) reduction to sulfide and reoxidation of sulfide by the oxygen in air are proposed to maintain the viability of the bacteria by providing substrates for energy production and by reducing oxygen tension. Scanning and transmission electron microscopy of colony and cellular changes are shown. Key words: Desulfovibrio, sulfate-reducing bacteria, oxygen tolerance, sulfate cycling, scanning electron microscopy.


Parasitology ◽  
1986 ◽  
Vol 93 (1) ◽  
pp. 17-32 ◽  
Author(s):  
I. W. Sherman ◽  
Jane R. T. Greenan

SUMMARYThe distribution of anionic residues on the surface of erythrocytes infected withPlasmodium falciparumwas studied using cationized ferritin (CF) and transmission electron microscopy. CF staining of uninfected erythrocytes or erythrocytes infected with a knobless variant resulted in a dense and uniform distribution of ferritin particles; however, when red cells infected with a knob-inducing variant were exposed to CF, aggregates of ferritin particles were observed in the region of membrane elevation. Lectin binding to the erythrocyte surface was visualized by transmission electron microscopy using ferritin-conjugated lectins and lectin-fetuin-gold. No differences were observed in the lectin-binding patterns of malaria-infected or uninfected erythrocytes using WGA (wheat-germ agglutinin), RCA (ricin), andLimax flavuslectin. In distinct contrast to the uniform distribution of ferritin particles seen with these lectins was the appearance of clusters of ferritin-ConA over the knobby regions. Localized aggregates of ConA were not seen in knob-free areas or on the surface of red cells infected with a knobless variant. No significant differences were found in the agglutination reactions of normal and infected cells with theCancer antennariuslectin specific forO-acylated sialic acids.


1987 ◽  
Vol 35 (8) ◽  
pp. 843-853 ◽  
Author(s):  
G B Birrell ◽  
K K Hedberg ◽  
O H Griffith

The immunogold method is widely used to localize, identify, and distinguish cellular antigens. There are, however, some pitfalls that can lead to nonspecific binding, particularly in cytoskeletal studies with gold probes prepared from small gold particles. We present a list of suggestions for minimizing nonspecific binding, with particular attention to two problems identified in this study. First, we find that the method used to prepare the colloidal gold particles affects the degree of nonspecific binding. Second, the standard BSA-stabilized small gold probes evidently possess exposed regions that bind to the proteins of cytoskeletal preparations. This was investigated in whole-mount cytoskeletal preparations of cultured cells by use of light microscopy, transmission electron microscopy, and photoelectron microscopy of silver-enhanced specimens. Gold probes were made from approximately 5-nm particles generated by reduction of HAuCl4 with three different reducing agents: white phosphorus, sodium borohydride, and citrate-tannic acid. All three preparations stabilized in the conventional way showed significant levels of nonspecific binding, which was highest with citrate-tannic acid. This problem was largely solved with all three types of probes by including fish gelatin in the probe buffer, by substituting fish gelatin for the BSA stabilizer used to prepare the probes, or by pre-adsorption methods. Application of these techniques resulted in clear immunogold labeling patterns with minimal nonspecific background.


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