Strains Of Sulfate-reducing Bacteria Research Articles

Гуанідінійвмісний олігомер як інгібітор мікробної корозії металу

Guanidinium-containing oligomers, a poorly studied class of organic compounds, have attracted attention due to their bactericidal properties. A guanidinium-containing oligomer based on an aliphatic oligoepoxide is a newly synthesized substance with bactericidal activity, which gives it the prospects for use as a microbial corrosion inhibitor. Aim. The goal of the study was to synthesize oligomeric alkyl substituted guanidinium bromide and study of its anticorrosive properties in the presence of steel under the influence of corrosive active sulfate-reducing bacteria. Methods. The guanidine-containing alkyl substituted oligomer was obtained by the reaction of the aliphatic oligoepoxide DEG-1 with guanidine, followed by interaction with alkyl bromide. The anticorrosive properties of the guanidine-containing oligomer were studied using collection strains of sulfate-reducing bacteria: UCM B-11503 Desulfovibrio sp.10, UCM B-11501 Desulfovibrio desulfuricans DSM642, and UCM B-11502 Desulfovibrio vulgaris DSM644. The sulfate-reducing bacteria were grown on Postgate В medium for 14 days at a temperature of 28 ± 2 °C. The number of bacteria was determined by the method of tenfold dilution. The corrosion rate was determined by the gravimetric method. The physicochemical parameters of pH and redox potential of the bacterial culture liquid were studied by the potentiometric method. The accumulation of hydrogen sulfide in the culture liquid was determined by the iodometric method. Lipolytic activity was studied spectrophotometrically using a KFK-3 device by reaction with p-nitrophenyl palmitate, catalase activity — using 0.03% hydrogen peroxide, which formed a stable colored complex with a 4% molybdenum diphosphate solution. The specific activity of the studied enzymes was expressed as unit ∙ mg-1 protein. Protein was determined in the supernatant by the conventional Lowry method. Results. It was shown that the oligomer based on aliphatic oligoepoxide has biocidal properties. A significant inhibition of the development of sulfate-reducing bacteria (SRB) was observed after the oligomer was added; only dozens of bacterial cells were detected in the medium after the exposure period. The corrosion rate of steel in the presence of SBR without addition of inhibitors was 0.15 — 0.35 mg/cm2∙h. The addition of DPC (a quaternary ammonium compound based on N-decylpyridinium chloride) (Kyiv Polytechnic Institute (KPI), Ukraine) to the culture medium led to a decrease in the steel corrosion rate to 0.032 — 0.047 mg ∙ cm-2 h-1 (by 6.5—10.6 times). In the presence of Armohib CI-28 inhibitor (based on Diamine Ethoxylate) (Akzonobel, Holland), the corrosion rate was reduced to 0.02—0.039 mg ∙ cm-2 h-1 (by 4.2 — 12.7 times). The addition of guanidinium-containing oligomer to the medium with bacteria reduced the corrosion rate to 0.075—0.079 mg/cm2 ∙ h (by 2.5—2.7 times). According to the mass loss of steel samples, the degree of metal protection against microbial corrosion in the presence of guanidinium-containing oligomer was 60.15—63.17%. Conclusions. The obtained data indicate that the guanidinium-containing oligomer based on aliphatic oligoepoxy has biocidal and anticorrosive properties and is promising for use as a means of combating microbially induced corrosion.

Study on the factors of hydrogen sulfide production from lignite bacterial sulfate reduction based on response surface method

Bacterial sulfate reduction (BSR) is one of the key factors leading to the anomalous accumulation of hydrogen sulphide in coal mines. Environmental factors such as temperature and pH play a crucial role in the metabolism and degradation of coal by sulfate-reducing bacteria (SRB). In this study, coal samples were selected from Shengli Coal Mine, and SRB strains were isolated and purified from mine water using a dilution spread-plate anaerobic cultivation method. Based on single-factor experiments and response surface methodology (RSM), the impact of temperature, pH, oxidation–reduction potential (ORP), chemical oxygen demand to sulfate ratio (COD/SO42−) on the generation of hydrogen sulphide during brown coal BSR was analyzed. The results showed that the anaerobic degradation of coal by SRB was inhibited by either too high or too low a temperature to produce hydrogen sulfide, and the greatest production of hydrogen sulfide occurred at a temperature of about 30 °C; The greatest production of hydrogen sulfide occurred at an initial ambient pH of 7.5; COD/SO42− ratio of around 2.0 is most conducive to hydrogen sulphide generation; the lower ORP value is more favorable for hydrogen sulfide generation. The optimal conditions obtained by RSM were: temperature of 30.37 °C, pH of 7.64 and COD/SO42− of 1.96. Under these conditions, the hydrogen sulfide concentration was 56.79 mg/L, the pH value was 8.40, the ORP value was −274 mV, and the SO42− utilization rate was 58.04%. The RSM results showed that temperature, ambient pH and COD/SO42− had a significant effect on hydrogen sulfide production, and the degree of effect was: ambient pH > temperature > COD/SO42−.

Mining-related multi-resistance genes in sulfate-reducing bacteria treatment of typical karst nonferrous metal(loid) mine tailings in China.

Management of tailings at metal mine smelter sites can reduce the potential hazards associated with exposure to toxic metal(loid)s and residual organic flotation reagents. In addition, microbes in the tailings harboring multi-resistance genes (e.g., tolerance to multiple antimicrobial agents) can cause high rates of morbidity and global economic problems. The potential co-selection mechanisms of antibiotic resistance genes (ARGs) and metal(loid) resistance genes (MRGs) during tailings sulfate-reducing bacteria (SRB) treatment have been poorly investigated. Samples were collected from a nonferrous metal mine tailing site treated with an established SRB protocol and were analyzed for selected geochemical properties and high throughput sequencing of 16S rRNA gene barcoding. Based on the shotgun metagenomic analysis, the bacterial domain was dominant in nonferrous metal(loid)-rich tailings treated with SRB for 12 months. KEGGs related to ARGs and MRGs were detected. Thiobacillus and Sphingomonas were the main genera carrying the bacA and mexEF resistance operons, along with Sulfuricella which were also found as the main genera carrying MRGs. The SRB treatment may mediate the distribution of numerous resistance genes. KOs based on the metagenomic database indicated that ARGs (mexNW, merD, sul, and bla) and MRGs (czcABCR and copRS genes) were found on the same contig. The SRB strains (Desulfosporosinus and Desulfotomaculum), and the acidophilic strain Acidiphilium significantly contributed to the distribution of sul genes. The functional metabolic pathways related to siderophores metabolism were largely from anaerobic genera of Streptomyces and Microbacterium. The presence of arsenate reductase, metal efflux pump, and Fe transport genes indicated that SRB treatment plays a key role in the metal(loid)s transformation. Overall, our findings show that bio-treatment is an effective tool for managing ARGs/MRGs and metals in tailings that contain numerous metal(loid) contaminants.

Stress corrosion cracking failure of X80 carbon steel U-bend caused by Desulfovibrio vulgaris biocorrosion

Sulfate reducing bacteria (SRB) are widely present in oil and gas industry, causing pitting corrosion on pipeline steel. Stress corrosion cracking (SCC) often occurs in the presence of mechanical stress before pitting perforation failure, leading to economic losses and even catastrophic accidents. In this study, stress distribution simulation using the finite element method (FEM), corrosion analysis techniques and electrochemical corrosion measurements were employed to investigate the SCC mechanism of X80 pipeline steel caused by Desulfovibrio vulgaris, which is a common SRB strain used in microbiologically influenced corrosion (MIC) studies. It was found that D. vulgaris MIC caused sharp microcracks on an X80 U-bend coupon after only 2 weeks of immersion at 37 °C in the deoxygenated ATCC 1249 culture medium inoculated with D. vulgaris. The X80 U-bend coupon's weight loss-based uniform corrosion rate for the 12 cm2 surface was 60% of that for the unstressed flat square coupon (2.3 mg cm−2 vs. 3.8 mg cm−2). This was likely because the square coupon had wide MIC pits, providing a larger effective surface area for more sessile cells (4.2×108 cells cm−2 on square coupon vs. 2.4×108 cells cm−2 on U-bend coupon) to attach and harvest more electrons. An SCC failure occurred on an X80 U-bend pre-cracked at the outer bottom after a 6-week immersion in the D. vulgaris broth. Apart from MIC damage, this could also be because D. vulgaris metabolism increased the availability hydrogen atoms on the steel surface, and promoted the diffusion of hydrogen atoms into the metal lattice, thus increasing the brittleness of the steel.

X80 U-bend stress corrosion cracking (SCC) crack tip dissolution by fast corroding Desulfovibrio ferrophilus IS5 biofilm

Stress corrosion cracking (SCC) is a serious problem threatening structural integrity and safety. Metal corrosion pits can develop into more harmful cracks under mechanical stress. It is known in abiotic corrosion that fast corrosion does not necessarily cause more severe SCC. This is because fast corrosion can prevent or dissolve crack tips and convert them into less harmful pits if the uniform corrosion rate is fast enough. This work experimentally proved the hypothesis that the same phenomenon occurs in biotic corrosion. It was found that a highly corrosive SRB (sulfate reducing bacterium) strain, namely Desulfovibrio ferrophilus (strain IS5), caused fast uniform corrosion and dull pits, but no SCC cracks against X80 carbon steel U-bend coupons (stressed). In comparison, Desulfovibrio vulgaris, a far less corrosive SRB strain caused less corrosion, but generated sharp pits and SCC cracks in a 3-month lab test. This work also revealed that the X80 U-bend had a 44% higher sessile cell count and 34% higher weight loss after a 14-d D. ferrophilus incubation in deoxygenated enriched artificial seawater culture medium at 28 °C when compared with the (flat) square coupons (no stress). This weight loss trend was corroborated by transient electrochemical measurements.

The Isolation of Anaerobic and Facultative Anaerobic Sulfate-Reducing Bacteria (SRB) and a Comparison of Related Enzymes in Their Sulfate Reduction Pathways.

Sulfate-reducing bacteria (SRB) are an important group of microorganisms that cause microbial corrosion. In this study, culturable SRB were isolated and identified from the inner rust layer of three kinds of steel and from sediments, and a comparison of amino acid sequences encoding related enzymes in the sulfate reduction pathway between anaerobic and facultative anaerobic SRB strains was carried out. The main results are as follows. (1) Seventy-seven strains were isolated, belonging to five genera and seven species, with the majority being Desulfovibrio marinisediminis. For the first time, Holodesulfovibrio spirochaetisodalis and Acinetobacter bereziniae were separated from the inner rust layer of metal, and sulfate reduction by A. bereziniae, Virgibacillus dokdonensis, and Virgibacillus chiguensis, etc., was also demonstrated for the first time. (2) Three strains of strictly anaerobic bacteria and four strains of facultative anaerobic bacteria were screened from seven bacterial strains. (3) Most of the anaerobic SRB only contained enzymes for the dissimilatory sulfate reduction pathway, while those of facultative anaerobic bacteria capable of producing hydrogen sulfide included two possible ways: containing the related enzymes from the dissimilatory pathway only, or containing enzymes from both dissimilatory and assimilation pathways. This study newly discovered that some bacterial genera exhibit sulfate reduction ability and found that there are differences in the distribution of enzymes related to the sulfate reduction pathway between anaerobic and facultative anaerobic SRB type trains, providing a basis for the development and utilization of sulfate-reducing bacterial resources and furthering our understanding of the metabolic mechanisms of SRB.

Variation of sulfate reducing bacteria communities in ionic rare earth tailings and the potential of a single cadmium resistant strain in bioremediation

Heap leaching ionic rare earth tailings might be prone to nourish sulfate reducing bacteria (SRB), but the SRB community in terrestrial ecosystems, such as tailings, has never been studied. This work was conducted to investigate the SRB communities in revegetated and bare tailings in Dingnan county, Jiangxi province, China, incorporating with indoor experiments to isolate SRB strain in bioremediation of Cd contamination. Significant increases in richness, accompanied by reductions in evenness and diversity, were found in the SRB community in revegetated tailings compared to bare tailings. At genus taxonomic level, two distinct dominant SRB were observed in samples from bare and revegetated tailings, with Desulfovibrio dominating in the former and Streptomyces dominating in the latter, respectively. A single SRB strain was screened out from the bare tailings (REO-01). The cell of REO-01 was rod-shaped and belonged to family Desulfuricans and genus Desulfovibrio. The Cd resistance of the strain was further examined, no changes in cell morphology were observed at 0.05 mM Cd, additionally, the atomic ratios of S, Cd, and Fe changed with the increase in Cd dosages, indicating FeS and CdS were produced simultaneously, XRD results further confirmed the production changed gradually from FeS to CdS with increasing Cd dosages from 0.05 to 0.2 mM. FT-IR analysis showed that functional groups containing amide, polysaccharide glycosidic linkage, hydroxyl, carboxy, methyl, phosphodiesters and sulfhydryl groups in extracellular polymeric substances (EPS) of REO-01 might have affinity with Cd. This study demonstrated the potential of a single SRB strain isolated from ionic rare earth tailings in bioremediation of Cd contamination.

In-Vitro Assessment of the Corrosion Potential of an Oral Strain of Sulfate-Reducing Bacteria on Metallic Orthodontic Materials.

Orthodontic literature is scant when it comes to microbial corrosion. The oral prevalence of many bacteria which are capable of causing microbial corrosion is reported in the dental literature. The aim of this study is to experimentally determine the corrosive potential of an oral strain of Sulfate-reducing bacteria. Stainless steel (SS) bracket, stainless steel archwire, NiTi archwire, Titanium molybdenum (TMA) archwire, and titanium miniscrew were immersed in five media which included Artificial saliva (group I), Sulfate rich artificial saliva (group II), API agar medium specific for SRB (group III), AS + API medium+ bacterial strain (group IV), SRAS+ API medium+ bacterial strain (group V). The materials were then subjected to Scanning electron microscopy and energy-dispersive X-ray analysis (EDX). Materials in groups I, II, and III did not show any surface changes whereas materials in groups IV and V which contained the bacteria showed surface changes which were erosive patches suggestive of corrosion. EDX analyses were in line with similar findings. This in vitro study suggested that the oral strain of Sulfate-reducing bacteria was able to induce corrosive changes in the experimental setup.

Steel rust layers immersed in the South China Sea with a highly corrosive Desulfovibrio strain

Although it is well known that microbes play a significant role in marine corrosion, few studies have systematically studied the relationship between microorganisms and corrosion products under long-term immersion. In this study, the corrosion characteristics of the rust layer formed on carbon steel immersed in the South China Sea for 5.5 years were investigated using various surface analysis and microbial community analysis techniques. Magnetite (Fe3O4), iron sulfide, and green rust were identified in the inner rust layer. The middle rust layer was composed of maghemite (γ-Fe2O3), and some Fe3O4 and mackinamite were also detected. The outer rust layer contained several Fe(III)-oxyhydroxides, and it had a large number of fouling organisms attached to it. In all of the rust layers, anaerobic sulfate-reducing bacteria (SRB) were the dominant bacteria, and they may have played a key role in the formation of the corrosion products. One SRB strain (Desulfovibrio bizertensis SY-1) with a highly corrosivity (13.561 mg/cm2) was isolated from these rust layers, and its physiological and metabolic characteristics were studied. These results expand the membership of corrosive SRB and establish a better understanding of marine microbiologically influenced corrosion (MIC).

Removal of Antimony in Wastewater by Antimony Tolerant Sulfate-Reducing Bacteria Isolated from Municipal Sludge.

Antimony (Sb), a global and priority controlled pollutant, causes severe environmental issues. Bioremediation by microbial communities containing sulfate-reducing bacteria (SRB) is considered to be among the safest, economical, and environmentally friendly methods to remove Sb from wastewater. However, the roles of SRB species in these communities remain uncertain, and pure cultures of bacteria that may be highly efficient have not yet been developed for Sb removal. In this study, an Sb tolerant community was enriched from municipal sludge, and molecular ecological analysis showed that Escherichia (40%) and Desulfovibrio (15%) were the dominant bacteria. Further isolation and identification showed that the enriched SRB strains were closely related to Cupidesulfovibrio oxamicus, based on the molecular analyses of 16S rRNA and dsrB genes. Among them, a strain named SRB49 exhibited the highest activity in removal of Sb(V). SRB49 was able to remove 95% of Sb(V) at a concentration of 100 mg/L within 48 h under optimum conditions: a temperature of 37–40 °C, an initial pH value of 8, 4 mM of sulfate, and an initial redox potential of 145–229 mV. SEM-EDX analysis showed that SRB49 did not adsorb Sb(V) but reduced and precipitated Sb(V) via the formation of Sb2S3. The results demonstrated the potential roles that pure cultures of SRB species may play in Sb removal and the use of Sb-tolerant SRB strains for Sb remediation.

Research on sulfate bacteriostatic effect of some metal nanos

Sulfate-reducing bacteria (Desulfomicrobium baculatum) is a group of anaerobic bacteria capable of producing H2S gas, which sours crude oil, corrodes metal equipment, pipeline systems, and even forms biofilms that cause blockages reservoir, reducing the reception capacity of pumped water in oil and gas exploitation. Currently, the biocides are mainly aldehydes or cyclic amines combined with cationic active substances, which are very toxic to humans and the environment. The development of materials technology has created new nano-sized materials that are capable of replacing traditional biocides and are environmentally friendly. This study has shown that silver and copper metal nanoparticles with an average size of 50 nm and concentration from 500 ppm can inhibit and kill both sulfate-reducing bacteria strains swimming and adhering in the water at normal and high temperature conditions.

The antibiofilm and antibacterial effects of medicinal plant extracts on isolated sulfate-reducing bacteria from orthodontic appliances

Sulfate-Reducing Bacteria (SRB) are a heterogeneous group of oral microflorae that cause dental caries, destructive periodontal infections, and biofilms on fixed orthodontic appliances. Plant extracts as a source of antibacterial components may have acceptable activity against SRB strains and their biofilm development. Our study aimed at determining the antibiofilm, antimicrobial efficacy, and cytotoxicity of Iran's endemic plants against pathogenic SRB. Plantago ovata, Terminalia chebula, Eugenia caryophyllata, and Aloe vera extracts were prepared according to standard methods. Patients and orthodontic appliances samples were collected, and SRB strains were isolated using standard tests and 16S rRNA analysis. The extracts' antimicrobial, antibiofilm activity, and cytotoxicity were determined using dilution broth micro, crystal violet staining, and MTT method, respectively. Biofilm involved gene expression of strains determined using Real-time PCR. Three SRB isolates harboring target genes were identified using 16S rDNA that showed a more than 90% of similarity to Desulfomicrobium orale spp. All investigated extracts showed good activity against SRB isolates. T. chebula, E. caryophyllata, and A. vera showed promising antibiofilm activity, and their subMIC concentrations downregulated biofilm-forming related genes in all isolates. T. Chebula had the lowest cytotoxicity, followed by E. caryophyllata and A. vera. T. Chebula also showed the best average selectivity index (SI) of 17.9, followed by E. caryophyllata (12.52). Extracts investigated in the present study had good antibacterial and antibiofilm activity against isolated sulfate-reducing bacteria isolated from patients' orthodontic appliances. Most of these extracts may be potential candidates for the development of antibacterial drugs.

Exogenous autoinducer-2 inhibits biofilm development of Desulfovibrio sp. Huiquan2017.

Sulfate-reducing bacteria (SRB) are culprits for microbiologically influenced corrosion, and biofilms are believed to play essential roles in the corrosion induced by SRB. However, little is known about the regulation of SRB biofilms. Quorum sensing signal molecules acyl-homoserine lactones (AHLs) and autoinducer-2 (AI-2) regulate biofilm formation of many bacteria. In this study, the production of AHLs and AI-2 by one SRB strain, Desulfovibrio sp. Huiquan2017, was detected, and the effect of exogenous AI-2 on bacterial biofilm formation was discussed. It was found that the cell-free supernatants of Desulfovibrio sp. Huiquan2017 induced luminescence in a ∆luxS mutant strain Vibrio harveyi BB170, indicating the production of functional AI-2 by the bacterium. In the presence of exogenous AI-2, the growth of Desulfovibrio sp. Huiquan2017 and early biofilm formation were not affected, but the later stage of biofilm development was inhibited significantly. The biofilms became looser, smaller, and thinner, and contained less bacteria and extracellular polymeric substances (EPS). The inhibition effect of AI-2 on the biofilm development of Desulfovibrio sp. Huiquan2017 was mainly achieved through reducing the amount of EPS in biofilms. These findings shed light on the biofilm regulation of SRB.

SRB-mediated corrosion of marine submerged AISI 1020 steel under impressed current cathodic protection

Metallic corrosion is a recurrent and costly problem to almost every industry; therefore, prevention strategies might be well-defined on a case-by-case basis. Commonly, cathodic protection (CP) is the world’s most widely-adopted technique to guarantee the integrity of buried or submerged structures from corrosion. However, as current potential values are dependent on metal-structure and environmental features, the target shall be well-identified; otherwise, the intended effect will not be reached. In seawater, a protective current potential of −800 mVAg/AgCl is recommended by technical standards, while a more negative potential (−900 mVAg/AgCl) is the suggested criterion for the control of corrosion induced by sulfate-reducing bacteria (SRB), even though without proper scientific support. Thus, this study focused on evaluating the efficiency of different cathodic protection potentials (−800, −900 and, −1000 mVAg/AgCl) on inhibiting SRB-mediated corrosion of AISI 1020 steel. Both unprotected and impressed current cathodically protected steel specimens were exposed to indigenous microorganisms in seawater for 7 days. The Most Probable Number (MPN) enumeration of sessile aerobic heterotrophic bacteria, acid-producing bacteria and, sulfate-reducing bacteria was performed at the beginning and at the end of the assays. Also, the reducing activity of hydrogenase-positive SRB strains was measured. Although the microbial colonization was greater on unprotected steel surfaces than on the cathodically protected ones, biofilm quantification of CP specimens did not show important differences regardless of the potential. However, hydrogenase-positive SRB counts increased with the reduction of CP potential value, promoting an increase in the number and depth of pits on specimens protected at −1000 mVAg/AgCl when compared with those protected at −800 mVAg/AgCl and unprotected ones.

Significance of lysogeny for the metabolism of Desulfovibrio spp. strains isolated from aquatic environments of Georgia.

Sulphate-reducing bacteria (SRB) are ecologically important group of anaerobic micro-organisms that can reduce sulphate to form hydrogen sulphide-a toxic gas causing iron corrosion on metal surfaces. In this work, SRB strains were isolated from aquatic environments in the country of Georgia to determine their lysogenicity and the role of temperate phages in host metabolism. SRB strains were isolated in samples from the Black Sea coast of Georgia. Based on their genetic, cytological and physiological properties of bacteria, 10 Georgian isolates were assigned to the genus Desulfovibrio. Temperate bacteriophages were induced from three out of ten strains by UV-exposure. Comparison of metal (Fe and Cr) reduction and utilization of various carbon sources by the wild-type (lysogenic) bacterial strains and their UV-irradiated counterparts was done. Temperate phage in the cells of SRB could alter significant functions of bacteria and may have a contribution in the acquisition of different traits by SRB. This article pointed to a significant role for temperate bacteriophages in the metabolism and metabolic potential of host strains of SRB, which were first isolated from the aquatic environment of Georgia.

Environmental Impact of Sulfate-Reducing Bacteria, Their Role in Intestinal Bowel Diseases, and Possible Control by Bacteriophages

Sulfate-reducing bacteria (SRB) represent a group of prokaryotic microorganisms that are widely spread in the anoxic environment (seabed, riverbed and lakebed sediments, mud, intestinal tract of humans and animals, metal surfaces). SRB species also have an impact on processes occurring in the intestinal tract of humans and animals, including the connections between their presence and inflammatory bowel disease (IBD). Since these SRB can develop antimicrobial resistance toward the drugs, including antibiotics and antimicrobial agents, bacteriophages could represent an additional potential effective treatment. The main objectives of the review were as follows: (a) to review SRB (both from intestinal and environmental sources) regarding their role in intestinal diseases as well as their influence in environmental processes; and (b) to review, according to literature data, the influence of bacteriophages on SRB and their possible applications. Since SRB can have a significant adverse influence on industry as well as on humans and animals health, phage treatment of SRB can be seen as a possible effective method of SRB inhibition. However, there are relatively few studies concerning the influence of phages on SRB strains. Siphoviridae and Myoviridae families represent the main sulfide-producing bacteria phages. The most recent studies induced, by UV light, bacteriophages from Desulfovibrio vulgaris NCIMB 8303 and Desulfovibrio desulfuricans ATCC 13541. Notwithstanding costly and medically significant negative impacts of phages on SRB, they have been the subject of relatively few studies. The current search for alternatives to chemical biocides and antibiotics has led to the renewed interest in phages as antibacterial biocontrol and therapeutic agents, including their use against SRB. Hence, phages might represent a promising treatment against SRB in the future.

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

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.

Desulfolutivibrio sulfoxidireducens gen. nov., sp. nov., isolated from a pyrite-forming enrichment culture and reclassification of Desulfovibrio sulfodismutans as Desulfolutivibrio sulfodismutans comb. nov

Two strains of sulfate-reducing bacteria (J.5.4.2-L4.2.8T and J.3.6.1-H7) were isolated from a pyrite-forming enrichment culture and were compared phylogenetically and physiologically to the closest related type strain Desulfovibrio sulfodismutans DSM 3696T. The isolated strains were vibrio-shaped, motile rods that stained Gram-negative. Growth occurred from 15 to 37°C and within a pH range of 6.5–8.5. Both strains used sulfate, thiosulfate, sulfite, and dimethyl sulfoxide (DMSO) as electron acceptor when grown with lactate. Lactate was incompletely oxidized to acetate. Formate and H2 were used as electron donor in the presence of acetate. Dismutation of thiosulfate and pyrosulfite was observed. The two new isolates differed from D. sulfodismutans by the utilization of DMSO as electron acceptor, 82% genome-wide average nucleotide identity (ANI) and 32% digital DNA-DNA hybridization (dDDH), thus representing a novel species. The type strain of the type species Desulfovibrio desulfuricans Essex6T revealed merely 88% 16S rRNA gene identity and 49% genome-wide average amino acid identity (AAI) to the new isolates as well as to D. sulfodismutans. Furthermore, the dominance of menaquinone MK-7 over MK-6 and the dominance of ai-C15:0 fatty acids were observed not only in the two new isolated strains but also in D. sulfodismutans. Therefore, the definition of a new genus is indicated for which the name Desulfolutivibrio is proposed. We propose for strains J.5.4.2-L4.2.8T and J.3.6.1-H7 the name Desulfolutivibrio sulfoxidireducens gen. nov. sp. nov. with strain J.5.4.2-L4.2.8T defined as type strain. In addition, we propose the reclassification of Desulfovibrio sulfodismutans as Desulfolutivibrio sulfodismutans comb. nov.

Evaluation of Physiological Parameters of Intestinal Sulfate-Reducing Bacteria Isolated from Patients Suffering from IBD and Healthy People.

Background: Inflammatory bowel diseases (IBDs) are multifactorial illnesses of the intestine, to which microorganisms are contributing. Among the contributing microorganisms, sulfate-reducing bacteria (SRB) are suggested to be involved in the process of bowel inflammation due to the production of hydrogen sulfide (H2S) by dissimilatory sulfate reduction. The aims of our research were to physiologically examine SRB in fecal samples of patients with IBD and a control group, their identification, the study of the process of dissimilatory sulfate reduction (sulfate consumption and H2S production) and biomass accumulation. Determination of biogenic elements of the SRB and evaluation of obtained parameters by using statistical methods were also included in the research. The material for the research consisted of 14 fecal samples, which was obtained from patients and control subjects. Methods: Microscopic techniques, microbiological, biochemical, biophysical methods and statistical analysis were included. Results: Colonies of SRB were isolated from all the fecal samples, and subsequently, 35 strains were obtained. Vibrio-shaped cells stained Gram-negative were dominant in all purified studied strains. All strains had a high percentage of similarity by the 16S rRNA gene with deposited sequences in GenBank of Desulfovibrio vulgaris. Cluster analysis of sulfate reduction parameters allowed the grouping of SRB strains. Significant (p < 0.05) differences were not observed between healthy individuals and patients with IBD with regard to sulfate reduction parameters (sulfate consumption, H2S and biomass accumulation). Moreover, we found that manganese and iron contents in the cell extracts are higher among healthy individuals in comparison to unhealthy individuals that have an intestinal bowel disease, especially ulcerative colitis. Conclusions: The observations obtained from studying SRB emphasize differences in the intestinal microbial processes of healthy and unhealthy people.

Adenosine-5'-Phosphosulfate- and Sulfite Reductases Activities of Sulfate-Reducing Bacteria from Various Environments.

A comparative study of the kinetic characteristics (specific activity, initial and maximum rate, and affinity for substrates) of key enzymes of assimilatory sulfate reduction (APS reductase and dissimilatory sulfite reductase) in cell-free extracts of sulphate-reducing bacteria (SRB) from various biotopes was performed. The material for the study represented different strains of SRB from various ecotopes. Microbiological (isolation and cultivation), biochemical (free cell extract preparation) and chemical (enzyme activity determination) methods served in defining kinetic characteristics of SRB enzymes. The determined affinity data for substrates (i.e., sulfite) were 10 times higher for SRB strains isolated from environmental (soil) ecotopes than for strains from the human intestine. The maximum rate of APS reductase reached 0.282–0.862 µmol/min×mg−1 of protein that is only 10 to 28% higher than similar initial values. The maximum rate of sulfite reductase for corrosive relevant collection strains and SRB strains isolated from heating systems were increased by 3 to 10 times. A completely different picture was found for the intestinal SRB Vmax in the strains Desulfovibrio piger Vib-7 (0.67 µmol/min × mg−1 protein) and Desulfomicrobium orale Rod-9 (0.45 µmol/min × mg−1 protein). The determinant in the cluster distribution of SRB strains is the activity of the terminal enzyme of dissimilatory sulfate reduction—sulfite reductase, but not APS reductase. The data obtained from the activity of sulfate reduction enzymes indicated the adaptive plasticity of SRB strains that is manifested in the change in enzymatic activity.