Influence of EPS isolated from thermophilic sulphate-reducing bacteria on carbon steel corrosion

Extracellular polymeric substances (EPS) were isolated by centrifugation of thermophilic sulphate-reducing bacteria (SRB) grown in API-RP38 culture medium. The protein and polysaccharide fractions were quantified and the highest concentrations were extracted from a 14-day old culture. The effect of EPS on carbon steel corrosion was investigated by electrochemical techniques. At 30°C, a small amount of EPS in 3% NaCl solution inhibited corrosion, whilst excessive amounts of EPS facilitated corrosion. In addition, the inhibition efficiency of EPS decreased with temperature due to thermal desorption of the EPS. The results suggest that adsorbed EPS layers could be beneficial to anti-corrosion by hindering the reduction of oxygen. However, the accumulation of an EPS film could stimulate the anodic dissolution of the underlying steel by chelation of Fe²⁺ ions.     

Influence of EPS isolated from thermophilic sulphate-reducing bacteria on carbon steel corrosion

Extracellular polymeric substances (EPS) were isolated by centrifugation of thermophilic sulphate-reducing bacteria (SRB) grown in API-RP38 culture medium. The protein and polysaccharide fractions were quantified and the highest concentrations were extracted from a 14-day old culture. The effect of EPS on carbon steel corrosion was investigated by electrochemical techniques. At 30°C, a small amount of EPS in 3% NaCl solution inhibited corrosion, whilst excessive amounts of EPS facilitated corrosion. In addition, the inhibition efficiency of EPS decreased with temperature due to thermal desorption of the EPS. The results suggest that adsorbed EPS layers could be beneficial to anti-corrosion by hindering the reduction of oxygen. However, the accumulation of an EPS film could stimulate the anodic dissolution of the underlying steel by chelation of Fe2+ ions.     

The assay of in-situ activities of sulphate-reducing bacteria in a laboratory marine corrosion model

A critical reappraisal of the [³⁵S] sulphate reduction assay as applied to studies of microbial corrosion has identified sulphate limitation and the production of both acid volatile and nonacid volatile sulphides as key factors which must be incorporated into the assay and into the interpretation of its findings. Application of the modified assay to a laboratory corrosion model gave data reinforcing the fundamental role of environmental factors, notably oxygen, in determining the nature and extent of the corrosion process.     

Neutrophilic iron-oxidizing "zetaproteobacteria" and mild steel corrosion in nearshore marine environments

Microbiologically influenced corrosion (MIC) of mild steel in seawater is an expensive and enduring problem. Little attention has been paid to the role of neutrophilic, lithotrophic, iron-oxidizing bacteria (FeOB) in MIC. The goal of this study was to determine if marine FeOB related to Mariprofundus are involved in this process. To examine this, field incubations and laboratory microcosm experiments were conducted. Mild steel samples incubated in nearshore environments were colonized by marine FeOB, as evidenced by the presence of helical iron-encrusted stalks diagnostic of the FeOB Mariprofundus ferrooxydans, a member of the candidate class "Zetaproteobacteria." Furthermore, Mariprofundus-like cells were enriched from MIC biofilms. The presence of Zetaproteobacteria was confirmed using a Zetaproteobacteria-specific small-subunit (SSU) rRNA gene primer set to amplify sequences related to M. ferrooxydans from both enrichments and in situ samples of MIC biofilms. Temporal in situ incubation studies showed a qualitative increase in stalk distribution on mild steel, suggesting progressive colonization by stalk-forming FeOB. We also isolated a novel FeOB, designated Mariprofundus sp. strain GSB2, from an iron oxide mat in a salt marsh. Strain GSB2 enhanced uniform corrosion from mild steel in laboratory microcosm experiments conducted over 4 days. Iron concentrations (including precipitates) in the medium were used as a measure of corrosion. The corrosion in biotic samples (7.4 ± 0.1 mM) was significantly higher than that in abiotic controls (5.0 ± 0.1 mM). These results have important implications for the role of FeOB in corrosion of steel in nearshore and estuarine environments. In addition, this work shows that the global distribution of Zetaproteobacteria is far greater than previously thought.     

Detection of abundant sulphate-reducing bacteria in marine oxic sediment layers by a combined cultivation and molecular approach

The depth distribution and diversity of sulphate-reducing bacteria (SRB) was analysed in the upper intertidal zone of a sandy marine sediment of the Dutch island Schiermonnikoog. The upper centimetre of the sediment included the oxic-anoxic interface and was cut into five slices. With each slice, most probable number (MPN) dilution series were set up in microtitre plates using five different substrates. In the deeper sediment layers, up to 1 x 10(8) cm(-3) lactate-utilizing SRB were counted, corresponding to 23% of the total bacterial count. From the highest positive dilutions of the MPN series, 27 strains of SRB were isolated in pure culture. Sequencing of a 580 bp fragment of the 16S rDNA revealed that 21 isolates had identical sequences, also identical with that of the previously described species Desulfomicrobium apsheronum. However, the diversity of the isolates was higher with respect to their physiological properties: a total of 11 different phenotypes could be distinguished. Genomic fingerprinting by enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR) revealed an even higher diversity of 22 different genotypes. A culture-independent analysis by PCR and denaturing-gradient gel electrophoresis (DGGE) revealed that the partial 16S rDNA sequence of the isolated D. apsheronum strains constituted a significant fraction of the Desulfovibrionaceae. The high subspecies diversity suggests that this abundant aggregate-forming species may have evolved adaptations to different ecological niches in the oxic sediment layers.     

Sulphate reduction and vertical distribution of sulphate-reducing bacteria quantified by rRNA slot-blot hybridization in a coastal marine sediment

In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization is a valuable tool for a more realistic assessment of SRB abundance in the natural environment. The distribution of SRB was investigated in a coastal marine sediment by hybridization of membrane-immobilized rRNA with oligonucleotide probes. As represented by general probe-target groups, SRB rRNA contributed between 18% and 25% to the prokaryotic rRNA pool. The dominant SRB were related to complete oxidizing genera (Desulphococcus, Desulphosarcina and Desulphobacterium), while Desulphobacter could not be detected. The vertical profile and quantity of rRNA from SRB was compared with sulphate reduction rates (SRR) measured with 35SO4(2-) tracer in whole-core incubations. While SRB abundance was highest near the surface, peaking at around 1.5 cm, measured sulphate reduction rates were lowest in this region. A second peak of SRB rRNA was observed at the transition zone from oxidized to reduced sediment, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis of these estimated cell numbers were between 0.01 and 0.09 fmol SO4(2-) cell(-1) day(-1), which is below the rates that have been determined for pure cultures (0.2-50 fmol SO4(2-) cell(-1) day(-1)) growing exponentially at nearoptimal temperature with a surplus of substrates.     

水环境中工程纳米颗粒物的生态毒理学机理及理想模式生物的筛选

随着纳米技术产业的高速发展,大量工程纳米颗粒物(Engineering nano-particles,ENPs)被排放到自然水环境中,因此对其进行生态毒性及环境风险的研究尤为迫切。综述了ENPs在水环境中的毒理学机理及理想模式生物筛选的研究进展。目前的研究表明ENPs的毒性作用机制主要包括两方面:一是影响细胞信号通路,二是氧化应激造成基因表达的变化。此外,光催化活性、细胞表面附着、溶解特性、表面特征、赋存形态、溶剂效应及与其他环境污染物的协同作用也是可能的毒性作用机理。模式生物的筛选与确定在纳米生态毒理学研究中极为重要。鱼类作为水环境中普遍存在的脊椎动物,群落庞大,其具有行为端点敏感性高、且在生物毒性实验中存在明显的量效关系等特征,被认为是研究ENPs生态毒理学最适合的水生模式生物。研究表明针对在ENPs影响下的未成年鱼类的行为特征研究比传统的胚胎发育及致死率研究更为有效。无脊椎动物和浮游植物同样在各种水环境中普遍存在,对环境污染物极为敏感,且对有害物质具有显著的富集放大效应,因此作为模式生物也具有一定的优势。     

Diversity and distribution of pigmented heterotrophic bacteria in marine environments

A systematic investigation of marine pigmented heterotrophic bacteria (PHB) based on the cultivation method and sequencing analysis of 16S rRNA genes was conducted in Chinese coastal and shelf waters and the Pacific Ocean. Both the abundance of PHB and the ratio of PHB to CFU decreased along trophic gradients from coastal to oceanic waters, with the highest values of 9.9 x 10(3) cell mL(-1) and 39.6%, respectively, in the Yangtze River Estuary. In contrast to the total heterotrophic bacteria (TB) and CFU, which were present in the whole water column, PHB were primarily confined to the euphotic zone, with the highest abundance of PHB and ratio of PHB to CFU occurring in surface water. In total, 247 pigmented isolates were obtained during this study, and the phylogenetic analysis showed a wide genetic diversity covering 25 genera of six phylogenetic classes: Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, Bacilli, Flavobacteria and Sphingobacteria. PHB belonging to Alphaproteobacteria, Flavobacteria and Sphingobacteria were obtained mainly from the South China Sea and East China Sea; PHB from the Pacific Ocean water were predominantly affiliated with Gammaproteobacteria, and most isolates from the Yangtze River Estuary fell into the classes Actinobacteria and Bacilli. The isolates exhibited various colours (e.g. golden, yellow, red, pink and orange), with genus or species specificity. Furthermore, the pigment of PHB cells absorbed light mainly in the wavelength range between 450 and 550 nm. In conclusion, our work has revealed that PHB with broad genetic diversity are widely distributed in the marine environment, and may account for up to 39.6% of culturable bacteria, equivalent to 1.4% of the total microbial community. This value might even be underestimated because it is probable that not all pigmented bacteria were isolated. Their abundance and genetic distribution are heavily influenced by environmental properties, such as light and nutrition, suggesting that they have important roles in the marine ecosystem, especially in the absorption of visible light.     

Isolation of new types of sulphate-reducing bacteria from estuarine and marine sediments using chemostat enrichments

Anaerobic chemostat enrichments have been successfully employed to isolate acetate-utilizing, sulphate-reducing bacteria from sediments in the Tay estuary. Several different morphologically and nutritionally versatile types of sulphate-reducing bacteria have been isolated by this means which has proved especially useful when they are present in low numbers. The principal advantage of this method is that strict anaerobes such as sulphate-reducing bacteria can be isolated in a controlled and reproducible enrichment system at low growth rates.     

海洋铁细菌对紫铜腐蚀行为的影响

【目的】研究海洋铁细菌(Iron-oxidizing bacteria,IOB)对铜的腐蚀影响。【方法】采用电化学极化曲线、电化学阻抗谱(EIS)研究紫铜在海洋环境铁细菌影响下的腐蚀行为。【结果】扫描电子显微镜SEM形貌分析结果表明,在紫铜上附着的铁细菌呈杆状。电化学测试结果表明紫铜在含IOB海水中的腐蚀过程主要受活化极化控制,有菌海水的活化电阻比无菌海水的小6 000.cm2,表明在含IOB的海水中铜发生溶解反应受到的阻滞较小。【结论】IOB的存在导致紫铜的自腐蚀电位变小,使得紫铜发生腐蚀的倾向变大;IOB也使紫铜的阳极极化曲线塔菲尔斜率和变小,从而加速了紫铜的腐蚀进程。     

Specific inhibition of sulphate-reducing bacteria in methanogenic co-culture

Various inhibitors of Desulfovibrio desulfuricans were examined for their selectivity in co-cultures with Methanobacterium formicicum . The inhibitors were chlorohexidine diacetate, m -dinitrobenzene, 2,4-dinitrophenol, potassium nitrate and sodium molybdate. Only sodium molybdate was able to inhibit D. desulfuricans selectively in the methanogenic co-culture. Sodium molybdate at all the concentrations used inhibited D. desulfuricans without adversely affecting M. formicicum .     

Nitrite reductase activity of sulphate-reducing bacteria prevents their inhibition by nitrate-reducing,sulphide-oxidizing bacteria

Sulphate-reducing bacteria (SRB) can be inhibited by nitrate-reducing, sulphide-oxidizing bacteria (NR-SOB), despite the fact that these two groups are interdependent in many anaerobic environments. Practical applications of this inhibition include the reduction of sulphide concentrations in oil fields by nitrate injection. The NR-SOB Thiomicrospira sp. strain CVO was found to oxidize up to 15 mM sulphide, considerably more than three other NR-SOB strains that were tested. Sulphide oxidation increased the environmental redox potential (Eh) from -400 to +100 mV and gave 0.6 nitrite per nitrate reduced. Within the genus Desulfovibrio, strains Lac3 and Lac6 were inhibited by strain CVO and nitrate for the duration of the experiment, whereas inhibition of strains Lac15 and D. vulgaris Hildenborough was transient. The latter had very high nitrite reductase (Nrf) activity. Southern blotting with D. vulgaris nrf genes as a probe indicated the absence of homologous nrf genes from strains Lac3 and Lac6 and their presence in strain Lac15. With respect to SRB from other genera, inhibition of the known nitrite reducer Desulfobulbus propionicus by strain CVO and nitrate was transient, whereas inhibition of Desulfobacterium autotrophicum and Desulfobacter postgatei was long-lasting. The results indicate that inhibition of SRB by NR-SOB is caused by nitrite production. Nrf-containing SRB can overcome this inhibition by further reducing nitrite to ammonia, preventing a stalling of the favourable metabolic interactions between these two bacterial groups. Nrf, which is widely distributed in SRB, can thus be regarded as a resistance factor that prevents the inhibition of dissimilatory sulphate reduction by nitrite.     

Isolation and characterization of sulfate-reducing bacteria from various marine environments

Summary Forty-one strains of non-sporulating sulfate-reducing bacteria were isolated from estuaries, deep sea and other saline environments. Their salinity requirements, utilization of significant carbon compounds, resistance against growth inhibition by Hibitane, optimal growth temperatures and growth temperature ranges were studied. The results include data on strains isolated from the Red Sea hot brine deep area. Basing on the determined characteristics the strains were identified as Desulfovibrio desulfuricans, D. vulgaris, D. salexigens, and D. desulfuricans var. aestuarii.     

海水富营养化对海洋细菌影响的研究进展

综述了海水富营养化对海洋细菌影响的研究进展。随着海水富营养化程度的增加,海洋细菌数量或生物量增加;反硝化细菌、大肠菌群尤其是厌氧性的硫酸盐还原菌和产甲烷菌等典型细菌生理群数量增加;浮游细菌群落结构随富营养化递增趋于简单,物种多样性降低;富营养化也明显导致细菌群落正常功能活性的紊乱。海水富营养化对细菌群落的结构和功能有着深远的影响。     

Iron-oxidizing bacteria in marine environments: recent progresses and future directions

Iron-oxidizing bacteria (FeOB) refers to a group of bacteria with the ability to exchange and accumulate divalent iron dissolved in water as trivalent iron inside and outside the bacterial cell. Most FeOB belong the largest bacterial phylum, Proteobacteria. Within this phylum, FeOB with varying physiology with regards to their response to oxygen (obligate aerobes, facultative and obligate anaerobes) and pH optimum for proliferation (neutrophiles, moderate and extreme acidophiles) can be found. Although FeOB have been reported from a wide variety of environments, most of them have not been isolated and their biochemical characteristics remain largely unknown. This is especially true for those living in the marine realm, where the properties of FeOB was not known until the isolation of the Zetaproteobacteria Mariprofundus ferrooxydans, first reported in 2007. Since the proposal of Zetaproteobacteria by Emerson et al., the detection and isolation of those microorganisms from the marine environment has greatly escalated. Furthermore, FeOB have also recently been reported from works on ocean drilling and metal corrosion. This review aims to summarize the current state of phylogenetic and physiological diversity in marine FeOB, the significance of their roles in their environments (on both global and local scales), as well as their growing importance and applications in the industry.     

Role of iron-reducing bacteria in corrosion and protection of carbon steel

The role of iron-reducing bacteria (IRB) in biocorrosion is under discussion. According to some reports, IRB are able to induce protection of carbon steel while others suggest an important enhancement of corrosion through the reduction and removal of passive films of ferric compounds on the metal surface. In this work, we review recent knowledge concerning microbial respiration, the ecology of IRB containing biofilms and the corrosive or protective effect of such biofilms on metal surfaces.     

Study of parameters implicated in the biodeterioration of mild steel in the presence of different species of sulphate-reducing bacteria

Two different species of sulphate-reducing bacteria, strain classified by NCIMB as Desulfovibrio desulfuricans subspecies desulfuricans New Jersey (8313) isolated from the corroding heat exchanger, and SRB species recovered from a corroding ship hull anchored off the Indonesian coast (Indo isolate) were grown as laboratory batch cultures. Several factors such as the surface finish of substratum, metabolic activity of planktonic and sessile bacterial populations, initial attachment of cells to surfaces and subsequent formation of biofilms on the process of biodeterioration of mild steel in the presence of these two different species of SRB were investigated. The corrosion rates of mild steel were estimated by weight loss measurements and correlated with the density of sessile SRB population. The yield and composition of extracellular polymers released into the bulk phase of culture media were determined and the amount of dissolved hydrogen sulphide was monitored. The results revealed differences between SRB species in their aggressiveness towards mild steel under identical growth conditions, emphasising the importance of biochemistry and physiology of SRB for the biocorrosion process. Biochemical and genetic characterisation of SRB isolates chosen for this study are currently in progress.     

Switching between apparently redundant iron-uptake mechanisms benefits bacteria in changeable environments

Bacteria often possess multiple siderophore-based iron uptake systems for scavenging this vital resource from their environment. However, some siderophores seem redundant, because they have limited iron-binding efficiency and are seldom expressed under iron limitation. Here, we investigate the conundrum of why selection does not eliminate this apparent redundancy. We focus on Pseudomonas aeruginosa, a bacterium that can produce two siderophores—the highly efficient but metabolically expensive pyoverdine, and the inefficient but metabolically cheap pyochelin. We found that the bacteria possess molecular mechanisms to phenotypically switch from mainly producing pyoverdine under severe iron limitation to mainly producing pyochelin when iron is only moderately limited. We further show that strains exclusively producing pyochelin grew significantly better than strains exclusively producing pyoverdine under moderate iron limitation, whereas the inverse was seen under severe iron limitation. This suggests that pyochelin is not redundant, but that switching between siderophore strategies might be beneficial to trade off efficiencies versus costs of siderophores. Indeed, simulations parameterized from our data confirmed that strains retaining the capacity to switch between siderophores significantly outcompeted strains defective for one or the other siderophore under fluctuating iron availabilities. Finally, we discuss how siderophore switching can be viewed as a form of collective decision-making, whereby a coordinated shift in behaviour at the group level emerges as a result of positive and negative feedback loops operating among individuals at the local scale.