Contemporary strategies and approaches for characterizing composition and enhancing biofilm penetration targeting bacterial extracellular polymeric substances

Extracellular polymeric substances(EPS)constitutes crucial elements within bacterial biofilms,facili-tating accelerated antimicrobial resistance and conferring defense against the host's immune cells.Developing precise and effective antibiofilm approaches and strategies,tailored to the specific charac-teristics of EPS composition,can offer valuable insights for the creation of novel antimicrobial drugs.This,in turn,holds the potential to mitigate the alarming issue of bacterial drug resistance.Current analysis of EPS compositions relies heavily on colorimetric approaches with a significant bias,which is likely due to the selection of a standard compound and the cross-interference of various EPS compounds.Considering the pivotal role of EPS in biofilm functionality,it is imperative for EPS research to delve deeper into the analysis of intricate compositions,moving beyond the current focus on polymeric materials.This ne-cessitates a shift from heavy reliance on colorimetric analytic methods to more comprehensive and nuanced analytical approaches.In this study,we have provided a comprehensive summary of existing analytical methods utilized in the characterization of EPS compositions.Additionally,novel strategies aimed at targeting EPS to enhance biofilm penetration were explored,with a specific focus on high-lighting the limitations associated with colorimetric methods.Furthermore,we have outlined the challenges faced in identifying additional components of EPS and propose a prospective research plan to address these challenges.This review has the potential to guide future researchers in the search for novel compounds capable of suppressing EPS,thereby inhibiting biofilm formation.This insight opens up a new avenue for exploration within this research domain.

Evaluation of Rhizobium tropici-Derived Extracellular Polymeric Substances on Selected Soil Properties, Seed Germination, and Growth of Black-Eyed Peas (Vigna unguiculata)

Rhizobium tropici-derived extracellular polymeric substances (EPS) have been used in soils to enhance soil structures and mitigate soil erosions. However, information on their use to improve soil health and fertility indicators, and plant growth is limited. In a greenhouse study, we investigated their effects on some soil health, soil fertility indices, and the growth of black-eyed peas (Vigna unguiculate). Results showed that soils incubated with EPS significantly increased basal soil respiration, soil microbial biomass, permanganate oxidizable carbon (POC), and potentially mineralizable nitrogen (PMN). The EPS shifted microbial populations from bacteria to fungi and Gram (−ve) to Gram ( ve) bacteria. However, it had little or no effects on soil pH, soil organic matter (SOM), and cation exchange capacity (CEC). The EPS decreased soil moisture loss, increased soil aggregate stability, but delayed blacked-eyed peas germinations in the soils. At 0.1% (w/w) concentrations in soils, there was increase in plant root nodulations and vegetative growth. This study was carried out within 40 days of incubating soils with EPS or growing the black-eyed peas in a greenhouse study. The plant growth parameters were taken before flowering and fruiting. Further studies of the effects of incubating soils with the extracellular polymeric substances on plant growth. Soil microbial biomass, microbial diversities, and other soil fertility indices are deemed necessary.

Role of extracellular polymeric substances in resistance to allelochemical stress on Microcystis aeruginsosa and its mechanism

Using allelochemicals to suppress cyanobacteria growth is a prospective method for its high efficiency and ecological safety.However,the suppression efficiency is affected inevitably by the extracellular polymeric substances(EPS)produced by cyanobacteria,and the knowledge about the roles of EPS in resistance to allelochemical stress is scarce.For the study,two typical anti-cyanobacterial allelochemicals were adopted to investigate the role of EPS in resistance to allelochemical stress on Microcystis aeruginosa.Results show that EPS was crucial in alleviating the toxicity of allelochemicals to algae,especially in stabilizing the metabolism and photosynthetic activity of algal cells.The aggregation rate of algal cells increased with the increase of EPS secretion,which alleviated the stress of allelopathy.Tryptophan proteins and humic acids in EPS provided a binding site for allelochemicals,and the EPS-allelochemicals complex were formed by chemical bonding.This study improved our comprehension of the role of EPS in algal inhibition by allelochemicals.

Effect of extracellular polymeric substances on Dolichospermum aggregation during temperature rise

Dolichospermum,a typical model filamentous of cyanobacteria,has the potential to cause severely bloom.Extracellular polymeric substances(EPSs)are considered to influence the aggregation of the algae,and temperature is a significant factor affecting EPSs secretion.However,the mechanism of how EPSs affects the aggregation of Dolichospermum is still unclear because the structure and composition of EPSs are complex.In this study,the effects of EPSs on the aggregation of Dolichospermum during the rise of temperature(7-37℃)were determined.The results showed that the concentration of extracellular polysaccharides and proteins changed significantly with increasing temperature(P<0.01).Firstly,during the increasing temperature,the polysaccharide content of EPSs increased from 20.34 to 54.64 mg/L,and the polysaccharides in the soluble EPS(S-EPS)layer changed significantly.The protein content reached maximum value at 21℃(14.52 mg/L)and varied significantly in S-EPS and loosely bound EPS(LB-EPS).In the EPSs matrix,humus substances and protein were main components of S-EPS and LB-EPS,and protein was the main component of tightly bound EPS(TB-EPS).Secondly,the cell density of Dolichospermum increased during the temperature rise while the aggregation ratio decreased.Moreover,zeta potential and surface thermodynamic analysis of Dolichospermum revealed that the interfacial free energy and electrostatic repulsion increased gradually with increasing temperature,which further reduced the aggregation of Dolichospermum.Finally,principal component analysis(PCA)analysis showed the aggregation of Dolichospermum was directly related to the changes of protein in EPSs(especially S-EPS and LB-EPS)and zeta potential,and polysaccharides in EPSs inhibited the aggregation of Dolichospermum.Based on these results,it was illustrated that the composition and concentration of EPSs affected the cell surface properties of Dolichospermum with the change of temperature and thus affected the aggregation of Dolichospermum.

Relationship and effect of redox potential,jarosites and extracellular polymeric substances in bioleaching chalcopyrite by acidithiobacillus ferrooxidans

The changes of pH,redox potential,concentrations of soluble iron ions and Cu^2＋ with the time of bioleaching chalcopyrite concentrates by acidithiobacillus ferrooxidans were investigated under the different conditions of initial total-iron amount as well as mole ratio of Fe（III） to Fe（II） in the solutions containing synthetic extracellular polymeric substances （EPS）.When the solution potential is lower than 650 mV （vs SHE）,the inhibition of jarosites to bioleaching chalcopyrite is not vital as EPS produced by bacteria can retard the contamination through flocculating jarosites even if concentration of Fe（III） ions is up to 20 g/L but increases with increasing the concentration of Fe（III） ions;jarosites formed by bio-oxidized Fe3＋ ions are more easy to adhere to outside surface of EPS space on chalcopyrite;the EPS layer with jarosites acts as a weak diffusion barrier to further rapidly create a high redox potential of more than 650 mV by bio-oxidizing Fe^2＋ ions inside and outside EPS space into Fe^3＋ ions,resulting in a rapid deterioration of ion diffusion performance of the EPS layer to inhibit bioleaching chalcopyrite severely and irreversibly.

Interaction mechanism of Cu^(2+),Fe^(3+) ions and extracellular polymeric substances during bioleaching chalcopyrite by Acidithiobacillus ferrooxidans ATCC2370

The extracellular polymeric substances（EPS） of Acidithiobacillus ferrooxidans ATCC 23270,and iron and copper enclosed in EPS were extracted by ultrasonication and centrifugation methods to determine the interaction mechanism of Cu2＋,Fe3＋ and EPS during bioleaching chalcopyrite.Generally,Cu2＋ ions can stimulate bacteria to produce more EPS than Fe3＋ ions.The mass ratio of Fe3＋/Cu2＋ enclosed in EPS decreased gradually from about 4：1 to about 2：1 when the concentration of Cu2＋ ions increased from 0.01 to 0.04 mol/L.The amount of iron and copper bound together by EPS in ferrous-free 9K medium containing 1% chalcopyrite was about 2 times of that in 9K medium containing 0.04 mol/L Cu2＋ ions.It was inferred that the EPS with jarosites on the surface of chalcopyrite gradually acted as a weak diffusion barrier for Cu2＋,Fe3＋ ions transference during bioleaching chalcopyrite.

Optimization of Extraction Conditions of Extracellular Polymeric Substances from Activated Sludge

[Objective] To explore the optimal extraction conditions of extracellular polymeric substances （EPS） from activated sludge. [Method] The efficiency of five methods （H2SO4, formaldehyde-NaOH, mixing, heating and NaOH） on the extraction of EPS was investigated comparatively. The optimal extraction conditions of the most suitable method were determined. [Result] NaOH method is most effective in extracting EPS with less DNA contamination and shortened extraction period. The optimal extraction condition was pH of 11, extraction time of 10 min and agitation speed of 80-120 r/min. [Conclusion] The determined optimal extraction condition provided theoretical basis for EPS study.

Effects of ultrasonic pretreatment on sludge dewaterability and extracellular polymeric substances distribution in mesophilic anaerobic digestion

Effect of ultrasonic pretreatment on sludge dewaterability was determined and the fate of extracellular polymeric substances （EPS） matrix in mesophilic anaerobic digestion after ultrasonic pretreatment was studied. Characteristics of proteins （PN）, polysaccharides （PS）, excitation-emission matrix （EEM） fluorescence spectroscopy and molecular weight （MW） distribution of dissolved organic matters （DOM） in different EPS fractions were evaluated. The results showed that after ultrasonic pretreatment, the normalized capillary suction time （CST） decreased from 44.4 to 11.1 （sec·L）/g total suspended solids （TSS） during anaerobic digestion, indicating that sludge dewaterability was greatly improved. The normalized CST was significantly correlated with PN concentration （R2 = 0.92, p 〈 0.01） and the PN/PS ratio （R2 = 0.84, p 〈 0.01） in the loosely bound EPS （LB-EPS） fraction. Meanwhile, the average MW of DOM in the LB- EPS and tightly bound EPS （TB-EPS） fractions also had a good correlation with the normalized CST （R2 〉 0.66, p 〈 0.01）. According to EEM fluorescence spectroscopy, tryptophan-like substances intensities in the slime, LB-EPS and TB-EPS fractions were correlated with the normalized CST. The organic matters in the EPS matrix played an important role in influencing sludge dewaterability.

Functions and behaviors of activated sludge extracellular polymeric substances (EPS): a promising environmental interest

Extracellular polymeric substances （EPS） are the predominant constituents of activated sludge and represent up to 80% of the mass of activated sludge. They play a crucial role in the flocculation, settling and dewatering of activated sludge. Furthermore, EPS also show great efficiency in binding heavy metals. So EPS are key factors influencing reduction in sludge volume and mass, as well as activity and utilization of sludge. EPS are of considerable environmental interest and hundreds of articles on EPS have been published abroad, while information on EPS in China is limited. In this paper, results of over 60 publications related to constituents and characteristics of EPS and their influences on flocculation, settling and dewatering of sludge are compiled and analyzed. Metal-binding ability of EPS is also discussed, together with a brief consideration of possible research interests in the future.

Key physiological traits and chemical properties of extracellular polymeric substances determining colony formation in a cyanobacterium

Colony formation of cyanobacteria is crucial for the formation of surface blooms in lakes.However,the underlying mechanisms of colony formation involving in physiological and cell surface characteristics remain to not well be established.Six cyanobacterial Microcystis strains(including both unicellular and colonial ones)were employed to estimate the influences of their physiological traits and the composition of extracellular polymeric substances(EPS)on colony or aggregate formation.Results show that raising the number of the photosynthetic reaction center and light-harvesting antenna in the PSII and reducing the growth rate were the major physiological strategies of Microcystis to produce excess EPS enhancing colony formation.Tightly bound EPS(T-EPS)was responsible for colony formation,which approximately accounted for 50%of the total amount of EPS.Five fluorescent components(protein-,tryptophan-,and tyrosine-like components and two humic-like components)were found in the T-EPS,although the amounts of these components varied with strains.Importantly,colonial strains contained much higher tyrosine-like substances than unicellular ones.We suggest that tyrosine-like substances might serve as a crosslinking agent to connect other polymers in EPS(e.g.,proteins or polysaccharides)for colony formation.Our findings identified key physiological traits and chemical components of EPS for colony formation in Microcystis,which can contribute to a better understanding on the formation of Microcystis blooms.

Evaluation of the Influence of Extracellular Polymeric Substances on the Mass Transport of Substrate within Multispecies Biofilms

A model, for evaluating the effect of porosity and volume fraction of extracellular polymeric substances (EPS) within multispecies biofilms on the effective diffusivity, is developed and experimentally validated, based on the extraction of EPS from intact biofilms. The amount of EPS in biofilms significantly affects the effective diffusivity. For biofilms with porosity of 77%—95% in the top layers and 54%—58% in the bottom layers, the value of De/Dw decreases from 0.52—0.83 in the top layers to 0.23—0.31 in the bottom layers. Generally, the effective diffusivity in the heterotrophic/autotrophic biofilms is slightly lower than that in the heterotrophic biofilms, due to the lower porosity in the heterotrophic/autotrophic biofilms.

Glycine betaine modulates extracellular polymeric substances to enhance microbial salinity tolerance

High salinity inhibits microbial activity in the bioremediation of saline wastewater.To alleviate osmotic stress,glycine betaine(GB),an osmoprotectant,is added to enhance the secretion of extracellular polymeric substances(EPS).These EPS are pivotal in withstanding environmental stressors,yet the intricate interplay between GB supplementation and microbial responses through EPS modificationsdencompassing composition,molecular architecture,and electrochemical featuresdremains elusive in hypersaline conditions.Here we show microbial strategies for salinity endurance by investigating the impact of GB on the dynamic alterations of EPS properties.Our findings reveal that GB supplementation at 3.5%salinity elevates the total EPS(T-EPS)content from 12.50±0.05 to 24.58±0.96 mg per g dry cell weight.The observed shift in zeta potential from-28.95 to-6.25 mV at 0%and 3.5%salinity,respectively,with GB treatment,indicates a reduction in electrostatic repulsion and compaction.Notably,the EPS protein secondary structure transition from b-sheet to a-helix,with GB addition,signifies a more compact protein configuration,less susceptible to salinity fluctuations.Electrochemical analyses,including cyclic voltammetry(CV)and differential pulse voltammetry(DPV),reveal GB's role in promoting the release of exogenous electron shuttles,such as flavins and c-type cytochromes(c-Cyts).The enhancement in DPV peak areas(Q_(DPV))with GB addition implies an increase in available extracellular electron transfer sites.This investigation advances our comprehension of microbial adaptation mechanisms to salinity through EPS modifications facilitated by GB in saline habitats.

Extracellular polymeric substances as paper coating biomaterials derived from anaerobic granular sludge

Recovering extracellular polymeric substances(EPS)from waste granular sludge offers a cost-effective and sustainable approach for transforming wastewater resources into industrially valuable products.Yet,the application potential of these EPS in real-world scenarios,particularly in paper manufacturing,remains underexplored.Here we show the feasibility of EPS-based biomaterials,derived from anaerobic granular sludges,as novel coating agents in paper production.We systematically characterised the rheological properties of various EPS-based suspensions.When applied as surface sizing agents,these EPS-based biomaterials formed a distinct,ultra-thin layer on paper,as evidenced by scanning electron microscopy.A comprehensive evaluation of water and oil penetration,along with barrier properties,revealed that EPS-enhanced coatings markedly diminished water absorption while significantly bolstering oil and grease resistance.Optimal performance was observed in EPS variants with elevated protein and hydrophobic contents,correlating with their superior rheological characteristics.The enhanced water-barrier and grease resistance of EPS-coated paper can be attributed to its non-porous,fine surface structure and the functional groups in EPS,particularly the high protein content and hydrophobic humic-like substances.This research marks the first demonstration of utilizing EPS from anaerobic granular sludge as paper-coating biomaterials,bridging a critical knowledge gap in the sustainable use of biopolymers in industrial applications.

The effect of extracellular polymeric substances (EPS) of iron-oxidizing bacteria (Ochrobactrum EEELCW01) on mineral transformation and arsenic (As) fate

Extracellular polymeric substances(EPS)are an importantmedium for communication and material exchange between iron-oxidizing bacteria and the external environment and could induce the iron(oxyhydr)oxides production which reduced arsenic(As)availability.The main component of EPS secreted by iron-oxidizing bacteria(Ochrobactrum EEELCW01)was composed of polysaccharides(150.76-165.33 mg/g DW)followed by considerably smaller amounts of proteins(12.98–16.12 mg/g DW).Low concentrations of As(100 or 500μmol/L)promoted the amount of EPS secretion.FTIR results showed that EPS was composed of polysaccharides,proteins,and a miniscule amount of nucleic acids.The functional groups including-COOH,-OH,-NH,-C=O,and-C-O played an important role in the adsorption of As.XPS results showed that As was bound to EPS in the form of As3+.With increasing As concentration,the proportion of As3+adsorbed on EPS increased.Ferrihydrite with a weak crystalline state was only produced in the system at 6 hr during the mineralization process of Ochrobactrum sp.At day 8,the minerals were composed of goethite,galena,and siderite.With the increasing mineralization time,the main mineral phases were transformed from weakly crystalline hydrous iron ore into higher crystallinity siderite(FeCO_(3))or goethite(α-FeOOH),and the specific surface area and active sites of minerals were reduced.It can be seen from the distribution of As elements that As is preferentially adsorbed on the edges of iron minerals.This study is potential to understand the biomineralizationmechanism of iron-oxidizing bacteria and As remediation in the environment.

Extracellular polymeric substances-antibiotics interaction in activated sludge:A review

Antibiotics,the most frequently prescribed drugs,have been widely applied to prevent or cure human and veterinary diseases and have undoubtedly led to massive releases into sewer networks and wastewater treatment systems,a hotspot where the occurrence and transformation of antibiotic resistance take place.Extracellular polymeric substances(EPS),biopolymers secreted via microbial activity,play an important role in cell adhesion,nutrient retention,and toxicity resistance.However,the potential roles of sludge EPS related to the resistance and removal of antibiotics are still unclear.This work summarizes the composition and physicochemical characteristics of state-of-the-art microbial EPS,highlights the critical role of EPS in antibiotics removal,evaluates their defense performances under different antibiotics exposures,and analyzes the typical factors that could affect the sorption and biotransformation behavior of antibiotics.Next,interactions between microbial EPS and antibiotic resistance genes are analyzed.Future perspectives,especially the engineering application of microbial EPS for antibiotics toxicity detection and defense,are also emphatically stressed.

Flocculating characteristic of activated sludge flocs:Interaction between Al^(3+) and extracellular polymeric substances

Aluminum flocculant can enhance the flocculating performance of activated sludge.However,the binding mechanism of aluminum ion（Al 3＋） and extracellular polymeric substances（EPS） in activated sludge is unclear due to the complexity of EPS.In this work,threedimensional excitation emission matrix fluorescence spectroscopy（3DEEM）,fluorescence quenching titration and Fourier transform infrared spectroscopy（FT-IR） were used to explore the binding behavior and mechanism between Al 3＋ and EPS.The results showed that two fluorescence peaks of tyrosineand tryptophan-like substances were identified in the loosely bound-extracellular polymeric substances（LB-EPS）,and three peaks of tyrosine-,tryptophanand humic-like substances were identified in the tightly boundextracellular polymeric substances（TB-EPS）.It was found that these fluorescence peaks could be quenched with Al 3＋ at the dosage of 3.0 mg/L,which demonstrated that strong interactions took place between the EPS and Al 3＋.The conditional stability constants for Al 3＋ and EPS were determined by the Stern-Volmer equation.As to the binding mechanism,the-OH,N-H,C=O,C-N groups and the sulfurand phosphorus-containing groups showed complexation action,although the groups in the LB-EPS and TB-EPS showed different behavior.The TB-EPS have stronger binding ability to Al 3＋ than the LB-EPS,and TB-EPS play an important role in the interaction with Al 3＋.

Effects of solution conditions on the physicochemical properties of stratification components of extracellular polymeric substances in anaerobic digested sludge

The composition and effects of solution conditions on the physicochemical properties of the stratification components of extracellular polymeric substances （EPS） in anaerobic digested sludge were determined. The total EPS in anaerobic digested sludge were extracted by the cation exchange resin method. Another EPS extraction method, the ceutrifugation and sonication technique was employed to stratify the EPS into three fractions： slime, loosely bound （LB）-EPS, and tightly bound （TB）-EPS from the outside to the inside of the anaerobic digested sludge. Proteins and polysaccharides were dispersed uniformly across the different EPS fractions, and humic-like substances were mainly partitioned in the slime, with TB-EPS second. Protein was the major constituent of the LB-EPS and TB-EPS, and the corresponding ratios ranged from 54.0% to 65.6%. The hydrophobic part in the EPS chemical components was primarily comprised of protein and DNA, while the hydrophilic part was mainly composed of polysaccharide. In the slime, the hydrophobic values of several EPS chemical components （protein, polysaccharide, humic-like substances and DNA） were all below 50%. The protein/polysaccharide ratio had a significant influence on the Zeta potentials and isoelectric point values of the EPS： the greater the protein/polysaccharide ratio of the EPS was, the greater the Zeta potential and the higher the isoelectric point value were. All Zeta potentials of the EPS showed a decreasing trend with increasing pH. The corresponding isoelectric point values （pH） were 2.8 for total EPS, 2.2 for slime, 2.7 for LB-EPS, and 2.6 for TB-EPS. As the ionic strength increased, the Zeta potentials sharply increased and then gradually became constant without charge reversal. In addition, as the temperature increased （〈 40℃）, the apparent viscosity of the EPS decreased monotonically and then gradually became stable between 40 and 60℃.

Effects of Al^3+on pollutant removal and extracellular polymeric substances(EPS)under anaerobic,anoxic and oxic conditions

Aluminum ions produced by aluminum mining,electrolytic industry and aluminum-based coagulants can enter wastewater treatment plants and interact with activated sludge.They can subsequently contribute to the removal of suspended solids and affect activated sludge flocculation,as well as nitrogen and phosphorus removal.In this study,the effects of Al^3+on pollutant removal,sludge flocculation and the composition and structure of extracellular polymeric substances(EPS)were investigated under anaerobic,anoxic and oxic conditions.Results demonstrated that the highest chemical oxygen demand(COD)and total nitrogen(TN)removal efficiencies were detected for an Al^3+concentration of 10 mg/L.In addition,the maximal dehydrogenase activity and sludge flocculation were also observed at this level of Al^3+.The highest removal efficiency of total phosphorus(TP)was achieved at an Ar+concentration of30 mg/L.The flocculability of sludge in the anoxic zone was consistently higher than that in the anaerobic and oxic zones.The addition of Al^3+promoted the secretion of EPS.Tryptophan-like fluorescence peaks were detected in each EPS layer in the absence of Al^3+.At the Al^3+concentration of 10 mg/L,fulvic acid and tryptophan fluorescence peaks began to appear while the majority of protein species and the highest microbial activity were also detected.Low Al^3+concentrations(<10 mg/L)could promote the removal efficiencies of COD and TN,yet excessive Al^3+levels(>10 mg/L)weakened microbial activity.Higher Al^3+concentrations(>30 mg/L)also inhibited the release of phosphorus in the anaerobic zone by reacting with PO4^3-.

Biosorption of Cu（II） to extracellular polymeric substances （EPS） from Synechoeystis sp.： a fluorescence quenching study

Biosorption of extracellular polymeric substances （EPS） from Synechocystis sp. （cyanobacterium） with Cu（II） was investigated using fluorescence spectroscopy. Three fluorescence peaks were found in the excitation-emission matrix （EEM） fluorescence spectra of EPS. Fluorescence of peak A （Ex/Em = 275/452 nm） and peak C （Ex/Em= 350/452nm） were originated from humic-like substances and fluorescence of peak B （Ex/ Em= 275/338nm） was attributed to protein-like sub- stances. Fluorescence of peaks A, B, and C could be quenched by Cu（II）. The effective quenching constants （lg Ka） were 2.8-5.84 for peak A, 6.4-9.24 for peak B, and 3.48-6.68 for peak C, respectively. The values of lg Ka showed a decreasing trend with increasing temperature, indicating that the quenching processes were static in nature. The binding constants （lg Kb） followed the order of peak A 〉 peak B 〉 peak C, implying that the humic-like substances in EPS have greater Cu（II） binding capacity than the protein-like substances. The binding site number, n, in EPS-Cu（II） complexes for peaks A, B, and C was less than 1. This suggests the negative cooperativity between multiple binding sites and the presence of more than one Cu binding site.

A Synergistic Acceleration of Corrosion of Q235 Carbon Steel Between Magnetization and Extracellular Polymeric Substances

In this work, surface characterization and electrochemical measurement were employed to investigate the effects of magnetic field（MF） on the corrosion of Q235 carbon steel in a NaCl solution containing sulphate-reducing bacteria（SRB） or extracellular polymeric substances（EPS）. Results demonstrated that a 150 mT MF enhanced steel corrosion in a SRB-containing NaCl solution by 202% calculated from weight loss with pitting corrosion as the main corrosion type.Either EPS or MF rendered steel corrosion, but a synergistic interaction between MF and EPS boosted up steel corrosion.This synergistic enhancement could be referred to the alteration in orientation of EPS induced by MF. The presence of higher percentage of chloride ions on the carbon steel surface manifested that MF initiated the erosion of chloride ions on the carbon steel coupon.