Treatment of high strength organic wastewater by a mixed culture of photosynthetic microorganisms

The growth characteristics and nutrient removal fromsynthetic wastewater by Rhodobacter sphaeroides,Chlorella sorokiniana and Spirulinaplatensis were investigated under aerobic dark(heterotrophic) and aerobic light (photoheterotrophic)conditions. Both in terms of economy and efficiency,aerobic dark conditions were the best for wastewatertreatment using R. sphaeroides and C.sorokiniana, but light was necessary with S.platensis. Neither growth nor nutrient removalcharacteristics of the cells were affected insynthetic wastewater with as high as 10 000 ppmacetate, 1000 ppm propionate, 700 ppm nitrate and 100 ppmphosphate. Although R. sphaeroides and C. sorokiniana showed good growth in syntheticwastewater containing 400 ppm of ammonia, S.platensis was completely inhibited.When grown as a monoculture, none of thestrains could simultaneously remove acetate,propionate, ammonia, nitrate and phosphate from thewastewater. R. sphaeroides could remove allthe above nutrients except nitrate, but the rate of removal was relatively low. The rate of nutrientsremoval by C. sorokiniana was higher, but theorganism could not remove propionate; S.platensis could efficiently remove nitrate, ammoniaand phosphate, but none of the organic acids. A mixedculture of R. sphaeroides and C.sorokiniana was therefore used for simultaneousremoval of organic acids, nitrate, ammonia andphosphate. The optimum ratio of the cells depended onthe composition of the wastewater.     

Bioremediation of wastewater from edible oil refinery factory using oleaginous microalga Desmodesmus sp. S1

Edible oil industry produced massive wastewater, which requires extensive treatment to remove pungent smell, high phosphate, carbon oxygen demand (COD), and metal ions prior to discharge. Traditional anaerobic and aerobic digestion could mainly reduce COD of the wastewater from oil refinery factories (WEORF). In this study, a robust oleaginous microalga Desmodesmus sp. S1 was adapted to grow in WEORF. The biomass and lipid content of Desmodesmus sp. S1 cultivated in the WEORF supplemented with sodium nitrate were 5.62 g·L^?1 and 14.49%, whereas those in the WEORF without adding nitrate were 2.98 g·L^?1 and 21.95%. More than 82% of the COD and 53% of total phosphorous were removed by Desmodesmus sp. S1. In addition, metal ions, including ferric, aluminum, manganese and zinc were also diminished significantly in the WEORF after microalgal growth, and pungent smell vanished as well. In comparison with the cells grown in BG-11 medium, the cilia-like bulges and wrinkles on the cell surface of Desmodesmus sp. S1 grown in WEORF became out of order, and more polyunsaturated fatty acids were detected due to stress derived from the wastewater. The study suggests that growing microalgae in WEORF can be applied for the dual roles of nutrient removal and biofuel feedstock production.     

The Potential of Canna lily for Wastewater Treatment Under Indian Conditions

Low cost treatment of polluted wastewater has become a serious challenge in most of the urban areas of developing countries. The present study was undertaken to investigate the potential of Canna lily towards removal of carbon, nitrogen, and phosphorus from wastewater under sub-tropical conditions. A constructed wetland (CW) cell supporting vegetative layer of Canna lily was used to treat wastewater having high strength of CNP. Removal of biological oxygen demand (BOD₃) and chemical oxygen demand (COD) varied between 69.8–96.4% and 63.6–99.1%, respectively. C. lily could efficiently remove carbon from a difficult to degrade wastewater at COD:BOD ratio of 24.4. Simultaneous reduction in TKN and nitrate pointed to good nitrification rates, and efficient plant assimilation as the dominant nutrient removal mechanism in the present study. Suitable Indian agro-climatic conditions favored plant growth and no evident stress over the Canna plant was observed. High removal rate of 809.8 mg/m²-day for TKN, 15.0 mg/m²-day for nitrate, and 164.2 mg/m²-day for phosphate suggests for a possible use of Canna-based CW for wastewater treatment for small, rural, and remote Indian communities.     

Simultaneous removal of chemical oxygen demand and nitrate in aerobic treatment of sewage wastewater using an immobilized photosynthetic bacterium of porous ceramic plates

A denitrifying photosynthetic bacterium (PSB), Rhodobacter sphaeroides S was immobilized on a porous ceramic plate (10×10×0.5cm) using a small amount of agar. In the bioreactor (3.5 l liquid) using this plate, repeated batch treatments of sewage wastewater (acetic acid as a carbon source) were carried out under aerobic conditions (DO 5mg/l) for two weeks. The simultaneous and effective removal of chemical oxygen demand (COD), ammonium, and nitrate was observed. In continuous treatment at a relatively high dilution rate of 0.20 h^–1 (retention time, five hours), the same simultaneous treatments were observed for about one month.     

Removal of nitrate, nitrite, ammonium and phosphate ions from water by the aerial microalga Trentepohlia aurea

The aerial microalga Trentepohlia aurea has beeninvestigated in relation to removal characteristics of nitrate, nitrite,ammonium and phosphate ions. When the alga was cultured in medium with veryhighconcentrations of ammonium, nitrate and phosphate ions, it showed relativelyhigh growth and removal rates. It also grew quite well with high nitriteconcentration (< 141 mg NO₂-N L^–1).The removal rate was 0.28 mg NO₂-N L^–1day^–1 in the 40-day culture, when it was cultured in modifiedBold's basal medium with added 51 mg NO₂-NL^–1. In addition, we examined simultaneous removal of nutrientions. The biomass was 1.5 times higher in medium which N- and P-sourcesufficient than in ordinary medium. Higher removal ratios of nitrite andnitratefrom medium were shown in a 30-day culture, reaching 37% and 32%, respectively.It is concluded that T. aurea has the potential for use inthe purification of wastewater.     

The influence of anaerobic pretreatment on the nitrogen removal from biosynthetic pharmaceutical wastewaters

The C:N ratio of the pharmaceutical wastewaters is usually suitable for a combination of the anaerobic pretreatment with the high COD removal and aerobic posttreatment with the efficient biological N removal. This kind of anaerobic-aerobic process was tested in semipilot scale by using a UASB reactor and an activated sludge system with a predenitrification (total volume 100 1). It was found that at a total HRT of 2.3 days an average of 97.5% of COD and 73.5% of total N was removed. The UASB reactor was operated at 30°C with a volumetric loading rate of 8.7 kg.m^-3.d^-1, the efficiency of COD removal was 92.2%. The processes, which take part in the biological removal of nitrogen, especially the nitrification, were running with lower rates than usually observed in aerobic treatment systems.Abbreviations AAO anaerobic anoxic oxic configuration - AOO anaerobic oxic oxic configuration - B _V volumetric organic loading rate (kg COD.m^-3. d^-1) - dB _x specific COD removal rate (mg COD. g^-1 VSS. d^-1) - DNR denitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - E_COD efficiency of COD removal (%) - HRT hydraulic retention time (d) - NR nitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - R recirculation ratio (%) - SBP specific biogas production (m³.kg^-1 removed COD) - SRT solids retention time; sludge age (d) - SS suspended solids (g.1^-1) - UASB upflow anaerobic sludge blanket reactor - VSS volatile suspended solids (g.1^-1)     

On-site removal of H<Subscript>2</Subscript>S from biogas produced by food waste using an aerobic sludge biofilter for steam reforming processing

H₂S in biogas was removed by sludge-loaded biofiltration, rendering the biogas suitable for catalytic reforming into a mixture of CO and H₂ syngas that was then applied for the generation of electricity using a solid oxide fuel cell or for the chemical synthesis of methanol. The biogas was anaerobically produced in a 2 m³ bioreactor at 35°C for 2 years using restaurant food waste from Korea Advanced Institute of Science and Technology (KAIST), and the concentration of H₂S in the biogas ranged from 612 to 1,500 ppmv (Avg. 1,060 ppmv). Two immobilized cell bioreactors 0.2 and 8.5 L in volume were loaded with aerobic sludge and used to study characteristics of H₂S removal from biogas. At a retention time of 400 sec, the removal efficiency of H₂S was over 99% following initial stabilization for 7 days in the 8.5 L bioreactor installed at the on-site biogas facility. The maximum rate of H₂S removal in this study was 359 g-H₂S/m³/h with an average mass loading rate of 14.7 g-H₂S/m³/h (kinetic analysis: V _m = 842.6 g-H₂S/m³/h and K _s = 2.2 mg/L). Therefore, purified biogas with a negligible concentration H₂S was efficiently reformed to syngas. This study demonstrates the feasibility of biogas purification as a part of high-quality syngas production.     

Simultaneous di-oxygenation and denitrification in an internal circulation baffled bioreactor

An internal circulation baffled bioreactor was employed to realize simultaneous di-oxygenation of phthalic acid (PA) and denitrification of nitrate, which require aerobic and anoxic conditions, respectively. Adding a small concentration of succinate as an exogenous electron donor stimulated PA di-oxygenation, which produced readily oxidizable downstream products whose oxidation also enhanced denitrification of nitrate; succinate addition also stimulated denitrification. Depending on the concentration of PA, addition of 0.17 mM succinate increased the PA removal rate by 25 and 42%, while the corresponding nitrate removal rate was increased by 73 and 51%. UV/H₂O₂ advanced oxidation of PA had the same effects as adding succinate, since succinate is generated by UV/H₂O₂; this acceleration effect was approximately equivalent to adding 0.17 mM succinate.     

Effect of alum on nitrification during simultaneous phosphorous removal in anoxic/oxic reactor

Phosphorus and nitrogen are the important eutrophication nutrients. They are removed in the anoxic/oxic reactor through simultaneous precipitation and biological nitrogen removal. The effect of alum a commonly used simultaneous precipitant on biological nitrification and denitrification are investigated in the present study. Simultaneous removal of phosphorus was carried out using the coagulant alum Al₂(SO₄)₃·14H₂O at 2.2 mol ratio. Before the start of simultaneous precipitation the nitrification rate of the A/O reactor was found to be 0.05 g N-NH₄ ⁺/g VSS/d. It starts to decrease with increase in coagulant dosage. The nitrification rate for alum dosage 97.13 mg/L was 0.38 g N- NH₄ ⁺/g VSS/d. There was no accumulation of nitrate in anoxic tank. The nitrogen removal efficiency of the reactor was affected and it fell from 88 to 78%. There was a slight decrease in effluent COD from 16∼20 mg/L to 8∼12 mg/L after the introduction of simultaneous precipitation into the reactor. The usage of alum as a simultaneous precipitant in the anoxic/oxic reactor was limited due to its inhibition on nitrification. Alum did not have any affect over denitrification process.     

Treatment of H2S using a horizontal biotrickling filter based on biological activated carbon: reactor setup and performance evaluation

Biological treatment is an emerging and prevalent technology for treating off-gases from wastewater treatment plants. The most commonly reported odorous compound in off-gases is hydrogen sulfide (H₂S), which has a very low odor threshold. A self-designed, bench-scale, cross-flow horizontal biotrickling filter (HBF) operated with bacteria immobilized activated carbon (termed biological activated carbon—BAC), was applied for the treatment of H₂S. A mixed culture of sulfide-oxidizing bacteria dominated by Acidithiobacillus thiooxidans acclimated from activated sludge was used as bacterial seed and the biofilm was developed by culturing the bacteria in the presence of carbon pellets in mineral medium. HBF performance was evaluated systematically over 120 days, depending on a series of changing factors including inlet H₂S concentration, gas retention time (GRT), pH of recirculation solution, upset and recovery, sulfate accumulation, pressure drop, gas-liquid ratio, and shock loading. The biotrickling filter system can operate at high efficiency from the first day of operation. At a volumetric loading of 900 m³ m^–3 h^–1 (at 92 ppmv H₂S inlet concentration), the BAC exhibited maximum elimination capacity (113 g H₂S/m^–3 h^–1) and a removal efficiency of 96% was observed. If the inlet concentration was kept at around 20 ppmv, high H₂S removal (over 98%) was achieved at a GRT of 4 s, a value comparable with those currently reported for biotrickling filters. The bacterial population in the acidic biofilter demonstrated capacity for removal of H₂S over a broad pH range (pH 1–7). A preliminary investigation into the different effects of bacterial biodegradation and carbon adsorption on system performance was also conducted. This study shows the HBF to be a feasible and economic alternative to physical and chemical treatments for the removal of H₂S.     

Sulphide effects on the physiology of Candidatus Accumulibacter phosphatis type I

Sulphate-rich wastewaters can be generated due to (i) use of saline water as secondary-quality water for sanitation in urban environments (e.g. toilet flushing), (ii) discharge of industrial effluents, (iii) sea and brackish water infiltration into the sewage and (iv) use of chemicals, which contain sulphate, in drinking water production. In the presence of an electron donor and absence of oxygen or nitrate, sulphate can be reduced to sulphide. Sulphide can inhibit microbial processes in biological wastewater treatment systems. The objective of the present study was to assess the effects of sulphide concentration on the anaerobic and aerobic physiology of polyphosphate-accumulating organisms (PAOs). For this purpose, a PAO culture, dominated by Candidatus Accumulibacter phosphatis clade I (PAO I), was enriched in a sequencing batch reactor (SBR) fed with acetate and propionate. To assess the direct inhibition effects and their reversibility, a series of batch activity tests were conducted during and after the exposure of a PAO I culture to different sulphide concentrations. Sulphide affected each physiological process of PAO I in a different manner. At 189 mg TS-S/L, volatile fatty acid uptake was 55% slower and the phosphate release due to anaerobic maintenance increased from 8 to 18 mg PO₄-P/g VSS/h. Up to 8 mg H₂S-S/L, the decrease in aerobic phosphorus uptake rate was reversible (Ic₆₀). At higher concentrations of sulphide, potassium (>16 mg H₂S-S/L) and phosphate (>36 mg H₂S-S/L) were released under aerobic conditions. Ammonia uptake, an indicator of microbial growth, was not observed at any sulphide concentration. This study provides new insights into the potential failure of enhanced biological phosphorus removal sewage plants receiving sulphate- or sulphide-rich wastewaters when sulphide concentrations exceed 8 mg H₂S-S/L, as PAO I could be potentially inhibited.

     

Effect of rhamnolipid on the aerobic removal of polyaromatic hydrocarbons (PAHs) and COD components from petrochemical wastewater

The removal efficiencies of 15 PAHs and some COD components (inert, readily degradable, slowly degradable and metabolic products) from a wastewater taken from a petrochemical industry treatment plant (İzmir, Turkey) have been determined using an aerobic completely stirred tank reactor (CSTR). Addition of rhamnolipid surfactant (15 mg l⁻¹) increased the removal efficiencies of PAHs and soluble COD from 72% and 90% to 80% and 99%, respectively. The rhamnolipid treatment caused a significant increase of 5- and 6-ring PAH degradation. The soluble COD removal efficiency was 93%, in CSTR reactors with rhamnolipid added. The inert COD removal efficiency was 60% in a CSTR reactor containing rhamnolipid. Batch tests showed that removal arising from the adsorption of the PAHs was low (between 1.88% and 4.84%) while the removal of PAHs from the petrochemical industry wastewater via volatilization varied between 0.69% and 5.92%. Low sorption capacity (K_p) values for refinery activated sludge (approximately 2.98 l g⁻¹) confirmed that bio-sorption was not an important mechanism controlling the fate of PAHs in aerobic CSTR reactors. Models proposed to simulate the PAH removal indicated that 94% of the PAHs were removed via biodegradation.     

Isolation of a carbon disulfide utilizing <Emphasis Type="Italic">Thiomonas</Emphasis> sp. and its application in a biotrickling filter

The carbon disulfide (CS₂)-oxidizing bacterium Thiomonas sp. WZW was enriched and isolated using activated sewage sludge as inoculum. Growth of Thiomonas sp. WZW was observed on CS₂, thiosulfate, dimethylsulfide (DMS), dimethyldisulfide (DMDS), and H₂S. No growth occurred on dimethylsulfoxide, methanol, acetate, and on complex media with glucose, yeast extract, or tryptone. DMDS-grown cells respired CS₂, DMS, and DMDS, while thiosulfate-grown cells did not respire CS₂. Chemostat cultures growing on thiosulfate could be rapidly adapted to growth on CS₂. Growth was observed between pH 6 and 8. The K _s values for CS₂, thiosulfate, and sulfide of CS₂-grown cells were between 5 and 10 μM. CS₂ was inhibitory above 0.3 mM. A lab-scale biotrickling filter with lava stone as carrier material for treatment of CS₂-polluted air was inoculated with Thiomonas sp. WZW. A rapid start up (95% removal in 1 week) was obtained at an inlet CS₂ concentration of 2 cmol l⁻¹ and an initial space velocity (SV) of 54 h⁻¹. Subsequent thiosulfate addition for a week during start up increased the removal to 99%. The step-wise increase of SV to 130 h⁻¹ and a CS₂ concentration to 3 μmol l⁻¹ resulted in a stable performance with a removal efficiency of 95%. Feeding mixtures of volatile sulfur compounds showed simultaneous conversion of H₂S, CS₂, dimethyldisulfide (DMDS), and DMS, with a preference in this order.     

H2 production by mixed cultures

Production of H₂ from glucose by an anoxygenic phototrophic bacterium (Rhodobacter sphaeroides), a cyanobacterium (Synechococcus cedrorum) and a heterotrophic bacterium (Pseudomonas fluorescens) was tested individually and in mixed cultures of various combinations in light. H₂ production was maximal with a mixed culture of R. sphaeroides and P. fluorescens, which could be further enhanced by immobilization of the bacteria in alginate gel. Inhibition of H₂ photoproduction was observed in a mixture of S. cedrorum and P. fluorescens and a co-culture of all the three organisms.Ch. Sasikala and Ch. V. Ramana are and G. S. Prasad was with the Microbial Biotechnology Laboratory, Department of Botany, Osmania University, Hyderabad-500 007, India. G. S. Prasad is now with the Microbial Type Culture Collection Centre (MTCC), IMTECH, Chandigar, India.     

Heterotrophic nitrogen removal by <Emphasis Type="Italic">Providencia rettgeri</Emphasis> strain YL

Providencia rettgeri strain YL was found to be efficient in heterotrophic nitrogen removal under aerobic conditions. Maximum removal of NH₄ ⁺–N occurred under the conditions of pH 7 and supplemented with glucose as the carbon source. Inorganic ions such as Mg²⁺, Mn²⁺, and Zn²⁺ largely influenced the growth and nitrogen removal efficiency. A quantitative detection of nitrogen gas by gas chromatography was conducted to evaluate the nitrogen removal by strain YL. From the nitrogen balance during heterotrophic growth with 180 mg/l of NH₄ ⁺–N, 44.5% of NH₄ ⁺–N was in the form of N₂ and 49.7% was found in biomass, with only a trace amount of either nitrite or nitrate. The utilization of nitrite and nitrate during the ammonium removal process demonstrated that the nitrogen removal pathway by strain YL was heterotrophic nitrification-aerobic denitrification. A further enzyme assay of nitrate reductase and nitrite reductase activity under the aerobic condition confirmed this nitrogen removal pathway.     

Isolation and Characterization of Pseudoxanthomonas sp. Strain YP1 Capable of Denitrifying Phosphorus Removal (DPR)

A bacterial strain (designated as YP1) was isolated from an aerobic granular sequence batch reactor (SBR) performing simultaneous nitrogen and phosphorus removal. Based on the morphological, biochemical characteristics, and phylogenetic analysis of 16S rRNA gene sequence, YP1 was identified as Pseudoxanthomonas sp. strain. Strain YP1 was confirmed to have the ability to conduct denitrifying phosphorus removal (DPR). The optimal conditions for YP1 were pH 8.0, phosphorus (PO₄^3?-P) concentration of 8.0 mg/L, sodium citrate as carbon source, and nitrate nitrogen (NO₃^?-N) concentration of 30 mg/L. The functional genes including ppk and ppx, narG and narA, nirS and nirK were amplified for understanding the DPR pathways. The results provide more information about denitrifying polyphosphate-accumulating organisms (DPAOs) in aerobic granular sludge (AGS) and lay the foundations for full-scale DPR.     

Simultaneous nitrification and denitrification by diverse <Emphasis Type="Italic">Diaphorobacter</Emphasis> sp.

Eight bacterial isolates closely related to Diaphorobacter sp. were isolated from activated biomass surviving on wastewater laden with dyes and nitro-substituted chemicals and were identified by 16S rDNA sequence analysis. The isolates showed sequence similarity of 99–100% to other Diaphorobacter strains such as ZY 2006b, F2, NA5, PCA039, D. nitroreducens KSP4, and KSP3 and 98–99% sequence homology to D. nitroreducens NA10B (type strain JCM 11421). Neighbor-joining tree revealed that all the eight strains formed tight cluster together and also showed close clustering with other Diaphorobacter strains. Isolates demonstrated the ability to perform simultaneous nitrification and denitrification under aerobic conditions. Strains HPC 805, 815, 821, and 856 gave highest chemical oxygen demand removal (85–93%) and ammonia removal (92–96%), which correlated well with higher growth rates of the cultures. Simultaneously, complete removal of nitrate supplied in the medium in presence of ammonium and acetate (electron donor) was observed in addition to aerobic nitrite release from ammonium. Thus, the above strains showed ability to perform partial nitrification followed by further aerobic removal of common intermediate nitrite, which indicated their potential application in treatment systems for treatment of high-nitrogen-containing wastewaters.     

Bioremediation of synthetic high–chemical oxygen demand wastewater using microalgal species Chlorella pyrenoidosa

The microalgal species Chlorella pyrenoidosa was cultivated in synthetic wastewater of initial chemical oxygen demand (COD), nitrate, and phosphate concentrations of 5000, 100, and 40 mg/L, respectively. The aim of the study was to find out the tolerance of microalgae to different COD concentrations and the extent of COD degradation at those concentrations. Three dilutions of wastewater (initial COD concentrations 5000, 3000, and 1000 mg/L) and three inoculum sizes (0.1, 0.2, and 0.3 g/L) were considered for the study. The experimental parameters such as total organic carbon, total inorganic carbon, COD, optical density, total solids, nitrate, and phosphate were measured on a daily basis. Biodegradation kinetics was determined for all cases using first-order reaction and Monod degradation equations. Optimal results showed that up to 90% reduction in TOC was obtained for 1000 COD wastewater while only 38% reduction in total organic carbon (TOC) was achieved for 5000 COD wastewater. Over 95% reduction in nitrate and nearly 90% removal of phosphate were obtained with the lowest microalgal inoculum concentration (i.e., 0.1 g/L) for all COD dilutions. This study showed that microalgal species C. pyrenoidosa can successfully degrade the organic carbon source (i.e., acetate) with significant removal efficiencies for nitrate and phosphate.     

Aerobic chemolithoautotrophic growth and RubisCO function in Rhodobacter capsulatus and a spontaneous gain of function mutant of Rhodobacter sphaeroides

Photosynthetic prokaryotes that assimilate CO₂ under anoxic conditions may also grow chemolithoautotrophically with O₂ as the electron acceptor. Among the nonsulfur purple bacteria, two species (Rhodobacter capsulatus and Rhodopseudomonas acidophilus), exhibit aerobic chemolithoautotrophic growth with hydrogen as the electron donor. Although wild-type strains of Rhodobacter sphaeroides grow poorly, if at all, with hydrogen plus oxygen in the dark, we report here the isolation of a spontaneous mutant (strain HR-CAC) of Rba. sphaeroides strain HR that is fully capable of this mode of growth. Rba. sphaeroides and Rba. capsulatus fix CO₂ via the reductive pentose phosphate pathway and synthesize two forms of ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO). RubisCO levels in the aerobic-chemolithoautotrophic-positive strain of Rba. sphaeroides were similar to those in wild-type strains of Rba. sphaeroides and Rba. capsulatus during photoheterotrophic and photolithoautotrophic growth. Moreover, RubisCO levels of Rba. sphaeroides strain HR-CAC approximated levels obtained in Rba. capsulatus when the organisms were grown as aerobic chemolithoautotrophs. Either form I or form II RubisCO was able to support aerobic chemolithoautotrophic growth of Rba. capsulatus strain SB 1003 and Rba. sphaeroides strain HR-CAC at a variety of CO₂ concentrations, although form II RubisCO began to lose the capacity to support aerobic CO₂ fixation at high O₂ to CO₂ ratios. The latter property and other facets of the physiology of this system suggest that Rba. sphaeroides and Rba. capsulatus strains may be effectively employed for the biological selection of RubisCO molecules of altered substrate specificity. Received: 8 August 1997 / Accepted: 26 December 1997     

The effect of long-term nitrate treatment on SRB activity,corrosion rate and bacterial community composition in offshore water injection systems

Biogenic production of hydrogen sulphide (H₂S) is a problem for the oil industry as it leads to corrosion and reservoir souring. Continuous injection of a low nitrate concentration (0.25–0.33 mM) replaced glutaraldehyde as corrosion and souring control at the Veslefrikk and Gullfaks oil field (North Sea) in 1999. The response to nitrate treatment was a rapid reduction in number and activity of sulphate-reducing bacteria (SRB) in the water injection system biofilm at both fields. The present long-term study shows that SRB activity has remained low at ≤0.3 and ≤0.9 μg H₂S/cm²/day at Veslefrikk and Gullfaks respectively, during the 7–8 years with continuous nitrate injection. At Veslefrikk, 16S rRNA gene based community analysis by PCR–DGGE showed that bacteria affiliated to nitrate-reducing sulphide-oxidizing Sulfurimonas (NR–SOB) formed major populations at the injection well head throughout the treatment period. Downstream of deaerator the presence of Sulfurimonas like bacteria was less pronounced, and were no longer observed 40 months into the treatment period. The biofilm community during nitrate treatment was highly diverse and relative stable for long periods of time. At the Gullfaks field, a reduction in corrosion of up to 40% was observed after switch to nitrate treatment. The present study show that nitrate injection may provide a stable long-term inhibition of SRB in sea water injection systems, and that corrosion may be significantly reduced when compared to traditional biocide treatment.