Predominance of ammonia-oxidizing archaea on granular activated carbon used in a full-scale advanced drinking water treatment plant

Ozonation followed by granular activated carbon (GAC) is one of the advanced drinking water treatments. During GAC treatment, ammonia can be oxidized by ammonia-oxidizing microorganisms associated with GAC. However, there is little information on the abundance and diversity of ammonia-oxidizing microorganisms on GAC. In this study, the nitrification activity of GAC and the settlement of ammonia-oxidizing archaea (AOA) and bacteria (AOB) in GAC were monitored at a new full-scale advanced drinking water treatment plant in Japan for 1 year after plant start-up. Prechlorination was implemented at the receiving well for the first 10 months of operation to treat ammonia in raw water. During this prechlorination period, levels of both AOA and AOB associated with GAC were below the quantification limit. After prechlorination was stopped, 10⁵ copies g-dry⁻¹ of AOA amoA genes were detected within 3 weeks and the quantities ultimately reached 10⁶-10⁷ copies g-dry⁻¹, while levels of AOB amoA genes still remained below the quantification limit. This observation indicates that AOA can settle in GAC rapidly without prechlorination. The nitrification activity of GAC increased concurrently with the settlement of AOA after prechlorination was stopped. Estimation of in situ cell-specific ammonia-oxidation activity for AOA on the assumption that only AOA and AOB determined can contribute to nitrification suggests that AOA may account for most of the ammonia-oxidation. However, further validation on AOB contribution is required.     

Ammonia-oxidizing archaea involved in nitrogen removal

Ammonia oxidation is critical to global nitrogen cycling and is often thought to be driven only by ammonia-oxidizing bacteria. The recent finding of new ammonia-oxidizing organisms belonging to the archaeal domain challenges this perception. Two major microbial groups are now believed to be involved in ammonia oxidation: chemolithotrophic ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA). Candidatus “Nitrosopumilus maritimus”, the first isolated ammonia-oxidizing archaeon from a tropical marine aquarium tank, representative of the ubiquitous marine group 1 Crenarchaeota, contains putative genes for all three subunits (amoA, amoB, and amoC) of ammonia monooxygenase, the key enzyme responsible for ammonia oxidation. In this article, important concepts of the nitrogen cycle, ammonia oxidation processes, ammonia-oxidizing organisms, and their physiology are described. AOA are found to thrive in various habitats including hot/thermal springs, marine and fresh waters, soils, and wastewater treatment systems, where they may outnumber their counterpart, AOB. Various molecular tools have been applied to study AOB and AOA and determine their abundance and community structure changes from natural and engineered systems. The presence of AOA in activated sludge opens new opportunities for elucidating its role of ammonia removal in wastewater treatment plants and wetlands. Several significant questions related to AOA research have been raised to evoke reader involvement for broadening future studies.     

Diversity of nitrifying bacteria in full-scale chloraminated distribution systems

Chloramination for secondary disinfection of drinking water often promotes the growth of nitrifying bacteria in the distribution system due to the ammonia introduced by chloramine formation and decay. This study involved the application of molecular biology techniques to explore the types of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) present in several full-scale chloraminated systems. The results of AOB community characterization indicated the ubiquitous detection of representatives from the Nitrosomonas genus, with Nitrosospira constituting a negligible or small fraction of the AOB community in all but one sample. Cloning and sequencing demonstrated the presence of AOB representatives within the Nitrosomonas oligotropha cluster, a phylogenetic subgroup of AOB from which isolates demonstrate a high affinity for ammonia. For the NOB communities, Nitrospira were detected in most of the samples, while Nitrobacter were only detected in a few samples. These results provide insight into the types of AOB responsible for nitrification episodes in full-scale chloraminated systems, which should help direct future studies aimed at characterizing relevant AOB growth and inactivation properties. Furthermore, the detection of NOB in most of the samples suggests a need to evaluate the contribution of biological nitrite oxidation relative to chemical oxidation in these systems.     

Monochloramine cometabolism by Nitrosomonas europaea under drinking water conditions

Chloramine is widely used in United States drinking water systems as a secondary disinfectant, which may promote the growth of nitrifying bacteria because ammonia is present. At the onset of nitrification, both nitrifying bacteria and their products exert a monochloramine demand, decreasing the residual disinfectant concentration in water distribution systems. This work investigated another potentially significant mechanism for residual disinfectant loss: monochloramine cometabolism by ammonia-oxidizing bacteria (AOB).Monochloramine cometabolism was studied with the pure culture AOB Nitrosomonas europaea (ATCC 19718) in batch kinetic experiments under drinking water conditions. Three batch reactors were used in each experiment: a positive control to estimate the ammonia kinetic parameters, a negative control to account for abiotic reactions, and a cometabolism reactor to estimate the cometabolism kinetic constants. Kinetic parameters were estimated in AQUASIM with a simultaneous fit to all experimental data. The cometabolism reactors showed a more rapid monochloramine decay than in the negative controls, demonstrating that cometabolism occurs. Cometabolism kinetics were best described by a pseudo first order model with a reductant term to account for ammonia availability. Monochloramine cometabolism kinetics were similar to those of ammonia metabolism, and monochloramine cometabolism was a significant loss mechanism (30–60% of the observed monochloramine decay). These results suggest that monochloramine cometabolism should occur in practice and may be a significant contribution to monochloramine decay during nitrification episodes in drinking water distribution systems.     

Effect of temperature and disinfection strategies on ammonia-oxidizing bacteria in a bench-scale drinking water distribution system

The establishment of ammonia-oxidizing bacteria (AOB), a group of autotrophic microorganisms responsible for nitrification in chloraminated distribution systems, was studied in a bench-scale distribution system. The potential significance of temperature and disinfectant residual associated with chloramination in full-scale drinking water distribution systems was assessed. Biofilm development was primarily monitored using AOB abundance and nitrite concentrations. The bench-scale system was initially operated under typical North American summer (22 degrees C) and fall (12 degrees C) temperatures, representing optimal and less optimal growth ranges for these microorganisms. Additional experimentation investigated AOB establishment at a suboptimal winter distribution system temperature of 6 degrees C. The effect of chloramine residual on AOB establishment was studied at higher (0.2-0.6mg/L) and lower (0.05-0.1mg/L) ranges, using a 3:1 (w/w) chlorine:ammonia dosing ratio. Conditions were selected to represent those typically found in a North American distribution system, in areas of low flow and longer retention times, respectively. Finally, the effect of a free chlorine residual on an established nitrifying biofilm was briefly examined. Results clearly indicate that AOB development occurs at all examined temperatures, as well as at selected monochloramine residuals. The maintenance of a disinfectant residual was difficult at times, but was more inhibitory to the nitrifying biofilm than the lower temperature. It can be concluded from the data that nitrification may not be adequately inhibited during the winter months, which may result in more advanced stages of nitrification the following season. Free chlorination can be effective in controlling AOB activity in the short term, but may not prevent reestablishment of a nitrifying biofilm upon return to chloramination.     

Mechanisms of N2O production in biological wastewater treatment under nitrifying and denitrifying conditions

Nitrous oxide (N₂O) is an important greenhouse gas and a major sink for stratospheric ozone. In biological wastewater treatment, microbial processes such as autotrophic nitrification and heterotrophic denitrification have been identified as major sources; however, the underlying pathways remain unclear. In this study, the mechanisms of N₂O production were investigated in a laboratory batch-scale system with activated sludge for treating municipal wastewater. This relatively complex mixed population system is well representative for full-scale activated sludge treatment under nitrifying and denitrifying conditions.Under aerobic conditions, the addition of nitrite resulted in strongly nitrite-dependent N₂O production, mainly by nitrifier denitrification of ammonia-oxidizing bacteria (AOB). Furthermore, N₂O is produced via hydroxylamine oxidation, as has been shown by the addition of hydroxylamine. In both sets of experiments, N₂O production was highest at the beginning of the experiment, then decreased continuously and ceased when the substrate (nitrite, hydroxylamine) had been completely consumed. In ammonia oxidation experiments, N₂O peaked at the beginning of the experiment when the nitrite concentration was lowest. This indicates that N₂O production via hydroxylamine oxidation is favored at high ammonia and low nitrite concentrations, and in combination with a high metabolic activity of ammonia-oxidizing bacteria (at 2 to 3 mgO₂/l); the contribution of nitrifier denitrification by AOB increased at higher nitrite and lower ammonia concentrations towards the end of the experiment.Under anoxic conditions, nitrate reducing experiments confirmed that N₂O emission is low under optimal growth conditions for heterotrophic denitrifiers (e.g. no oxygen input and no limitation of readily biodegradable organic carbon). However, N₂O and nitric oxide (NO) production rates increased significantly in the presence of nitrite or low dissolved oxygen concentrations.     

Modeling the decay of ammonium oxidizing bacteria

A bench-scale sequencing batch reactor was used to study factors affecting the endogenous decay of the ammonium oxidizing biomass (AOB) in different operating conditions. AOB decay was very sensitive to oxygen concentration, and increased up to 0.4 d⁻¹ for oxygen concentration of 7 mg O₂ L⁻¹. The decay in anaerobic conditions was shown to be very low (0.03 d⁻¹) when compared to literature data.The effect of nitrite and nitrate on AOB decay was also studied. The correlation was quite weak suggesting that both nitrate and nitrite absence had little impact on decay which is contrary to what is typically assumed in some of the existing process models. A simple expression for the decay of AOB was proposed, calibrated and validated using the results of batch kinetic tests and of the continuous sequencing batch reactor monitoring.     

Phosphorus limitation in nitrifying groundwater filters

Phosphorus limitation has been demonstrated for heterotrophic growth in groundwater, in drinking water production and distribution systems, and for nitrification of surface water treatment at low temperatures. In this study, phosphorus limitation was tested, in the Netherlands, for nitrification of anaerobic groundwater rich in iron, ammonium and orthophosphate. The bioassay method developed by Lehtola et al. (1999) was adapted to determine the microbially available phosphorus (MAP) for nitrification. In standardized batch experiments with an enriched mixed culture inoculum, the formation of nitrite and nitrate and ATP and the growth of ammonia-oxidizing bacteria (AOB; as indicated by qPCR targeting the amoA-coding gene) were determined for MAP concentrations between 0 and 100 μg PO₄-P L⁻¹. The nitrification and microbial growth rates were limited at under 100 μg PO₄-P L⁻¹ and virtually stopped at under 10 μg PO₄-P L⁻¹. In the range between 10 and 50 μg PO₄-P L⁻¹, a linear relationship was found between MAP and the maximum nitrification rate. AOB cell growth and ATP formation were proportional to the total ammonia oxidized. Contrary to Lehtola et al. (1999), biological growth was very slow for MAP concentrations less than 25 μg PO₄-P L⁻¹. No full conversion nor maximum cell numbers were reached within 19 days. In full-scale groundwater filters, most of the orthophosphate was removed alongside with iron. The remaining orthophosphate appeared to have only limited availability for microbial growth and activity. In some groundwater filters, nitrification was almost totally prevented by limitation of MAP. In batch experiments with filtrate water from these filters, the nitrification process could be effectively stimulated by adding phosphoric acid.     

联氨对HABR全程自养脱氮系统的影响

为考察联氨作为自养脱氮系统菌群调节剂的可行性,以实验室内运行的HABRCANON反应器为试验装置,研究不同浓度联氨对自养脱氮系统脱氮效能和功能微生物的影响。结果表明,低浓度(1~4 mg/L)联氨可以抑制亚硝酸盐氧化菌(NOB)的活性,促进厌氧氨氧化菌(AnAOB)的活性,从而提高脱氮效能;高浓度(10 mg/L)联氨对好氧氨氧化菌(AOB)和NOB的抑制作用明显;停止投加联氨后,CANON系统的脱氮效能可迅速恢复;高浓度(10 mg/L)联氨对HABR全程自养脱氮工艺的影响是可逆的,但对NOB的抑制不可逆。对生物膜样品中的优势菌种进行分析发现,AOB和AnAOB为主要的功能微生物。采用低-高-低的联氨投加方式,可以有效抑制自养脱氮反应器内NOB的生长,保证自养脱氮系统的稳定运行。     

Application of bacteriophages to selectively remove Pseudomonas aeruginosa in water and wastewater filtration systems

Water and wastewater filtration systems often house pathogenic bacteria, which must be removed to ensure clean, safe water. Here, we determine the persistence of the model bacterium Pseudomonas aeruginosa in two types of filtration systems, and use P. aeruginosa bacteriophages to determine their ability to selectively remove P. aeruginosa. These systems used beds of either anthracite or granular activated carbon (GAC), which were operated at an empty bed contact time (EBCT) of 45 min. The clean bed filtration systems were loaded with an instantaneous dose of P. aeruginosa at a total cell number of 2.3 (±0.1 [standard deviation]) × 10⁷ cells. An immediate dose of P. aeruginosa phages (1 mL of phage stock at the concentration of 2.7 × 10⁷ PFU (Plaque Forming Units)/mL) resulted in a reduction of 50% (±9%) and >99.9% in the effluent P. aeruginosa concentrations in the clean anthracite and GAC filters, respectively. To further evaluate the effects of P. aeruginosa phages, synthetic stormwater was run through anthracite and GAC biofilters where mixed-culture biofilms were present. Eighty five days after an instantaneous dose of P. aeruginosa (2.3 × 10⁷ cells per filter) on day 1, 7.5 (±2.8) × 10⁷ and 1.1 (±0.5) × 10⁷ P. aeruginosa cells/g filter media were detected in the top layer (close to the influent port) of the anthracite and GAC biofilters, respectively, demonstrating the growth and persistence of pathogenic bacteria in the biofilters. A subsequent 1-h dose of phages, at the concentration of 5.1 × 10⁶ PFU/mL and flow rate of 1.6 mL/min, removed the P. aeruginosa inside the GAC biofilters and the anthracite biofilters by 70% (±5%) and 56% (±1%), respectively, with no P. aeruginosa detected in the effluent, while not affecting ammonia oxidation or the ammonia-oxidizing bacterial community inside the biofilters. These results suggest that phage treatment can selectively remove pathogenic bacteria with minimal impact on beneficial organisms from attached growth systems for effluent quality improvement.     

Occurrence of nitrifiers and diversity of ammonia-oxidizing bacteria in developing drinking water biofilms

We studied the population dynamics of nitrifying bacteria during the development of biofilms up to 233 or 280 days on polyvinylchloride pipes connected to two full-scale drinking water distribution networks supplying processed and chloraminated surface water. The numbers of nitrifiers in biofilms were enumerated at intervals of 10–64 days by the most probable number (MPN) method at waterworks and at several study sites in distribution network areas. The numbers of nitrifiers increased towards the distal sites. The highest detected MPN counts of ammonia-oxidizing bacteria (AOB) for study areas 1 and 7 were 500 MPN cm⁻² and 1.0×10⁶ MPN cm⁻², and those of nitrite-oxidizing bacteria (NOB) 96 MPN cm⁻² and 2.2×10³ MPN cm⁻², respectively. The diversity of AOB was determined by PCR amplifying, cloning and sequencing the partial ammonia monooxygenase (amoA) gene of selected biofilm samples presenting different biofilm ages. The PCR primers used, A189 and A682, also amplified a fragment of particulate methane monooxygenase (pmoA) gene of methane-oxidizing bacteria. The majority of biofilm clones (24 out of 30 studied) contained Nitrosomonas amoA-like sequences. There were only two pmoA-like sequences of Type I methanotrophs, and four sequences positioned in amoA/pmoA sequence groups of uncultured bacteria. From both study area very similar or even completely identical Nitrosomonas amoA-like sequences were obtained despite of high difference in AOB numbers. The results show that the conditions in newly formed biofilms in drinking water distribution systems favor the growth of Nitrosomonas-type AOB.     

Dynamic and distribution of ammonia-oxidizing bacteria communities during sludge granulation in an anaerobic-aerobic sequencing batch reactor

The structure dynamic of ammonia-oxidizing bacteria (AOB) community and the distribution of AOB and nitrite-oxidizing bacteria (NOB) in granular sludge from an anaerobic-aerobic sequencing batch reactor (SBR) were investigated. A combination of process studies, molecular biotechniques and microscale techniques were employed to identify and characterize these organisms. The AOB community structure in granules was substantially different from that of the initial pattern of the inoculants sludge. Along with granules formation, the AOB diversity declined due to the selection pressure imposed by process conditions. Denaturing gradient gel electrophoresis (DGGE) and sequencing results demonstrated that most of Nitrosomonas in the inoculating sludge were remained because of their ability to rapidly adapt to the settling-washing out action. Furthermore, DGGE analysis revealed that larger granules benefit more AOB species surviving in the reactor. In the SBR were various size granules coexisted, granule diameter affected the distribution range of AOB and NOB. Small and medium granules (d < 0.6 mm) cannot restrict oxygen mass transfer in all spaces of the sludge. Larger granules (d > 0.9 mm) can result in smaller aerobic volume fraction and inhibition of NOB growth. All these observations provide support to future studies on the mechanisms responsible for the AOB in granules systems.     

Biological oxidation of arsenite in synthetic groundwater using immobilised bacteria

Biological oxidation of arsenite (As(III)) in synthetic groundwater was examined by using arsenite oxidising bacteria (AOB) isolated from an activated sludge. The phylogenetic analysis indicated that the isolated AOB was closely related to Ensifer adhaerens. Batch experiments showed that for an As(III) oxidation with the isolated AOB the optimum ratio of nitrogen source (NH₄-N) concentration to As(III) concentration was 0.5 (52 mg/L-110 mg/L) and the isolated AOB preferred pH values ranging from 6 to 8, and water temperatures greater than 20 °C. Further continuous experiments were conducted using a bioreactor with immobilised AOB. With an initial As(III) concentration of 1 mg/L at a hydraulic retention time (HRT) of 1 h, an As(III) oxidation rate was around 1 × 10⁻⁹ μg/cell/min and an As(III) oxidation efficiency of 92% was achieved. Although the maximum oxidation rate measured at an HRT of 0.5 h was 2.1 × 10⁻⁹ μg/cell/min, the oxidation efficiency decreased to 87%.These results advocate that a biological process involving immobilised AOB may be useful as an economical and environmentally friendly pre-treatment step for As removal from groundwater.     

Performance of conventional multi-barrier drinking water treatment plants for the removal of four artificial sweeteners

Due to incomplete removal of artificial sweeteners in wastewater treatment plants some of these compounds end up in receiving surface waters, which are used for drinking water production. The sum of removal efficiency of single treatment steps in multi-barrier treatment systems affects the concentrations of these compounds in the provided drinking water. This is the first systematic study revealing the effectiveness of single treatment steps in laboratory experiments and in waterworks. Six full-scale waterworks using surface water influenced raw water were sampled up to ten times to study the fate of acesulfame, saccharin, cyclamate and sucralose. For the most important treatment technologies the results were confirmed by laboratory batch experiments. Saccharin and cyclamate proved to play a minor role for drinking water treatment plants as they were eliminated by nearly 100% in all waterworks with biologically active treatment units like river bank filtration (RBF) or artificial groundwater recharge. Acesulfame and sucralose were not biodegraded during RBF and their suitability as wastewater tracers under aerobic conditions was confirmed. Sucralose proved to be persistent against ozone and its transformation was <20% in lab and field investigations. Remaining traces were completely removed by subsequent granular activated carbon (GAC) filters. Acesulfame readily reacts with ozone (pseudo first-order rate constant k = 1.3 × 10⁻³ s⁻¹ at 1 mg L⁻¹ ozone concentration). However, the applied ozone concentrations and contact times under typical waterworks conditions only led to an incomplete removal (18-60%) in the ozonation step. Acesulfame was efficiently removed by subsequent GAC filters with a low throughput of less than 30 m³ kg⁻¹, but removal strongly depended on the GAC preload. Thus, acesulfame was detected up to 0.76 μg L⁻¹ in finished water.     

Influence of manganese and ammonium oxidation on the removal of 17 alpha-ethinylestradiol (EE2)

Flow-through reactors with manganese oxides were examined for their capacity to remove 17α-ethinylestradiol (EE2) at μg L⁻¹ and ng L⁻¹ range from synthetic wastewater treatment plant (WWTP) effluent. The mineral MnO₂ reactors removed 93% at a volumetric loading rate (B_V) of 5 μg EE2 L⁻¹ d⁻¹ and from a B_V of 40 μg EE2 L⁻¹ d⁻¹ on, these reactors showed 75% EE2 removal. With the biologically produced manganese oxides, only 57% EE2 was removed at 40 μg EE2 L⁻¹ d⁻¹. EE2 removal in the ng L⁻¹ range was 84%. The ammonium present in the influent (10 mg N L⁻¹) was nitrified and ammonia-oxidizing bacteria (AOB) were found to be of prime importance for the degradation of EE2. Remarkably, EE2 removal by AOB continued for a period of 4 months after depleting NH₄⁺ in the influent. EE2 removal by manganese-oxidizing bacteria was inhibited by NH₄⁺. These results indicate that the metabolic properties of nitrifiers can be employed to polish water containing EE2 based estrogenic activity.     

Development of biomass in a drinking water granular active carbon (GAC) filter

Indigenous bacteria are essential for the performance of drinking water biofilters, yet this biological component remains poorly characterized. In the present study we followed biofilm formation and development in a granular activated carbon (GAC) filter on pilot-scale during the first six months of operation. GAC particles were sampled from four different depths (10, 45, 80 and 115 cm) and attached biomass was measured with adenosine tri-phosphate (ATP) analysis. The attached biomass accumulated rapidly on the GAC particles throughout all levels in the filter during the first 90 days of operation and maintained a steady state afterward. Vertical gradients of biomass density and growth rates were observed during start-up and also in steady state. During steady state, biomass concentrations ranged between 0.8-1.83 x 10⁻⁶ g ATP/g GAC in the filter, and 22% of the influent dissolved organic carbon (DOC) was removed. Concomitant biomass production was about 1.8 × 10¹² cells/m²h, which represents a yield of 1.26 × 10⁶ cells/μg. The bacteria assimilated only about 3% of the removed carbon as biomass. At one point during the operational period, a natural 5-fold increase in the influent phytoplankton concentration occurred. As a result, influent assimilable organic carbon concentrations increased and suspended bacteria in the filter effluent increased 3-fold as the direct consequence of increased growth in the biofilter. This study shows that the combination of different analytical methods allows detailed quantification of the microbiological activity in drinking water biofilters.     

缺氧生物膜滤池的自养脱氮性能研究

在成功实现亚硝酸盐自养脱氮(厌氧氨氧化)的基础上,探讨了亚硝酸盐浓度对缺氧生物膜滤池脱氮性能的影响。结果显示,在一定范围内提高亚硝酸盐浓度可加快氨氮去除速率,当NO2--N为118.4 mg/L时氨氮去除速率达到最大;此后,进一步提高进水NO2--N浓度会对氨氮的去除产生明显的抑制作用,导致反应速率下降,但此时的厌氧氨氧化菌仍具有较高的活性;为获得良好的脱氮效果,应控制进水NO2--N/NH4 -N值为1.3。     

Azolla–Anabaena's behaviour in urban wastewater and artificial media – Influence of combined nitrogen

The results of using the nitrogen fixing symbiotic system Azolla–Anabaena to improve the quality of treated urban wastewater, particularly on what concerns phosphorus removal efficiencies (40–65%), obtained in continuous assays performed during the past few years and presented earlier, were very promising. Nevertheless, the presence of combined nitrogen in some wastewaters can compromise the treatment efficiency. The main goal of this work was to compare plants behaviour in wastewater and in mineral media with and without added nitrogen.Azolla filiculoides's specific growth rates in wastewater and in mineral media without added nitrogen or with low nitrate concentration were very similar (0.122 d⁻¹–0.126 d⁻¹), but decreased in the presence of ammonium (0.100 d⁻¹). The orthophosphate removal rate coefficients were similar in all the growth media (0.210 d⁻¹–0.232 d⁻¹), but ammonium removal rate coefficient in wastewater was higher (0.117 d⁻¹) than in mineral medium using that source of nitrogen (0.077 d⁻¹).The ammonium present in wastewater, despite its high concentration (34 mg N L⁻¹), didn't seem to inhibit growth and nitrogen fixation, however, in mineral media, ammonium (40 mg N L⁻¹) was found to induce, respectively, 18% and 46% of inhibition.     

Evaluation of nitrifying activity in sediments by dark 14C-bicarbonate incorporation

A method has been developed for measuring the activity of nitrifying bacteria in sediment samples within very short incubation times (a few hours). Biological dark ¹⁴C-bicarbonate incorporation is measured with and without addition of 5 mg 1^?1 N-serve, a substance which was proved to be a specific inhibitor of autotrophic CO₂-fixation by nitrifying bacteria. The value of N-serve sensitive bicarbonate incorporation can be quantitatively related to the rate of nitrogen oxidation by autotrophic nitrifiers.First application of this method on marine sediments gives results in good agreement with information provided by other methods. Nitrification rates in the range 0–20 μmole nitrogen oxidized per dm³ sediment and per hour are found in the upper cm of the sediments of the coastal regions of the Southern Bight of the North Sea.