The relationship between dissimilatory nitrate reduction and oxygen uptake by cells of an Alcaligenes strain in flocs and in suspension and by activated sludge flocs

The oxygen uptake and the dissimilatory nitrate reduction by anaerobically grown cells of a denitrifying Alcaligenes strain, occurring in floc form or in suspension, were studied at different oxygen concentrations in the surrounding medium. When the oxygen concentration in the medium fell below 1·5 mg l⁻¹, the nitrate reduction by the cells within flocs increased considerably. The cells in suspension showed an increased nitrate reduction when the oxygen concentration was below 0·1 mg l⁻¹. When anaerobically grown cells had been aerated for 24 h in a nitrogen-free medium, the cells became sensitive to respiration inhibition by nitric oxide, resulting from nitrate reduction. This gave rise to an increased nitrate reduction below 2·5 mg oxygen l⁻¹ when the cells were aggregated in flocs and below 0·1 mg oxygen per litre when the cells were in suspension. The nitrate reduction by the denitrifying, floc-forming pure culture was compared with that of activated sludge flocs.     

Factors affecting the denitrification rate in two water-sediment systems

Effects of temperature, oxygen and nitrate concentrations of the overlying water, and the thickness of the sediment layer on the rate of denitrification in the sediment were investigated in two water-sediment systems, A and B. At 4°C, denitrification started after a prolonged lag period in contrast to nitrification which did not occur significantly. At 15°C, and particularly at 25°C, both processes proceeded readily. The disappearance of NO₂⁻ - N from the overlying water was more rapidly than that of NO₃⁻ - N.The denitrification rate was slightly reduced by increasing the dissolved oxygen concentration in the overlying water from 0 to approximately 2 mgl⁻¹. A further rise of the dissolved oxygen concentration had no further decreasing effect on the denitrification rate.The denitrification rate in sediment was dependent on the nitrate concentration in the overlying water approximating first order kinetics at lower concentrations, gradually becoming independent of the nitrate concentration at higher nitrate contents (zero order kinetics).When starting with a nitrate-nitrogen concentration of 25.2 mgl⁻¹, a sediment layer of 7 mm with A and 14 mm with B was roughly found to be involved in denitrification.Denitrification rates found in the present laboratory experiments were supposed to be considerably lower than those occurring under natural conditions as additional mechanisms for the transport of nitrate into sediments occurred in natural environments.     

Gas production in aquatic sediments in the presence and absence of nitrate

In the overlying water of a water-sediment system the pH was controlled at 7.0, the nitrate-nitrogen concentration at 25.0 mg 1⁻¹ and the dissolved oxygen concentration above 6.1 mg t⁻¹. The temperature of the whole system was kept at 15°C. The average rate of nitrate removal from the system as a result of denitrification amounted to 160 mg NO₃⁻ -N m⁻² day⁻¹. By means of Eh measurements at various depths in the sediment, it was attempted to figure out the course of the penetration fronts of nitrate and oxygen in the sediment during 241 days of incubation. From these results the layer in which denitrification occurred was derived. The course of the denitrification zone was followed during the incubation period. As a result of the depletion of the available hydrogen donors in the sediment, oxygen reached the bottom of the sediment after 235 days of incubation.     

Effect of temperature and dissolved oxygen on biodegradation of nitrilotriacetate

The effect of temperature and dissolved oxygen on the rate of biodegradation of nitrilotriacetate (NTA) was examined in water samples collected from the Rur River. Biodegradation of NTA was first order with respect to NTA concentration over a concentration range of 50–1000 μg l⁻¹. First order rate constants showed a typical temperature dependency (temperature coefficient, Q₁₀ = 2) and biodegradation of NTA was observed over a temperature range of 2–24°C. The effect of temperature on the rate of NTA biodegradation was described by the Arrhenius equation, with calculated activation energies in the range reported for ordinary enzyme reactions. Biodegradation of NTA was also observed at low dissolved oxygen concentrations (0.3 mg l⁻¹), although at reduced rates compared to high oxygen concentrations (13 mg l⁻¹). Biodegradation of NTA was oxygen-dependent, suggesting an obligate oxygen requirement for the initial steps in NTA metabolism by natural microbial communities in surface waters. In general, our results indicate that NTA biodegradation will occur in natural waters under conditions of low temperature and low dissolved oxygen and also at low NTA concentrations.     

Nitrification kinetics in activated sludge at various temperatures and dissolved oxygen concentrations

Activated sludge from a domestic sewage works was enriched with nitrifying bacteria by running a laboratory fermenter on ammonia-supplemented sewage. This enriched culture was used to determine respirometrically the kinetics of microbial nitrification. It was demonstrated that the reaction fits the Michaelis-Menten model for temperatures from 10 to 35°C, having a temperature optimum at 15°C (K₃ 0.72 mg 1⁻¹ NH₃). Nitrification is unaffected by high dissolved oxygen concentration 38 mg 1⁻¹ O₂ at 30°C) after acclimatisation. Nitrite concentrations > 20 mg 1⁻¹ are inhibitory to the reaction.     

The effect of dissolved oxygen concentration on nitrification

The effect of dissolved oxygen concentration on the rate of nitrification has been investigated by a number of researchers using both pure and mixed cultures, and cultures found in wastewater treatment systems. The maximum growth rate of both nitrification reactions are reported to be affected by dissolved oxygen concentration over the range of 0.3 mg l⁻¹ to as much as 4.0 mg l⁻¹. In some instances, it has been reported that a dissolved oxygen concentration in excess of 4.0 mg l⁻¹ is required to achieve maximum nitrification rates, while other investigators have found that only 0.5 to 1.0 mg l⁻¹ is required.It has been proposed that several factors are responsible for the wide range of reported nitrification rates with varying dissolved oxygen concentrations. Among these factors are the effects of oxygen diffusion in flocs, variation between measured results due to steady-state and dynamic measuring techniques, and double-substrate limited kinetics. This paper reviews the nitrification literature with respect to the effects of dissolved oxygen concentration, and shows that double-substrate limiting kinetics could account for the variation in the reported results.     

Concurrent denitrification and oxygen uptake in microbial films

Mixed microbial films were grown on the inner walls of a tubular reactor with recirculation of the reactor contents and continuous flow-through of nutrient solution. The loss of total oxidized nitrogen was correlated to the film population, the nitrite concentration and the dissolved oxygen concentration in the reactor. When film population was greater than 0.5 × 10⁹ cells cm⁻², reactor dissolved oxygen concentration greater than 1 mg l⁻¹ had little effect on the nitrate loss. Nitrate loss declined for film populations greater than 2 × 10⁹ cells cm⁻². Models based on Monod and zero-order microbial kinetics were calibrated using these data and a nonlinear least squares method. There was little difference in the residual errors with these methods.     

Detoxification of seawater used for gas scrubbing in the steel industry

Cyanide ion present in seawater after scrubbing blast furnace and coke ovens gases can be removed by sedimentation of hexacyanoferrate complexes followed by oxidation of residual cyanide with Caro's acid. Zinc ion is removed at the same time by adsorption on the hexacyanoferrate/hydrous ferric oxide precipitate.Sulphide is precipitated as ferrous sulphide, then oxidised by atmospheric oxygen. At 25°C and using an Fe/CN ratio of 1·00, initial concentrations of 50 mg l⁻¹ of CN⁻ and 10 mg l⁻¹ of Zn²⁺ in seawater are reduced to 5–7 mg l⁻¹ and 0·1 mg l⁻¹. Subsequent treatment with H₂SO₅/CN = 1·2 reduces the [CN⁻] to 0·1 mg l⁻¹.Treatment of a combined blast furnace/coke ovens effluent ([CN⁻] = 24 mgl⁻¹, [Zn²⁺] = 4·0 mgl⁻¹) with Fe/CN = 1·5 reduced [CN⁻] to 0·2 mg l⁻¹ and [Zn²⁺] to <0·1 mgl⁻¹. Subsequent treatment with H₂SO₅/CN = 2·0 reduced [CN⁻] to 0·2 mg l⁻¹. The process operates best in the pH range 7–9 and so is not affected by the buffer characteristics of seawater.     

Distribution of halomethanes in potable waters of Kuwait

The distribution of bromine containing trihalomethanes in the water distribution system of Kuwait has been studied. Total halomethanes in the drinking water averaged 25.6 ± 9.1 μg l⁻¹ with a maximum of 50.5 μg l⁻¹. Average concentrations (μg l⁻¹) of individual compounds were: CHBr₃, 13.6 ± 4.6; CHBr₂Cl, 8.8 ± 3.7; CHCl₂Br, 3.3 ± 1.5. Water from roof top storage tanks contained significantly less halomethanes than that from underground reservoirs.     

Restoration of heavily polluted branches of the Shatt Al-Arab river, Iraq

In view of the desire to improve the water quality of the heavily polluted branches of the Shatt al-Arab River at the City of Basrah, it was proposed to maintain effective flushing as well as contracting sewerage system. The present study was conducted in order to examine the water quality of these branches in an attempt to evaluate the effectiveness of the proposed flushing system. It has been found that their waters contained very low levels of dissolved oxygen and relatively high amounts of both COD and BOD₅. The annual average water quality parameters for Basrah Branches were: dissolved oxygen 3.4 ppm; pH 7.67; hydrogen sulphide 1.4 ppm; ammonia 97 μg-at. N l⁻¹; COD 15.9 mg l⁻¹; BOD₅ 12.7 mg l⁻¹; dissolved silicates 202 μg-at. Si l⁻¹; dissolved reactive phosphate 13.4 μg-at. P-PO₄³⁻ l⁻¹; nitrate 10.4 μg-at. N-NO₃⁻ l⁻¹; nitrite 2.1 μg-at. N-NO₂⁻ l⁻¹ and chlorophyll-α 14.3 mg m⁻³. Based on our calculations, it has been concluded that the proposed system is effective, thus within a flushing cycle all of the above mentioned parameters will become within the acceptable values of the Shatt al-Arab water quality. Moreover, this system has no appreciable effect upon the water quality characteristics of the Shatt al-Arab River due to the fact that it discharges a high volume of water annually. However, It has been recommended to dredge the deposited sludge to a minimum depth of 50 cm.     

The use of uric acid as a method of tracing domestic sewage discharges in riverine, estuarine and coastal water environments

The chemical tracking of sewage effluents discharged into fresh and saline waters presents difficulties, especially in estuaries. The main difficulty is caused by the dissolved constituents being used to monitor the effluent also occurring naturally at similar levels. Uric acid is present at significant levels in untreated sewage and is not normally found in unpolluted waters. Until now no suitable routine method has been available for uric acid estimation in fresh and saline waters at levels normally encountered in the environment. In this paper we describe a recently developed technique using high-performance liquid chromatography which estimates uric acid in both fresh and saline waters in the range 1–10,000 μg l⁻¹ with a precision (2σ) of ±20% at 2 μg l⁻¹, ±4% at 40 μg l⁻¹ and ±2% at 10 mg l⁻¹.     

Potential hazards of nitrate and fluoride in underground waters

Underground waters of the semi-arid tract of Agra region (India) were examined for their nitrate and fluoride concentration along with other chemical constituents. The results indicated that 40 and 49% of the waters contained injurious concentrations of nitrate and fluoride respectively for drinking water. Only 28.4% of the waters were useful for some crops because of excessively high nitrate concentrations (> 100 mg l⁻¹) and 23.1% were harmful for common crops with respect to fluoride concentration (> 10 mg l⁻¹). Generally, high salinity waters had a high nitrate and fluoride content.     

Acute toxicity and behavioral responses of coho salmon (Oncorhynchus kisutch) and shiner perch (Cymatogaster aggregata) to chlorine in heated sea-water

The acute toxicity and behavioral response to chlorinated and heated sea-water was determined for coho salmon smolts and 1–3 month old shiner perch. LC₅₀'s were determined for 7.5, 15, 30 and 60 min exposure times; 13, 16 and 20°C (Δt = 0, 3, 7°C) temperatures and total residual oxidant (TRO) concentrations ranging from 0.077 to 1.035 mg l⁻¹. The mean 60 min LC₅₀ for shiner perch was significantly reduced (P ≤ 0.05) from 308 μg l⁻¹ TRO at 13°C to 230 μg l⁻¹ TRO at 20°C. The 60 min LC₅₀ for coho salmon decreased from 208 μg l⁻¹ TRO at 13°C to 130 μg l⁻¹ at 20°C. The LC₅₀'s for coho salmon in chlorinated sea-water averaged 55% of those for shiner perch. The relationship between TRO concentration, exposure time, and percent survival in chlorinated sea-water at 13°C is presented for both species.A significant (P ≤ 0.01) avoidance threshold for coho salmon occurred at 2 μg l⁻¹ TRO and was reinforced with increasing temperature. A significant (P ≤ 0.01) avoidance threshold for shiner perch occurred at 175 μg l⁻¹ TRO, while a significant preference (P ≤ 0.05 or 0.01) response at 16°C and 20°C occurred at 10, 25, 50 and 100 μg l⁻¹ TRO. The ecological implications of the toxicity tests and the behavioral responses are discussed.     

Experimental simulation of biodegradation in rivers: Oxygen, organic matter and biomass concentration changes

Dissolved oxygen (DO) and biochemical oxygen demand (BOD) concentration changes after an organic matter discharge into a river have been studied in the absence of oxygen transfer. According to these laboratory experiments, biodegradation of various organic compounds (glucose, glutamic acid, starch, ovalbumin and ethanol) in Seine river samples incubated at 15 30°C follow a biphasic behaviour. During a lag-phase of 10–20 h, DO decreases linearly (0.12 ppm h⁻¹ at 20°C), whereas BOD is constant. During a subsequent aerobic exponential phase, DO and BOD uptake are proportional and increase exponentially with time (0.13 h⁻¹ at 20°C). Using cell ATP as biomass indicator, the latter phase was shown to correspond to a cell division step. A kinetic model was developed for stimulating DO and BOD concentration changes after a waste water discharge at temperatures ranging between 15 and 30°C.     

Nitrate reduction by denitrifying bacteria in single and two stage continuous flow reactors

Denitrification by a mixed bacterial population of medium containing 1000 mg NO₃⁻-N1⁻¹ and acetate as carbon source was studied in batch, a single stage continuous flow stirred reactor (CFSTR) and a two stage CFSTR at 30°C. The optimum pH for denitrification, nitratase, nitrite reductase activities and growth was found to be 7.5 in batch culture. A single stage CFSTR growth limited by nitrate had an optimum denitrification rate of 0.13 mg NO₃⁻-N mg⁻¹ cells h at a residence time of 8 h. The experimentally observed carbon to nitrate ratio (mg CH₃ COO⁻-C mg⁻¹ NO₃⁻-N)was 1.7 for the dilution rates of 0.02–0.18 h⁻¹. For the second stage CFSTR, bacteria growing at the maximum rate of 0.25 h⁻¹ and not limited by nitrate had a denitrification rate of 0.24 mg NO₃⁻-N mg⁻¹ h. Dissolved oxygen (up to 9.5 mg 1⁻¹) did not effect denitrification rates in the second stage CFSTR. As the second stage CFSTR runs progressed extensive wall growth occurred and concurrently the output gas contained increasing quantities of nitrous oxide. A development from this study would be a two stage CFSTR with wall growth in the second stage which would make an efficient nitrate removal process.     

Comparative disinfection of treated sewage with chlorine and ozone: Effect of nitrification

Chlorine and ozone were compared in pilot plants (capacity about 3.2 m³ h⁻¹), which were fed with the same activated sludge treated and filtered water. Together with physico-chemical analysis the water was analysed for different types of microorganisms, including vegetative bacteria (total and thermotolerant coliforms, faecal streptococci and Pseudomonas aeruginosa), bacterial spores (spores of aerobic bacteria at 37°C and sulphite reducing clostridia) and bacterial viruses (somatic coliphages and F-specific bacteriophages).The average chlorine and ozone dose were, respectively, 3.65 and 15.3 mg l⁻¹ of water, while after a contact time for both of about 25 min the average residual concentrations were 1.79 and 0.35 mg l⁻¹ of water. These residuals were measured with the DPD-method. The ammonia-N concentration varied greatly (0.06–72.0 mg l⁻¹) and was used to group the data into four classes: (1) non-nitrified water, defined as water in which nitrate-N was smaller than ammonia-N; (2) moderately nitrified water, in which nitrate-N was larger than ammonia-N and the ammonia-N was higher than 2 mg l⁻¹; (3) well nitrified water, defined as water in which ammonia-N was lower than 2 mg l⁻¹; (4) very well nitrified water, in which ammonia-N was smaller than 0.5 mg l⁻¹.This classification indicated that the concentrations of most other impurities decreased with a better nitrification. Statistical analysis of the data showed also that ozone was a better disinfectant than chlorine in the case where the disinfection is based upon their residual content. The degree of nitrification had a greater effect on chlorine disinfection than on ozone disinfection.During chlorination the total residual chlorine decreased, with better nitrification; the chlorine demand increased; the composition of the residual chlorine changed very much and the inactivation of bacterial viruses improved. The vegetative bacteria showed a varying pattern; most were inactivated in moderately nitrified water, when the dichloramine concentration was highest and false positive FAC concentration was lowest of the four classes. Reduction of bacterial spores was not observed.During ozonization other effects were indicated. Reductions of most organims increased slightly with better nitrification; only reductions of F-specific bacteriophages decreased. There was also a small decrease of bacterial spores. The treated effluent had a high ozone consumption and the inactivation of the organisms was low in relation to ozone dose and residual ozone.The bromide concentration (0.3–2.9 mg l⁻¹) effected the chemistry of chlorine and ozone and had a positive effect on chlorine and ozone disinfection of total coliforms.For most types of micoorganisms the disinfection coefficients of the Selleck model and the germicidal efficiencies could be determined.     

An improved determination of chemical oxygen demand in water and wastes by a simplified acid dichromate digestion

A simplified and improved chemical oxygen demand (COD) procedure is suitable for water and waste water samples containing up to 1500 mg Cl l⁻¹. Samples are digested with sulfuric acid, potassium dichromate, silver sulfate, mercury(II) sulfate and sulfamic acid in open glass tubes for 2 h at 140 ± 2°C without boiling, and excess dichromate is determined colorimetrically at 440 nm. The use of 49-position digestion racks, a large oven and a spectrophotometer fitted with a 1-cm flow cell allow analysis of ca. 12 samples h⁻¹. A correction for chloride interference is not required in the 0–500 mg Cl l⁻¹ range. The detection limit is 3 mg l⁻¹, and the relative standard deviation at the 112 mg COD l⁻¹ level is 4.3%. Thirty-five waste water samples were analyzed by the standard dichromate reflux method and by the proposed procedure. There was no significant difference between the two sets of data (P > 0.25). Recovery data for 15 major water pollutants including benzene, toluene and pyridine are presented.     

Hypolimnetic aeration: field test of the empirical sizing method

A hypolimnetic aeration system was recently installed in a small (16 ha S_α) eutrophic lake and a comparison made between measured performance and predicted performance from an empirical sizing method. The design variables used to size the system were: hypolimnetic volume 451,600 m³; maximum hypolimnetic oxygen consumption 0.2 mg l⁻¹ d⁻¹; aerator input rate 2 mg l⁻¹; water velocity 0.76 m s⁻¹ and depth of air release 12.2 m. A 3.7 kW compressor (0.57 m³ min⁻¹) generated a water velocity of 0.46 m s⁻¹, a water flow of 17.7 m³ min⁻¹ and a theoretical hypolimnetic circulation period of 18 days. Dissolved oxygen increased by an average of 1.6 mg l⁻¹ on each cycle through the aerator, and aerator input rates ranged from 0.6 to 2.6 mg l⁻¹. Hypolimnetic oxygen consumption averaged 0.12 mg l⁻¹ d⁻¹ and ranged between 0.02 and 0.21 mg l⁻¹ d⁻¹. The aeration system was unable to meet the daily oxygen demand (90 kg) as the water velocity was slower than expected (0.46 m s⁻¹). To avoid undersizing future aeration installations the following recommendations should be considered when using the empirical sizing formula: (1) estimates of oxygen consumption should be annual maximums from aerobic hypolimnia; (2) aerator input rates should be conservative (e.g. 1–4 mg l⁻¹) and increase with depth; (3) water velocity of 0.45–0.50 m s⁻¹ should initially be used when no information on actual bubble size or velocity is available; (4) aeration start-up should be timed to avoid periods of accumulated oxygen demands.     

Inactivation of Naegleria gruberi cysts by chlorine dioxide

Investigations were carried out to determine the kinetics of chlorine dioxide inactivation of Naegleria gruberi cysts and the influence of pH from 5 to 9, temperature from 5 to 30°C, cyst age from 3 to 12 days, and cyst clumping. Inactivation was characterized by an initial concave downward shoulder followed by an essentially linear portion but the process obeyed Watson's Law. At 25°C and pH 7 the mean concentration time product for 99% inactivation was 5.5 mg·min l⁻¹. These products varied inversely with temperature and pH.     

Phthalic acid esters in water

Two phthalic acid esters, di(n-butyl)phthalate and di (2-ethylhexyl)phthalate, were surveyed in the river water, well water and city water of Tokyo metropolice area. In the river water these esters were found in the range of 0·4–6·8 μ gl⁻¹. The concentration was low in the upper reaches of the stream but increased downstream, giving no seasonal variation. The esters were not found in the well water examined. In the household tap water they were contained in the range of 1·2–3·3 μg l⁻¹ while in the raw water of water supply they were contained in the range of 1·9–8·2 μg l⁻¹. Results of jar test revealed that these esters were efficiently removed from water by using activated carbon or aluminium sulphate.