Denitrification with a bacterial disc unit

An enclosed rotating disc unit was operated anaerobically as a denitrifying system, with methanol as the hydrogen donor. As the bacterial population became established, denitrification rate increased by 1·5 mg NO₃⁻—N reduced m⁻² h⁻², to a maximum rate of 260 mg NO₃⁻—N reduced m⁻²h⁻¹. The C:N ratio necessary for complete denitrification was found to be 2·6:1. Optimum pH for denitrification lay in the range between pH 7·0 and 8·5. Q₁₀ values were 1·38 between 10 and 30°C, −2·66 above 30°C and 13·06 below 10°C.     

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.     

The effect of temperature on the removal of non-ionic surfactants during small-scale activated-sludge sewage treatment—I. Comparison of alcohol ethoxylates with a branched-chain alkyl phenol ethoxylate

The degree of biodegradability of three non-ionic surfactants has been assessed using small-scale activated-sludge sewage treatment plants. The effect of temperature on biodegradability was studied by operation at 15°, 11° and 8°C.The two alcohol ethoxylate surfactants tested were well degraded at all three operating temperatures. The alkyl phenol ethoxylate surfactant tested was well degraded at 15°C but at lower temperatures the biodegradability was dependent on concentration. At 5 mg 1⁻¹, greater than 90 per cent removal was achieved but at 20 mg 1⁻¹ the degree of removal fluctuated between 40 and 95 per cent at 11°C and between 20 and 80 per cent at 8°C.The present results have been compared with those obtained using the same surfactants in community trials at a small sewage works employing biological filtration. Although a similar temperature effect was observed with the alkyl phenol ethoxylate, the biodegradabilities obtained in the laboratory were consistently greater than those obtained at the sewage works, perhaps because of the constant conditions of the laboratory test.     

Low temperature removal of nitrate by bacterial denitrification

The efficiency of two denitrifying sludges enriched at 5 and 20°C were compared using methanol as an electron donor. Both sludges were exposed to the same hydraulic and chemical conditions using an influent containing methanol and mineral salts. The low temperature sludge seemed to have several advantages over the sludge selected for at the higher temperature. In the range 0–17°C, the specific denitrification rate was 1.5–4 times the rate for the high temperature sludge, temperatures below 8°C being the most favourable. At 2°C, under nitrate limiting conditions, 98% nitrate reduction was obtained at a hydraulic residence time of 3.5 h, with an effluent concentration of 0.8 mg NO₃---Nl⁻¹. Sedimentation characteristics were always better for the low temperature sludge, and the utilization of methanol equally good as the high temperature sludge. The low temperature sludge appeared to be biochemically and microbiologically stable to temperature changes within the range 0–17°C, the latter temperature being close to the limit for maintaining the psychrophilic characteristics of the sludge. Studies on pure culture isolates of the denitrifying bacteria showed >90% dominance of one bacterial strain in both sludges. Studies of the isolates also showed that the low-temperature sludge consisted predominantly of psychrotrophs/psychrophiles, and not well-adapted mesophiles, which were only present in low concentrations. The dominant strain in both sludges was unable to grow on methanol in pure culture without access to nutrient growth factors. Only a few minor strains were obligate methylotrophs.Low temperature sludges were tested in a 3-stage biological process receiving domestic sewage. Each stage; carbon oxidation, nitrification and denitrification had separate sludge recycle, and methanol was added to the denitrification stage. These sludges were grown and selected for at temperatures 5°C. At 5°C the laboratory scale process gave 90% removal of total nitrogen at hydraulic residence times of 1.5, 9 and 4 h for the two aeration stages and the anaerobic stage respectively. Overall nitrification/denitrification was 95%, while denitrification separately was 98%. The effluent contained 0.4 mg NO₃---Nl⁻¹. The critical step in the process was unquestionably nitrification. Oxidation of ammonium was satisfactory at low temperature, but the reaction was somewhat vulnerable to changes in external conditions. The low temperature denitrifying sludge was originally enriched on synthetic waste but did not appear to change its microbial composition or characteristics by exposure to municipal wastewater.     

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.     

Water quality criteria for European freshwater fish: Report on ammonia and inland fisheries

In establishing water quality criteria for European inland fisheries, the effect of ammonia is an important factor to be considered. Sewage effluent, effluents from certain industries and from agriculture are common sources of ammonia in water.The harmful effects of ammonia on fish are related to the pH value and the temperature of the water due to the fact that only the un-ionized fraction of ammonia is poisonous. The un-ionized fraction increases with rising pH value, and with rising temperature.Fish differ slightly in their tolerance to ammonia depending on species. The difference in tolerance being more significant for short periods of exposure. The difference in tolerance is, however, not great enough to justify different criteria for different species.The lowest toxic concentration found for salmonids is 0·2 mg NH₃ 1⁻¹ (un-ionized), but other adverse effects caused by prolonged exposure are only absent at concentrations lower than 0·025 mg NH₃ 1⁻¹ (un-ionized). Concentrations of total ammonia which contain this amount of un-ionized ammonia vary from 19·6 mg 1⁻¹ (pH 7·0, 5°C) to 0·12 mg 1⁻¹ (pH 8·5, 30°C).The criterion should not be applied to temperatures below 5°C or to pH values above 8·5 when other factors have to be taken into consideration.     

Treatment of low strength domestic wastewater using the anaerobic filter

A laboratory scale anaerobic filter packed with synthetic high surface area trickling filter media was used to treat a low strength domestic wastewater averaging 288 mg 1⁻¹ COD. The filter was operated for 60 days after reaching steady-state at 20, 25, 35°C at a loading rate of 0.02 lb COD ft⁻³ day⁻¹ and 24 h hydraulic retention time. Filter effluent BOD₅ averaged 38 mg 1⁻¹ providing an average removal rate of 79%, and effluent COD averaged 78 mg 1⁻¹, corresponding to a 73% removal rate. Removal efficiencies showed very little sensitivity to daily fluctuations in influent wastewater quality. The filter performance at 25 and 35°C was not significantly different, but BOD and TSS removal efficiency declined a: 20°C. Gas production averaged 0.027 ft⁻³ of gas per ft³ of influent wastewater, or 1.875 ft³ of gas per pound of influent COD. Gas composition averaged 30% nitrogen, 65% methane, and 5% carbon dioxide. Ammonia nitrogen and sulfides both increased during treatment. It is concluded that the anaerobic filter is a promising candidate for treatment of low strength wastewaters and that post treatment for sulfides and ammonia may be necessary.     

Ozonation of water containing chlorine or chloramines. Reaction products and kinetics

Ozone reacts with free aqueous chlorine when present as hypochlorite ion (OCl⁻) with a second order rate constant of 120 ± 15 M⁻¹ s⁻¹ at 20°C. About 77% of the chlorine reacts to produce Cl⁻ and 23% is oxidized to ClO⁻₃. No ClO⁻₄ is formed. Conversion of chlorine to monochloramine reduces the ozone reaction rate to 26 ± 4 M⁻¹ s⁻¹, independent of pH, NH₂Cl is transformed quantitatively to NO⁻₃ and Cl⁻ by O₃. Rate data for other chloramines are also presented. The direct reaction of ozone with chlorine accounts for a significant amount of the chlorine and ozone demand found when the two oxidants are used in combination under water works conditions.     

A modified version of the sodium salicylate method for analysis of wastewater nitrates

The German sodium salicylate method for nitrate determination has been modified and improved by utilizing flocculating effect of the preservative HgCl₂ (1000 mg 1⁻¹). The method gives highly reproducible results in the range of 0·2–16 mg 1⁻¹ nitrate nitrogen and is applicable for routine analysis of untreated sewage. Interfering substances are chloride ions above 1500 mg 1⁻¹ and nitrite ions above 10 mg 1⁻¹N. The yellow color produced by sodium salicylate with nitrate ions obeys Beer's law and remains stable for several hours.     

Kinetics of microbial nitrification: nitrite-nitrogen oxidation

The kinetics of nitrite-nitrogen oxidation by enriched Nitrobacter cultures were examined under various temperatures (9°–35°C), pH (6.0–8.5), and microbial concentrations. For the substrate concentrations examined (100–1100 mg N1⁻¹) the reaction was zero order and correlations between temperature, pH, microbial concentrations and reaction rates were formulated. Inhibition by free ammonia and nitrous acid is discussed. This study of enriched cultures of nitrobacter organisms also provide some insight into the growth characteristics of this fastideous organism under conditions of dense population and severe environmental conditions (temperature and pH).     

Effect of copper and hexavalent chromium on the specific growth rate of ciliata isolated from activated-sludge

After growing mass and monoxenic cultures of three species of Ciliata (Vorticella microstoma, Colpidium campylum, and Opercularia sp.) with Alcaligenes faecalis as the sole protozoan food, and confirming that copper and hexavalent chromium did not affect the bacterial growth, the effect of these metals on the specific growth rate of each protozoa was examined. The term IL_m (median inhibitory limit) was used to define the metal concentration required to reduce the specific growth rate of protozoa to one-half of that of a control. The IL_m for copper was: 0·25 mg 1⁻¹ (V. microstoma), 0·32 mg 1⁻¹ (C. campylum), and 0·27 mg 1⁻¹ (Opercularia sp.). Regarding hexavalent chromium, IL_m for V. microstoma, C. campylum, and Opercularia sp. were 0·53, 12·9 and 20·2 mg 1⁻¹, respectively. Acclimation of each protozoa to these metals for 96 h resulted in IL_m enhanced values of 1·2–2·2 times as large as that for the control.     

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.     

The influence of substrate, pH, diet and temperature upon cadmium accumulation in the asiatic clam (Corbicula fluminea) in laboratory artificial streams

The influence of substrate, pH, diet and temperature upon the accumulation of cadmium (0.05 mg l⁻¹ dose < 0.001 mg l⁻¹ control) in the visceral mass of the Asiatic clam Corbicula fluminea was studied in laboratory artificial stream systems at intervals of 0, 3, 5, 7, 10 and 14-day exposures. Four substrate conditions, sand; sand, silt and clay (SSC); sand, clay and organic matter (SCO); and no substrate (NoS), were considered. The greatest tissue accumulation of cadmium in C. fluminea occurred at 0.05 mg l⁻¹ Cd in NoS and the lowest in clams occupying SCO. Complexation of available metals, lower clam filtering rates and physical protection by the substrate were attributed to the depressed cadmium accumulation of clams exposed in the SCO substrate. Lower pH exposures (5.0 vs 7.8) significantly (P 0.05 level) reduced cadmium uptake at 21°C but had little effect at 9°C. Uptake was higher in clams fed with cadmium-exposed Chlamydomonas reinhardt at 21°C but not at 9°C. In temperature exposures alone at 0.05 mg l⁻¹ Cd, accumulation was significantly higher in C. fluminea exposed at 21°C than at 9°C. The results are discussed relative to the importance of standardized laboratory protocol and the use of C. fluminea as a bioindicator of heavy metal stress.     

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.     

pH adjustment in anaerobic digestion

In the pH range 6·0–7·5, the pH in anaerobic processes is controlled by the interaction of the carbonic system and a net strong base. The acid-base state of a digestor can be monitored by only measuring pH and CO₂ partial pressure. Shock doses of strong bases and carbonates causes temporary undersaturated CO₂ conditions and excessively high pH. Bicarbonate dosing leaves the CO₂ solubility equilibrium unchanged. In the absence of a CaCO₃ precipitation inhibiting agent. CaCO₃ solubility limits the pH, and Ca(OH)₂ dosing is unable to raise the pH significantly. Orthophosphates inhibit CaCO₃ precipitation. With [PO₄] > 1·0 × 10⁻³mole·1⁻¹. CaCO₃. precipitation is partially inhibited. Ca(OH)₂ dosing being approximately 45 per cent effective for doses up to 15000 mg 1⁻¹ as CaCO₃. At [PO₄] < 1·0 × 10⁻³moles·1⁻¹ orthophosphates eventually precipitate out during Ca(OH)₂ dosing, thus removing the inhibition mechanism: pH is then limited by the CaCO₃ solubility. Most wastes contain [PO₄] > 2·0 × 10⁻³moles·1⁻¹ making pH adjustment with Ca(OH)₂ possible to a pH of about 7·2 although the dosages will be very high. The pH changes in a process following dosing can be predicted by the graphical representation of the carbonic and net strong base systems.     

Enhanced uptake of phosphorus by activated sludge-effect of substrate addition

Studies were undertaken to examine the effect of substrate addition upon the release and subsequent uptake of phosphorous by a biological wastewater treatment laboratory scale system.A train of six reactors were fed at a rate of 16 ml min⁻¹ raw wastewater using a sludge recycle ratio of 0.75 (12 ml min⁻¹). The first two reactors were simply stirred (anoxic) without the addition of air and the remaining four were aerated with excess air.Various substrates were added to the first reactor (anoxic) at different concentrations. It was determined that all the short chain substrates tested enhanced the release of phosphorus in the anoxic stages and subsequently led to increased biological uptake (removal) of phosphorus. The substrates tested included sodium acetate, acetic acid, butyric acid, ethanol and methanol.It was determined that 30 mg l⁻¹ sodium acetate was the optimum dose for biological release and uptake of phosphorus and the addition of any greater concentration had marginal effect on the ultimate removal of phosphorus.The effect of these substrates showed some specificity regarding phosphorus release, with butyric acid causing the greatest release and sodium acetate the least. However as far as phosphorus uptake (removal) was concerned, this phenomenon of substrate-specificity was less significant. For all substrates, effluent phosphorus concentration was within ± 1 mg l⁻¹ with an approximate mean concentration of 1 mg l⁻¹ residual.Phosphorus released (approx. 14 mg l⁻¹) at higher temperature (29°C) was 75% greater than at the lower temperature (24°C). Similarly the final residual phosphorus at 29°C was 33% lower than at 24°C.     

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.     

Mechanism of NTA degradation by a bacterial mutant

Transport of nitrilotriacetic acid (NTA) into the cytoplasm of a bacterial mutant was an active process. The mutant was able to degrade NTA from an initial concentration of 290,000 μg l⁻¹ of NTA to less than 50 μg l⁻¹ in 45 min, representing a rate of 486 μg NTA degraded per hour per mg dry weight of cells. This extremely fast rate of NTA utilization was substantiated by kinetic studies in which a Km of 82 μg l⁻¹ and a V_max of 370 μg h⁻¹ (mg dry weight cells)⁻¹ were found. The maximal temperature for NTA degradation was 50°C. The ability of the mutant to metabolize NTA resided mainly in the cell membrane fraction. Exchange diffusion technique showed that glycine and acetic acid were the metabolic products of NTA degradation. No iminodiacetic acid (IDA), succinate or citrate could be detected.     

Influence de sediments argileux incorpores au milieu d'insemination sur le succes de la fecondation chez la truite arc-en-ciel (Salmo Gairdneri)

The aim of this paper is to determine whether clay sediments suspended in water can prevent trout eggs from being fertilized. Kaolinite-rich clays (granulometric fraction: <2μm) (Fig. 1) were suspended in an artificial insemination diluent in doses ranging between 0 and 20 g l⁻¹. The eggs were exposed for 1, 10 or 20 min (experiment A) or inseminated (experiment B) in the diluent-sediment mixture. In experiment (C), the eggs were exposed to this mixture at three different temperatures (10, 15, 20°C). After insemination, the eggs were incubated for 10 days at 10°C and the percentage of eyed-eggs was used as an approximation of the fertility rate. The presence of clay sediments in the medium in which artificial insemination was carried out did not affect fertilization rate after the ovules had been exposed during 1 min to clay suspensions, at any of the temperatures used (8°C: Fig. 3; 10–15 or 20°C: Fig. 5) or at any of the sperm dilution rates (10⁻², 10⁻³, 10⁻⁴) (Fig. 3). On the contrary, there was a significant decline (P < 0.01) in the fertilization rate after the eggs had been exposed for 10 min at 8°C to doses of sediment exceeding 1.2 g l⁻¹ (Fig. 2). The fertilization rate also decreased significantly (P < 0.05) when the dose of sediment in the medium increased after 20 min at 20°C and 40 min at 15°C (Fig. 4). The 15 and 20°C temperatures were unfavourable for the eggs anyway. It is probable that fertility decreased due to micropyle clogging when the eggs were exposed longer than 10 min to the sediments. It is concluded that presence of sediments in the medium in which the gametes meet does not prevent fertilization.