Bioremediation of crystal violet using air bubble bioreactor packed with Pseudomonas aeruginosa

Seven water and sediment samples were collected and tested for decolorizing crystal violet. Pseudomonas aeruginosa was the most effective isolate for dye decolorization. The LC₅₀ of the crystal violet (115 mg/l) was measured using Artemia salina as a biomarker. The effect of different heavy metals on crystal violet decolorization was investigated. Cd²⁺ and Fe³⁺ ions showed marginal enhancement of the decolorization process, the rate was 1.35 mg/l/h compared to 1.25 mg/l/h for the control. Phenol and m-cresol showed no effect on crystal violet decolorization, meanwhile p-cresol and p-nitrophenol reduced the decolorization rate to 1.07 and 0.01 mg/l/h, respectively. P. aeruginosa cells were immobilized by entrapment in agar–alginate beads. The beads were cultivated and reused in Erlenmeyer flask and in an air bubble column bioreactor and they enhanced the crystal violet decolorization rate to 3.33 and 7.5 mg/l/h, respectively.     

UV light assisted decolorization of dark brown colored coffee effluent by photo-Fenton reaction

The photochemical decolorization of coffee effluent has been examined by photo-Fenton (UV/Fe²⁺/H₂O₂) process. Effects of UV light intensity, initial coffee concentration, iron dose and H₂O₂ dose on the color removal of model coffee effluent have been investigated. The rate of decolorization increased with decreasing initial coffee effluent concentration. It was found that the Fe ion dose and UV light intensity enhanced the decolorization rate. The decolorization process of coffee effluent could be divided into three established phases. At the beginning of the photo-Fenton process, the instantaneous and significant increase in color of the solution was found (Phase-I). In the subsequent phase (Phase-II), the decolorization rate was initially fast and subsequently decreased. In Phase-III, the rate was accelerated and then the complete decolorization of model coffee effluent was achieved. In order to elucidate the mechanisms of coffee effluent color removal process, the concentration changes in Fe³⁺ and Fe²⁺ besides H₂O₂ were measured during the course of the photo-Fenton process. The rate-determining step in Phase-II was the photo-Fenton reaction or photoreduction of Fe³⁺. On the other hand, the decolorization process in Phase-III was highly affected by Fenton reaction or decomposition of H₂O₂ with Fe²⁺. About 93% mineralization of 250 mg L⁻¹ model coffee effluent was achieved after 250 min. A comparative study for TiO₂, ZnO and photo-Fenton oxidation processes has been also carried out and the photo-Fenton process was found to be the most effective for color removal of coffee effluent.     

Sorption of cadmium(II) and zinc(II) ions from aqueous solutions by cassava waste biomass (Manihot sculenta Cranz)

The sorption of two divalent metal ions, Cd(II) and Zn(II), onto untreated and differentially acid-treated cassava waste biomass over a wide range of reaction conditions was studied at 30°C. The metal ion removal from the spent biomass was also measured. The batch experiments show that pH 4.5–5.5 was the best range for the sorption of the metal ions for untreated and acid-treated biomass. Time-dependent experiments for the metal ions showed that for the two metals examined, binding to the cassava waste biomass was rapid and occurred within 30 min and completed within 1 h. High sorption capacities were observed for the two metals. The binding capacity experiments revealed the following amounts of metal ions bound per gram of biomass: 86.68 mg/g Cd, 55.82 mg/g Zn and 647.48 mg/g Cd, 559.74 mg/g Zn for untreated and acid-treated biomass, respectively. It was further found that the rate of sorption was particle-diffusion controlled, and the sorption rate coefficients were determined to be 2.30×10⁻¹ min⁻¹ (Cd²⁺), 4.0×10⁻³ min⁻¹ (Zn²⁺) and 1.09×10⁻¹ min⁻¹ (Cd²⁺), 3.67×10⁻² min⁻¹ (Zn²⁺) for 0.5 and 1.00 M differential acid treatment, respectively. Desorption studies showed that acid treatment inhibited effective recovery of metal ions already bound to the biomass as a result of stronger sulfhydryl-metal bonds formed. Less than 25% of both metals were desorbed as concentration of acid treating reagent increases. However, over 60% Cd and 40% Zn were recovered from untreated biomass during the desorption study. The results from these studies indicated that both untreated and acid-treated cassava waste biomass could be employed in the removal of toxic and valuable metals from industrial effluents.     

Production, composition and Pb2+ adsorption characteristics of capsular polysaccharides extracted from a cyanobacterium Gloeocapsa gelatinosa

Pb²⁺ adsorption by the living cells of the cyanobacterium Gloeocapsa gelatinosa was studied. Cyanobacterial cells with intact capsular polysaccharide (CPS) showed 5.7 times higher Pb adsorption capacity than that of cells without CPS. The adsorbed Pb was desorbed by EDTA, indicating that Pb²⁺ adsorption occurred mainly on cell surface. Production, sugar content and ability of CPS to remove Pb²⁺ were then studied in details. CPS production by G. gelatinosa increased when culture time was prolonged. The maximum CPS production was 35.43 mg g⁻¹ dry weight after 30-day cultivation. Xylose, arabinose, ribose, rhamnose, galactose, glucose, mannose and fructose were the neutral sugars presented in CPS of G. gelatinosa. Acidic sugars including galacturonic and glucuronic acids were also found in CPS. The amount and composition of G. gelatinosa's CPS varied according to its growth phase and culture conditions. The highest amount of acidic sugars was produced when cultured under low light intensity. The extracted CPS rapidly removed Pb²⁺ from the solution (82.22±4.82 mg Pb²⁺ per g CPS), directly demonstrating its roles in binding Pb²⁺ ions. Its ability to remove Pb²⁺ rapidly and efficiently, to grow under sub-optimal conditions (such as low pH and low light intensity), and to produce high amount of CPS with acidic sugars, leads us to conclude that G. gelatinosa is a potential viable bioadsorber for mildly acidic water contaminated with Pb²⁺.     

Effects of nitrobenzene and zinc on acetate utilizing methanogens

Determination of anaerobic degradation rates and toxic effects of nitrobenzene (NB) on acetate utilizing methanogens was the first objective of this research. Serum bottles were used for anaerobic toxicity assays with an acetate enrichment culture of methanogens. Ten mg/l of nitrobenzene did not inhibit total gas production in the acetate enrichment methanogenic culture. Twenty and thirty mg/l of nitrobenzene caused reversible inhibition of methanogenesis. Batch kinetic experiments showed that 20 mg/l of nitrobenzene was degraded with a first-order rate constant, k, of 0.37 d⁻¹. Acetate was not degraded during the first 7 days when the measured nitrobenzene concentration was higher than about 1 mg/l. The second objective was to determine the effect of zinc on nitrobenzene degradation in methanogenic systems. Ten mg/l of spiked zinc caused a reduction of gas production in the systems with 10 mg/l of nitrobenzene; 20 mg/l of zinc led to failures of systems with 10 and 20 mg/l of nitrobenzene. With 10 and 20 mg/l of added zinc, the k value for nitrobenzene degradation decreased to 0.18 d⁻¹ and 0.14 d⁻¹, respectively. With 20 mg/l of Zn, acetate was not degraded at all even after nitrobenzene concentration reached 0.1 mg/l, indicating toxicity of Zn to methanogenesis. Abiotic control tests with autoclaved culture showed that adsorption alone could remove 60–70% of spiked nitrobenzene in 36 days. However, the samples extracted from solids in the methanogenic test systems showed that nitrobenzene was below the detection limit of 0.1 mg/l, indicating biodegradation of nitrobenzene in these systems. Traces of benzene were seen as an intermediate in the liquid samples. Headspace analysis showed that nitrobenzene and benzene were below detection limits.     

Evaluation of kinetic parameters of a sulfur-limestone autotrophic denitrification biofilm process

In this study, four kinetic parameters of autotrophic denitrifiers in fixed-bed sulfur–limestone autotrophic denitrification (SLAD) columns were evaluated. The curve-matching method was used by conducting 22 non-steady-state tests for estimation of half-velocity constant, K_s and maximum specific substrate utilization rate, k. To estimate the bacteria yield coefficient, Y and the decay coefficient, k_d, two short term batch tests (before and after the starvation of the autotrophic denitrifiers) were conducted using a fixed-bed SLAD column where the biofilm was fully penetrated by nitrate-N. It was found that K_s=0.398 mg NO₃⁻–N/l, k=0.15 d⁻¹, k_d=0.09–0.12 d⁻¹, and Y=0.85–1.11 g VSS/g NO₃⁻–N. Our results are consistent with those obtained from SLAD biofilm processes, but different from those obtained from suspended-growth systems with thiosulfate or sulfur powders as the S source. The method developed in this study might be useful for estimation of four Monod-type kinetic parameters in other biofilm processes. However, cautions must be given when the estimated parameters are used because the measurements of the biomass and the biofilm thickness could be further improved, and the assumption of sulfur being a non-limiting substrate needs to be proved.     

Removal of cadmium and manganese by a non-toxic strain of the freshwater cyanobacterium Gloeothece magna.

The ability of both living and dry cells of Gloeothece magna, a non-toxic freshwater cyanobacterium, to adsorb cadmium and manganese is demonstrated in this study. Chlorophyll a content of living cells was not influenced by either cadmium or manganese concentrations, indicating that adsorption of both Cd²⁺ and Mn²⁺ by living cells of G. magna, was independent of the metabolic state of the organism. Moreover, the adsorption of both Cd²⁺ and Mn²⁺ to living cells and dry cells, was dependent on the metal concentrations, and fitted the Freundlich adsorption isotherm. However, dry cells had larger binding capacity for both Cd²⁺ (K_f=912.6) and Mn²⁺ (K_f=2398) than living cells (K_f=151.4 & 63, respectively). The role of the capsular polysaccharides, the main constituents of the cyanobacterial envelope, in binding these two metals was also studied. Polysaccharide extracts of this organism adsorbed high amounts of both Cd²⁺ (115–425 μg mg⁻¹) and Mn²⁺ (473–906 μg mg⁻¹). This study suggests that G. magna would probably be cultured in water bodies contaminated by heavy metals to ameliorate their toxicity. Also dry material of this cyanobacterium being a non-toxic species, could be used as a safe biofilter to remove toxic metals from drinking water.     

The effect of different drinking water treatment processes on the rate of chloroform formation in the reactions of natural organic matter with hypochlorite

This study is concerned with the changes in the rate of chloroform formation during the reactions of groundwater natural organic matter (NOM) and sodium hypochlorite caused by different drinking water treatments schemes: coagulation with FeCl₃, Al₂(SO₄)₃ and polyaluminum chloride (PACl), filtration of the raw water through granular activated carbon (GAC) and filtration through the columns filled with strong base macroporous ion-exchange resins (Purolite A501P and A500P) with and without pretreatment by coagulation process. It was found that the change of the concentration of chloroform, within 2 h and c(Cl₂)=100 mg l⁻¹, can be described by a kinetic equation of the form [CHCl₃]=a+bt^c. On the basis of this equation rates of the reaction were calculated. All processes applied decreased the rate of chloroform formation (process of coagulation moderately decreased the rate of reactions, while GAC adsorption caused dramatically drop of the rate). Also, it was found that the resins have had a higher affinity towards slow-reacting chloroform precursors.

The value of the chloroform formation potential was conventionally determined after a 7-day reaction at pH 7. In addition, the same parameter was estimated with a satisfactory deviation for raw water and for treated water on the basis of the kinetic constant (k) or by determining the chloroform concentration after 2 h (ChFP₂) under rigourous chlorination conditions at pH 8.4.     

Uptake and release of zinc by aquatic bryophytes (Fontinalis antipyretica L. ex. Hedw.)

The zinc uptake and posterior release by an aquatic bryophyte—Fontinalis antipyretica L. Ex Hedw.—was experimentally studied in laboratory exposing the plants to different zinc concentrations in the range, 1.0–5.0 mg l⁻¹, for a 144 h contamination period, and then exposed to metal-free water for a 120 h decontamination period. The experiments were carried out in perfectly mixed contactors at controlled illumination, using mosses picked out in February 1997, with a background initial zinc concentration of 263 mg g⁻¹ (dry wt.). A first-order mass transfer kinetic model was fitted to the experimental data to determine the uptake and release constants, k₁ and k₂, the zinc concentration in mosses at the end of the uptake period, C_mu, and at the equilibrium, for the contamination and decontamination stages, C_me and C_mr, respectively. A bioconcentration factor, BCF=k₁/k₂ (zinc concentration in the plant, dry wt./zinc concentration in the water) was determined. A biological elimination factor defined as BEF=1−C_mr/C_mu was also calculated. BCF decreases from about 4500 to 2950 as Zn concentration in water increases from 1.05 to 3.80 mg l⁻¹. BEF is approximately constant and equal to 0.80. Comparing Zn and Cu accumulation by Fontinalis antipyretica, it was concluded that the uptake rate for Zn (145 h⁻¹) is much lower than for Cu (628 h⁻¹) and the amount retained by the plant decreased by a factor of about seven.     

Interaction of light metals and protons with seaweed biosorbent

Based on their displacement by protons, the following ascending order of light metals affinity toward Sargassum fluitans biomass was observed: Na⁺ K⁺ < Mg²⁺ < Ca²⁺ < Al³⁺. Higher biomass-metal affinities resulted in lowering the leaching tendency of (polysaccharidic) biomass components during the sorption process. More than 12% weight loss of protonated biomass occurred during 2 min of sorption, which should be considered in all stoichiometric and sorption calculations. The rate of proton uptake for Na-loaded and Ca-loaded S. fluitans biomass at pH 4.5 was lower than the rate of desorption at pH 1.1. In the case of Na-loaded S. fluitans, the predominant proton uptake rate was of a zero order. Na uptake by protonated S. fluitans biomass did not take place at all in the presence of Ca and Al ions. However, it increased in the absence of other light metals. Since the affinity of Ca²⁺ for the biomass was much greater than that of Na⁺, the three-component system of Na⁺, H⁺ and Ca²⁺ could be simplified into proton-calcium ion exchange. The initial rate of Ca uptake with protonated S. fluitans biomass at pH 4.5 was lower than that of proton release from biomass. Approximately 20% of Ca²⁺ sorbed was bound to the binding site as a mono-valent ion. Titration and stoichiometric indications pointed to the fact that Al was being sorbed as a hydroxyl complex.     

Effect of water hardness on the toxicity of cadmium to micro-organisms

A study was made of the effect of water hardness at different concentrations (viz. 0, 80, 120, 160, 240, 320, 400 and 480 mg l⁻¹ as CaCO₃) on the toxicity of cadmium metal (5 mg 1⁻¹) as sulphate to saprophytic and nitrifying bacteria, with respect to the rate constant (K) and ultimate biochemical oxygen demand (L) which were calculated from BOD data (15 days) using the Thomas Graphical Method. Glucose was used as a source of carbon for micro-organisms. It was observed that the toxicity of cadmium to micro-organisms (both saprophytic and nitrifying) decreased with increasing hardness and reached a maximum at 320 mg 1⁻¹ as CaCO₃ for nitrifying and 400 mg l⁻¹ as CaCO₃ for saprophytic bacteria. After these hardness levels, the ultimate BOD (L) and rate constant (K) showed a decrease. Nitrifying bacteria were found to be more sensitive to the metal as well as to its complexation with calcium or with other ions as they retained their normal activity at a lower hardness level as compared to saprophytic bacteria.     

The bioavailability of copper in wastewater to Lemna minor with biological and electrochemical measures of complexation

Lemna minor (duckweed) was grown in treated domestic wastewater containing added Cu to study the relationship between complexation and bioavailability. Growth was not inhibited until total Cu exceeded 0.079–0.119 mg/l or internal Cu exceeded 81 to 235 μ/g. Cu²⁺ was detectable in the wastewater when total Cu measured ≥ 0.378 mg/l. There was a significant (P < 0.05) relationship between Cu bioconcentration and total Cu in wastewater that was attributed, in part, to heterotrophy of duckweed. Results indicated that, in addition to the bioavailable Cu²⁺ species, some complexes of Cu formed with ligands in wastewater may also be bioavailable. Measurement of the Cu Complexing Capacity (CC) of the wastewater by Ion-Selective Electrode (ISE) gave values of 0.26 to 0.29 mg/l and indicated the presence of ligands with a continuum of binding strengths which eliminated two of three commonly used methods for calculating CC. The biologically-measured CuCC based on growth was 0.077–0.125 mg/l when calculated by two out of three measures of Effective Concentration (EC) and was two to three times more sensitive than the ISE method for measurement of CC. ECs based on tissue concentration of Cu rather than solution concentration were 81–471 μ/g and have been proposed as an alternative for work in complex solutions like wastewater.     

Empirical rate equation for trihalomethane formation with chlorination of humic substances in water

Trihalomethane (THM) in drinking water is formed by chlorination of humic substances. In this study, the rates of THM formation in aqueous solution of humic acid were examined under various conditions. The following rate equation was obtained empirically. [THM] = k (pH − a)[TOC][Cl₂]₀^mtⁿ.

Here, [THM] is the concentration of total THM after t h, [TOC] and [Cl₂]₀ are the concentrations of total organic carbon and chlorine dose, k is the rate constant and a, m and n are parameters. The values of k, a, m and n for humic acid as reagent were obtained as 8.2 × 10⁻⁴ (l^mmg^−mh⁻ⁿ), 2.8, 0.25 and 0.36, respectively. The activation energy was obtained as 37 kJ mol⁻¹. Further, it was proved that the above equation could be applied to the rates of THM formation from precursors in actual river and lake waters.     

Decolorization of an azo dye by unacclimated activated sludge under anaerobic conditions

Studies on the reductive decolorization of a complex azo dye, Reactive Red 3.1, were made as part of the development of a practical approach to better exploit the metabolic potential of biomass in wastewater treatment. Decolorization was achieved at low and variable rates by mixed microbial cultures under various environmental conditions, including low pH and high salt concentration. It was caused by reductive cleavage of the azo bond to yield two aromatic amines. More reliable and effective decolorization rates, of up to 20–30 mg l⁻¹ h⁻¹, were given by unadapted activated sludge, (6 g l⁻¹) incubated with 400 mg l⁻¹ of Reactive Red 3.1 under anaerobic conditions. Decolorization also occurred best in static conditions.     

Fish toxicity of jet fuels—I. The toxicity of the synthetic fuel JP-9 and its components

The toxicity of the jet fuel JP-9 and its components RJ-4, RJ-5 and MCH was assessed in static bioassays on the warm water fish, golden shiner (Notemigonus chrysoleucas). The 96-h LC₅₀ of emulsions was 2.0 mg 1⁻¹ for JP-9, 0.51 mg 1⁻¹ for RJ-4, 0.61 mg 1⁻¹ for RJ-5 and 72 mg 1⁻¹ for MCH. As determined by the 96-h LC₅₀ values unemulsified fuel (pools of fuel) JP-9 was 235 times less toxic, RJ-4 was 196 times less toxic, RJ-5 was 7700 times less toxic and MCH was 3.3 times less toxic than the corresponding emulsified materials.

In continuous flow bioassays with the water soluble fraction of the fuel and its components the effect on egg hatchability and fry development of flagfish (Jordanella floridae) and rainbow trout (Salmo gairdneri) was studied. The no effect of level on flagfish egg hatchability was 0.23 mg 1⁻¹ for JP-9 and 0.05 mg 1⁻¹ for RJ-5. Concentrations of 0.83 mg 1⁻¹ MCH and 0.2 mg 1⁻¹ RJ-4 did not affect egg hatchability. In rainbow trout studies 97-day LC₅₀ values for RJ-4 and RJ-5 were 0.045 mg 1⁻¹ and 0.072 mg 1⁻¹, respectively, and 23-day LC₅₀ values for JP-9 and MCH were 0.26 mg 1⁻¹ and 1.3 mg 1⁻¹, respectively.

The accumulation of fuels in fish bodies was studied and it was found that flagfish can tolerate a total body burden of 0.5 mg MCH g⁻¹ wet weight without lethality. It was also found that body burdens of 0.43 mg RJ-4 g⁻¹ and of 0.27 mg RJ-5 g⁻¹ on a wet weight basis will produce 50% mortality in rainbow trout.

Voiding of MCH from fish bodies occurs readily in fuel-free water, but RJ-4 and RJ-5 are retained in the tissues.     

A comparison of fenuron degradation by hydroxyl and carbonate radicals in aqueous solution

A comparative study of the transformation of the herbicide fenuron (1,1-dimethyl-3-phenylurea) by hydroxyl radicals and carbonate radicals in aqueous solution (pH 7.2-phosphate buffer) has been undertaken. Hydroxyl radical was generated by the well-known photolysis of hydrogen peroxide at 254 nm and carbonate radical was formed by photolysis of Co(NH₃)₅CO₃⁺ at 254 nm. Competitive kinetic experiments were performed with atrazine used as the main competitor for both processes. Accordingly, the second-order rate constant of reaction between fenuron and carbonate radical was found to be (7−12±3)×10⁶ M⁻¹ s⁻¹ [(7±1)×10⁹ M⁻¹ s⁻¹ for hydroxyl radical]. The formation of degradation products was studied by LC-MS in the two cases and a comparison has been performed. The reaction with carbonate radical leads to the formation of a quinone-imine derivative which appears as the major primary product together with ortho and para hydroxylated compounds. These two compounds represent the major products in the reaction with hydroxyl radicals. The reaction of both radicals also leads to the transformation of the dimethylurea moiety.     

Comparative studies on potential of consortium and constituent pure bacterial isolates to decolorize azo dyes

The decolorization potential of the consortium HM-4 constituted by mixing four laboratory isolates identified as Bacillus cereus (BN-7), Pseudomonas putida (BN-4), Pseudomonas fluorescens (BN-5) and Stenotrophomonas acidaminiphila (BN-3) was compared with that of individual isolates. Six different azo dyes viz., C.I. Acid Red 88 (AR-88), C.I. Acid Red 119 (AR-119), C.I. Acid Red 97 (AR-97), C.I. Reactive Red 120 (RR-120), C.I. Acid Blue 113 (AB-113) and C.I. Acid Brown 100 (AB-100) were used in this study. The individual bacterial isolates were not able to completely decolorize these dyes, except for dyes AR-119 and AB-113. The consortium HM-4 was able to decolorize all the dyes used at an initial dye concentration of 20 mg L⁻¹ at a significantly higher rate as compared to that achieved by individual isolates.     

Utilisation de l''algue oocystis sp. pour le traitement tertiaire des eaux usées—I. Culture en vrac de cellules préalablement conditionnées en cyclostat

With the aim of developing an efficient and economic method for the tertiary treatment of wastewater, a two-phased culture system of Oocystis alga is presented. During the first phase, a unialgal strain grows in a cyclostat supplied with secondary effluents diluted to a low concentration (50 μM NH₄⁺, i.e. 0.7 mg N 1⁻¹) of inorganic ions. Once the equilibrium is reached (i.e. the cell population is conditioned and the nutrient concentration is zero), in a second phase, the starved cells are mixed with a secondary effluent which has a higher nutrient content (200–400 μM NH₄⁺, i.e., 2.8–5.6 mg N 1⁻¹). Ion depletion (NH₄⁺, NO₃^-, NO₂^- and PO₄^3-) follows specific kinetics; successive identifiable stages related to photoperiod lead to a complete stripping of nutrients. In addition to ion concentrations, pH and cell population were determined every 2 h during the experiment. Results and conclusions are presented.     

Algal and invertebrate communities in three subarctic lakes receiving mine wastes

The effects of liquid mine wastes on the density, species composition and diversity of phytoplankton and zooplankton were determined between May 1977 and May 1978 in a series of three small, shallow lakes (Meg, Keg, Peg Lakes), situated in the Canadian subarctic. The concentrations of the most common toxicants fluctuated widely during the study in each of the lakes. Meg Lake was the most polluted, with the concentrations of arsenic, lead, cyanide and copper reaching 3.0, 0.65, 0.60 and 0.65 g m⁻³ respectively. Although the levels of arsenic and lead in the other two lakes were roughly similar to those recorded in Meg Lake, cyanide and copper usually fell below detectable limits. Overall, changes in the level of contamination were poorly correlated with variations in the abundance of the flora and fauna. While phytoplankton (Chlamydomonas lapponica, Aphanothece microspora) were common in Meg Lake, reaching densities of 200 mg m⁻³, zooplankton were either rare or absent. Algal populations in Keg and Peg Lakes, which were dominated by Chlamydomonas lapponica and Oocystis parva, attained maximum densities of 200 and 150 mg m⁻³ respectively. The most common zooplankter, Keratella quadrata, maintained populations of up to 20–40 × 10⁺ animals m⁻³, and was followed in abundance by Daphnia middendorfiana and Cyclops bicuspidatus thomasi. It is concluded that: (1) The prevalent species may have developed a resistance to metals, (2) taxa not previously associated with heavy metal contamination may have the ability to adapt to pollution, and (3) high arsenic and lead levels probably had less impact on zooplankton than on phytoplankton, under field conditions.     

Denitrification with natural gas and various new growth media

Biological denitrification was investigated in an attached growth reactor system using several growth media, denitrifying cultures and natural gas (95% methane) as a carbon source. In order to establish a baseline of operation, initial experiments were conducted with a bed of 2–3 mm sand and methanol as a carbon source using a methylotrophic denitrifying culture and then the system was compared with natural gas using various methane utilising cultures. Compared to methanol, performance with methane was considerably lower. In order to improve denitrification with methane a plastic medium (Etapak, surface area 200 m²/m³) was placed above the sand bed (which increased the surface area for bacteria growth in the upper part of the bed), and a new methanotrophic mixed culture (NCIMB-code 11085) was introduced to the system. This combination resulted in a 27% higher denitrification efficiency. Experiments were continued by systematically varying the operating conditions to obtain highest denitrification using methane gas and replacing the Etapak media with different plastic media of higher surface area, but keeping the NCIMB culture unchanged. Other media tested were Pall-rings (surface area 319 m²/m³), IP-spacers (surface area 500 m²/m³) and granular activated carbon (GAC-code: Norit PK 1–3). Best results were obtained with IP-spacers which, surprisingly, are designed for use in the concrete industry rather than as a bacterial support medium. These produced nitrate removal efficiencies of up to 93% at 0.6 m/h or 55% at 1.6 m/h water filtration rates. Run times of 10 days or more to a limiting headloss of about 1.0 m,were usually achieved before “bumping” or back-washing to reduce headloss. Effluent turbidities were generally below 1.0 NTU. Tests for bacteria present with GAC media and COD removal with IP spacers were also carried out. Results are discussed with operational conditions and denitrification efficiencies achieved.