Removal and detoxification of pentahalogenated phenols using a photocatalytically induced enzymatic process.

Poly-halogenated phenols generated from a range of industrial processes can find their way into rivers and ground water. Here we report on a potential treatment for reducing the toxicity of these aqueous pollutants using two highly toxic penta-halogenated phenols (pentachlorophenol (PCP) and pentabromophenol (PBP)) as surrogates. Solutions were passed through a glass column packed with a silica support fused with titanium dioxide (TiO2) and horseradish peroxidase (HRP) immobilized on its TiO2/glass surface (HRP-Tglass). TiO2 photocatalysis was activated through irradiation with UVB (320 nm) which in turn activated the HRP. Two operational flow rates (0.5 and 1.25 mL min-1; hydraulic retention times (HRTs) of 20 and 8 min, respectively), tested the effect of retention time on the extent of degradation and reduction in toxicity of the treated effluent. Microtox® was used to measure the toxicity of the substrate and its by-products at both flow rates. At the highest flow rate, dehalogenation was limited (removal of 37 % chlorine and 22 % bromine) and the toxicity of the reaction products increased. At the lowest flow rate, the longer exposure time resulted in approximately 97 % and 96 % transformation of PCP and PBP, respectively, a greater degree of dehalogenation (removal of 65 % chlorine and 70 % bromine) and a substantial decrease in toxicity of the treated solutions. The higher toxicity of effluent from the higher flow rate was attributed to the initial degradation products being more toxic than the substrates. With a longer HRT, these were then further broken down to less toxic products. Additional toxicity tests (Hydra hexactinella (Hydra) and Chinese Hamster Ovary (CHO) cell toxicity were conducted on the effluent from the lowest flow rate. Both were less sensitive than the Microtox test, with Hydra proving more sensitive than CHO. The novelty of this work is the toxicity risk assessment of the products resulting from the use of a spatially separated immobilized enzyme and photooxidation system. The system was robust and showed no decrease in treatment efficacy over 10 h.

Impact of Microcystis aeruginosa on membrane fouling in a biologically treated effluent.

Microcystis aeruginosa was cultured in biologically treated municipal effluent to simulate blue-green algal bloom conditions in a treatment lagoon. The effect of algae in the early, mid and late phases of growth on membrane fouling, chemical coagulation (alum or aluminium chlorohydrate (ACH)) and hydraulic cleaning on the microfiltration of this effluent was investigated. The effect of M. aeruginosa in the early phase was negligible and gave a similar flux profile and permeate volume to that of effluent alone. The increase in M. aeruginosa concentration for the mid and late phases caused a significant reduction in permeate volume compared with the early phase. Full flux recovery was achieved with an alum dose of 1 mg Al3+ L(-1) (early phase) and 10 mg Al3+ L(-1) (mid phase), demonstrating that membrane fouling was hydraulically reversible. For the late phase, the highest flux recovery was 89%, which was achieved with an alum dose of 5 mg Al3+ L(-1). Higher alum dosages resulted in a reduction in flux recovery. The use of 1.5 pm pre-filtration after alum treatment showed little improvement in water quality but led to a drastic reduction in flux recovery, which was attributed to diminishing the protective layer on the membrane surface, thus enabling internal fouling. The performance of ACH was comparable to alum at low dissolved organic carbon (DOC) and cell concentration, but was not as effective as alum at high DOC and cell concentration due to the formation of more compact ACH flocs, which resulted in a higher cake layer specific resistance, leading to the deterioration of performance.

Potential of UV/H2O2 oxidation for enhancing the biodegradability of municipal reverse osmosis concentrates.

UVC/H2O2 and VUV/H2O2 oxidation processes were evaluated for the degradation of organic pollutants in reverse osmosis concentrate (ROC) produced from a municipal secondary effluent. It was found that the oxidation by UVC/H2O2 and VUV/H2O2 processes could be described as a pseudo first-order reaction. For UVC increased oxidation occurred with increasing H2O2 dosage up to 2 mM above which improvement in oxidation performance decreased. At the same H2O2 dosage, VUV irradiation gave better overall oxidation performance. Compared with UVC/6 mM H2O2, VUV/2 mM H2O2 gave a greater rate of reduction of chemical oxygen demand, but a lower rate for reduction of dissolved organic carbon, suggesting that oxidation of organics by the two methods followed different pathways. The change of absorbance at 254 nm and fluorescence excitation emission matrix spectra of irradiated samples indicated that the large and complex compounds were fragmented rapidly by the *OH, resulting in rapid decolourisation. The biodegradability of the organics in the ROC was increased from 11% to 35% after 1 h treatment by UVC/3 mM H2O2, whereas a greater increase (41%) was obtained with VUV/2 mM H2O2. This increase in biodegradability indicates the potential for employing a subsequent biological treatment process.

Membrane foulants and fouling mechanisms in microfiltration and ultrafiltration of an activated sludge effluent.

Membrane fouling in microfiltration (MF) and ultrafiltration (UF) of an activated sludge (AS) effluent was investigated. It was found that the major membrane foulants were polysaccharides, proteins, polysaccharide-like and protein-like materials and humic substances. MF fouling by the raw effluent was governed by pore adsorption of particles smaller than the pores during the first 30 minutes of filtration and then followed the cake filtration model. UF fouling could be described by the cake filtration model throughout the course of filtration. Coagulation with alum and (poly)aluminium chlorohydrate (ACH) altered the MF fouling mechanism to follow the cake filtration model from the beginning of filtration. The MF and UF flux improvement by coagulation was due to the removal of some of the foulants in the raw AS effluent by the coagulants. The MF flux improvement was greater for alum than for ACH whereas the two coagulants performed equally well in UF. Coagulation also reduced hydraulically irreversible fouling on the membranes and this effect was more prominent in MF than in UF. The unified membrane fouling index (UMFI) was used to quantitatively evaluate the effectiveness of coagulation on membrane flux enhancement.

Continuous enzymatic treatment of 4-bromophenol initiated by UV irradiation.

Horseradish peroxidase (HRP) can be used for the treatment of halogenated phenolic substances. In the presence of hydrogen peroxide phenols are oxidized to form polymers which undergo partial dehalogenation. However, when immobilized, the peroxidase is subject to inactivation due to blockage of the active sites by the growing polymers and to deactivation by elevated levels of hydrogen peroxide. When HRP immobilized on a novel glass-based support incorporating titanium dioxide is subjected to UV irradiation, hydrogen peroxide is produced and the nascent polymer is removed. In this work a reactor was constructed that utilized HRP immobilized on the novel support and the in situ production of hydrogen peroxide to treat 4-bromophenol as a model substrate. The system was operated for almost 17 hours with no apparent decline in activity.

Reducing the effect of cyanobacteria in the microfiltration of secondary effluent.

Cyanobacterial blooms in the lagoons of sewage treatment plants can severely impact the performance of membrane plants treating the effluent. This paper investigates the impact of Microcystis aeruginosa in a secondary effluent on the microfiltration filterability and cleaning of the membrane. Alum coagulation and dissolved air flotation (DAF) were investigated to remove the algae and so enhance the volume of effluent processed, and their influence on reversible and irreversible fouling. Degree of fouling due to the algal components was found to be in decreasing order of algal cells, algal organic matter and extracellular organic matter. Alum coagulation with 5 mg L⁻¹ as Al³(+) led to a substantial increase in permeate volume, an increase in dissolved organic carbon removal, and a foulant layer which protected the membrane from internal fouling but which was hydraulically removable resulting in full flux recovery. Pre-treatment by DAF or 1.5 µm filtration following alum coagulation enhanced the flux rate and permeate volume but exposed the membrane to internal irreversible fouling.

Identification of key water quality characteristics affecting the filterability of biologically treated effluent in low-pressure membrane filtration.

There are many water quality characteristics which could influence the filterability of biologically treated effluent from Melbourne's Western Treatment Plant (WTP). Statistical correlation was used to identify the key water characteristics affecting the microfiltration (MF) and ultrafiltration (UF) filterability in terms of permeate volume of the treated effluent. The models developed showed that turbidity, dissolved organic carbon (DOC) and total suspended solids (TSS) were the key factors which influenced the MF and UF filterability. Turbidity was the dominant factor affecting the accuracy of the model for MF filterability while DOC was the major factor affecting the accuracy of the model for UF filterability. A prediction accuracy of 85% was obtained for MF and 86% for UF filterability of the WTP effluent. The characteristics of the organic components of the wastewater were demonstrated by EEM spectra to have seasonal variation which would have reduced the prediction accuracy. As turbidity, DOC and TSS can be determined on-line, the models would be useful for rapid prediction of the filterability of WTP effluent and this may assist the control of low-pressure membrane filtration processes.

Greywater treatment by UVC/H2O2.

Greywater treatment by UVC/H(2)O(2) was investigated with regard to the removal of chemical oxygen demand (COD). A COD reduction from 225 to 30 mgl(-1) (overall removal of 87%) was achieved after settling overnight and subsequent irradiation for 3h with 10mM H(2)O(2). Most of the contaminants were removed by oxidation since only 13% COD was removed by settlement. The removal of COD in the greywater followed a second-order kinetic equation, r=0.0637[COD][H(2)O(2)], up to 10mM H(2)O(2). A slightly enhanced COD removal was observed at the initial pH of 10 compared with pH 3 and 7. This was attributed to the dissociation of H(2)O(2) to O(2)H(-). The treatment was not affected by total concentration of carbonate (c(T)) of at least 3 mM, above which operation between pH 3 and 5 was essential. The initial biodegradability of the settled greywater (as BOD(5):COD) was 0.22. After 2h UVC/H(2)O(2) treatment, a higher proportion of the residual contaminants was biodegradable (BOD(5):COD=0.41) which indicated its potential as a pre-treatment for a biological process.

Chemical and biological oxidation of NOM surrogates and effect on HAA formation.

Formation of disinfection by-products (DBPs) can be controlled by removal of disinfection by-product precursors before disinfection. Variable success has been reported, depending on the treatment used and water tested. Chemical and biological oxidations are candidate technologies to control DBP formation. Given the uncertainty over the identity of DBP precursors, the use of surrogates of natural organic matter (NOM) allows fundamental probing of the links between compound character, removal and DBP formation. Nine compounds were chosen to represent NOM and their removal by two advanced oxidation processes (AOPs), UV-C irradiation and biological treatment compared while haloacetic acid (HAA) formation before and after treatment was measured. Although AOPs were able to fully remove all compounds, incomplete mineralisation led to increased HAA levels, dramatically in the case of two amino acids. Biological treatment was effective in removing amino acids but also moderately increased the HAA formation potential (HAAFP) of hydrophilic compounds. These findings indicate waters with high amino acid concentrations will be susceptible to raised HAA levels following AOP treatment and careful process selection for HAA control is required in such cases.

Isolation and screening of natural organic matter-degrading fungi.

Drinking water quality and its treatment are negatively impacted by the presence of coloured natural organic matter (NOM) derived from the breakdown of animal and plant materials. Ligninolytic fungi (i.e., white rot fungi - WRF) secrete non-specific oxidative enzymes that can oxidise a wide range of recalcitrant organic compounds. The potential for these organisms to decolourise concentrated aquatic NOM was investigated. Twenty-one isolates from diverse fungal genera were screened using NOM plate assays. Four WRF strains: Trametes sp., Polyporus sp., Trametes versicolor ATCC 7731 and Bjerkandera adusta, which displayed good NOM decolourisation on solid medium were further investigated in shake-flask culture using concentrated NOM as the only source of nutrients. Of these, B. adusta demonstrated the greatest decolourisation (65% for 100 mg C L(-1) NOM). NOM decolourisation coincided with ligninolytic enzyme activity and decrease in average molecular weight of NOM. The expression of the oxidative enzymes (manganese peroxidase (MnP), lignin peroxidase and laccase (Lac)) varied with fungal strain. The enzyme activities of Polyporus sp. and the two Trametes strains were significantly greater than those of B. adusta, although their decolourisation was less. For the Trametes and Polyporus sp., Lac activity was greatest, whereas for B. adusta MnP activity was greatest, suggesting its predominant role in the decolourisation process. This research demonstrates the significant potential for WRF in NOM removal so long as the enzyme activity can be controlled.

Removal of VUV pre-treated natural organic matter by biologically activated carbon columns.

A potential alternative water treatment process using VUV (185 nm+254 nm) irradiation followed by a biological treatment is described. The system uses sufficient VUV radiation (16J cm(-2)) to significantly enhance the production of biologically degradable moieties prior to treatment with biologically activated carbon (BAC). Two similar activated carbons were used, one virgin and one taken from a water treatment plant with an established biofilm. The VUV-BAC process decreased the overall dissolved organic carbon (DOC) concentration of a natural water sample by 54% and 44% for the virgin carbon and previously used BAC, respectively. Furthermore, VUV-BAC treatment decreased the trihalomethane (THM) formation potential (THMFP) by 60-70% and the haloacetic acid (HAA) formation potential (HAAFP) by 74%. The BAC systems effectively removed the hydrogen peroxide residual produced by VUV irradiation. Although nitrite formation can result from VUV treatment of natural organic matter (NOM), none was detected before or after BAC treatment.

Formation of hazardous by-products resulting from the irradiation of natural organic matter: comparison between UV and VUV irradiation.

The use of ultraviolet (UV) or vacuum ultraviolet (VUV) photo-oxidation followed by biological treatment for the removal of natural organic matter (NOM) in drinking water is a potential water treatment technique under investigation. This paper reports on the trihalomethane formation potential (THMFP), the haloacetic acid formation potential (HAAFP), and formation of nitrite and peroxide following both UV and VUV irradiation of NOM prior to biological treatment. The total THMFP was found to decrease with increasing UV and VUV irradiation dose, although there was a linear increase in bromoform formation. Determination of the THMFP of NOM fractions obtained after irradiation, showed that the hydrophobic fraction was dominated by chlorinated species which accounted for the majority of the total THMFP, while bromoform was observed only in the hydrophilic fraction of NOM. VUV irradiation reduced the HAAFP with increasing dose, in contrast, UV irradiation had a limited effect on the overall HAAFP. Following UV or VUV irradiation, the chlorinated species accounted for the majority of HAAFP; however, significant formation of brominated haloacetic acid (HAA) was observed. The nitrate concentration of the untreated water directly influenced the concentration of nitrite produced as a consequence of UV and VUV irradiation. Hydrogen peroxide formation was greater during VUV irradiation than during UV irradiation. Samples exposed to various doses of UV or VUV irradiation (up to 138 J cm(-2)) were deemed non-cytotoxic (African green monkey kidney cells) and non-mutagenic (Ames test).

Fractionation of UV and VUV pretreated natural organic matter from drinking water.

Recent studies have examined the potential of ultraviolet (UV, 254 nm) and vacuum ultraviolet (VUV, 185 nm + 254 nm) irradiation as either a pretreatment for a biological process or as a sole treatment for the removal of natural organic matter as dissolved organic carbon from drinking water. To understand the potential of UV and VUV irradiation followed by subsequent biological treatment, treated water was fractionated into four components: very hydrophobic acid (VHA), slightly hydrophobic acid (SHA), hydrophilic charged (CHA), and hydrophilic neutral (NEU). The VHA fraction was found to be very susceptible to both UV and VUV irradiation, and the fragmentation products of the high molecular weight VHA and SHA molecules contributed to the CHA and NEU fractions to form a pool of biodegradable, non-UV-absorbing, low molecular weight moieties. The NEU fraction was the most difficult to remove, as most of the components in this fraction were refractory to both the biological and photo-oxidative processes. Therefore, enhanced removal of the NEU fraction is required to increase the effectiveness and potential of the treatment process.

Preliminary toxicity assessment of water after treatment with UV-irradiation and UVC/H2O2.

Photooxidation (UV radiation) and enhanced photooxidation (UVC/H2O2) are water treatment technologies which remove aquatic natural organic matter (NOM) by photodegradation, producing lower molecular weight components and CO2. Since these technologies are being investigated for the treatment of drinking water, knowledge of the potential toxicity of the photooxidation by-products is vital. The potential toxicity of UVA-, UVB-, UVC-irradiated, and UVC/H2O2-treated aquatic NOM in two spot samples from two Australian reservoirs was analysed in two spot samples using Vibriofischeri in the Microtox test, African green monkey kidney cells (AS/NZS 4020:1999), and Daphnia carinata in an acute immobilisation test. Toxicity was not apparent for both the Microtox procedure and cytotoxicity analyses for the UVC-irradiated and UVC/H2O2-treated NOM samples, while UVA- and UVB-irradiated water samples were non-toxic to D. carinata. In contrast, acute toxicity was observed for UVC- and UVC/H2O2-treated water samples. The observed toxicity was attributed to photooxidative degradation of NOM-metal binding sites, which resulted in the release of bioavailable copper ions, as evidenced by higher concentrations of free copper ions in photooxidised water. As the total copper concentrations of the two raw water samples were well below the Australian Water Quality Guidelines for metals in domestic supplies, the release of copper from photooxidised NOM is unlikely to cause health concerns in these samples.

Removal of dissolved organic compounds in fixed-bed columns: evaluation of low-rank coal adsorbents.

Characterisation of a range of Victorian low-rank coal (VLRC) based and commercial adsorbents under continuous flow conditions was conducted in down flow fixed-bed columns. The effect of bed depth, hydraulic loading and initial concentration of the adsorbate 4-nitrophenol (4-NP) was studied. Prediction of the performance of the columns using bed-depth/service time analysis showed good agreement with the experimental results. The VLRC-based activated carbons gave lower service times for removal of 4-NP compared with the coconut-based commercial activated carbon Picactif. However, they showed comparable efficiency in terms of bed volumes treated and carbon usage rate (CUR). The VLRC-activated power station char (APSC) showed the lowest CUR of 0.57 g/L in comparison with 0.62 and 3.61 g/L exhibited by the commercial carbons Picactif and Hydraffin, respectively. The power station char (PSC) and Auschar gave poor CUR of 11.23 and 75.36 g/L, respectively. Three adsorbents were evaluated for the removal of natural organic matter (NOM) from aqueous solution. The breakthrough behaviour indicated that the pore size distribution of the adsorbents is an important physical characteristic for the adsorption of the NOM. There was a non-adsorbable fraction of the NOM that was a function of the type of adsorbent. In the presence of NOM, Picactif gave longer service time for the removal of 4-NP compared with APSC. consistent with the trend obtained in the absence of NOM. However, lower breakthrough times (at 10% C0) of 44 and 47% were obtained for APSC and Picactif, respectively. Further removal (35%) of 4-NP was achieved over extended operation of the beds, however more frequent backwashing was required as a result of the biological growth exhibited in the presence of NOM.

Influence of the characteristics of natural organic matter on the fouling of microfiltration membranes.

Natural organic matter (NOM) plays a significant role in fouling microfiltration membranes in drinking water treatment processes even though the NOM is retained only to a small extent. The aim of this study was to obtain a better understanding of the interactions between the fractional components of NOM and microfiltration membranes. Filtration experiments were performed using 0.22 microm hydrophobic and hydrophilic polyvinylidene fluoride (PVDF) membranes in a stirred-cell system on the NOM isolated from three Australian surface waters. As expected, the fouling rate for the hydrophobic membrane was considerably greater than for the hydrophilic membrane. Focusing on the hydrophobic membrane, it was shown that the high molecular weight fraction of NOM ( > 30 kDa) was responsible for the major flux decline. Filtration tests on the four fractions of NOM isolated on the basis of hydrophobicity and charge using non-functionalised and anionic resins revealed that the fouling potential for the three waters was hydrophilic neutral > hydrophobic acids > transphilic acids > hydrophilic charged. The low-aromatic hydrophilic neutral compounds were the main determinant of the rate and extent of flux decline. This was linked to the colloidal size fraction ( > 30 kDa) and to the selective concentration of calcium in the fraction leading to organics-Ca2+ bridging. It was also shown that the higher the aromaticity of the NOM the greater the flux decline, and the aromatics mainly resided in the hydrophobic acids fraction. Overall, the fouling mechanism controlling the flux decline involved the combined effects of adsorptive and colloidal fouling by the hydrophilic neutral fraction in the internal pore structure of the membrane.

Factors affecting the production of L-phenylacetylcarbinol by yeast: a case study.

L-Phenylacetylcarbinol (L-PAC) is the precursor for L-ephedrine and D-pseudoephedrine, alkaloids possessing alpha- and beta-adrenergic activity. The most commonly used method for production of L-PAC is a biological method whereby the enzyme pyruvate decarboxylase (PDC) decarboxylates pyruvate and then condenses the product with added benzaldehyde. The process may be undertaken by either whole cells or purified PDC. If whole cells are used, the biomass may be grown and allowed to synthesize endogenous pyruvate, or the cells may be used as a catalyst only, with both pyruvate and benzaldehyde being added. Several yeast species have been investigated with regard to L-PAC-producing potential; the most commonly used organisms are strains of Saccharomyces cerevisiae and Candida utilis. It was found that initial high production rates did not necessarily result in the highest final yields. Researchers then examined ways of improving the productivity of the process. The substrate, benzaldehyde, and the product, L-PAC, as well as the by-products, were found to be toxic to the biomass. Methods examined to reduce toxicity include modification of benzaldehyde dosing regimes, immobilization of biomass or purified enzymes, modification of benzaldehyde solubility and the use of two-phase reaction systems. Various means of modifying metabolism to enhance enzyme activity, relevant metabolic pathways and yield have been examined. Methods investigated include the use of respiratory quotient to influence pyruvate production and induce fermentative activity, reduced aeration to increase PDC activity, and carbohydrate feeding to modify glycolytic enzyme activity. The effect of temperature on L-PAC yield has been examined to identify conditions which provide the optimal balance between L-PAC and benzyl alcohol production, and L-PAC inactivation. However, relatively little work has been undertaken on the effect of medium composition on L-PAC yield.

Biochemical and immunochemical characterisation of strains of Treponema hyodysenteriae.

The protein composition of 18 clinical isolates of Treponema hyodysenteriae from pigs with swine dysentery in Australia were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. Coomassie Blue stained SDS-PAGE-profiles of whole cell and outer membrane (OM) proteins demonstrated the same gel pattern among the T. hyodysenteriae isolates, particularly the OM proteins in the molecular mass (Mr) range of 30 kDa to 40 kDa. The T. hyodysenteriae isolates were categorised into two distinct groups (A and B) based on the strain-variability in the 37 kDa OM protein. Immunoblotting of whole cell proteins after SDS-PAGE using serum from rabbits and pigs immunised with known T. hyodysenteriae serotypes revealed a number of common immunoreactive bands in all isolates. LPS typing of the T. hyodysenteriae isolates by immunoblotting with the rabbit antiserum revealed one additional serotype emphasising the LPS heterogeneity among the strains isolated from geographic locations in Australia, Great Britain and the U.S.A. Immunoblotting of the OM preparations revealed several common immunoreactive polypeptides corresponding to Mr values of 34 kDa to 30 kDa among the T. hyodysenteriae and T. innocens isolates but a distinct 39 kDa found only in the T. hyodysenteriae isolates. Trypsin proteolysis of intact. T. hyodysenteriae cells caused selective loss of these and other major abundant proteins identifying the location of the 39 kDa, 36 kDa, 34 kDa and 30 kDa proteins on the cell surface and suggesting a possible role of these proteins in the pathogenesis of swine dysentery.