Biotreatment of pulp mill bleachery effluents with the coelomycetous fungusStagonospora gigaspora

Summary The coelomyceteStagonospora gigaspora degrades lignin derivatives within pulp mill bleachery effluents. Besides dechlorination, 90% of the color was removed from CEH bleachery effluents. Lignin derivatives in the effluents of the EOP bleaching stages revealed more persistent against fungal attack. Toxicity of both effluents was diminished significantly byS. gigaspora.     

Decolorization of practical textile industry effluents by white rot fungus Coriolus versicolor IBL-04

Textile industry discharges a vast amount of unused synthetic dyes in effluents. The discharge of these effluents into rivers and lakes leads to a reduction in sunlight penetration in natural water bodies, which, in turn, decreases both photosynthetic activity and dissolved oxygen concentration rendering it toxic to living beings. This paper describes the decolorization potential of a local white rot fungus, Coriolus versicolor IBL-04 for practical industrial effluents collected from five different textile industries of Faisalabad, Pakistan. Screening of C. versicolor IBL-04 on five effluents showed best decolorization results (36.3%) for Arzoo Textile Industry (ART) effluent in 6 days followed by Crescent Textile Industry (CRT), Itmad Textile Industry (ITT), Megna Textile Industry (MGT) and Ayesha Textile Industry (AST) effluents. Optimization of different process parameters for ART effluent decolorization by C. versicolor IBL-04 showed that manganese peroxidase (MnP) (486 U/mL) was the lignolytic enzyme present in the culture filtrates with undetectable lignin peroxidase (LiP) and laccase. The MnP synthesis and effluent decolorization could be enhanced to 725 U/mL and 84.4%, respectively, with a significant time reduction to 3 days by optimizing pH and temperature and using 1% starch as a supplementary carbon source.     

Biological treatment of a pulp and paper industry effluent by Fomes lividus and Trametes versicolor

White rot fungi Fomes lividus and Trametes versicolor, isolated from the Western Ghats region of Tamil Nadu, India, were used to treat pulp and paper industry effluents on a laboratory scale and in a pilot scale. On the laboratory scale a maximum decolourization of 63.9% was achieved by T. versicolor on the fourth day. Inorganic chloride at a concentration of 765 mg/l, which corresponded to 227% of that in the untreated effluent, was liberated by F. lividus on the 10th day. The chemical oxygen demand (COD) was also reduced to 1984 mg/l (59.3%) by each of the two fungi. On the pilot scale, a maximum decolourization of 68% was obtained with the 6-day incubation by T. versicolor, inorganic chloride 475 mg/l (103%) was liberated on the seventh day by T. versicolor, and the COD was reduced to 1984 mg/l corresponding to 59.32% by F. lividus. These results suggested that F. lividus seems to be another candidate efficient for dechlorination of wastewater.     

Degradation of chlorinated lignin compounds in a bleach plant effluent by the white-rot fungus Trametes versicolor

Summary Chlorinated lignin derivatives in a combined bleach plant effluent from sulphite pulping were degraded by several white-rot fungi among which Trametes versicolor (Coriolus versicolor) strains were the most efficient. With glucose as co-substrate, about 90% colour reduction was achieved within 3 days. Simultaneously, the concentration of chloro-organic compounds measured as adsorbable organic halogens decreased by about 45%. As shown by gel chromatography, the high-molecular-weight fraction in the effluent was completely depolymerized while over 50% of total aromatic compounds were degraded. The presence of a co-substrate was necessary for all these activities of the fungus. The residue obtained after degradation was extremely recalcitrant and not further degradable. Offprint requests to: M. Bergbauer     

In vitro genotoxic evaluation of conventional bleached and biobleached softwood pulp mill effluents

The effluents of pulp and paper mills contain about 300 different chemical compounds; many of them are mutagens and clastogens. Genotoxic studies have shown that chlorination stage liquors are significantly more genotoxic, in the Ames Salmonella assay, than the other process of lignin extraction, and that lyophilized effluents are genotoxic in cultured mammalian cells. Since these effluents from conventional bleaching stages are genotoxic, Chilean industries are interested in changing this process to a less toxic one, such as biobleaching using enzymes. In this study, we tested the in vitro genotoxicity of two types of effluents: an effluent obtained from a conventional radiata pine kraft-bleaching process (effluent D) and one derived from a biobleaching process with hemicellulase (effluent B). Both effluents were tested without any concentration or purification steps in the Ames Salmonella assay (TA100) and in the micronuclei (MN) and sister chromatid exchange (SCE) tests in CHO cells. The results showed that neither effluent induced base pair substitution mutations in the Ames Salmonella assay, and neither increased the micronucleus frequency in CHO cells. But, both increased the SCE frequencies in CHO cells, showing that this assay is more sensitive than the other ones, and that the two effluents contained chemical compounds in amounts enough to induce in vitro genotoxicity measured by the SCE induction.     

Decolorization of phenolic effluents by soluble and immobilized phenol oxidases

Summary Colour removal from phenplic industrial effluents by phenol oxidase enzymes and white-rot fungi was compared. Soluble laccase and horseradish peroxidase (HRP) removed colour from pulp mill (E), cotton mill hydroxide (OH) and cotton mill sulphide (S) effluents, but rapid and irreversible enzyme inactivation took place. Entrapment of laccase in alginate beads improved decolorization by factors of 3.5 (OH) and 2 (E); entrapment of HRP improved decolorization by 36 (OH), 20 (E) and 9 (S). Beads were unsuitable for continuous use because the enzymes were rapidly released into solution. Co-polymerization of laccase or HRP with L-tyrosine gave insoluble polymers with enzyme activity. Entrapment of the co-polymers in gel beads further increased the efficiency of decolorization of E by 28 (laccase) and by 132 (HRP) compared with soluble enzymes. Maximum decolorization of all three effluents by batch cultures of Coriolus versicolor (70%–80% in 8 days) was greater than the maximum enzymic decolorization (48% of OH in 3 days by entrapped laccase). Soluble laccase (222 units ml^–1) precipitated 1.2 g l^–1 phenol from artificial coal conversion effluent at pH 6.0 and the rate of precipitation and enzyme inactivation was faster at pH 6.0 than at pH 8.5.Offprint requests to: R. G. Burns     

Effective bioremoval of reactive dye and heavy metals by Aspergillus versicolor

In this study, bioaccumulation of heavy metal and dye by Aspergillus versicolor was investigated. Optimum pH values of the maximum heavy metal bioaccumulation was found as 6 for 50 mg/L Cr(VI), Ni(II) and 5 for Cu(II) ions with the 99.89%, 30.05% and 29.06% removal yield, respectively. The bioremoval of the dye up to 800 mg/L at pH 5 and 6 was investigated and 27.15% and 28.95% removal rates were measured respectively. The presence of Cr(VI) with dye, decreased the uptake yield for both pollutants. In the medium with Cu(II) and dye, dye removal was not affected by Cu(II), but Cu(II) removal rate increased from 29.06% to 37.91% by the existence of the dye. When Ni(II) and dye were combined, neither pollutant affected the other’s removal yield. These results indicate that the isolated A. versicolor strain deserves attention as a promising bioaccumulator of heavy metal ions and reactive dyes in wastewater effluents.     

Effects of Crude and Bioremediated Thermal Power Plant Effluents in Brassica Juncea

Crude phenol and cyanides are the major components of the effluents discharged by industries involved in the manufacture of many synthetic inorganic and organic compounds, pharmaceuticals, electroplating units, and thermal power plants.

In this study, an effort was made to use effluents as manure. The effect of various amendments (10, 40, 70, and 100%) of bioremediated and crude effluents was checked on plants of Brassica juncea. Roots, shoot lengths, and the mean dry weights were maximum in the plants irrigated with 10% treated and 10% crude effluent as compared to the control plants. Total plant phenolics increased with an increase in the amendment of the effluents in the plants irrigated with bioremediated as well as crude effluents. Applications of lower amendments of treated and untreated effluents in Brassica juncea improved the chlorophyll levels in the experimental plants and the values obtained were similar to the control plants. Effect of bioremediated and crude effluents on antioxidant enzymes of Brassica juncea showed a differential effect. The activity of antioxidant enzymes peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) was observed to increase with an increase in the effluent concentration.

Here we demonstrate that the industrial effluents, when bioremediated of their harmful components, can also serve as a nourishment for plants, which can further help in the rehabilitation of wastelands.     

Kinetic studies of laccase enzyme of Coriolus versicolor MTCC 138 in an inexpensive culture medium

An attempt was made to use cyanobacterial biomass of water bloom, groundnut shell (GNS) and dye effluent as culture medium for laccase enzyme production by Coriolus versicolor. Laccase production was found to be 10.15 ± 2.21 U/ml in the medium containing groundnut shell and cyanobacterial bloom in a ratio of 9:1 (dry weight basis) in submerged fermentation at initial pH 5.0 and 28 ± 2 °C temperature. Half life of enzyme was found to be 74 min at 60 °C. Kinetic analysis of laccase when made with substrate ABTS, K_m and V_max were found to be 0.29 mM and 9.49 μmol/min respectively. Azide and hydroxylamine were found to exert significant inhibition on thermostable laccase. Inhibitor constant (k_i) for azide and hydroxylamine were 1.33 and 0.18 mM respectively. This study forms the first report on the potential application of waste water cyanobacterial bloom and dyeing effluent as a medium for laccase production by C. versicolor MTCC138.     

Endophytic bacteria enhance remediation of tannery effluent in constructed wetlands vegetated with Leptochloa fusca

The use of constructed wetlands (CWs) is a promising approach for the remediation of wastewater. The present study aims to develop a plant–bacteria system within CWs for the efficient remediation of tannery effluent. In a vertical-flow CW vegetated with Leptochloa fusca (Kallar grass), a consortium of three different endophytic bacteria, Pantoea stewartii ASI11, Microbacterium arborescens HU33, and Enterobacter sp. HU38, was used for bioaugmentation. CWs vegetated with only L. fusca had the potential to remediate tannery effluent, but augmentation with endophytic bacteria enhanced the growth of L. fusca while aiding in the removal of both organic and inorganic pollutants from the tannery effluent. Moreover, the bacterial augmentation decreased toxicity in the effluent as well. A higher number of chromium (Cr)-resistant bacteria were isolated from the rhizosphere and endosphere of L. fusca inoculated with the endophytes than from uninoculated plants. Due to promising bioremediation and detoxification potential of L. fusca, it is reported for the first time as a potential candidate to develop effective CWs for the remediation of polluted effluents in combination with pollutant-degrading endophytic bacteria.     

A comparison of organochlorine removal from bleached kraft pulp and paper-mill effluents by dehalogenating Pseudomonas,Ancylobacter and Methylobacterium strains

The relative importance of each of three dechlorinating species to overall organochlorine removal from bleached kraft-mill effluents (BKME) was assessed. Ancylobacter aquaticus A7, Pseudomonas P1, and Methylobacterium CP13, strains indigenous to a BKME treatment system, were tested for growth on chlorinated acetic acids and alcohols, and for adsorbable organic halogen (AOX) reduction in batch cultures of sterile BKME from three sources. A. aquaticus A7 exhibited the broadest substrate range, but could only affect significant AOX reduction in softwood effluents. Methylobacterium CP13 exhibited a limited substrate range, but was capable of removing significant amounts of AOX from both hardwood and softwood effluents. By contrast, Pseudomonas sp. P1 exhibited a limited substrate range and poor to negligible reductions in AOX levels from both effluent types. Mixed inocula of all three species combined and inocula of sludge from mill treatment systems removed as much AOX from softwood effluents as did pure populations of Methylobacterium CP13. When BKME was hydrolysed prior to AOX analysis, the subsequent estimates of recalcitrant, or non-hydrolysable, AOX levels were far less variable than their counterpart total AOX measures. It is suggested that this is a relevant and useful measure of AOX for biodegradation studies.     

Decolorizing Reactive Textile Dyes with White‐Rot Fungi by Temporary Immersion Cultivation

The decontamination of effluents from textile industries is problematic due to the fact that textile dyes are resistant to degradation in the environment. Enzymes from white rot fungi, especially laccase, are able to degrade various complex aromatic structures, and are therefore able to decolorize textile dyes. The white‐rot fungi Trametes versicolor and Phanerochaete chrysosporium were immobilized, separately, on both pine wood chips and palm oil fiber, and cultivated in the temporary immersion RITA® (Récipient à Immersion Temporaire Automatique) System, which was adapted to serve as a fungal bioreactor in a series of four experiments to determine optimal conditions for decolorizing the textile dyes Levafix Blue and Remazol Brilliant Red. The maximum rate of decolorization of both dyes occurred within 24 h of incubation, and laccase was detected in the system.     

Degradation of diclofenac,trimethoprim, carbamazepine,and sulfamethoxazole by laccase from Trametes versicolor: Transformation products and toxicity of treated effluent

Abstract

The degradation of diclofenac (DCF), trimethoprim (TMP), carbamazepine (CBZ), and sulfamethoxazole (SMX) by laccase from Trametes versicolor was investigated. Experiments were conducted using the pharmaceuticals individually, or as a mixture at different initial concentrations (1.25 and 5?mg/L each). The initial enzymatic activity of all the treated samples was around 430–460?U_(DMP)/L. The removal of the four selected pharmaceuticals tested individually was more effective than when tested in mixtures under the same conditions. For example, 5?mg DCF/L was completely removed to below its detection limit (1?µg/L) within 8?h in the individual experiment vs. after 24?h when dosed as a mixture with the other pharmaceuticals. A similar trend was visible with other three pharmaceuticals, with 95 vs. 39%, 82 vs. 34% and 56 vs. 49% removal after 48?h with 5?mg/L of TMP, CBZ, and SMX tested individually or as mixtures, respectively. In addition, at the lower initial concentration (1.25?mg/L each), the removal efficiency of TMP, CBZ, and SMX in mixtures was lower than that obtained at the higher initial concentrations (5?mg/L each) during both the individual and combined treatments. Four enzymatic transformation products (TPs) were identified during the individual treatments of DCF and CBZ by T. versicolor. For TMP and SMX, no major TPs were observed under the experimental conditions used. The toxicity of the solution before and after enzymatic treatment of each pharmaceutical was also assessed and all treated effluent samples were verified to be non-toxic.     

13C NMR cross polarization and magic angle spinning (CPMAS) and gas chromatography/mass spectrometry analysis of the products from a soda pulp mill effluent decolourised by two Streptomyces strains

Two Streptomyces strains, UAH 30 and UAH 51, have been shown to decolourise a paper-mill effluent obtained after semichemical alkaline pulping of wheat straw. Fractionation of the effluent decolourised by strains UAH 30 and UAH 51 showed that 60% and 80% respectively of the alkali-lignin fraction have been removed from the effluent after 7 days of growth. ¹³C NMR cross polarization and magic angle spinning (CPMAS) spectra of the alkali-lignin remaining in the effluent after decolourisation revealed a decrease in the relative amount of aromatic lignin units compared to that obtained from the untreated effluent along with a reduction in the ratio of syringyl:guaiacyl units. Gas chromatography/mass spectrometry analysis of the low-molecular-mass compounds extracted from the decolourised effluent revealed the presence of new aromatic lignin-related compounds that were not present in the untreated control effluent. This was linked to a general depolymerization of larger lignin molecules occurring during decolourisation by the two Streptomyces strains. Identification of low-molecular-mass aromatic compounds extracted from the decolourised effluent revealed only the presence of p-hydroxyphenyl units in effluents decolourised by the strain UAH 30 while p-hydroxyphenyl, guaiacyl and syringyl units were detected in effluents decolourised by Streptomyces strain UAH 51. The study indicates that, while decolourisation is a common feature of the two Streptomyces strains, the mechanisms involved in the degradation of the lignin fractions may be different and strain-specific. Received: 8 July 1996 / Received revision: 9 October 1996 / Accepted: 14 October 1996     

Dechlorination of high-molecular-mass compounds in spent sulphite bleach effluents by free and immobilized cells of streptomycetes

Streptomyces chromofuscus and S. rochei dechlorinated high-molecular-mass compounds (HMM) from industrial bleach effluents. Compounds of the effluents from the first chlorination [(C + D)_red] and the subsequent alkaline extraction stage (E₁O) of a sulphite cellulose pulp mill were used as substrates for the microbial transformations. HMM bleach effluent fractions obtained by ultrafiltration were treated by free and immobilized cells in two types of bioreactors. The dechlorination was followed by measuring the reduction of activated-carbon-adsorbable organic-bound halogen (AOX) and the release of inorganic chloride. While 38–45% of the organic bound chlorine was released from a mixture of (C + D)_red and E₁O stage effluents within 20 days of incubation with S. chromofuscus, only 11–16% were liberated from E₁O-stage HMM bleach-effluent compounds by S. chromofuscus and S. rochei. Immobilized cells of both strains remained active for over 100 days in successive batches, each releasing 12–24% of chloride from HMM bleach-effluent compounds.     

Investigation of the sublethal effects of some petroleum refinery effluents

In Canada, environmental regulations for protection of the biota from the adverse effects of effluents from petroleum refineries have tended to focus on acute toxicity. There is concern those effluents may have other subtle, but still deleterious, long-term effects on aquatic ecosystems. We have used a battery of toxicity tests to assess the acute toxicity, genotoxicity, and chronic toxicity of effluent samples from two Ontario refineries. The test organisms included representatives of the bacterial, algal, plant, cladoceran, and fish communities. The results of our preliminary study indicate that the effluent samples had little acute toxicity to the test organisms. There were indications of some sublethal toxicity to Ceriodaphnia dubia, Panagrellus redivivus, and Pimephales promelas. One of the effluents inhibited the growth of Selanastrum capricornutum (IC₅₀ of 59.9%) and Lemna gibba (IC₂₅ of 73.3%) and also caused a 15 percent reduction in the germination of Lactuca sativa seeds. The SOS-Chromotest, a commercially available test that measures the activity of a bacterial DNA repair system, detected genotoxic effects in a single effluent that had been concentrated ten fold. There was no apparent relationship between several chemical parameters and the observed sublethal effects. Further research is needed to establish whether or not the observed toxic effects are typical of effluents from Ontario refineries.     

Decolorization of a dye industry effluent by <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> XC6

The strain Aspergillus fumigatus XC6 isolated from mildewing rice straw was evaluated for its ability to decolorize a dye industry effluent. The strain was capable of decolorizing dyes effluent over a pH range 3.0–8.0 with the dyes as sole carbon and nitrogen sources. The optimum pH was 3.0; however, supplemented with either appropriate nitrogen sources (0.2% NH₄Cl or (NH₄)₂SO₄ ) or carbon sources (1.0% sucrose or potato starch), the strain decolorized the effluent completely at the original pH of the dyes effluent. Therefore, A. fumigatus XC6 is an efficient strain for the decolorization of reactive textile dyes effluents, and it might be a practical alternative in dyeing wastewater treatment.     

On the dual localization of lipoprotein lipase in rat heart. Studies with a modified perfusion technique

Rat hearts were perfused $\text{in vitro}$ using a modified $\text{Langendorff}$ technique, allowing the separate collection of coronary- and interstitial effluents. When heparin was added to the perfusion medium lipoprotein lipase was found in the coronary, as well as in the interstitial effluents. The relative amounts of lipase activity in both effluents varied with the feeding conditions of the animals, being high in the coronary effluent during fasting and high in the interstitial effluent during feeding. When glucagon (2.10^?7 M) was included in the perfusion medium, no differences between fasted and fed animals were obtained. The apparent K_m of the interstitial lipase was lower than that of the lipase found in the coronary effluent. The results are discussed in the light of the localization of lipoprotein lipase in rat hearts $\text{in situ}$.     

Phytoremediation potential of Lemna minor L. for heavy metals

Phytoremediation potential of L. minor for cadmium (Cd), copper (Cu), lead (Pb), and nickel (Ni) from two different types of effluent in raw form was evaluated in a glass house experiment using hydroponic studies for a period of 31 days. Heavy metals concentration in water and plant sample was analyzed at 3, 10, 17, 24, and 31 day. Removal efficiency, metal uptake and bio-concentration factor were also calculated. Effluents were initially analyzed for physical, chemical and microbiological parameters and results indicated that municipal effluent (ME) was highly contaminated in terms of nutrient and organic load than sewage mixed industrial effluent (SMIE). Results confirmed the accumulation of heavy metals within plant and subsequent decrease in the effluents. Removal efficiency was greater than 80% for all metals and maximum removal was observed for nickel (99%) from SMIE. Accumulation and uptake of lead in dry biomass was significantly higher than other metals. Bio-concentration factors were less than 1000 and maximum BCFs were found for copper (558) and lead (523.1) indicated that plant is a moderate accumulator of both metals. Overall, L. minor showed better performance from SMIE and was more effective in extracting lead than other metals.     

Reuse of Textile Wastewater after Treatment with Isolated Bacteria from Oued Hamdoun River

Industrialization is a boon for developing countries such as Tunisia. However, textile effluents that are being discharged are environmental pollutants, extremely toxic to plants and other living organisms, including humans. The current study was conducted to determine the phytotoxic effect of textile effluents, collected near an industrial zone in the center of Tunisia (Ksar Helal), on the germination and various growth indices of durum wheat (Triticumaestivum L.). Results showed that textile effluent treatments reduced significantly the percentage of seed germination and slowed its kinetic as compared with control. Roots and leaves were also significantly reduced. The phytotoxicity was highly reduced from textile effluents after aerobic biotreatment with bacteria. It can be concluded that the biological treatment process of textile wastewater might serve as a fertilizer production that is able to improve the growth of plants. These results are encouraging in the context of developing a low-budget technology for the effective management of these effluents.