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1.  Quality of effluents from Hattar Industrial Estate 
Of 6634 registered industries in Pakistan, 1228 are considered to be highly polluting. The major industries include textile, pharmaceutical, chemicals (organic and inorganic), food industries, ceramics, steel, oil mills and leather tanning which spread all over four provinces, with the larger number located in Sindh and Punjab, with smaller number in North Western Frontier Province (NWFP) and Baluchistan. Hattar Industrial Estate extending over 700 acres located in Haripur district of NWFP is a new industrial estate, which has been developed with proper planning for management of industrial effluents. The major industries located in Hattar are ghee industry, chemical (sulfuric acid, synthetic fiber) industry, textile industry and pharmaceuticals industry. These industries, although developed with proper planning are discharging their effluents in the nearby natural drains and ultimately collected in a big drain near Wah. The farmers in the vicinity are using these effluents for growing vegetables and cereal crops due to shortage of water. In view of this discussion, there is a dire need to determine if these effluents are hazardous for soil and plant growth. So, effluents from different industries, sewage and normal tap water samples were collected and analysed for pH, electrical conductivity (EC), total soluble salts (TSS), biological oxygen demand (BOD), chemical oxygen demand (COD), total nitrogen, cations and anions and heavy metals. The effluents of ghee and textile industries are highly alkaline. EC and TSS loads of ghee and textile industries are also above the National Environmental Quality Standards (NEQS), Pakistan. All the effluents had residual sodium carbonates (RSCs), carbonates and bicarbonates in amounts that cannot be used for irrigation. Total toxic metals load in all the effluents is also above the limit i.e. 2.0 mg/L. Copper in effluents of textile and sewage, manganese in ghee industry effluents and iron contents in all the effluents were higher than NEQS. BOD and COD values of all the industries are also above the NEQS. On the whole, these effluents cannot be used for irrigation without proper treatment otherwise that may cause toxicity to soil, plants and animals as well add to the problems of salinity and sododicity. Similarly, these effluents cannot be used for fish farming.
PMCID: PMC1661672  PMID: 17111466
Industrial effluents; Biological oxygen demand (BOD); Chemical oxygen demand (COD); pH; Residual salts; Lead; Zinc; Copper; Nickel; Manganese; Sewage
2.  Removal of Mercury from Chloralkali Electrolysis Wastewater by a Mercury-Resistant Pseudomonas putida Strain 
Applied and Environmental Microbiology  1999;65(12):5279-5284.
A mercury-resistant bacterial strain which is able to reduce ionic mercury to metallic mercury was used to remediate in laboratory columns mercury-containing wastewater produced during electrolytic production of chlorine. Factory effluents from several chloralkali plants in Europe were analyzed, and these effluents contained total mercury concentrations between 1.6 and 7.6 mg/liter and high chloride concentrations (up to 25 g/liter) and had pH values which were either acidic (pH 2.4) or alkaline (pH 13.0). A mercury-resistant bacterial strain, Pseudomonas putida Spi3, was isolated from polluted river sediments. Biofilms of P. putida Spi3 were grown on porous carrier material in laboratory column bioreactors. The bioreactors were continuously fed with sterile synthetic model wastewater or nonsterile, neutralized, aerated chloralkali wastewater. We found that sodium chloride concentrations up to 24 g/liter did not inhibit microbial mercury retention and that mercury concentrations up to 7 mg/liter could be treated with the bacterial biofilm with no loss of activity. When wastewater samples from three different chloralkali plants in Europe were used, levels of mercury retention efficiency between 90 and 98% were obtained. Thus, microbial mercury removal is a potential biological treatment for chloralkali electrolysis wastewater.
PMCID: PMC91717  PMID: 10583977
3.  Catalytic ozonation-biological coupled processes for the treatment of industrial wastewater containing refractory chlorinated nitroaromatic compounds*  
A treatability study of industrial wastewater containing chlorinated nitroaromatic compounds (CNACs) by a catalytic ozonation process (COP) with a modified Mn/Co ceramic catalyst and an aerobic sequencing batch reactor (SBR) was investigated. A preliminary attempt to treat the diluted wastewater with a single SBR resulted in ineffective removal of the color, ammonia, total organic carbon (TOC) and chemical oxygen demand (COD). Next, COP was applied as a pretreatment in order to obtain a bio-compatible wastewater for SBR treatment in a second step. The effectiveness of the COP pretreatment was assessed by evaluating wastewater biodegradability enhancement (the ratio of biology oxygen demand after 5 d (BOD5) to COD), as well as monitoring the evolution of TOC, carbon oxidation state (COS), average oxidation state (AOS), color, and major pollutant concentrations with reaction time. In the COP, the catalyst preserved its catalytic properties even after 70 reuse cycles, exhibiting good durability and stability. The performance of SBR to treat COP effluent was also examined. At an organic loading rate of 2.0 kg COD/(m3·d), with hydraulic retention time (HRT)=10 h and temperature (30±2) °C, the average removal efficiencies of NH3-N, COD, BOD5, TOC, and color in a coupled COP/SBR process were about 80%, 95.8%, 93.8%, 97.6% and 99.3%, respectively, with average effluent concentrations of 10 mg/L, 128 mg/L, 27.5 mg/L, 25.0 mg/L, and 20 multiples, respectively, which were all consistent with the national standards for secondary discharge of industrial wastewater into a public sewerage system (GB 8978-1996). The results indicated that the coupling of COP with a biological process was proved to be a technically and economically effective method for treating industrial wastewater containing recalcitrant CNACs.
PMCID: PMC2833402  PMID: 20205304
Industrial wastewater; Catalytic ozonation; Sequencing batch reactor; Chlorinated nitroaromatic compounds
4.  The Enzymatic Decolorization of Textile Dyes by the Immobilized Polyphenol Oxidase from Quince Leaves 
The Scientific World Journal  2014;2014:685975.
Water pollution due to release of industrial wastewater has already become a serious problem in almost every industry using dyes to color its products. In this work, polyphenol oxidase enzyme from quince (Cydonia Oblonga) leaves immobilized on calcium alginate beads was used for the successful and effective decolorization of textile industrial effluent. Polyphenol oxidase (PPO) enzyme was extracted from quince (Cydonia Oblonga) leaves and immobilized on calcium alginate beads. The kinetic properties of free and immobilized PPO were determined. Quince leaf PPO enzyme stability was increased after immobilization. The immobilized and free enzymes were employed for the decolorization of textile dyes. The dye solutions were prepared in the concentration of 100 mg/L in distilled water and incubated with free and immobilized quince (Cydonia Oblonga) leaf PPO for one hour. The percent decolorization was calculated by taking untreated dye solution. Immobilized PPO was significantly more effective in decolorizing the dyes as compared to free enzyme. Our results showed that the immobilized quince leaf PPO enzyme could be efficiently used for the removal of synthetic dyes from industrial effluents.
PMCID: PMC3918733  PMID: 24587743
5.  Experimental study of dye removal from industrial wastewater by membrane technologies of reverse osmosis and nanofiltration 
Currently, biological method has been utilized in the treatment of wastewater -containing synthetic dyes used by textile industries in Iraq. The present work was devoted to study the operating feasibility using reverse osmosis (RO) and nanofiltration (NF) membrane systems as an alternative treatment method of wastewater discharged from Iraqi textile mills. Acid red, reactive black and reactive blue dyes were selected, based on the usage rate in Iraq. Effects of dye concentration, pH of solution, feed temperature, dissolved salts and operating pressure on permeate flux and dye rejection were studied. Results at operating conditions of dye concentration = 65 mg/L, feed temperature = 39°C and pressure = 8 bar showed the final dye removal with RO membrane as 97.2%, 99.58% and 99.9% for acid red, reactive black and reactive blue dyes, respectively. With NF membrane, the final dye removal were as 93.77%, 95.67%, and 97% for red, black and blue dyes, respectively. The presence of salt (particularly NaCl) in the dye solution resulted in a higher color removal with a permeate flux decline. It was confirmed that pH of solution had a positive impact on dye removal while feed temperature showed a different image. A comparison was made between the results of dye removal in biological and membrane methods. The results showed that membrane method had higher removal potential with lower effective cost. The present study indicates that the use of NF membrane in dye removal from the effluent of Iraqi textile mills is promising.
PMCID: PMC3570404  PMID: 23369335
Membrane separation; Synthetic dyes; Reverse osmosis; Nanofiltration; Wastewater reuse
6.  Combined Industrial Wastewater Treatment in Anaerobic Bioreactor Posttreated in Constructed Wetland 
BioMed Research International  2013;2013:957853.
Constructed wetland (CW) with monoculture of Arundo donax L. was investigated for the posttreatment of anaerobic bioreactor (ABR) treating combined industrial wastewater. Different dilutions of combined industrial wastewater (20, 40, 60, and 80) and original wastewater were fed into the ABR and then posttreated by the laboratory scale CW. The respective removal efficiencies of COD, BOD, TSS, nitrates, and ammonia were 80%, 78–82%, 91.7%, 88–92%, and 100% for original industrial wastewater treated in ABR. ABR was efficient in the removal of Ni, Pb, and Cd with removal efficiencies in the order of Cd (2.7%) > Ni (79%) > Pb (85%). Posttreatment of the ABR treated effluent was carried out in lab scale CW containing A. donax L. CW was effective in the removal of COD and various heavy metals present in ABR effluents. The posttreatment in CW resulted in reducing the metal concentrations to 1.95 mg/L, 0 mg/L, and 0.004 mg/L for Ni, Pb, and Cd which were within the permissible water quality standards for industrial effluents. The treatment strategy was effective and sustainable for the treatment of combined industrial wastewater.
PMCID: PMC3876802  PMID: 24396832
7.  Assessing the resistance and bioremediation ability of selected bacterial and protozoan species to heavy metals in metal-rich industrial wastewater 
BMC Microbiology  2013;13:28.
Heavy-metals exert considerable stress on the environment worldwide. This study assessed the resistance to and bioremediation of heavy-metals by selected protozoan and bacterial species in highly polluted industrial-wastewater. Specific variables (i.e. chemical oxygen demand, pH, dissolved oxygen) and the growth/die-off-rates of test organisms were measured using standard methods. Heavy-metal removals were determined in biomass and supernatant by the Inductively Couple Plasma Optical Emission Spectrometer. A parallel experiment was performed with dead microbial cells to assess the biosorption ability of test isolates.
The results revealed that the industrial-wastewater samples were highly polluted with heavy-metal concentrations exceeding by far the maximum limits (in mg/l) of 0.05-Co, 0.2-Ni, 0.1-Mn, 0.1-V, 0.01-Pb, 0.01-Cu, 0.1-Zn and 0.005-Cd, prescribed by the UN-FAO. Industrial-wastewater had no major effects on Pseudomonas putida, Bacillus licheniformis and Peranema sp. (growth rates up to 1.81, 1.45 and 1.43 d-1, respectively) compared to other test isolates. This was also revealed with significant COD increases (p < 0.05) in culture media inoculated with living bacterial isolates (over 100%) compared to protozoan isolates (up to 24% increase). Living Pseudomonas putida demonstrated the highest removal rates of heavy metals (Co-71%, Ni-51%, Mn-45%, V-83%, Pb-96%, Ti-100% and Cu-49%) followed by Bacillus licheniformis (Al-23% and Zn-53%) and Peranema sp. (Cd-42%). None of the dead cells were able to remove more than 25% of the heavy metals. Bacterial isolates contained the genes copC, chrB, cnrA3 and nccA encoding the resistance to Cu, Cr, Co-Ni and Cd-Ni-Co, respectively. Protozoan isolates contained only the genes encoding Cu and Cr resistance (copC and chrB genes). Peranema sp. was the only protozoan isolate which had an additional resistant gene cnrA3 encoding Co-Ni resistance.
Significant differences (p < 0.05) observed between dead and living microbial cells for metal-removal and the presence of certain metal-resistant genes indicated that the selected microbial isolates used both passive (biosorptive) and active (bioaccumulation) mechanisms to remove heavy metals from industrial wastewater. This study advocates the use of Peranema sp. as a potential candidate for the bioremediation of heavy-metals in wastewater treatment, in addition to Pseudomonas putida and Bacillus licheniformis.
PMCID: PMC3575345  PMID: 23387904
Industrial wastewater; Heavy metal; Bioremediation; Bacteria; Protozoa; Metal toxicity; Pollution; Metal-resistance
8.  Dual application of duckweed and azolla plants for wastewater treatment and renewable fuels and petrochemicals production 
Shortages in fresh water supplies today affects more than 1 billion people worldwide. Phytoremediation strategies, based on the abilities of aquatic plants to recycle nutrients offer an attractive solution for the bioremediation of water pollution and represents one of the most globally researched issues. The subsequent application of the biomass from the remediation for the production of fuels and petrochemicals offers an ecologically friendly and cost-effective solution for water pollution problems and production of value-added products.
In this paper, the feasibility of the dual application of duckweed and azolla aquatic plants for wastewater treatment and production of renewable fuels and petrochemicals is explored. The differences in absorption rates of the key wastewater nutrients, ammonium and phosphorus by these aquatic macrophytes were used as the basis for optimization of the composition of wastewater effluents. Analysis of pyrolysis products showed that azolla and algae produce a similar range of bio-oils that contain a large spectrum of petrochemicals including straight-chain C10-C21 alkanes, which can be directly used as diesel fuel supplement, or a glycerin-free component of biodiesel. Pyrolysis of duckweed produces a different range of bio-oil components that can potentially be used for the production of “green” gasoline and diesel fuel using existing techniques, such as catalytic hydrodeoxygenation.
Differences in absorption rates of the key wastewater nutrients, ammonium and phosphorus by different aquatic macrophytes can be used for optimization of composition of wastewater effluents. The generated data suggest that the composition of the petrochemicals can be modified in a targeted fashion, not only by using different species, but also by changing the source plants’ metabolic profile, by exposing them to different abiotic or biotic stresses. This study presents an attractive, ecologically friendly and cost-effective solution for efficient bio-filtration of swine wastewater and petrochemicals production from generated biomass.
PMCID: PMC3944989  PMID: 24576349
Algae; Biofuel; Bioremediation; Pyrolysis; Swine wastewater; Thermochemical conversion
9.  Bioremediation and Detoxification of Synthetic Wastewater Containing Triarylmethane Dyes by Aeromonas hydrophila Isolated from Industrial Effluent 
Economical and bio-friendly approaches are needed to remediate dye-contaminated wastewater from various industries. In this study, a novel bacterial strain capable of decolorizing triarylmethane dyes was isolated from a textile wastewater treatment plant in Greece. The bacterial isolate was identified as Aeromonas hydrophila and was shown to decolorize three triarylmethane dyes tested within 24 h with color removal in the range of 72% to 96%. Decolorization efficiency of the bacterium was a function of operational parameters (aeration, dye concentration, temperature, and pH) and the optimal operational conditions obtained for decolorization of the dyes were: pH 7-8, 35°C and culture agitation. Effective color removal within 24 h was obtained at a maximum dye concentration of 50 mg/L. Dye decolorization was monitored using a scanning UV/visible spectrophotometer which indicated that decolorization was due to the degradation of dyes into non-colored intermediates. Phytotoxicity studies carried out using Triticum aestivum, Hordeum vulgare, and Lens esculenta revealed the triarylmethane dyes exerted toxic effects on plant growth parameters monitored. However, significant reduction in toxicity was obtained with the decolorized dye metabolites thus, indicating the detoxification of the dyes following degradation by Aeromonas hydrophila.
PMCID: PMC3145198  PMID: 21808740
10.  Are Endocrine Disrupting Compounds a Health Risk in Drinking Water? 
There has been a great deal of international discussion on the nature and relevance of endocrine disrupting compounds in the environment. Changes in reproductive organs of fish and mollusks have been demonstrated in rivers downstream of sewage discharges in Europe and in North America, which have been attributed to estrogenic compounds in the effluent. The anatomical and physiological changes in the fauna are illustrated by feminization of male gonads. The compounds of greatest hormonal activity in sewage effluent are the natural estrogens 17β-estradiol, estrone, estriol and the synthetic estrogen ethinylestradiol. Androgens are also widely present in wastewaters. Investigations of anthropogenic chemical contaminants in freshwaters and wastewaters have shown a wide variety of organic compounds, many of which have low levels of estrogenic activity. In many highly populated countries the drinking water is sourced from the same rivers and lakes that are the recipients of sewage and industrial discharge. The River Thames which flows through London, England, has overall passed through drinking water and sewage discharge 5 times from source to mouth of the river. Under these types of circumstance, any accumulation of endocrine disrupting compounds from sewage or industry potentially affects the quality of drinking water. Neither basic wastewater treatment nor basic drinking water treatment will eliminate the estrogens, androgens or detergent breakdown products from water, due to the chemical stability of the structures. Hence a potential risk to health exists; however present data indicate that estrogenic contamination of drinking water is very unlikely to result in physiologically detectable effects in consumers. Pesticide, detergent and industrial contamination remain issues of concern. As a result of this concern, increased attention is being given to enhanced wastewater treatment in locations where the effluent is directly or indirectly in use for drinking water. In some places at which heavy anthropogenic contamination of drinking water sources occurs, advanced drinking water treatment is increasingly being implemented. This treatment employs particle removal, ozone oxidation of organic material and activated charcoal adsorption of the oxidation products. Such processes will remove industrial organic chemicals, pesticides, detergents, pharmaceutical products and hormones. Populations for which only basic wastewater and drinking water treatment are available remain vulnerable.
PMCID: PMC3807508  PMID: 16823090
Endocrine disruptors; estrogens; feminization; wastewater; drinking water
11.  Biological treatment of tannery wastewater by using salt-tolerant bacterial strains 
High salinity (1–10% w/v) of tannery wastewater makes it difficult to be treated by conventional biological treatment. Salt tolerant microbes can adapt to these saline conditions and degrade the organics in saline wastewater.
Four salt tolerant bacterial strains isolated from marine and tannery saline wastewater samples were identified as Pseudomonas aeruginosa, Bacillus flexus, Exiguobacterium homiense and Staphylococcus aureus. Growth factors of the identified strains were optimized. Tannery saline wastewater obtained from a Common Effluent Treatment Plant (CETP) near Chennai (southern India) was treated with pure and mixed consortia of four salt tolerant bacterial strains. Experiments with optimized conditions and varying salt content (between 2 and 10% (w/v) were conducted. Salt inhibition effects on COD removal rate were noted. Comparative analysis was made by treating the tannery saline wastewater with activated sludge obtained from CETP and with natural habitat microbes present in raw tannery saline wastewater.
Salt tolerant bacterial mixed consortia showed appreciable biodegradation at all saline concentrations (2%, 4%, 6%, 8% and 10% w/v) with 80% COD reduction in particular at 8% salinity level the consortia could be used as suitable working cultures for tannery saline wastewater treatment.
PMCID: PMC2397378  PMID: 18445252
12.  Microalgae and wastewater treatment 
Organic and inorganic substances which were released into the environment as a result of domestic, agricultural and industrial water activities lead to organic and inorganic pollution. The normal primary and secondary treatment processes of these wastewaters have been introduced in a growing number of places, in order to eliminate the easily settled materials and to oxidize the organic material present in wastewater. The final result is a clear, apparently clean effluent which is discharged into natural water bodies. This secondary effluent is, however, loaded with inorganic nitrogen and phosphorus and causes eutrophication and more long-term problems because of refractory organics and heavy metals that are discharged. Microalgae culture offers an interesting step for wastewater treatments, because they provide a tertiary biotreatment coupled with the production of potentially valuable biomass, which can be used for several purposes. Microalgae cultures offer an elegant solution to tertiary and quandary treatments due to the ability of microalgae to use inorganic nitrogen and phosphorus for their growth. And also, for their capacity to remove heavy metals, as well as some toxic organic compounds, therefore, it does not lead to secondary pollution. In the current review we will highlight on the role of micro-algae in the treatment of wastewater.
PMCID: PMC4052567  PMID: 24936135
Microalgae; Wastewater treatment
13.  A pilot plant for removing chromium from residual water of tanneries. 
Environmental Health Perspectives  1995;103(Suppl 1):63-65.
The purpose of this study is to develop a technical process for removing trivalent chromium from tannery wastewater via precipitation. This process can be considered an alternative that avoids a remediation procedure against the metal presence in industrial wastes. This process was verified in a treatment pilot plant located in León, México handling 10 m3/day of three types of effluents. The effluent streams were separated to facilitate the elimination of pollutants from each one. The process was based on in situ treatment and recycle to reduce problems associated with transportation and confinement of contaminated sludges. Two types of treatment were carried out in the pilot plant: The physical/chemical and biological treatments. Thirty-five experiments were conducted and the studied variables were the pH, type of flocculant, and its dose. The statistical significance of chromium samples was 94.7% for its precipitation and 99.7% for recovery. The objectives established for this phase of the development were accomplished and the overall efficiencies were measured for each stage in the pilot plant. The results were: a) chromium precipitation 99.5% from wastewater stream, b) chromium recovery 99% for recycling, and c) physical/chemical treatment to eliminate grease and fat at least 85% and 65 to 70% for the biological treatment. The tanning of a hide lot (350 pieces) was accomplished using 60% treated and recycled water without affecting the product quality. The recovered chromium liquor was also used in this hide tanning. This technical procedure is also applicable for removing heavy metals in other industrial sectors as well as in reducing water consumption rates, if pertinent adjustments are implemented.
PMCID: PMC1519324  PMID: 7621802
14.  Performances and microbial features of an aerobic packed-bed biofilm reactor developed to post-treat an olive mill effluent from an anaerobic GAC reactor 
Olive mill wastewater (OMW) is the aqueous effluent of olive oil producing processes. Given its high COD and content of phenols, it has to be decontaminated before being discharged. Anaerobic digestion is one of the most promising treatment process for such an effluent, as it combines high decontamination efficiency with methane production. The large scale anaerobic digestion of OMWs is normally conducted in dispersed-growth reactors, where however are generally achieved unsatisfactory COD removal and methane production yields. The possibility of intensifying the performance of the process using a packed bed biofilm reactor, as anaerobic treatment alternative, was demonstrated. Even in this case, however, a post-treatment step is required to further reduce the COD. In this work, a biological post-treatment, consisting of an aerobic biological "Manville" silica bead-packed bed aerobic reactor, was developed, tested for its ability to complete COD removal from the anaerobic digestion effluents, and characterized biologically through molecular tools.
The aerobic post-treatment was assessed through a 2 month-continuous feeding with the digested effluent at 50.42 and 2.04 gl-1day-1 of COD and phenol loading rates, respectively. It was found to be a stable process, able to remove 24 and 39% of such organic loads, respectively, and to account for 1/4 of the overall decontamination efficiency displayed by the anaerobic-aerobic integrated system when fed with an amended OMW at 31.74 and 1.70 gl-1day-1 of COD and phenol loading rates, respectively. Analysis of 16S rRNA gene sequences of biomass samples from the aerobic reactor biofilm revealed that it was colonized by Rhodobacterales, Bacteroidales, Pseudomonadales, Enterobacteriales, Rhodocyclales and genera incertae sedis TM7. Some taxons occurring in the influent were not detected in the biofilm, whereas others, such as Paracoccus, Pseudomonas, Acinetobacter and Enterobacter, enriched significantly in the biofilter throughout the treatment.
The silica-bead packed bed biofilm reactor developed and characterized in this study was able to significantly decontaminate anaerobically digested OMWs. Therefore, the application of an integrated anaerobic-aerobic process resulted in an improved system for valorization and decontamination of OMWs.
PMCID: PMC1501037  PMID: 16595023
15.  Determination of phenols and pharmaceuticals in municipal wastewaters from Polish treatment plants by ultrasound-assisted emulsification–microextraction followed by GC–MS 
A method combining ultrasound-assisted emulsification–microextraction (USAEME) with gas chromatography–mass spectrometry (GC–MS) was developed for simultaneous determination of four acidic pharmaceuticals, ibuprofen, naproxen, ketoprofen, and diclofenac, as well as four phenols, 4-octylphenol, 4-n-nonylphenol, bisphenol A, and triclosan in municipal wastewaters. Conditions of extraction and simultaneous derivatization were optimized with respect to such aspects as type and volume of extraction solvent, volume of derivatization reagent, kind and amount of buffering salt, location of the test tube in the ultrasonic bath, and extraction time. The average correlation coefficient of the calibration curves was 0.9946. The LOD/(LOQ) values in influent and effluent wastewater were in the range of 0.002–0.121/(0.005–0.403) μg L−1 and 0.002–0.828/(0.006–2.758) μg L−1, respectively. Quantitative recoveries (≥94 %) and satisfactory precision (average RSD 8.2 %) were obtained. The optimized USAEME/GC–MS method was applied for determination of the considered pharmaceuticals and phenols in influents and treated effluents from nine Polish municipal wastewater treatment plants. The average concentration of acidic pharmaceuticals in influent and effluent wastewater were in the range of 0.06–551.96 μg L−1 and 0.01–22.61 μg L−1, respectively, while for phenols were in the range of 0.03–102.54 μg L−1 and 0.02–10.84 μg L−1, respectively. The removal efficiencies of the target compounds during purification process were between 84 and 99 %.
PMCID: PMC3877427  PMID: 23818073
Ultrasound-assisted emulsification–microextraction; Gas chromatography–mass spectrometry; Acidic pharmaceuticals; Phenols; Influents; Effluents; Municipal wastewater treatment plant
16.  Reclaimed water as a reservoir of antibiotic resistance genes: distribution system and irrigation implications 
Treated wastewater is increasingly being reused to achieve sustainable water management in arid regions. The objective of this study was to quantify the distribution of antibiotic resistance genes (ARGs) in recycled water, particularly after it has passed through the distribution system, and to consider point-of-use implications for soil irrigation. Three separate reclaimed wastewater distribution systems in the western U.S. were examined. Quantitative polymerase chain reaction (qPCR) was used to quantify ARGs corresponding to resistance to sulfonamides (sul1, sul2), macrolides (ermF), tetracycline [tet(A), tet(O)], glycopeptides (vanA), and methicillin (mecA), in addition to genes present in waterborne pathogens Legionella pneumophila (Lmip), Escherichia coli (gadAB), and Pseudomonas aeruginosa (ecfx, gyrB). In a parallel lab study, the effect of irrigating an agricultural soil with secondary, chlorinated, or dechlorinated wastewater effluent was examined in batch microcosms. A broader range of ARGs were detected after the reclaimed water passed through the distribution systems, highlighting the importance of considering bacterial re-growth and the overall water quality at the point of use (POU). Screening for pathogens with qPCR indicated presence of Lmip and gadAB genes, but not ecfx or gyrB. In the lab study, chlorination was observed to reduce 16S rRNA and sul2 gene copies in the wastewater effluent, while dechlorination had no apparent effect. ARGs levels did not change with time in soil slurries incubated after a single irrigation event with any of the effluents. However, when irrigated repeatedly with secondary wastewater effluent (not chlorinated or dechlorinated), elevated levels of sul1 and sul2 were observed. This study suggests that reclaimed water may be an important reservoir of ARGs, especially at the POU, and that attention should be directed toward the fate of ARGs in irrigation water and the implications for human health.
PMCID: PMC3664959  PMID: 23755046
antibiotic resistance genes; water reuse; reclaimed water distribution systems; irrigation
17.  “NiCo Buster”: engineering E. coli for fast and efficient capture of cobalt and nickel 
Metal contamination is widespread and results from natural geogenic and constantly increasing anthropogenic sources (mainly mining and extraction activities, electroplating, battery and steel manufacturing or metal finishing). Consequently, there is a growing need for methods to detoxify polluted ecosystems. Industrial wastewater, surface water and ground water need to be decontaminated to alleviate the contamination of soils and sediments and, ultimately, the human food chain. In nuclear power plants, radioactive metals are produced; these metals need to be removed from effluents before they are released into the environment, not only for pollution prevention but also for waste minimization. Many physicochemical methods have been developed for metal removal from aqueous solutions, including chemical coagulation, adsorption, extraction, ion exchange and membrane separation; however, these methods are generally not metal selective. Bacteria, because they contain metal transporters, provide a potentially competitive alternative to the current use of expensive and high-volume ion-exchange resins.
The feasibility of using bacterial biofilters as efficient tools for nickel and cobalt ions specific remediation was investigated. Among the factors susceptible to genetic modification in Escherichia coli, specific efflux and sequestration systems were engineered to improve its metal sequestration abilities. Genomic suppression of the RcnA nickel (Ni) and cobalt (Co) efflux system was combined with the plasmid-controlled expression of a genetically improved version of a specific metallic transporter, NiCoT, which originates from Novosphingobium aromaticivorans. The resulting strain exhibited enhanced nickel (II) and cobalt (II) uptake, with a maximum metal ion accumulation of 6 mg/g bacterial dry weight during 10 min of treatment. A synthetic adherence operon was successfully introduced into the plasmid carrying the improved NiCoT transporter, conferring the ability to form thick biofilm structures, especially when exposed to nickel and cobalt metallic compounds.
This study demonstrates the efficient use of genetic engineering to increase metal sequestration and biofilm formation by E. coli. This method allows Co and Ni contaminants to be sequestered while spatially confining the bacteria to an abiotic support. Biofiltration of nickel (II) and cobalt (II) by immobilized cells is therefore a promising option for treating these contaminants at an industrial scale.
PMCID: PMC4124493  PMID: 25104972
Bioremediation; Cobalt; Nickel; Biofilm; Biofilter; NiCoT; Synthetic biology
18.  Dyeing Industry Effluent System as Lipid Production Medium of Neochloris sp. for Biodiesel Feedstock Preparation 
BioMed Research International  2014;2014:529560.
Microalgae lipid feedstock preparation cost was an important factor in increasing biodiesel fuel hikes. This study was conducted with the concept of implementing an effluent wastewater as lipid production medium for microalgae cultivation. In our study textile dyeing industry effluent was taken as a lipid production medium for Neochloris sp. cultivation. The changes in physicochemical analysis of effluent before and after Neochloris sp. treatment were recorded using standard procedures and AAS analysis. There was especially a reduction in heavy metal like lead (Pb) concentration from 0.002 ppm to 0.001 ppm after Neochloris sp. treatment. Neochloris sp. cultivated in Bold Basal Medium (BBM) (specific algal medium) produced 41.93% total lipid and 36.69% lipid was produced in effluent based cultivation. Surprisingly Neochloris sp. cultivated in effluent was found with enhanced neutral lipid content, and it was confirmed by Nile red fluorescence assay. Further the particular enrichment in oleic acid content of the cells was confirmed with thin layer chromatography (TLC) with oleic acid pure (98%) control. The overall results suggested that textile dyeing industry effluent could serve as the best lipid productive medium for Neochloris sp. biodiesel feedstock preparation. This study was found to have a significant impact on reducing the biodiesel feedstock preparation cost with simultaneous lipid induction by heavy metal stress to microalgae.
PMCID: PMC4163492  PMID: 25247176
19.  Fungal Laccases and Their Applications in Bioremediation 
Enzyme Research  2014;2014:163242.
Laccases are blue multicopper oxidases, which catalyze the monoelectronic oxidation of a broad spectrum of substrates, for example, ortho- and para-diphenols, polyphenols, aminophenols, and aromatic or aliphatic amines, coupled with a full, four-electron reduction of O2 to H2O. Hence, they are capable of degrading lignin and are present abundantly in many white-rot fungi. Laccases decolorize and detoxify the industrial effluents and help in wastewater treatment. They act on both phenolic and nonphenolic lignin-related compounds as well as highly recalcitrant environmental pollutants, and they can be effectively used in paper and pulp industries, textile industries, xenobiotic degradation, and bioremediation and act as biosensors. Recently, laccase has been applied to nanobiotechnology, which is an increasing research field, and catalyzes electron transfer reactions without additional cofactors. Several techniques have been developed for the immobilization of biomolecule such as micropatterning, self-assembled monolayer, and layer-by-layer techniques, which immobilize laccase and preserve their enzymatic activity. In this review, we describe the fungal source of laccases and their application in environment protection.
PMCID: PMC4052089  PMID: 24959348
20.  Quantitative Detection of Human Adenoviruses in Wastewater and Combined Sewer Overflows Influencing a Michigan River▿  
Enteric viruses are important pathogens found in contaminated surface waters and have previously been detected in waters of the Great Lakes. Human adenoviruses were monitored because of their high prevalence and persistence in aquatic environments. In this study, we quantified adenoviruses in wastewater, surface water, and combined sewer overflows (CSOs) by real-time PCR. Between August 2005 and August 2006, adenovirus concentrations in raw sewage, primary-treated effluent, secondary-treated effluent, and chlorinated effluent from a wastewater treatment plant in Michigan were examined. CSO samples (n = 6) were collected from a CSO retention basin in Grand Rapids, MI. Adenoviruses were detected in 100% of wastewater and CSO discharge samples. Average adenovirus DNA concentrations in sewage and CSOs were 1.15 × 106 viruses/liter and 5.35 × 105 viruses/liter, respectively. Adenovirus removal was <2 log10 (99%) at the wastewater treatment plant. Adenovirus type 41 (60% of clones), type 12 (29%), type 40 (3%), type 2 (3%), and type 3 (3%) were isolated from raw sewage and primary effluents (n = 28). Six of 20 surface water samples from recreational parks at the lower Grand River showed virus concentrations above the real-time PCR detection limit (average, 7.8 × 103 viruses/liter). This research demonstrates that wastewater effluents and wastewater-impacted surface waters in the lower Grand River in Michigan contain high levels of viruses and may not be suitable for full-body recreational activities. High concentrations of adenovirus in these waters may be due to inefficient removal during wastewater treatment and to the high persistence of these viruses in the environment.
PMCID: PMC2813034  PMID: 19948848
21.  Ozone oxidation of antidepressants in wastewater –Treatment evaluation and characterization of new by-products by LC-QToFMS 
The fate of 14 antidepressants along with their respective N-desmethyl metabolites and the anticonvulsive drug carbamazepine was examined in a primary sewage treatment plant (STP) and following advanced treatments with ozone (O3). The concentrations of each pharmaceutical compound were determined in raw sewage, effluent and sewage sludge samples by LC-MS/MS analysis. The occurrence of antidepressant by-products formed in treated effluent after ozonation was also investigated.
Current primary treatments using physical and chemical processes removed little of the compounds (mean removal efficiency: 19%). Experimental sorption coefficients (Kd) of each studied compounds were also calculated. Sorption of venlafaxine, desmethylvenlafaxine, and carbamazepine on sludge was assumed to be negligible (log Kd ≤ 2), but higher sorption behavior can be expected for sertraline (log Kd ≥ 4). Ozonation treatment with O3 (5 mg/L) led to a satisfactory mean removal efficiency of 88% of the compounds. Screening of the final ozone-treated effluent samples by high resolution-mass spectrometry (LC-QqToFMS) did confirm the presence of related N-oxide by-products.
Effluent ozonation led to higher mean removal efficiencies than current primary treatment, and therefore represented a promising strategy for the elimination of antidepressants in urban wastewaters. However, the use of O3 produced by-products with unknown toxicity.
PMCID: PMC3564768  PMID: 23351428
Antidepressants; Ozone; LC-MS/MS; Sewage treatment plants; Biosolids; Side-products
22.  Decolorization applicability of sol–gel matrix immobilized manganese peroxidase produced from an indigenous white rot fungal strain Ganoderma lucidum 
BMC Biotechnology  2013;13:56.
An eco-friendly treatment of industrial effluents is a major environmental concern of the modern world in the face of stringent environmental legislations. By keeping in mind the extensive industrial applications of ligninolytic enzymes, this study was performed to purify, and immobilize the manganese peroxidase (MnP) produced from an indigenous strain of Ganoderma lucidum. The present study was also focused on investigating the capability of immobilized MnP for decolorization of dye containing textile effluents.
A large magnitude of an indigenous MnP (882±13.3 U/mL) was obtained from white rot fungal strain G. lucidum in solid state bio-processing of wheat straw under optimized fermentation conditions (moisture, 50%; substrate, 5 g; pH, 5.5; temperature, 30°C; carbon source, 2% fructose; nitrogen source, 0.02% yeast extract; C: N ratio, 25:1; fungal spore suspension, 5 mL and fermentation time period, 4 days). After ammonium sulfate fractionation and Sephadex-G-100 gel filtration chromatography, MnP was 4.7-fold purified with specific activity of 892.9 U/mg. G. lucidum MnP was monomeric protein as evident by single band corresponding to 48 kDa on native and denaturing SDS-PAGE. The purified MnP (2 mg/mL) was immobilized using a sol–gel matrix of tetramethoxysilane (TMOS) and proplytrimethoxysilane (PTMS). The oxidation of MnSO4 for up to 10 uninterrupted cycles demonstrated the stability and reusability of the immobilized MnP. Shelf life profile revealed that enzyme may be stored for up to 60 days at 25°C without losing much of its activity. To explore the industrial applicability of MnP produced by G. lucidum, the immobilized MnP was tested against different textile effluents. After 4 h reaction time, the industrial effluents were decolorized to different extents (with a maximum of 99.2%). The maximally decolorized effluent was analyzed for formaldehyde and nitroamines and results showed that the toxicity parameters were below the permissible limits.
In conclusion, G. lucidum MnP was immobilized by sol–gel matrix entrapment with an objective to enhance its practical efficiencies. The MnP was successfully entrapped into a sol- gel matrix of TMOS and PTMS with an overall immobilization efficiency of 93.7%. The sol- gel entrapped MnP seems to have prospective capabilities which can be useful for industrial purposes, especially for bioremediation of industrial effluents.
PMCID: PMC3717284  PMID: 23849469
Bio-catalysis; G. lucidum; MnP; PAGE; Sol–gel; Immobilization; Textile effluents; Decolorization; Toxicity reduction
23.  Response surface methodology for optimization of culture conditions for dye decolorization by a fungus, Aspergillus niger HM11 isolated from dye affected soil 
Iranian Journal of Microbiology  2010;2(4):213-222.
Background and Objectives
Discharge of wastewater from textile dyeing industries has been a problem in terms of pollution and treatment of these waters is a great task. Keeping this in mind, the aim of our current research is to study the effect of various bioprocess variables on decolorization of an azo dye, Congo red, by a fungal isolate, Aspergillus niger HM11.
Materials and Methods
Central composite design (CCD) and response surface methodology (RSM) have been applied to design experiments to evaluate the interactive effects of the operating variables: on the decolorization of Congo red. A total of 30 experiments were conducted in the present study and a regression coefficient between the variables was generated.
The RSM indicated that pH 6.0, 150 rpm agitation, incubation time of 36 hrs and a glucose concentration of 1.0% were optimal for maximum decolorization of Congo red and the response indicated excellent evaluation of experimental data.
From this study, it is very obvious that the fungal isolate, Aspergillus niger HM11 can be used as a promising microbial strain for decolorization of textile dyeing effluent containing similar dyes.
PMCID: PMC3279788  PMID: 22347575
Decolorization; Congo red; Aspergillus niger HM11; RSM; CCD
24.  Enhancing the Decolorizing and Degradation Ability of Bacterial Consortium Isolated from Textile Effluent Affected Area and Its Application on Seed Germination 
The Scientific World Journal  2015;2015:628195.
A bacterial consortium BMP1/SDSC/01 consisting of six isolates was isolated from textile effected soil, sludge, and textile effluent from Hudiara drain near Nishat Mills Limited, Ferozepur Road, Lahore, Pakistan. It was selected because of being capable of degrading and detoxifying red, green, black, and yellow textile dyes. The pH and supplements were optimized to enhance the decolorization ability of the selected consortium. The results indicated that decolorizing ability of consortium for the red, green, black, and yellow dyes was higher as compared to individual strains. The consortium was able to decolorize 84%, 84%, 85%, 85%, and 82% of 200 ppm of red, green, black, yellow, and mixed dyes within 24 h while individual strain required 72 h. On supplementing urea, the consortium decolorized 87, 86, 89, 86, and 83%, respectively, while on supplementing sodium chloride the consortium decolorized 93, 94, 93, 94, and 89% of red, green, black, yellow, and mixed dyes, respectively, which was maximum while in the presence of ascorbic acid and ammonium chloride it showed intermediate results. The effect of untreated and treated dyes was investigated on Zea mays L. (maize) and Sorghum vulgare Pers. (sorghum). This study will help to promote an efficient biotreatment of textile effluents.
PMCID: PMC4306213  PMID: 25654132
25.  Pharmaceutical Formulation Facilities as Sources of Opioids and Other Pharmaceuticals to Wastewater Treatment Plant Effluents 
Environmental Science & Technology  2010;44(13):4910-4916.
Facilities involved in the manufacture of pharmaceutical products are an under-investigated source of pharmaceuticals to the environment. Between 2004 and 2009, 35 to 38 effluent samples were collected from each of three wastewater treatment plants (WWTPs) in New York and analyzed for seven pharmaceuticals including opioids and muscle relaxants. Two WWTPs (NY2 and NY3) receive substantial flows (>20% of plant flow) from pharmaceutical formulation facilities (PFF) and one (NY1) receives no PFF flow. Samples of effluents from 23 WWTPs across the United States were analyzed once for these pharmaceuticals as part of a national survey. Maximum pharmaceutical effluent concentrations for the national survey and NY1 effluent samples were generally <1 μg/L. Four pharmaceuticals (methadone, oxycodone, butalbital, and metaxalone) in samples of NY3 effluent had median concentrations ranging from 3.4 to >400 μg/L. Maximum concentrations of oxycodone (1700 μg/L) and metaxalone (3800 μg/L) in samples from NY3 effluent exceeded 1000 μg/L. Three pharmaceuticals (butalbital, carisoprodol, and oxycodone) in samples of NY2 effluent had median concentrations ranging from 2 to 11 μg/L. These findings suggest that current manufacturing practices at these PFFs can result in pharmaceuticals concentrations from 10 to 1000 times higher than those typically found in WWTP effluents.
Concentrations of pharmaceuticals in two wastewater effluents receiving discharges from pharmaceutical formulation facilities are 10−1000 times higher than concentrations measured in typical wastewater effluents.
PMCID: PMC2893609  PMID: 20521847

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