Monitoring of university wastewater within the sewage system and its performance evaluation through integrated constructed wetlands.

Rapid increase in population and industrialization has not only improved the lifestyle but adversely affected the quality and availability of water leading to ample amount of wastewater generation. The major contribution towards wastewater production is from sewage. Regular monitoring and treatment of sewage water is necessary to conserve and enhance the quality of water. The present study focuses on monitoring of sewage water within the sewage system of a residential university. A total of 16 samples from different manholes were collected for physicochemical and heavy metals analysis and compared with final effluent collected from integrated constructed wetlands (ICWs) to assess its removal efficiency. The mean concentrations of influent and effluent were compared with national environmental quality standards (NEQS) for municipal discharge (pH 6-9, COD 150 mg/L, TSS 200 mg/L and TDS 3500 mg/L) and international agricultural reuse standards (IARS) (pH 6-8, COD <150 mg/L, TSS < 100 mg/L) respectively. Among all physicochemical parameters, influent values for chemical oxygen demand (COD) (169.56-258.36) mg/L exceeded the limit of NEQS for discharge into inland waters, whereas for total suspended solids (TSS) the concentration exceeded for discharge into STP (406 mg/L) and inland waters (202.33 mg/L). However, effluent concentrations for all the parameters were found within the permissible limit set by IARS. The removal efficiency for different parameters such as phosphate- phosphorus (PO43-P), COD, TSS, total dissolved solids (TDS) and total kjeldahl nitrogen (TKN) were 52, 53, 54, 35, and 36%, respectively. Heavy metal concentrations were compared with WHO guidelines among which lead (Pb) in effluent and chromium (Cr) in influent exceeded the limit (Pb 0.01 and Cr 0.05 mg/L). Interpolation results showed that zone 2 was highly contaminated in comparison to zone 1 & 3. Statistical analysis showed that correlation of physicochemical parameters and heavy metals was found significant (p < 0.05).

Sulfamethoxazole (SMX) Alters Immune and Apoptotic Endpoints in Developing Zebrafish (Danio rerio).

Sulfamethoxazole (SMX) is a broad-range bacteriostatic antibiotic widely used in animal and fish farming and is also employed in human medicine. These antibiotics can ultimately end up in the aquatic ecosystem and affect non-target organisms such as fish. To discern the effect of SMX on developing zebrafish embryos and larvae, we investigated a broad range of sub-lethal toxicity endpoints. Higher concentrations of SMX affected survivability, caused hatch delay, and induced malformations including edema of the yolk sac, pericardial effusion, bent tail, and curved spine in developing embryos. Lower levels of SMX provoked an inflammatory response in larvae at seven days post fertilization (dpf), as noted by up-regulation of interferon (ifn-γ) and interleukin 1ß (il-1ß). SMX also increased the expression of genes related to apoptosis, including BCL2-Associated Agonist of Cell Death (bad) and BCL2 Associated X, Apoptosis Regulator (bax) at 50 µg/L and decreased caspase 3 (casp3) expression in a dose-dependent manner. SMX induced hyperactivity in larval fish at 500 and 2500 µg/L based upon the light/dark preference test. Collectively, this study revealed that exposure to SMX can disrupt the immune system by altering host defense mechanisms as well as transcripts related to apoptosis. These data improve understanding of antibiotic chemical toxicity in aquatic organisms and serves as a baseline for in-depth environmental risk assessment of SMX and antibiotics.

Comprehensive analysis of spatial distribution of microplastics in Rawal Lake, Pakistan using trawl net and sieve sampling methods.

Occurrence of microplastics (MPs) in freshwater environments, particularly reservoir and lakes, is an emerging concern. There are limited studies in Pakistan on microplastic pollution in the lacustrine environments and those that exist do not provide sufficient information on the spatial distribution of MPs in offshore surface water. The aims of this study were to determine microplastic abundance in Rawal Lake, Pakistan and to ascertain if sampling methodology influences microplastic counts. Surface water samples were collected from 10 sites; 5 tributaries, 2 human settlement and 3 fishing and boating areas using two different sampling techniques: 100 µm mesh trawl and 20 L sample through a 45 µm mesh sieve. A significant difference was observed in the abundance of MPs across two methods with the sieve method yielding 2.8 ± 1.44 particles/L and trawl yielding 0.025 ± 0.024 particles/L. Tributaries and boating/fishing area had higher microplastic abundance than the residential area regardless of sampling method. Filaments were the dominant shape of MPs in both type of samples followed by fragments in trawl samples and films in sieved samples. Microbeads were only detected in trawl samples. MPs within size range 0.1-0.9 mm were mostly fragments (82%). MPs were diverse in colors with white/transparent and black MPs common. Polypropylene was the main type of microplastic in Rawal Lake (40-74%). Scanning Electron Microscopy (SEM) of MPs showed cracks, roughness and striations on the particles. Energy Dispersive Spectroscopy (EDS) detected heavy metals (Fe, Cu, Ni, Pb, Zn, Co and Cr) in MPs. Findings suggest that microplastic pollution in Rawal Lake may pose great risk to aquatic and human life through leaching of inherent/adsorbed heavy metals and therefore requires future investigation.

Toxic effects of pristine and aged polystyrene microplastics on selective and continuous larval culture of acorn barnacle Amphibalanus amphitrite.

This study evaluates the toxicity of pristine (Unwashed) and aged, clean (Biofilm-) or fouled (Biofilm+), PS microspheres (3 µm,10 µm), using Washed particles as a reference material, on selective and continuous larval culture of Amphibalanus amphitrite. Exposure to 3 µm Unwashed and Biofilm+ particles for 24 h induced significant mortality (60 % and 57 % respectively) in stage II larvae. Stage II and VI nauplii showed greater uptake of 3 µm Biofilm- particles. Accumulative exposure to microplastics in continuous larval culture significantly affected the naupliar survival, particularly of stage III and IV. Cumulative mortality was > 70% after exposure to 3 µm Unwashed and 10 µm Biofilm+ particles. Unwashed particles with increasing concentration and aged particles with increasing size, delayed the development of nauplii to cyprids. Though,> 50% cyprids showed successful settlement however the highest concentration of 3 µm Biofilm+ microspheres inhibited the settlement and induced precocious metamorphosis in 9 % of the cyprids.

Detection in influx sources and estimation of microplastics abundance in surface waters of Rawal Lake, Pakistan.

The ever-growing production, usage and poor waste management practices of plastics are causing microplastics intrusion in freshwater environments all over the world. The identification of inflow processes and sources is equally important as the assessment of microplastic concentrations in freshwater. This study reports microplastic presence in the influx sources and provides an overall estimation of microplastic concentration in the surface water of a freshwater reservoir, Rawal Lake, Islamabad. In the current study, six major tributaries of Rawal lake were assessed for microplastic presence, out of which four tributaries showed microplastic contamination. Microplastics concentration in the lake ranged from 6.4 ± 0.5 particles/m³ to 8.8 ± 0.5 particles/m³. All the identified microplastics in tributaries and lake were secondary except granules. The prominent shape of microplastics among the studied waters was film, with transparent being the most frequent plastic-type according to color. Polyethylene (LDPE and HDPE) were the dominant type of microplastics found in the lake and the tributaries. More than 72% of microplastics had a size of 0.3-0.1 mm. This study provides a better understanding of the extent of microplastic pollution assessment in a freshwater lake with equal emphasis on microplastic presence in influx sources and the relationship of microplastics with fundamental water quality indicators (pH, temperature, dissolved oxygen, and biological oxygen demand), which may be beneficial in impeding the introduction of microplastics at sources.

Multi-biomarkers approach to determine the toxicological impacts of sulfamethoxazole antibiotic on freshwater fish Cyprinus carpio.

Antibiotics are increasingly detected in the aquatic environment and are raising severe concerns for human and ecological health. Sulfamethoxazole being a widely recommended antibiotic in both human and veterinary medicine is consequently found in the aquatic environment. The current research was aimed to investigate the potential bioaccumulation and sub-lethal toxicity in terms of oxidative stress and histopathology of targeted antibiotic sulfamethoxazole in Cyprinus carpio at environmentally relevant concentrations over a prolonged period. Fish were exposed for 28 days to environmentally realistic concentrations (25-200 µg/L) of sulfamethoxazole. HPLC analysis revealed an inverse relationship between dosages applied and bioaccumulation in fish muscle tissues. The highest concentrations of sulfamethoxazole in the muscle tissues exposed to 25, 50, 100 and 200 µg/L were 124, 202,104.5, and 123.2 ng/g, respectively at different sampling times. Moreover, exposure to sulfamethoxazole enabled ROS (Reactive oxygen species) production in both brain and gill tissues of fish, where the increase in ROS formation was dose and time dependent. Furthermore, histopathological analysis of gills and liver revealed various alterations including pycnotic nuclei, bile stagnation, vacuolization in the liver, and partial and complete fusion of lamella and blood congestion in gills, respectively. Organ pathological index also revealed that the intensity of tissue damage increased as sulfamethoxazole dosage was increased. Quantitative evaluation of gills morphometric parameters divulged that severity of histopathological changes increased with time for all the exposed groups at the end of exposure time (28th day) indicating physio-metabolic turmoil caused by molecular and biochemical action of sulfamethoxazole. In conclusion, the assessment of bioaccumulative potential, measurement of ROS, and histopathology of Cyprinus carpio appeared to be a useful biomarker to evaluate the toxic impacts of antibiotics on the health of fish.

Clearing the Clutter: Antiatherosclerotic activity of Eucalyptus camaldulensis crude extract.

Plant components have been extensively evaluated for their pharmacological activities. This study provides scientific rationale towards the therapeutic effect of Eucalyptus camaldulensis aqueous bark extract against induced atherosclerosis and hyperlipidemia in pigeons. Phytochemical components of Eucalyptus bark extract possess a great antioxidant activity that potentially reduced the risk of heart diseases. A total of 42 Pigeons of both sexes were distributed into negative control (fed normal grain diet), hyperlipidemic control (fed HFD 1% animal fat oil and 0.1% cholesterol for 3 months), test groups of variable doses (0.05, 0.1, 0.2 to 0.4 gms/kg BW for 21 days) and the group received atorvastatin daily after induction used. At the end of the experiment biochemical and histological evaluation has been performed. After HFD induction the serum levels of liver enzyme AST, glucose, urea, cholesterol, LDL, VLDL, and TG were significantly increased with the reduction in HDL levels. The atherogenic index was also found significantly raised. Microscopic examination of the liver and aorta showed the appearance of lipid-filled foam cells all over the liver parenchyma and intima after the HFD induction. Thus it was concluded that Eucalyptus aqueous bark extract can be effective against atherosclerosis and hyperlipidemia.

Combined pretreatment of sugarcane bagasse using alkali and ionic liquid to increase hemicellulose content and xylanase production.

BACKGROUND: Lignin in sugarcane bagasse (SB) hinders its utilization by microorganism, therefore, pretreatment methods are employed to make fermentable components accessible to the microbes. Multivariate analysis of different chemical pretreatment methods can aid to select the most appropriate strategy to valorize a particular biomass. RESULTS: Amongst methods tested, the pretreatment by using sodium hydroxide in combination with methyltrioctylammonium chloride, an ionic liquid, (NaOH+IL) was the most significant for xylanase production by Bacillus aestuarii UE25. Investigation of optimal levels of five significant variables by adopting Box-Behnken design (BBD) predicted 20 IU mL- 1 of xylanase and experimentally, a titer of 17.77 IU mL- 1 was obtained which indicated the validity of the model. The production kinetics showed that volumetric productivity of xylanase was much higher after 24 h (833.33 IU L- 1 h- 1) than after 48 h (567.08 IU L- 1 h- 1). The extracted xylan from SB induced more xylanase in the fermentation medium than pretreated SB or commercially purified xylan. Nuclear Magnetic Resonance, Fourier transform infrared spectroscopy and scanning electron microscopy of SB indicated removal of lignin and changes in the structure of SB after NaOH+IL pretreatment and fermentation. CONCLUSION: Combined pretreatment of SB with alkali and methyltrioctylammonium chloride appeared better than other chemical methods for bacterial xylanase production and for the extraction of xylan form SB.

Cellulose extraction from methyltrioctylammonium chloride pretreated sugarcane bagasse and its application.

Cellulose, the most abundant feedstock of chemicals and energy is extracted from various agro-industrial wastes, such as sugarcane bagasse (SB). Pretreatment of SB with ionic liquids improves extraction of cellulose, yet the use of ionic liquid is hindered by its high cost. In this study, cellulose was extracted from SB pretreated with methyltrioctylammonium chloride under relatively mild conditions. The extracted cellulose from pretreated SB (PTB) and untreated SB (UTB) was characterized by scanning electron microscopy and FTIR. Fermentation of cellulose extracted from PTB by a thermophilic bacterium, Bacillus aestuarii UE25, yielded 245.16% higher titers of cellulase than cellulose extracted from UTB. The recyclability of the IL was assessed to make the pretreatment process cost effective and was monitored through TLC and FTIR. The results of this research demonstrated the potential of ionic liquid pretreated SB for cellulose extraction and for its subsequent utilization in thermostable cellulase production.

Utilization of methyltrioctylammonium chloride as new ionic liquid in pretreatment of sugarcane bagasse for production of cellulase by novel thermophilic bacteria.

Fermentation of carbohydrates present in lignocellulosic (LC) biomass is facilitated by lignin removal, which is usually achieved by adopting various pretreatment methods to provide the enzymes proper access to their respective substrates. Pretreatment using ionic liquid (IL) is relatively recent advancement and considered as mild and green process. ILs can dissolve extensive quantities of biomass and depolymerize the cellulose. In this context, an abundantly available LC biomass, sugarcane bagasse (SB), was pretreated using alkali or with an IL, methyltrioctylammonium chloride, and was used for cellulase production from thermophilic bacteria. In all, 26 indigenously isolated thermophilic bacterial strains were quantitatively screened for cellulase production. 16S rDNA sequences of the promising isolates UE10 and UE27 revealed relatedness with Brevibacillus borstelensis, while the strain UE1 belonged to Aneurinibacillus thermoaerophilus. Cellulase production was compared by utilizing alkali pretreated and IL pretreated SB and the later was found more appropriate. UE1, UE10 and UE27 yielded 22.2, 22.18 and 33.3 IU mL-1 of endoglucanase, respectively, by fermenting IL pretreated SB. The changes in SB structure after pretreatment were evaluated by scanning electron microscopy. This study demonstrated the potential of novel thermophilic bacterial strains to utilize IL pretreated SB for production of industrially important enzyme, cellulase.

The selective pressure of quorum quenching on microbial communities in membrane bioreactors.

In conventional membrane bioreactor (MBR) treatment systems, Gram-negative bacterial population appears to be always outnumbered Gram-positive community. Thereby, acyl homoserine lactones (AHLs), major signaling molecules utilized by Gram-negative bacteria, have been targeted for biofouling control in quorum quenching (QQ) based studies. This study investigated the impact of AHL and autoinducer-2 (AI-2)-degrading QQ consortium on the selective accumulation of microbial communities in a QQ MBR (MBR-QQb). The results show that addition of the QQ consortium (in the form of beads) increased the filtration time of MBR-QQb by 3.5 times. The distribution of mixed liquor extracellular polymeric substances (EPS), especially the tightly bound (TB) proteinous EPS and the floc size were strongly affected by the QQ activity, and the endless 'battle' between QQ and quorum sensing (QS). More importantly, QQ induced the significant suppression of Gram-negative bacterial community. The average abundance of Gram-positive bacteria at the genus level in the biocake of MBR-QQb (51%) was significantly higher than that of the control MBR (11%) and the MBR with vacant beads (28%). These findings suggest that an unintended condition is created to favor the growth of Gram-positive bacteria in QQ MBRs, resulting in a distinct microbial social network in both bulk sludge and biocake.

Antibacterial behaviour of surface modified composite polyamide nanofiltration (NF) membrane by immobilizing Ag-doped TiO2 nanoparticles.

Modification of active membrane surface is an auspicious way to enhance the membrane performance. In our study, a commercially available composite polyamide Nanofiltration (NF) membrane was modified by immobilizing silver doped TiO2 (Ag-TiO2) nanoparticles. Ag-TiO2 with different nanoparticles concentration (0.05, 0.1, and 0.5 wt. %) were coated on the surface of the membrane by a dip coating method. The evidence of successful coating was evaluated by Scanning Electron Microscopy coupled with Energy Dispersive Spectroscopy and Atomic Force Microscopy images. Moreover, the Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), contact angle measurement and permeation tests were carried out in order to evaluate the membrane performance after coating. The antifouling property of the modified membrane was evaluated for Gram-positive (Bacillus subtilis) and Gram-negative (Escherichia coli) bacteria by colony counting method. The results indicated that the modified membranes keep efficient antibacterial efficacy against both types of bacteria. The bacterial growth reduced approximately 93% and 91% on the modified membrane as compared to the unmodified membrane for E.coli and B.subtilis, respectively. Ag-TiO2 nanoparticles imbedded nanofiltration membranes inhibit the biofilm formation and facilitate in cleaning membrane surface without using excessive chemical agents.

Evaluation of genotoxicity and hematological effects in common carp (Cyprinus carpio) induced by disinfection by-products.

Disinfection is intended to improve drinking water quality and human health. Although disinfectants may transform organic matter and form disinfection by-products (DBPs), many are branded as cyto- and genotoxic. Traditionally, research focuses on the effects of DBPs on human health, but cytogenic impacts on aquatic organisms still remain ill defined. The current study examines the potential toxic effect of chloroform and iodoform (DBPs) on Cyprinus carpio, selected as a model organism. Fish specimens were exposed to various concentrations of DBPs primarily based on LD50 values, where acute toxicity was monitored for 96 h. Headspace SPME extraction through gas chromatography was employed to assess the effects of spiked DBPs doses in fish blood. Cytotoxicity was monitored using Comet assay. Tail length, tail DNA, and olive tail moment values were quantified to be significant (P < 0.05) as compared to control. A statistically significant (P < 0.05) decrease in all blood parameters (hematology) was observed. Changes in biochemical indices (glucose, total protein, and alanine aminotransferase (ALT)) were also significant. ALT secretion was significantly increased (93 ± 0.05 and 82.8 ± 0.1 U/L) at higher concentration compared to control (56 ± 0.1 U/L), suggesting liver damage. Results demonstrated that iodoform was statistically more damaging as compared to chloroform.

Plant uptake and leaching potential upon application of amendments in soils spiked with heavy metals (Cd and Pb).

The aim of this study was to assess the effects of soil amendments (organic and inorganic) on phytoavailability and leaching of cadmium (Cd) and lead (Pb) during enhanced phytoextraction. The vertical column study was carried out to investigate the metal leaching meanwhile studying plant biomass and metals uptake for phytoextraction by Pelargonium hortroum. For this purpose, soil amendments at several concentration levels, such as ethylene diaminetetraacetic acid (EDTA 0, 4, 5 mmol kg-1), ammonium nitrate (0, 8, 10 mmol kg-1), citric acid (0, 8, 10 mmol kg-1), compost (0, 8%, 10%) and titanium dioxide nanoparticles (TNPs, 0, 80, 100 mg kg-1) were used. Results revealed that EDTA efficiently improved Cd and Pb accumulation (mg kg-1) in shoots and roots. Cd accumulation was significantly increased by 270%, 44%, 145%, and 6.4% in shoot and 94%, 19%, 48% and 14% in root upon EDTA, ammonium nitrate, citric acid and TNPs application, respectively compared to the control without amendment (WA). Similarly, Pb accumulation was significantly increased by 71%, 58% and 52% in shoot and 88.8%, 70.6% and 64.6% in root upon exposure to higher levels of EDTA, citric acid and ammonium nitrate, respectively when compared to the WA control. Application of TNPs reduced the Pb-accumulation in shoots and roots by 33% and 28%, respectively. Similarly, the Pb-accumulation in shoots and roots was decreased by 39% and 35%, respectively upon compost addition. Plant biomass was significantly increased upon application of soil amendments, with the exception of EDTA. The maximum Cd and Pb uptake was found in citric acid amended soil. Leachate analysis revealed that the concentrations of Cd and Pb were increased by 109% and 101% in leachates upon EDTA application as compared to other amendments. In comparison with other amendments, citric acid may be recommended as an environmentally friendly alternative for non-biodegradable EDTA for enhanced phytoextraction of Cd and Pb.

Methyltrioctylammonium chloride mediated removal of lignin from sugarcane bagasse for themostable cellulase production.

The present study was aimed to evaluate the Methyltrioctylammonium Chloride (IL) and Sodium hydroxide effect on sugarcane bagasse (SB) structure and its subsequent utilization to produce cellulase from a thermophilic bacterium Bacillus aestuarii UE25. The strain was isolated from a crocodile pond of Manghopir, Karachi. Ten different factors affecting IL pretreatment of SB and cellulase production by UE25 were evaluated by Plackett-Burman design and three significant factors were optimized by employing Box Behnken design. Under optimum conditions, the strain produced 118.4 IU mL-1 of EG and 75.01 IU mL-1 of BGL that corroborated well with the predicted values by the model. Scanning electron microscopy, gravimetric analysis, Fourier transform infrared spectroscopy and NMR of SB revealed removal of lignin, decrease in cellulose content and structural changes in the SB after pretreatment and fermentation. The data provide prospects of utilizing this IL in comparison to imidazolium based IL for pretreatment of biomass.

Metal tolerant bacteria enhanced phytoextraction of lead by two accumulator ornamental species.

Microbially enhanced availability and phytoextraction is a promising technique for phytoremediation of lead (Pb). In this study, Pb resistant strains were isolated and investigated for potential effects on plant growth and Pb phytoextraction. Incubation experiments were carried for inoculated and un-inoculated soil containing different levels of Pb. Results revealed that 20% of the isolated bacteria could tolerate Pb up to 800 mg L-1. Five Pb resistant strains with plant growth promoting (PGP) abilities were evaluated for possible influence on water soluble Pb through soil incubation experiments and significant increase i.e. 1.85- and 1.49-folds in water soluble Pb was observed for NCCP-1848 and NCCP-1862 strains, respectively. Pot experiments indicated significantly higher uptake by Pelargonium hortorum than that by Mesembryanthemum criniflorrum at all levels of soil Pb concentrations with the highest increase (1.9-folds) in plants inoculated with NCCP-1848 followed by NCCP-1862 (1.8-folds increase) compared to the control (Pb and without bacterial strain). The strains NCCP-1848 and NCCP-1862 were identified by 16S rRNA gene sequencing as Microbacterium sp. and Klebsiella sp. The results of present study suggest that Pb resistant plant growth promoting bacteria can serve as an effective bio-inoculant through wide action spectrum for maximizing efficiency of phytoremediation system.

Impact of meteorological conditions on the water quality of wastewater treatment systems: a comparative study of phytoremediation and membrane bioreactor system.

Two demonstration wastewater treatment systems, i.e. a phytoremediation system and a membrane bioreactor (MBR) system, were studied for a six-month period from August 2016 to January 2017. The phytoremediation system consists of wetland cells implanted with diverse phytoremediation macrophyte species at NUST H-12 sector Islamabad, Pakistan, while the MBR system comprises primary clarifiers, membrane tanks and bio tanks that treat domestic wastewater through hybridization of biological and biomechanical techniques. The phytoremediation system receives domestic wastewater at the rate of 283 m3/d, and greater hydraulic efficiencies were achieved because of compartmentalization and higher aspect ratios, whereas the MBR system receives 50 m3/d. The present study was conducted to analyze and compare the correlation between water quality parameters of wastewater treatment systems and meteorological conditions. Statistically significant correlation was exhibited between eight water quality parameters (pH, EC, turbidity, dissolved oxygen, total suspended solids (TSS), chemical oxygen demand (COD), biological oxygen demand (BOD) and total coliforms (TC)) and meteorological factors (ambient temperature and relative humidity). Predominant species isolated and identified through polymerase chain reaction and 16S rRNA sequencing from wastewater of the phytoremediation system and sludge of the MBR system belong to the phylum Proteobacteria with relatively higher abundance of Enterobacter, Shigella, Escherichia and Salmonella genera.

Strengthening calcium alginate microspheres using polysulfone and its performance evaluation: Preparation, characterization and application for enhanced biodegradation of chlorpyrifos.

Bacterial cell immobilization offer considerable advantages over traditional biotreatment systems using free cells. Calcium alginate matrix usually used for bacterial immobilization is susceptible to biodegradation in harsh environment. Current study aimed to produce and characterize stable macrocapsules (MCs) of Chlorpyrifos (CP) degrading bacterial consortium using biocompatible calcium alginate matrix coupled with environmentally stable polysulfone. In current study bacterial consortium capable of CP biodegradation was immobilized using calcium alginate in a form of microcapsule (MC) reinforced by being coated with a synthetic polymer polysulfone (PSf) through phase inversion. Consortium comprised of five bacterial strains was immobilized using optimized concentration of sodium alginate (2.5gL-1), calcium chloride (6gL-1), biomass (600mgL-1) and polysulfone (10gL-1). It has been observed that MCs have high thermal, pH and chemical stability than CAMs. In synthetic media complete biodegradation of CP (100-600mgL-1) was achieved using macrocapsules (MCs) within 18h. CAMs could be reused effectively only upto 5cycles, contrary to this MCs could be used 13 times to achieve more than >96% CP degradation. Shelf life and reusability studies conducted for MCs indicated unaltered biomass retention and CP biodegradation activity (95%) over 16weeks of storage. MCs achieved complete biodegradation of CP (536mgL-1) in real industrial wastewater and reused several times effectively. Metabolites (3,5,6-trichloro-2-pyridinol (TCP), 3,5,6-trichloro-2-methoxypyridine (TMP) and diethyl-thiophosphate (DETP) were traced using GC-MS and possible metabolic pathway was constructed. Study indicated MCs could be used for cleanup of CP contaminated wastewater repeatedly, safely, efficiently for a longer period of time.

Biodegradation of phenol and benzene by endophytic bacterial strains isolated from refinery wastewater-fed Cannabis sativa.

The presence of benzene and phenol in the environment can lead to serious health effects in humans and warrant development of efficient cleanup strategies. The aim of the present work was to assess the potential of indigenous endophytic bacterial strains to degrade benzene and phenol. Seven strains were successfully isolated from Cannabis sativa plants irrigated with oil refinery wastewater. Molecular characterization was performed by 16S rRNA gene sequencing. Phenol was biodegraded almost completely with Achromobacter sp. (AIEB-7), Pseudomonas sp. (AIEB-4), and Alcaligenes sp. (AIEB-6) at 250, 500, and 750 mg L-1; however, the degradation was only 81%, 72%, and 69%, respectively, when exposed to 1000 mg L-1. Bacillus sp. (AIEB-1), Enterobacter sp. (AIEB-3), and Acinetobacter sp. (AIEB-2) degraded benzene significantly at 250, 500, and 750 mg L-1. However, these strains showed 80%, 72%, and 68% benzene removal at 1000 mg L-1 exposure, respectively. Rates of degradation could be modeled with first-order kinetics with rate constant values of 1.86 × 10-2 for Pseudomonas sp. (AIEB-4) and 1.80 × 10-2 h-1 for Bacillus sp. (AIEB-1) and half-lives of 1.5 and 1.6 days, respectively. These results establish a foundation for further testing of the phytoremediation of hydrocarbon-contaminated soils in the presence of these endophytic bacteria.

Species-specific interaction of trihalomethane (THM) precursors in a scaled-up distribution network using response surface methodology (RSM).

Response surface methodology (RSM) with central composite design (CCD) was used to monitor and optimize species-specific interaction of trihalomethane (THM) precursors in a scaled-up distribution network (DN). Independent variables such as applied chlorine (Cl2), contact time (t), humic acid (HA) and bromide ions (Br-) were analyzed using full factorial CCD. Analysis of variance revealed a good agreement between experimental data and proposed a two-factor interaction model (p = .04, R2 = 0.7983). As a precursor, Cl- and Br- interaction with HA affected THMs' speciation. These precursor molecules were perceived least significant as discrete elements but HA: Br- and pH product significantly impacted total trihalomethane (TTHM) formation (r = 0.998, p = .007). This mutual interactive fraction was observed pH-dependent and influenced TTHM yield. Dibromochloromethane and bromoform formation was observed pH-dependent provided sufficient Br- in the system. Applied chlorine had significant (p = .01), while time had insignificant (p = .75) effect. Multiple response optimization suggested pH range between 6.0 and 7.6 and HA: Br- ratio between 1.3 and 5.9 were satisfactory for maintaining TTHM below ≤80 µg/L in DN with 0.88 desirability function (D). Their respective concentration may be minimized by changing precursor's individual concentration and possible combinations.