Improved degradation of azo dyes by lignin peroxidase following mutagenesis at two sites near the catalytic pocket and the application of peroxidase-coated yeast cell walls

• Mutations in Lignin peroxidase Trp171 environment improved azo dyes degradation. • Expression on yeast cell surface and cell lysis allowed reusability of biocatalyst. • Aga2-LiP chimeric variants were characterized. The enzymatic degradation of azo dyes is a promising alternative to ineffective chemical and physical remediation methods. Lignin peroxidase (LiP) from Phanerochaete chrysosporium is a heme-containing lignin-degrading oxidoreductase that catalyzes the peroxide-dependent oxidation of diverse molecules, including industrial dyes. This enzyme is therefore ideal as a starting point for protein engineering. Accordingly, we subjected two positions (165 and 264) in the environment of the catalytic Trp171 residue to saturation mutagenesis, and the resulting library of 10⁴ independent clones was expressed on the surface of yeast cells. This yeast display library was used for the selection of variants with the ability to break down structurally-distinct azo dyes more efficiently. We identified mutants with up to 10-fold greater affinity than wild-type LiP for three diverse azo dyes (Evans blue, amido black 10B and Guinea green) and up to 13-fold higher catalytic activity. Additionally, cell wall fragments displaying mutant LiP enzymes were prepared by toluene-induced cell lysis, achieving significant increases in both enzyme activity and stability compared to a whole-cell biocatalyst. LiP-coated cell wall fragments retained their initial dye degradation activity after 10 reaction cycles each lasting 8 h. The best-performing mutants removed up to 2.5-fold more of each dye than the wild-type LiP in multiple reaction cycles.     

Effect of coumaphos on cholinesterase activity,hematology, and biochemical blood parameters of bovines in tropical regions of Mexico

To assess the effect of coumaphos [O-(3-chloro-4-methyl-2-oxo-2H-1-benzopyran-7-yl) O,O-diethyl phosphorothioate] exposure on physiological responses during bovine production, acetylcolinesterase (AChE) and butyrylcholinesterase (BuChE) activities were measured in whole blood, erythrocytes, and plasma of healthy male steers (Bos Taurus x Bos indicus) sprayed with coumaphos at a non-lethal dose of 1 mg kg^{? 1} body weight per day once every 14 (in vivo group) or 21 days (southern and central groups). Coumaphos topically administered at 1 mg/kg body weight per day to cattle under normal management practices in tropical areas produced a significant inhibition in erythrocyte (RBC) AChE and BuAChE activities when compared to baseline levels. RBC-AChE activity for the in vivo group decreased 71.3% (P < 0.05) and BuChE activity 59.1% (P < 0.05); RBC-AChE activity decreased 55.1% (P < 0.05) (southern group) and 43.4% (P < 0.05) (central group). Compared to the control specimens, steers from in vivo, southern, and central groups after 150 days of exposure had lower (P < 0.05) leukocyte count, absolute lymphocyte, erythrocyte, and platelet counts. Decreases in RBC-AChE activities correlated with decreased lymphocyte (r = 1.000, p = 0.01), erythrocyte (r = 1.000, p = 0.003), and platelet counts (r = 0.841, p = 0.036). Significantly increased BUN levels (P < 0.05) correlated with the decrease in RBC-AChE activities (r = ? 0.997, p = 0.047) and with the decrease in absolute red blood cell (r = ? 0.883, p = 0.020) and lymphocyte (r = ? 0.825, p = 0.043) counts; increased (P < 0.05) total plasma protein levels correlated with the decrease in RBC-AChE activities (r = ?0.998, p = 0.043), absolute red blood cell (r = ? 0.998, p = 0.040), lymphocyte (r = ? 0.893, p = 0.017), and platelet (r = -0.855, p = 0.030) counts. The physiological responses correlated with the erythrocyte acetylcholinesterase inhibition could be considered as early indicators or warning responses of bovine exposures to organophosphorus pesticides (OPs).     

Assessing changes in the Presque Isle Bay watershed fish community using a modified index of biotic integrity: before and after the elimination of combined sewer overflows

An index of biotic integrity and species richness were used to assess changes in the Presque Isle Bay watershed fish community before and after the elimination of combined sewer overflows (CSOs). The fish community was sampled with a backpack electrofisher in 2011 at 12 stream locations on 4 tributaries of Presque Isle Bay, Erie County, Pennsylvania. All sites were previously sampled in 2001. Significant increases in species richness and index of biotic integrity (IBI) scores were observed in 2011 compared to 2001. The significant increases in species richness and IBI scores occurred following the elimination of 10 CSOs to Garrison Run, 7 CSOs to Cascade Creek, and 37 CSOs to Mill Creek. Despite the increased richness and IBI scores, the fish community remains in poor condition, which may be related to the highly urbanized land use of the watershed. Urban land uses comprise 77 % of the Presque Isle Bay watershed, and in both 2011 and 2001, the watershed as a whole did not meet warm-water habitat criteria. It is unlikely that the fish community will continue to recover without addressing urbanization throughout the watershed.     

Estimating trace gas and aerosol emissions over South America: Relationship between fire radiative energy released and aerosol optical depth observations

Contemporary human activities such as tropical deforestation, land clearing for agriculture, pest control and grassland management lead to biomass burning, which in turn leads to land-cover changes. However, biomass burning emissions are not correctly measured and the methods to assess these emissions form a part of current research area. The traditional methods for estimating aerosols and trace gases released into the atmosphere generally use emission factors associated with fuel loading and moisture characteristics and other parameters that are hard to estimate in near real-time applications. In this paper, fire radiative power (FRP) products were extracted from Moderate Resolution Imaging Spectroradiometer (MODIS) and from the Geostationary Operational Environmental Satellites (GOES) fire products and new South America generic biomes FRE-based smoke aerosol emission coefficients were derived and applied in 2002 South America fire season. The inventory estimated by MODIS and GOES FRP measurements were included in Coupled Aerosol-Tracer Transport model coupled to the Brazilian developments on the Regional Atmospheric Modeling System (CATT-BRAMS) and evaluated with ground truth collected in Large Scale Biosphere–Atmosphere Smoke, Aerosols, Clouds, rainfall, and Climate (SMOCC) and Radiation, Cloud, and Climate Interactions (RaCCI). Although the linear regression showed that GOES FRP overestimates MODIS FRP observations, the use of a common external parameter such as MODIS aerosol optical depth product could minimize the difference between sensors. The relationship between the PM_2.5μm (Particulate Matter with diameter less than 2.5 μm) and CO (Carbon Monoxide) model shows a good agreement with SMOCC/RaCCI data in the general pattern of temporal evolution. The results showed high correlations, with values between 0.80 and 0.95 (significant at 0.5 level by student t test), for the CATT-BRAMS simulations with PM_2.5μm and CO.     

Records of polychlorinated biphenyls (PCBs) in sediments of four remote Chilean Andean Lakes

Sediment cores from four Chilean lakes along the Andes Chain (Chungará, Laja, Castor and Venus) were analysed in order to investigate PCB concentrations and distributions in sediment samples. Sediment cores were analysed for PCBs using gas chromatography (GC-ECD/MS) and radioisotopically dated using 210Pb. Organic carbon content (OC) and 210Pb fluxes were also measured. Results showed that sediment PCB concentrations (ngg(-1) d.w.) at Lake Chungará (1.2 +/- 1) in northern Chile, Laja (5 +/- 4) in central, and in Lake Castor (3.5 +/- 4) in southern Chile (the eastern side of the Andes Mountain) were lower than sediments collected from Lake Venus (64 +/- 30) located in southern Chile (the western side) which contained 15-fold higher concentrations of PCBs. The percentage (%) of organic carbon was variable and showed a high range of values in the sediment fluctuating from 2% (Lake Laja) to 22% (Lake Chungará). Analysis of 210Pb fluxes, presented a decrease trend following Lake Laja>Castor>Chungará with a positive correlation with rainfall at each site. Sedimentation rates in Lake Castor (1846 gm(-2)yr(-1)) were higher than at Chungará (748 gm(-2)yr(-1)) and Lake Laja (508 gm(-2)yr(-1)). Focusing factor (FF) is used as a tool to elucidate PCB input in the aquatic ecosystem. FF were lower (<1) for the shallower lakes (Lakes Chungará and Castor). This study provides background levels of PCBs at remote lakes in Chile. Differences in geographical characteristics (orographic effect) might play an important role in the arrival of PCBs, particularly into the southern lakes. PCB fluxes indicated deposition of PCBs in recent sediments is higher than in previous years with peaks of PCB between 1991 and 1998. The continuing increase of PCB inputs in remote Chilean lakes, could be associated with long range atmospheric transport (LRAT).     

Performance and stability of an anaerobic fixed bed reactor subjected to progressive increasing concentrations of influent organic matter and organic shock loads

Data on the performance of a horizontal-flow anaerobic immobilized biomass (HAIB) reactor subjected to step increases of organic loading rates (OLR) and to organic shock loads (OSL) are presented and discussed. The tubular reactor (100 cm long and 5 cm diameter) with a useful volume of 1995 mL was filled with polyurethane foam cubic matrices holding immobilized biomass and fed with synthetic wastewater. The reactor was operated at the controlled temperature of 30+/-1 degrees C and hydraulic retention time of 7 h. After about 15 days, the HAIB reactor attained operating stability. Thereafter, it was subjected to step increases of the applied OLR that ranged from 6.8 to 18.8 kg COD/m(3)d. After steady state had been achieved at each step, OSL corresponding to approximately three times the operating OLR were applied for 7 h. No disturbance was observed due to the step increase in OLR. An increase in effluent chemical oxygen demand (COD) and volatile fatty acids (VFA) concentrations and a decrease in the percentage of methane in the biogas were observed due to OSL applications. However, stability of the monitoring parameters was always restored approximately 17 h after the application of OSL for all conditions tested.     

Primary prevention for worker health and safety: cleaner production and toxics use reduction in Massachusetts

Worker health and safety and environmental protection are not always considered simultaneously when attempting to reduce or eliminate hazardous materials from our environment. Cleaner Production-Pollution Prevention (CPPP), as primary prevention, has the ability to shift worker health and safety strategies from control to prevention, where exposure prevention precedes exposure control. This paper evaluates the effect of Cleaner Production-Pollution Prevention in the form of toxics use reduction (TUR) on worker health and safety at three printed wire board facilities covered under the Massachusetts Toxics Use Reduction Act. In-depth case study analysis, including an assessment of each facility’s health and safety status, explores the root causes of the worker health and safety changes attributable to the TUR interventions. By exploring the relationship between worker health and safety and environmental protection within the corporate structure; we can identify the factors driving companies to reduce toxics both inside and outside of their plants, as a single concern.While traditionally there have been divergent paths of practice for worker health and safety and environmental protection, the two are closely connected. It is important, however, to consider the implications of risk transfer/shifting between the general and work environment. In order to avoid this risk shifting, worker health and safety perspectives and goals must be more clearly incorporated into the Cleaner Production-Pollution Prevention/TUR management system. This study opens a dialog around the effects of environmental intervention programs on worker health and safety. We realize now that while CPPP/TUR reduces exposure to toxic substances in the general environment, it also offers unique opportunities to reaffirm primary prevention principles in the work environment.     

Taking advantage of storm and waste water retention basins as part of water use minimization in industrial sites

A methodology for water use minimization has been developed by the Clean Technology Network of Bahia over a 10 year period in joint cooperative programs with the chemical, petrochemical and copper metallurgy industries located in the largest Industrial Complex in Latin America, in Camaçari, Bahia, Brazil. The methodology comprises a set of tools including reconciled aqueous stream balances, database of aqueous streams; large scale training leading to the identification of water minimization alternatives in the processes, water reuse optimization approaches; geographical information systems as well as, consideration of the region's hydro and hydro-geological characteristics. The results of a study carried out to assess the possibility of using storm and wastewaters for industrial use is presented in this paper. The inorganic system is composed by three water reservoirs (basins) receiving stormwater contaminated with inorganic effluents, and occasionally with organics. These basins have been operated to control water flow inputs according to the capacity of the pumping outlet systems before their discharge to a submarine outfall. A mass balance was performed with historical updated data to assess water availability from the basins based on the daily volume variation and flow rate of inorganic effluent from 2001 to 2007. The study identified the possibility of recovering about 1140 m³/h of the overall 5400 m³/h consumed by the Industrial Complex at the moment. Organizational changes in the present effluent disposal and stormwater harvest systems will be required in order to maximize water recovery for industrial use.