In vitro Cr(VI) reduction by cell-free extracts of chromate-reducing bacteria isolated from tannery effluent irrigated soil

Four efficient Cr(VI)-reducing bacterial strains were isolated from rhizospheric soil of plants irrigated with tannery effluent and investigated for in vitro Cr(VI) reduction. Based on 16S rRNA gene sequencing, the isolated strains SUCR44, SUCR140, SUCR186, and SUCR188 were identified as Bacillus sp. (JN674188), Microbacterium sp. (JN674183), Bacillus thuringiensis (JN674184), and Bacillus subtilis (JN674195), respectively. All four isolates could completely reduce Cr(VI) in culture media at 0.2 mM concentration within a period of 24–120 h; SUCR140 completely reduced Cr(VI) within 24 h. Assay with the permeabilized cells (treated with Triton X-100 and Tween 80) and cell-free assay demonstrated that the Cr(VI) reduction activity was mainly associated with the soluble fraction of cells. Considering the major amount of chromium being reduced within 24–48 h, these fractions could have been released extracellularly also during their growth. At the temperature optima of 28 °C and pH?7.0, the specific activity of Cr(VI) reduction was determined to be 0.32, 0.42, 0.34, and 0.28 μmol Cr(VI)?min^?1?mg^?1 protein for isolates SUCR44, SUCR140, SUCR186, and SUCR188, respectively. Addition of 0.1 mM NADH enhanced the Cr(VI) reduction in the cell-free extracts of all four strains. The Cr(VI) reduction activity in cell-free extracts of all the isolates was stable in presence of different metal ions tested except Hg²⁺. Beside this, urea and thiourea also reduced the activity of chromate reduction to significant levels.     

Pentachlorophenol dechlorination and simultaneous Cr6+ reduction by Pseudomonas putida SKG-1 MTCC (10510): characterization of PCP dechlorination products, bacterial structure, and functional groups

It is the first report in which a novel psychrotrophic Pseudomonas putida SKG-1 strain was evaluated for simultaneous bioremediation of pentachlorophenol and Cr⁶⁺ under various cultural and nutritional conditions. Pentachlorophenol (PCP) dechlorination products, bacterial structure, and functional groups were characterized by gas chromatography and mass spectrometry (GC–MS), scanning electron microscope and energy dispersive X-ray spectroscopy (SEM–EDS), and Fourier-transform infrared (FTIR) techniques. The strain was extremely tolerant to excessively higher individual concentration of PCP (1,400 mg l^?1) and Cr⁶⁺ (4,300 mg l^?1). Increasing concentration of PCP and Cr⁶⁺ exerted inhibitory effect on bacterial growth and toxicants’ removal. The strain exhibited growth, and concomitantly remediated both the pollutants simultaneously over a broad pH (7.0–9.0) and temperature (28–32 °C) range; maximum growth, PCP dechlorination (87.5 %), and Cr⁶⁺ removal (80.0 %) occurred at optimum pH 8.0 and 30 °C (from initial PCP 100 mg l^?1 and Cr⁶⁺ 500 mg l^?1) under shaking (150 rpm) within 72 h incubation. Optimization of agitation (125 rpm) and aeration (0.4 vvm) in bioreactor further enhanced PCP dechlorination by ~10 % and Cr⁶⁺ removal by 2 %. A direct correlation existed between growth and bioremediation of both the toxicants. Among other heavy metals, mercury exerted maximum and cobalt minimum inhibitory effect on PCP dechlorination and Cr⁶⁺ removal. Chromate reductase activity was mainly associated with the supernatant and cytosolic fraction of bacterial cells. GC–MS analysis revealed the formation of tetrachloro-p-hydroquinone, 2,4,6-trichlorophenol, and 2,6-dichlorophenol as PCP dechlorination products. FTIR spectrometry indicated likely involvement of carbonyl and amide groups in Cr³⁺ adsorption, and SEM–EDS showed the presence of chromium on P. putida surface. Thus, our promising isolate can be ecofriendly employed for biotreatment of various industrial wastes contaminated with high PCP and Cr⁶⁺ concentrations.     

Arsenite tolerance in rice (Oryza sativa L.) involves coordinated role of metabolic pathways of thiols and amino acids

Thiolic ligands and several amino acids (AAs) are known to build up in plants against heavy metal stress. In the present study, alteration of various AAs in rice and its synchronized role with thiolic ligand was explored for arsenic (As) tolerance and detoxification. To understand the mechanism of As tolerance and stress response, rice seedlings of one tolerant (Triguna) and one sensitive (IET-4786) cultivar were exposed to arsenite (0–25 μM) for 7 days for various biochemical analyses using spectrophotometer, HPLC and ICPMS. Tolerant and sensitive cultivars respond differentially in terms of thiol metabolism, essential amino acids (EEAs) and nonessential amino acids (NEEAs) vis-á-vis As accumulation. Thiol biosynthesis-related enzymes were positively correlated to As accumulation in Triguna. Conversely, these enzymes, cysteine content and GSH/GSSG ratio declined significantly in IET-4786 upon As exposure. The level of identified phytochelatin (PC) species (PC₂, PC₃ and PC₄) and phytochelatin synthase activity were also more pronounced in Triguna than IET-4786. Nearly all EAAs were negatively affected by As-induced oxidative stress (except phenylalanine in Triguna), but more significantly in IET-4786 than Triguna. However, most of the stress-responsive NEAAs like glutamic acid, histidine, alanine, glycine, tyrosine, cysteine and proline were enhanced more prominently in Triguna than IET-4786 upon As exposure. The study suggests that IET-4786 appears sensitive to As due to reduction of AAs and thiol metabolic pathway. However, a coordinated response of thiolic ligands and stress-responsive AAs seems to play role for As tolerance in Triguna to achieve the effective complexation of As by PCs.     

Health care industries: potential generators of genotoxic waste

Health care waste includes all the waste generated by health care establishments, research facilities, and laboratories. This constitutes a variety of chemical substances, such as pharmaceuticals, radionuclides, solvents, and disinfectants. Recently, scientists and environmentalists have discovered that wastewater produced by hospitals possesses toxic properties due to various toxic chemicals and pharmaceuticals capable of causing environmental impacts and even lethal effects to organisms in aquatic ecosystems. Many of these compounds resist normal wastewater treatment and end up in surface waters. Besides aquatic organisms, humans can be exposed through drinking water produced from contaminated surface water. Indeed, some of the substances found in wastewaters are genotoxic and are suspected to be potential contributors to certain cancers. The aim of this study was to evaluate the genotoxic and cytotoxic potential of wastewaters from two hospitals and three clinical diagnostic centers located in Jaipur (Rajasthan State), India using the prokaryotic Salmonella mutagenicity assay (Ames assay) and the eukaryotic Saccharomyces cerevisiae respiration inhibition assay. In the Ames assay, untreated wastewaters from both of the health care sectors resulted in significantly increased numbers of revertant colonies up to 1,000–4,050 as measured by the Salmonella typhimurium TA98 and TA100 strains (with and without metabolic activation) after exposure to undiluted samples, which indicated the highly genotoxic nature of these wastewaters. Furthermore, both hospital and diagnostic samples were found to be highly cytotoxic. Effective concentrations at which 20 % (EC₂₀) and 50 % (EC₅₀) inhibition of the respiration rate of the cells occurred ranged between ～0.00 and 0.52 % and between 0.005 and 41.30 % (calculated with the help of the MS excel software XLSTAT 2012.1.01; Addinsoft), respectively, as determined by the S. cerevisiae assay. The results indicated that hospital wastewaters contain genotoxic and cytotoxic components. In addition, diagnostic centers also represent small but significant sources of genotoxic and cytotoxic wastes.     

Phototransformation of herbicide metsulfuron methyl

Phototransformation of the herbicide metsulfuron methyl was investigated on glass surface under sunlight and ultraviolet (UV) light and compared with dark condition. The half-lives of metsulfuron methyl under UV light and sunlight were found to be 0.5 and 7.8 days respectively. Rate of phototransformation followed first order kinetics with significant correlation coefficient. The major photoproducts were identified as methyl-2-sulfonyl-amino-benzoate, 2-amino-6-methoxy-4-methyltriazine and saccharin (O-sulfobenzoimide). Various metabolites from this study were identified by high performance liquid chromatography (HPLC). Authentic samples required for HPLC comparison were prepared in laboratory and characterized on the basis of nuclear magnetic resonance (NMR) and infra red (IR) spectral data. These metabolites were also identified from metsulfuron methyl treated wheat field soil.     

Reduced downward mobility of metribuzin in fly ash-amended soils

Metribuzin, a triazine herbicide, is poorly sorbed in the soils, therefore leaches to lower soil profile. Fly ash amendment, which enhanced metribuzin sorption in soils, may play a significant role in reducing the downward mobility of herbicide. Therefore, the present study reports the effect of Inderprastha fly ash amendment on metribuzin leaching in three soil types. Fly ash was amended at 1, 2 and 5% levels in the upper 15 cm of 30 cm long packed soil columns. Results suggested a significant reduction in the leaching losses of metribuzin in fly ash-amended columns of all the three soil types and effect increased with increase in the level of fly ash. Even after percolating water equivalent to 362 mm rainfall no metribuzin was recovered in the leachate of 5% fly ash-amended columns. Fly ash application affected both metribuzin breakthrough time and its maximum concentration in the leachate. Further, it resulted in greater retention of metribuzin in the application zone and better effect was observed in the organic carbon poor soils.     

Persistence,degradation and leaching of coumaphos in soil

Abstract

Dissipation, degradation and leaching of fresh ¹⁴C coumaphos, alkylated ¹⁴C coumaphos and aged residues of ¹⁴C coumaphos from vats were studied in alkaline sandy loam soil in soil columns in the field under subtropical conditions in Delhi for a year. Dissipation, degradation and bound residue formation was more in case of alkali treated coumaphos than fresh coumaphos. After 365 days total residues of fresh coumaphos accounted for 33.25% while that of alkali treated coumaphos was 19.12%. Bound residue formation was almost double in case of alkali treated coumaphos (18.95%) than fresh coumaphos (9.53%) after 150 days followed by release of bound residue in both the cases. The proportion of metabolites 4 ‐ methylumbelliferone, chlorferon and potasan collectively was 86.05% in fresh coumaphos extractable residues while the same was 91.74% in alkali treated coumaphos after 365 days. Aged residues from vats containing copper sulphate and buffer were found to be more persistent in soil as total residues remained were 95.58% in comparison with 83.09% total residues of aged residues from vats containing only buffer after 150 days of treatment. Copper sulphate seems to inhibit the degradatiion of coumaphos in soil by microorganisms. Chlorferon was the major metabolite in generally all the samples. Coumaphos did not leach below 10 cm in all the cases.     

Effect of UV radiation and temperature on mineralization and volatilization of coumaphos in water

This study was undertaken to determine the dissipation and degradation of coumaphos [O-(3-chloro-4-methyl-2-oxo-2H-1-benzopyran-7-yl) O,O-diethyl phosphorothioate] under different sunlight conditions and at different temperatures. The effect of the ultra violet (UV) component of solar radiation was also studied using quartz tubes in addition to other radiation in the visible range using glass tubes and the results were compared with those obtained under the dark light conditions. Water suspensions of coumaphos were incubated at three temperatures viz. 22°C, 37°C and 53°C in closed systems to study the effect of temperature. Volatilization, mineralization and degradation of coumaphos increased with an increase in temperature and exposure to solar radiation, particularly under the UV component of the solar radiation. Major loss of the pesticide occurred through volatilization. The optimum temperature for the degradation of coumaphos was found to be at 37°C. The data obtained from the mineralization and degradation studies indicated that 53°C crosses the biological range for suitable growth of microorganism. UV radiation exposure along with maintaining temperature at 37°C may prove useful in the dissipation and/or degradation of coumaphos prior to its disposal as waste from cattle dipping vats.     

Fate of parathion in soil under sub‐tropical field conditions

Abstract

Persistence, metabolism and binding of ¹⁴C‐parathion in alkaline sandy loam soil under sub‐tropical conditions of Delhi were studied for 545 days. After 3 days of treatment, ¹⁴C‐residues declined to 41% of the amount applied. The dissipation curve was biphasic; an initial rapid phase (up to 7 days) followed by slow dissipation. The half life of dissipation was only 3.36 days for the first phase and 84 days for the slow phase. The overall half life was 64.5 days. The total residues at zero‐time were 10.65 μg/g dry soil and were almost totally extractable. The extracts consisted of parathion, 4‐aminophenol, 4‐nitrophenol and paraoxon. The bound residues gradually increased and accounted for the total residue at the end of one year (0.7 μg/g).     

Development of controlled release nanoformulations of carbendazim employing amphiphilic polymers and their bioefficacy evaluation against Rhizoctonia solani

Controlled release nanoformulations of carbendazim (Methyl 1H-benzimidazol-2-ylcarbamate), a systemic fungicide, have been prepared using laboratory synthesized poly(ethylene glycols) (PEGs)-based functionalized amphiphilic copolymers. The release kinetics of carbendazim from developed controlled release (CR) formulations was studied and compared with that of the commercially available 50% Wettable Powder (WP). Further, the bioefficacy evaluation of developed formulations was done against plant pathogenic fungi Rhizoctonia solani by the poison food technique method. The release of maximum amount of carbendazim from developed formulations was dependent on the molecular weight of PEGs and was found to increase with increasing molecular weights. The range of carbendazim release was found to be between 10th to 35th day as compared to commercial formulation which was up to 7th day. The diffusion exponent (n value) of carbendazim in water ranged from 0.37 to 0.52 in the tested formulations. The half-release (t_1/2) values ranged between 9.47 and 24.20 days, and the period of optimum availability (POA) of carbendazim ranged from 9.15 to 26.63 days. Also, ED₅₀ values of the developed formulations vary from 0.40 to 0.74 mg L^?1. These formulations can be used to optimize the release of carbendazim to achieve disease control for the desired period depending on the matrix of the polymer used.