Bioremediation potential of select bacterial species for the neonicotinoid insecticides, thiamethoxam and imidacloprid.

Thiamethoxam (THM) and imidacloprid (IMI), are environmentally persistent neonicotinoid insecticides which have become increasingly favored in the past decade due to their specificity as insect neurotoxicants. However, neonicotinoids have been implicated as a potential contributing factor in Colony Collapse Disorder (CCD) which affects produce production on a global scale. The present study characterizes the bioremediation potential of six bacterial species: Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas aeruginosa, Alcaligenes faecalis, Escherichia coli, and Streptococcus lactis. In Phase I, we evaluated the utilization of IMI or THM as the sole carbon or nitrogen source by P. fluorescens, P. putida, and P. aeruginosa. All three species were better able to utilize THM over IMI as their sole carbon or nitrogen source. Thus, further studies proceeded with THM only. In Phase II, we assessed the kinetics of THM removal from aqueous media by the six species. Significant (p < 0.0001) reductions in 70 mg/L THM concentration were observed for P. fluorescens (67%), P. putida (65%), P. aeruginosa (52%), and A. faecalis (39%) over the 24-day study period, and for E. coli (60%) and S. lactis (12%) over the 14-day study period. The THM removal by all species followed a first-order kinetic reaction. HPLC chromatograms of P. fluorescens, P. putida, and E. coli cultures revealed that as the area of the THM peak decreased over time, the area of an unidentified metabolite peak increased. In Phase III, we examined the effect of temperature on the transformation capacity of the bacterial species which was observed at 2 â„ƒ, 22 â„ƒ, and 30 â„ƒ. Maximal THM removal occurred at 30 °C for all bacterial species assessed. Identification of the metabolite is currently underway. If the metabolite is found to be less hazardous than THM, further testing will follow to evaluate the use of this bioremediation technique in the field.

Phytoremediation potential of switchgrass (Panicum virgatum), two United States native varieties, to remove bisphenol-A (BPA) from aqueous media.

Plastic wastes burdening Earth's water and accumulating on land, releasing toxic leachates, are one of the greatest global threats of our time. Bisphenol-A (BPA), a potent endocrine disrupting compound, is a synthetic ingredient of the polycarbonate plastics and epoxy resins used in food containers, cans, and water bottles. Bisphenol-A's rising concentrations in the environment require a sustainable alternative to current removal practices, which are expensive and/or ecologically unsafe. Switchgrass offers a safe alternative. To investigate its potential for BPA removal, two United States native switchgrass varieties where tested in hydroponic media. Results show minimal hydrolysis and photolysis of BPA over 55 days, confirming its persistence. Both generic and heavy metal switchgrass exhibited statistically significant (p < 0.0001) BPA removal (40% and 46%, respectively) over approximately 3 months, underscoring switchgrass's effectiveness for BPA removal. Significantly higher (p < 0.005) BPA accumulation in roots than shoots and nonsignificant variances in biomass, chlorophyll (p > 0.19), and peroxidase between BPA-treated and untreated plants indicates substantial BPA tolerance in both varieties. Kinetic parameters of BPA removal and translocation factors were also determined, which will inform the design of BPA removal phytotechnologies for a variety of soil conditions, including landfills where BPA accumulation is greatest.

Proteomic profiling of vetiver grass (Chrysopogon zizanioides) under 2,4,6-trinitrotoluene (TNT) stress.

Vetiver grass is an ideal plant for 2,4,6-trinitrotoluene (TNT) phytoremediation, due to its ability to tolerate and metabolize TNT as previously reported. The current study is the first attempt to investigate the changes in the proteomic profile of a plant under TNT stress. Vetiver plants were grown in nutrient media with varying concentrations of TNT (0, 25, 50, and 100 mg L-1) for 10 days. Although the plants appeared healthy, significant biomass reductions (p = 0.0008) were observed in treated plants. Total proteins in the root decreased significantly (p = 0.0003). Proteomic analysis of root proteins revealed the downregulation of functional proteins involved in key cellular mechanisms such as transcription, ribosome biogenesis, nucleo-cytoplasmic transport of proteins, protein glycosylation, and translation. Growth-related proteins were downregulated; plant defense proteins were upregulated at lower TNT concentrations but downregulated at higher concentrations. Comprehensive understanding of changes in the proteomic profile provides important clues to the mechanism of TNT stress response and tolerance in vetiver.

Evidence for exocellular Arsenic in Fronds of Pteris vittata.

The arsenic (As) hyperaccumulating fern species Pteris vittata (PV) is capable of accumulating large quantities of As in its aboveground tissues. Transformation to AsIII and vacuolar sequestration is believed to be the As detoxification mechanism in PV. Here we present evidence for a preponderance of exocellular As in fronds of Pteris vittata despite numerous reports of a tolerance mechanism involving intracellular compartmentalization. Results of an extraction experiment show that 43-71% of the As extruded out of the fronds of PV grown in 0.67, 3.3 and 6.7 mM AsV. SEM-EDX analysis showed that As was localized largely on the lower pinna surface, with smaller amounts on the upper surface, as crystalline deposits. X-ray fluorescence imaging of pinna cross-sections revealed preferential localization of As on the pinna surface in the proximity of veins, with the majority localized near the midrib. Majority of the As in the pinnae is contained in the apoplast rather than vacuoles. Our results provide evidence that exocellular sequestration is potentially a mechanism of As detoxification in PV, particularly at higher As concentrations, raising concern about its use for phytoremediation.

Tetracycline uptake and metabolism by vetiver grass (Chrysopogon zizanioides L. Nash).

Environmental contamination by antibiotics not only perturbs the ecological balance but also poses a risk to human health by promoting the development of multiantibiotic-resistant bacteria. This study focuses on identifying the biochemical pathways associated with tetracycline (TC) transformation/degradation in vetiver grass that has the potential to be used as a biological remediation system in TC-contaminated water sources. A hydroponic experimental setup was used with four initial TC concentrations (0, 5, 35, 75 ppm), and TC uptake was monitored over a 30-day period. Results show that TC transformation in the media occurred during the first 5 days, where a decrease in the parent compound and an increase in the concentration of the isomers such as epitetracycline (ETC) and anhyrotetracycline (ATC) occurred, and TC disappeared in 20 days in tanks with vetiver grass. However, the isomers ETC and ATC remained in the control tanks for the duration of the trial. Transformation products of TC in plant tissue were analyzed by using ultra HPLC high-resolution Orbitrap mass spectrometery (HRMS/MS), which indicates amide hydrolysis of TC in vetiver roots. Metabolic profiling revealed that glyoxylate metabolism, TCA cycle, biosynthesis of secondary metabolites, tryptophan metabolism, and inositol phosphate metabolism were impacted in vetiver root by TC treatment.

Phytoremediation potential of vetiver grass [Chrysopogon zizanioides (L.)] for tetracycline.

The presence of veterinary and human antibiotics in soil and surface water is an emerging environmental concern. The current study was aimed at evaluating the potential of using vetiver grass as a phytoremediation agent in removing Tetracycline (TC) from aqueous media. The study determined uptake, translocation, and transformation of TC in vetiver grass as function of initial antibiotic concentrations and exposure time. Vetiver plants were grown for 60 days in a greenhouse in TC contaminated hydroponic system. Preliminary results show that complete removal of tetracycline occurred within 40 days in all TC treatments. Initial concentrations of TC had significant effect (p < 0.0001) on the kinetics of removaL Tetracycline was detected in the root as well as shoot tissues, confirming uptake and root-to-shoot translocation. Liquid-chromatography-tandem-mass-spectrometry analysis of plant tissue samples suggest presence of metabolites of TC in both root and shoot tissues of vetiver grass. The current data is encouraging and is expected to aid in developing a cost-effective, in-situ phytoremediation technique to remove TC group of antibiotics from wastewater.

Effectiveness of urea in enhancing the extractability of 2,4,6-trinitrotoluene from chemically variant soils.

One of the major challenges in developing an effective phytoremediation technology for 2,4,6-trinitrotoluene (TNT) contaminated soils is limited plant uptake resulting from low solubility of TNT. The effectiveness of urea as a solubilizing agent in increasing plant uptake of TNT in hydroponic systems has been documented. Our preliminary greenhouse experiments using urea were also very promising, but further characterization of the performance of urea in highly-complex soil-solution was necessary. The present study investigated the natural retention capacity of four chemically variant soils and optimized the factors influencing the effectiveness of urea in enhancing TNT solubility in the soil solutions. Results show that the extent of TNT sorption and desorption varies with the soil properties, and is mainly dependent on soil organic matter (SOM) content. Hysteretic desorption of TNT in all tested soils suggests irreversible sorption of TNT and indicates the need of using an extractant to increase the release of TNT in soil solutions. Urea significantly (p<0.0001) enhanced TNT extraction from all soils, by increasing its solubility at the solid/liquid interface. Soil organic matter content and urea application rates showed significant effects, whereas pH did not exert any significant effect on urea catalysis of TNT extraction from soil. The optimum urea application rates (125 or 350 mg kg(-1)) for maximizing TNT extraction were within the limits set by the agronomic fertilizer-N rates used for major agricultural crops. The data obtained from this batch study will facilitate the optimization of a chemically-catalyzed phytoremediation model for cleaning up TNT-contaminated soils.

Vetiver grass is capable of removing TNT from soil in the presence of urea.

The high affinity of vetiver grass for 2,4,6 trinitrotoluene (TNT) and the catalytic effectiveness of urea in enhancing plant uptake of TNT in hydroponic media we earlier demonstrated were further illustrated in this soil-pot-experiment. Complete removal of TNT in urea-treated soil was accomplished by vetiver at the low initial soil-TNT concentration (40 mg kg(-1)), masking the effect of urea. Doubling the initial TNT concentration (80 mg kg(-1)) significantly (p<0.002) increased TNT removal by vetiver, in the presence of urea. Without vetiver grass, no significant (p=0.475) change in the soil-TNT concentrations was observed over a period of 48 days, suggesting that natural attenuation of soil TNT could not explain the documented TNT disappearance from soil.

Symbiotic role of Glomus mosseae in phytoextraction of lead in vetiver grass [Chrysopogon zizanioides (L.)].

Lead (Pb) has limited solubility in the soil environment owing to complexation with various soil components. Although total soil Pb concentrations may be high at a given site, the fraction of soluble Pb that plants can extract is very small, which is the major limiting factor for Pb phytoremediation. The symbiotic effect of arbuscular mycorrhizal (AM) fungus, Glomus mosseae was examined on growth and phytoextraction of lead (Pb) by vetiver grass [Chrysopogon zizanioides (L.)]. A hydroponic study, Phase I (0, 1, 2, and 4mM Pb) was conducted followed by an incubation pot study, Phase II (0, 400, 800, and 1200 mg kg(-1) Pb) where vetiver plants were colonized with G. mosseae. The results obtained indicate that plants colonized by the AM fungi not only exhibit better growth (increase in plant biomass), but also significantly increase Pb uptake in root and higher translocation to the shoot at all given treatments. Moreover, plants colonized with AM fungi had higher chlorophyll content and reduced levels of low molecular weight thiols, indicating the ability to better tolerate metal-induced stress. Results from this study indicate that vetiver plants in association with AM fungi can be used for improved phytoextraction of Pb from contaminated soil.