Analysis of the Genome of the Heavy Metal Resistant and Hydrocarbon-Degrading Rhizospheric Pseudomonas qingdaonensis ZCR6 Strain and Assessment of Its Plant-Growth-Promoting Traits.

The Pseudomonas qingdaonensis ZCR6 strain, isolated from the rhizosphere of Zea mays growing in soil co-contaminated with hydrocarbons and heavy metals, was investigated for its plant growth promotion, hydrocarbon degradation, and heavy metal resistance. In vitro bioassays confirmed all of the abovementioned properties. ZCR6 was able to produce indole acetic acid (IAA), siderophores, and ammonia, solubilized Ca3(PO4)2, and showed surface active properties and activity of cellulase and very high activity of 1-aminocyclopropane-1-carboxylic acid deaminase (297 nmol α-ketobutyrate mg-1 h-1). The strain degraded petroleum hydrocarbons (76.52% of the initial hydrocarbon content was degraded) and was resistant to Cd, Zn, and Cu (minimal inhibitory concentrations reached 5, 15, and 10 mM metal, respectively). The genome of the ZCR6 strain consisted of 5,507,067 bp, and a total of 5055 genes were annotated, of which 4943 were protein-coding sequences. Annotation revealed the presence of genes associated with nitrogen fixation, phosphate solubilization, sulfur metabolism, siderophore biosynthesis and uptake, synthesis of IAA, ethylene modulation, heavy metal resistance, exopolysaccharide biosynthesis, and organic compound degradation. Complete characteristics of the ZCR6 strain showed its potential multiway properties for enhancing the phytoremediation of co-contaminated soils. To our knowledge, this is the first analysis of the biotechnological potential of the species P. qingdaonensis.

Environmental risk assessment of metformin and its transformation product guanylurea: II. Occurrence in surface waters of Europe and the United States and derivation of predicted no-effect concentrations.

Metformin (MET), CAS 1115-70-4 (Metformin hydrochloride), is an antidiabetic drug with high usage in North America and Europe and has become the subject of regulatory interest. A pharmaceutical industry working group investigated environmental risks of MET. Environmental fate and chronic effects data were collated across the industry for the present risk assessment. Predicted environmental concentrations (PECs) for MET were modeled for the USA and Europe using the PhATE and GREAT-ER models, respectively. PECs were compared with measured environmental concentrations (MECs) for the USA and Europe. A predicted no effect concentration (PNEC) of 1 mg/L for MET was derived by deterministic procedures, applying an assessment factor of 10 to the lowest no observed effect concentration (i.e., 10 mg/L) from multiple chronic studies with algae, daphnids and fish. The PEC/PNEC and MEC/PNEC risk characterization ratios were <1, indicating no significant risk for MET with high Margins of Safety (MOS) of >868. MET is known to degrade during wastewater treatment to guanylurea (GUU, CAS 141-83-3), which we have shown to further degrade. There are no GUU toxicity data in the literature; hence, chronic studies for GUU were conducted to derive a PNEC of 0.16 mg/L. PECs were derived for GUU as for MET, plus MECs were retrieved from the literature. The PEC/PNEC and MEC/PNEC risk characterization ratios for GUU were also <1, with an MOS of >6.5. Based on standard risk assessment procedures for both MET and its transformation product GUU, there is no significant risk to aquatic life.

Environmental risk assessment of metformin and its transformation product guanylurea. I. Environmental fate.

Metformin (MET) is a pharmaceutical with very high use worldwide that is excreted in unchanged form, leading to concern about potential aquatic life impacts associated with MET, and its primary transformation product guanylurea (GUU). This study presents, in two companion papers, a risk assessment following internationally accepted guidelines of MET and GUU in surface water based on literature data, previously unpublished studies, and a new degradation test that resolves conflicting earlier results. Previous studies have shown that MET is removed during sewage treatment, primarily through transformation to GUU. In addition, measurements in WWTPs suggest that MET is not only transformed to GUU, but that GUU is further biodegraded. A prolonged inherent biodegradation test strongly suggests not only primary transformation of MET to GUU, but also subsequent full mineralization of GUU, with both degradation phases starting after a clear lag phase. MET may partition from surface water to sediment, where both transformation to GUU and in part mineralization is possible, depending on the presence of competent degrading microorganisms. In addition, MET may form non-extractable residues in sediments (12.8-73.5%). Both MET and GUU may be anaerobically degraded during sludge digestion, in soils or in sediments. Bioconcentration factor (BCF) values in crops and most plants are close to 1 suggesting low bioaccumulation potential, moreover, at least some plants can metabolize MET to GUU; however, in aquatic plants higher BCFs were found, up to 53. Similarly, neither MET nor GUU are expected to bioaccumulate in fish based on estimated values of BCFs ≤3.16.

Multi-method assessment of the intrinsic biodegradation potential of an aquifer contaminated with chlorinated ethenes at an industrial area in Barcelona (Spain).

The bioremediation potential of an aquifer contaminated with tetrachloroethene (PCE) was assessed by combining hydrogeochemical data of the site, microcosm studies, metabolites concentrations, compound specific-stable carbon isotope analysis and the identification of selected reductive dechlorination biomarker genes. The characterization of the site through 10 monitoring wells evidenced that leaked PCE was transformed to TCE and cis-DCE via hydrogenolysis. Carbon isotopic mass balance of chlorinated ethenes pointed to two distinct sources of contamination and discarded relevant alternate degradation pathways in the aquifer. Application of specific-genus primers targeting Dehalococcoides mccartyi species and the vinyl chloride-to-ethene reductive dehalogenase vcrA indicated the presence of autochthonous bacteria capable of the complete dechlorination of PCE. The observed cis-DCE stall was consistent with the aquifer geochemistry (positive redox potentials; presence of dissolved oxygen, nitrate, and sulphate; absence of ferrous iron), which was thermodynamically favourable to dechlorinate highly chlorinated ethenes but required lower redox potentials to evolve beyond cis-DCE to the innocuous end product ethene. Accordingly, the addition of lactate or a mixture of ethanol plus methanol as electron donor sources in parallel field-derived anoxic microcosms accelerated dechlorination of PCE and passed cis-DCE up to ethene, unlike the controls (without amendments, representative of field natural attenuation). Lactate fermentation produced acetate at near-stoichiometric amounts. The array of techniques used in this study provided complementary lines of evidence to suggest that enhanced anaerobic bioremediation using lactate as electron donor source is a feasible strategy to successfully decontaminate this site.

Determining soil enzyme activities for the assessment of fungi and citric acid-assisted phytoextraction under cadmium and lead contamination.

Microorganism or chelate-assisted phytoextraction is an effective remediation tool for heavy metal polluted soil, but investigations into its impact on soil microbial activity are rarely reported. Consequently, cadmium (Cd)- and lead (Pb)-resistant fungi and citric acid (CA) were introduced to enhance phytoextraction by Solanum nigrum L. under varied Cd and Pb pollution levels in a greenhouse pot experiment. We then determined accumulation of Cd and Pb in S. nigrum and the soil enzyme activities of dehydrogenase, phosphatase, urease, catalase, sucrase, and amylase. Detrended canonical correspondence analysis (DCCA) was applied to assess the interactions between remediation strategies and soil enzyme activities. Results indicated that the addition of fungi, CA, or their combination enhanced the root biomass of S. nigrum, especially at the high-pollution level. The combined treatment of CA and fungi enhanced accumulation of Cd about 22-47 % and of Pb about 13-105 % in S. nigrum compared with the phytoextraction alone. However, S. nigrum was not shown to be a hyperaccumulator for Pb. Most enzyme activities were enhanced after remediation. The DCCA ordination graph showed increasing enzyme activity improvement by remediation in the order of phosphatase, amylase, catalase, dehydrogenase, and urease. Responses of soil enzyme activities were similar for both the addition of fungi and that of CA. In summary, results suggest that fungi and CA-assisted phytoextraction is a promising approach to restoring heavy metal polluted soil.

A preliminary assessment of the potential of using an acacia--biochar system for spent mine site rehabilitation.

Mining activities result in extensive soil degradation by removing the top soil, disturbing soil structure and altering microbial communities. Rehabilitation of spent mine sites through revegetation thus requires proper soil amendments. In this study, a pot trial was conducted to investigate the effects of a jarrah biochar on the growth and nutrient status of a native legume, Acacia tetragonophylla, grown in a mixture of topsoil and mine rejects. Two biochar application rates (37 and 74 t ha(-1)) and two types of biochar, namely nutrient-enriched and non-enriched, were tested. We measured the soil pH and electrical conductivity, the carbon (C) and nitrogen (N) contents and C and N isotope composition (δ(13)C and δ(15)N) of soil and plants, the foliar phosphorus content and the growth and leaf biomass of the plants. Whilst no significant effect of biochar was observed on plant growth, biochar amendment affected soil properties and plant nutritional status. The highest rate of biochar application increased soil pH, C content and C/N ratio, and decreased soil δ(13)C. Biochar application also enhanced photosynthetic N use efficiency, as showed by the increase in foliar C/N ratio, and biological N fixation rates, as indicated by foliar δ(15)N. These positive effects were not observed when biochar was nutrient-enriched due to the associated increase in soil N. Revegetation of mine sites with acacia in combination with biochar amendment constitutes a plausible alternative to the wide use of N fertiliser through the supply of additional N to the system, even though other nutrients may be required in order to enhance plant early growth.

Assessment of methane biodegradation kinetics in two-phase partitioning bioreactors by pulse respirometry.

Biological methane biodegradation is a promising treatment alternative when the methane produced in waste management facilities cannot be used for energy generation. Two-phase partitioning bioreactors (TPPBs), provided with a non-aqueous phase (NAP) with high affinity for the target pollutant, are particularly suitable for the treatment of poorly water-soluble compounds such as methane. Nevertheless, little is known about the influence of the presence of the NAP on the resulting biodegradation kinetics in TPPBs. In this study, an experimental framework based on the in situ pulse respirometry technique was developed to assess the impact of NAP addition on the methane biodegradation kinetics using Methylosinus sporium as a model methane-degrading microorganism. A comprehensive mass transfer characterization was performed in order to avoid mass transfer limiting scenarios and ensure a correct kinetic parameter characterization. The presence of the NAP mediated significant changes in the apparent kinetic parameters of M. sporium during methane biodegradation, with variations of 60, 120, and 150% in the maximum oxygen uptake rate, half-saturation constant and maximum specific growth rate, respectively, compared with the intrinsic kinetic parameters retrieved from a control without NAP. These significant changes in the kinetic parameters mediated by the NAP must be considered for the design, operation and modeling of TPPBs devoted to air pollution control.

Removal and fate of endocrine disruptors chemicals under lab-scale postreatment stage. Removal assessment using light, oxygen and microalgae.

The aim of this study was to assess the effect of light, oxygen and microalgae on micropollutants removal. The studied micropollutants were 4-(1,1,3,3-tetramethylbutyl)phenol (OP), technical-nonylphenol (t-NP), 4-n-nonylphenol (4-NP), Bisphenol-A (BPA). In order to study the effect of the three variables on the micropollutants removal, a factorial design was developed. The experiments were carried out in four batch reactors which treated the effluent of an anaerobic membrane bioreactor. The gas chromatography mass spectrometry was used for the measurement of the micropollutants. The results showed that light, oxygen and microalgae affected differently to the degradation ratios of each micropollutant. The results showed that under aerated conditions removal ratios higher than 91% were achieved, whereas for non-aerated conditions the removal ratios were between 50% and 80%, except for 4-NP which achieved removal ratios close to 100%. Besides, mass balance showed that the degradation processes were more important than the sorption processes.

Production of novel microbial flocculants by Klebsiella sp. TG-1 using waste residue from the food industry and its use in defecating the trona suspension.

A microbial-flocculants-producing (MBF-producing) bacterium, named TG-1, was isolated from waste water of a starch factory, and identified as Klebsiella sp. TG-1. The microbial flocculants (MBF) produced by TG-1, named as MBF-TG-1, was applied to defecating the strong basic trona suspension in the trona industry. After optimizing medium and culturing conditions with single-factor and orthogonal designs, the highest flocculation rate of 86.9% was achieved. Chemical analysis showed that the purified microbial flocculants (MBF-TG-1) was mainly composed of polysaccharides (84.6%), with a small amount of protein or amino acid (11.1%). Bridging mechanism was supposed as the main flocculation mechanism by analyzing the flocculation process and the biochemistry properties of MBF-TG-1. The high flocculation rate (84%) was also achieved with a low-cost medium (the solid residue of tofu production from food industry).

Adsorption of dyestuff from aqueous solutions through oxalic acid-modified swede rape straw: adsorption process and disposal methodology of depleted bioadsorbents.

Swede rape straw (Brassica napus L.) was modified by oxalic acid under mild conditions producing an efficient dye adsorbent (SRSOA). This low-cost and environmental friendly bioadsorbent was characterized by various techniques and then applied to purify dye-contaminated aqueous solutions. Equilibrium study showed that the Langmuir model demonstrated the best fit to the equilibrium data and the methylene blue (MB) adsorption capacity calculated by this model was 432mgg(-1). The adsorption process and mechanism is also discussed. To properly deal with the dye-loaded bioadsorbents, the disposal methodology is discussed and a biochar based on depleted bioadsorbents was for the first time produced and examined. This method both solved the disposal problem of contaminant-loaded bioadsorbents and produced an useful adsorbent thereafter. The study indicates that SRSOA is a promising substitute for ACs in purifying dye-contaminated wastewater and that producing biochars from contaminant-loaded bioadsorbents maybe a feasible disposal method.

Application of integrated ozone biological aerated filters and membrane filtration in water reuse of textile effluents.

A combined process including integrated ozone-BAFs (ozone biological aerated filters) and membrane filtration was first applied for recycling textile effluents in a cotton textile mill with capacity of 5000 m(3)/d. Influent COD (chemical oxygen demand) in the range of 82-120 mg/L, BOD5 (5-day biochemical oxygen demand) of 12.6-23.1 mg/L, suspended solids (SSs) of 38-52 mg/L and color of 32-64° were observed during operation. Outflows with COD≤45 mg/L, BOD5≤7.6 mg/L, SS≤15 mg/L, color≤8° were obtained after being decontaminated by ozone-BAF with ozone dosage of 20-25 mg/L. Besides, the average removal rates of PVA (polyvinyl alcohol) and UV254 were 100% and 73.4% respectively. Permeate water produced by RO (reverse osmosis) could be reused in dyeing and finishing processes, while the RO concentrates could be discharged directly under local regulations with COD≤100 mg/L, BOD5≤21 mg/L, SS≤52 mg/L, color≤32°. Results showed that the combined process could guarantee water reuse with high quality, and solve the problem of RO concentrate disposal.

Excess sludge reduction using pilot-scale lysis-cryptic growth system integrated ultrasonic/alkaline disintegration and hydrolysis/acidogenesis pretreatment.

A pilot-scale lysis-cryptic growth system was built and operated continuously for excess sludge reduction. Combined ultrasonic/alkaline disintegration and hydrolysis/acidogenesis were integrated into its sludge pretreatment system. Continuous operation showed that the observed biomass yield and the sludge reduction efficiency of the lysis-cryptic growth system were 0.27 kg VSS/kg COD consumed and 56.5%, respectively. The water quality of its effluent was satisfactory. The sludge pretreatment system performed well and its TCOD removal efficiency was 7.9% which contributed a sludge reduction efficiency of 2.1%. The SCOD, VFA, TN, NH(4)(+)-N, TP and pH in the supernatant of pretreated sludge were 1790 mg/L, 1530 mg COD/L, 261.1mg/L, 114.0mg/L, 93.1mg/L and 8.69, respectively. The total operation cost of the lysis-cryptic growth system was $ 0.186/m(3) wastewater, which was 11.4% less than that of conventional activated sludge (CAS) system without excess sludge pretreatment.

Cadmium and Zn availability as affected by pH manipulation and its assessment by soil extraction, DGT and indicator plants.

Manipulation of soil pH by soil additives and / or rhizosphere processes may enhance the efficiency of metal phytoextraction. Here we report on the effect of nitric acid additions to four polluted soils on Cd and Zn concentrations in soil solution (C(soln)) and 0.005M Ca(NO(3))(2) extracts, and related changes in the diffusive fluxes and resupply of the metals as assessed by diffusive gradients in thin films (DGT). The responses of these chemical indicators of bioavailability were compared to metal uptake in two indicator plant species, common dandelion (Taraxacum officinale F.H. Wigg) and narrow leaf plantain (Plantago lanceolata L.) grown for 75days in a pot experiment. Lowering soil pH increased C(soln), the 0.005M Ca(NO(3))(2)-soluble fractions and the DGT-measured Cd and Zn concentrations (C(DGT)) in the experimental soils. This was associated with enhanced uptake of Cd and Zn on soils acidified to pH 4.5 whereas plants did not survive at pH 3.5. Toxicity along with decreased kinetics of metal resupply (calculated by the 2D DIFS model) in the strong acidification treatment suggests that moderate acidification is more appropriate to enhance the phytoextraction process. Each of the chemical indicators of bioavailability predicted well (R(2)>0.70) the Cd and Zn concentrations in plantain shoots but due to metal toxicity not for dandelion. Concentration factors, i.e. the ratio between metal concentrations in shoots and in soil solution (CF) indicate that Cd and Zn uptake in plantain was not limited by diffusion which may explain that DGT did not perform better than C(soln). However, DGT is expected to predict plant uptake better in diffusion-limited conditions such as in the rhizosphere of metal-accumulating phytoextraction crops.

Assessment of the efficacy of Aspergillus sp. EL-2 in textile waste water treatment.

Fungal biomass has the ability to decolorize a wide variety of dyes successfully through a number of mechanisms. A brown rot isolate, previously identified as Aspergillus sp. EL-2, was used in the aerobic treatment of textile waste water efficiently. In the current work, the treated waste water was tested chemically using more than one combined treatment. Microbial toxicity, phytotoxicity, genotoxicity and cytotoxicity were also studied to assess the toxicity level for each treatment. The obtained data suggest that the contribution of more than one mode of treatment is essential to ensure complete destruction of the by-products. The use of gamma irradiation (25 kGy) after the bioremediation step led to the decrease of the by-products of biodegradation as observed by visible spectrum and Fourier transfer infra red spectroscopy (FT-IR). The toxicity assessment presented variable results indicating the need for more than one toxicity test to confirm the presence or absence of hazardous compounds. Brown rot fungus could be used efficiently in the treatment of textile waste water without the risk of obtaining high carcinogenic or genotoxic compounds, especially if combined treatment is employed.

Phytoextraction of heavy metals from contaminated soil by co-cropping with chelator application and assessment of associated leaching risk.

Phytoextraction using hyperaccumulating plants is generally time-consuming and requires the cessation of agriculture. We coupled chelators and a co-cropping system to enhance phytoextraction rates, while allowing for agricultural production. An experiment on I m3 lysimeter beds was conducted with a co-cropping system consisting of the hyperaccumulator Sedum alfredii and low-accumulating corn (Zea Mays, cv. Huidan-4), with addition ofa mixture of chelators (MC), to assess the efficiency of chelator enhanced co-crop phytoextraction and the leaching risk caused by the chelator. The results showed that the addition of MC promoted the growth of S. alfredii in the first crop (spring-summer season) and significantly increased the metal phytoextraction. The DTPA-extractable and total metal concentrations in the topsoil were also reduced more significantly with the addition of MC compared with the control treatments. However, mono-cropped S. alfredii without MC was more suitable for maximizing S. alfredii growth and therefore phytoextraction of Zn and Cd during the autumn-winter seasons. No adverse impact to groundwater due to MC application was observed during the experiments with three crops and three MC applications. But elevated total Cd and Pb concentrations among subsoils compared to the initial subsoil concentrations were found for the co-crop + MC treatment after the third crop.

Assessment of Fenton's reagent and ozonation as pre-treatments for increasing the biodegradability of aqueous diethanolamine solutions from an oil refinery gas sweetening process.

The aim of this work was to evaluate the efficiency of three chemical oxidation processes for increasing the biodegradability of aqueous diethanolamine solutions (aqueous DEA solutions), to be used as pre-treatments before a biological process. The raw aqueous DEA solution, sourced from a sour gas sweetening plant at a Mexican oil refinery, was first characterized by standardized physico-chemical methods. Then experiments were conducted on diluted aqueous DEA solutions to test the effects of Fenton's reagent, ozone and ozone-hydrogen peroxide on the removal of some physicochemical parameters of these solutions. Lastly, biodegradability tests based on Dissolved Organic Carbon Die Away OECD301-A, were carried out on a dilution of the raw aqueous DEA solution and on the treated aqueous DEA solutions, produced by applying the best experimental conditions determined during the aforementioned oxidation tests. Experimental results showed that for aqueous DEA solutions treated with Fenton's reagent, the best degradation rate (70%) was obtained at pH 2.8, with Fe(2+) and H(2)O(2) at doses of 1000 and 10,000 mg/L respectively. In the ozone process, the best degradation (60%) was observed in aqueous DEA solution (100 mg COD/L), using 100 mg O(3)/L at pH 5. In the ozone-hydrogen peroxide process, no COD or DOC removals were observed. The diluted spent diethanolamine solution showed its greatest increase in biodegradability after a reaction period of 28 days when treated with Fenton's reagent, but after only 15 days in the case of ozonation.

A chemically enhanced biological process for lowering operative costs and solid residues of industrial recalcitrant wastewater treatment.

An innovative process based on ozone-enhanced biological degradation, carried out in an aerobic granular biomass system (SBBGR--Sequencing Batch Biofilter Granular Reactor), was tested at pilot scale for tannery wastewater treatment chosen as representative of industrial recalcitrant wastewater. The results have shown that the process was able to meet the current discharge limits when the biologically treated wastewater was recirculated through an adjacent reactor where a specific ozone dose of 120 mg O3/L(influent) was used. The benefits produced by using ozone were appreciable even visually since the final effluent of the process looked like tap water. In comparison with the conventional treatment, the proposed process was able to reduce the sludge production by 25-30 times and to save 60% of operating costs. Molecular in situ detection methods were employed in combination with the traditional measurements (oxygen uptake rate, total protein content, extracellular polymeric substances and hydrophobicity) to evaluate microbial activity and composition, and the structure of the biomass. A stable presence of active bacterial populations was observed in the biomass with the simultaneous occurrence of distinctive functional microbial groups involved in carbon, nitrogen and sulphate removal under different reaction environments established within the large microbial aggregates. The structure and activity of the biomass were not affected by the use of ozone.

Application of the OECD 301F respirometric test for the biodegradability assessment of various potential endocrine disrupting chemicals.

The biodegradability of several potential endocrine disrupting compounds, namely 4-n-nonylphenol (4-n-NP), nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), bisphenol A (BPA), triclosan (TCS), di-(2-ethylhexyl)-phthalate (DEHP), perfluorooctanoate (PFOA) and perfluorononanoate (PFNA) was evaluated in this study, using OECD method 301F (manometric respirometry test) and activated sludge as inoculum. According to the results, 4-n-NP and BPA meet the strict definition of ready biodegradability and they are not expected to be persistent during the activated sludge process. Partial biodegradation was observed for DEHP (58.7+/-5.7%, n=3), TCS (52.1+/-8.5%, n=3) and NP1EO (25.9+/-8.1%, n=3), indicating their possible biodegradation in wastewater treatment systems, while no biodegradation was observed for NP2EO, PFOA and PFNA. Experiments in the co-presence of a readily biodegradable compound showed the absence of co-metabolic phenomena during 4-n-NP, BPA and TCS biodegradation. Using first order kinetics to describe biodegradation of the target compounds, half-lives of 4.3+/-0.6, 1.3+/-0.2, 1.8+/-0.5, 6.9+/-2.6 days were calculated for 4-n-NP, BPA, TCS and DEHP, respectively. Toxicity tests using marine bacterium Vibrio fischeri showed that biodegradation of 4-n-NP, NP1EO, BPA and TCS is a simultaneous detoxification process, while possible abiotic or biotic transformations of NP2EO, DEHP, PFOA and PFNA during respirometric test resulted to significant increase of their toxicities.

The effect of lithium chloride on the biooxidation of aqueous methanol/acetone mixtures.

Lithium chloride, more specifically the lithium cation, has been implicated in interference in biological systems. In the case of Escherichia coli, interference involves the Na+(Li+)/H+ antiporter transport system. The study reported here concerns the effects of LiCl on a mixed enrichment culture that is able to biodegrade both methanol and acetone under aerobic conditions. The results obtained using unsteady state continuous flow culture techniques demonstrate a significant disruptive effect of LiCl on culture performance. In addition, a reduction in the substrate-based biomass yield coefficient, which is a clear advantage as far as biotreatment process performance is concerned, also occurs. The ultimate fate of the LiCl was not determined.