Bioaugmentation of polyethylene succinate-contaminated soil with Pseudomonas sp. AKS2 results in increased microbial activity and better polymer degradation

Pseudomonas sp. AKS2 isolated from soil degrades polyethylene succinate (PES) efficiently in the laboratory. However, this organism may not be able to degrade PES with similar efficiency in a natural habitat. Since in situ remediation is preferred for the effective removal of recalcitrant materials like plastic, in the current study, bioaugmentation potential of this organism was investigated. To investigate the potential of the AKS2 strain to bioaugment the PES-contaminated soil, a microcosm-based study was carried out wherein naturally attenuated, biostimulated, and AKS2-inoculated (bioaugmented) soil samples were examined for their ability to degrade PES. The results showed better degradation of PES by bioaugmented soil than other microcosms. Consistent with it, a higher number of PES-degrading organisms were found in the bioaugmented microcosm. The bioaugmented microcosm also exhibited a higher level of average well color development in BiOLOG ECO plate assay than the other two. The corresponding Shannon–Weaver index and Gini coefficient revealed a higher soil microbial diversity of bioaugmented microcosm than the others. This was further supported by community-level physiological profile of three different microcosms wherein we have observed better utilization of different carbon sources by bioaugmented microcosms. Collectively, these results demonstrate that bioaugmentation of PES-contaminated soil with AKS2 not only enhances polymer degradation but also increases microbial diversity. Bioaugmentation of soil with AKS2 enhances PES degradation without causing damage to soil ecology. Thus, Pseudomonas sp. AKS2 has the potential to be implemented as a useful tool for in situ bioremediation of PES.     

Integrated Bacillus sp. immobilized cell reactor and Synechocystis sp. algal reactor for the treatment of tannery wastewater

The wastewater discharged from leather industries lack biodegradability due to the presence of xenobiotic compounds. The primary clarification and aerobic treatment in Bacillus sp. immobilized Chemo Autotrophic Activated Carbon Oxidation (CAACO) reactor removed considerable amount of pollution parameters. The residual untreated organics in the wastewater was further treated in algal batch reactor inoculated with Synechocystis sp. Sodium nitrate, K₂HPO₄, MgSO₄.7H₂O, NH₄Cl, CaCl₂·2H₂O, FeCl₃ (anhydrous), and thiamine hydrochloride, rice husk based activated carbon (RHAC), immobilization of Bacillus sp. in mesoporous activated carbon, sand filter of dimensions diameter, 6 cm and height, 30 cm; and the CAACO reactor of dimensions diameter, 5.5 cm and height, 30 cm with total volume 720 ml, and working volume of 356 ml. In the present investigation, the CAACO treated tannery wastewater was applied to Synechocystis sp. inoculated algal batch reactor of hydraulic residence time 24 h. The BOD₅, COD, and TOC of treated wastewater from algal batch reactor were 20?±?7, 167?±?29, and 78?±?16 mg/l respectively. The integrated CAACO system and Algal batch reactor was operated for 30 days and they accomplished a cumulative removal of BOD₅,COD, TOC, VFA and sulphide as 98 %, 95 %, 93 %, 86 %, and 100 %, respectively. The biokinetic constants for the growth of algae in the batch reactor were specific growth rate, 0.095(day^?1) and yield coefficient, 3.15 mg of algal biomass/mg of COD destructed. The degradation of xenobiotic compounds in the algal batch reactor was confirmed through HPLC and FT-IR techniques. The integrated CAACO–Algal reactor system established a credible reduction in pollution parameters in the tannery wastewater. The removal mechanism is mainly due to co-metabolism between algae and bacterial species and the organics were completely metabolized rather than by adsorption.     

Toxicological response of a bioluminescent microbial assay to Zn, Pb and Cu in an artificial soil solution: relationship with total metal concentrations and free ion activities.

The relationship between toxicological response and both total concentrations and free ion activities of Pb, Cu and Zn in an artificial soil solution has been investigated using lux-marked Escherichia coli HB101 (pUCD607) as a bioassay. SO4(2-) (as K2SO4) was added as an inorganic complexing agent up to 0.01 M representing the range of ionic strengths found in soil solutions and giving a wide range of free metal ion activities. EC50 values expressed in terms of concentration, varied significantly with K2SO4 molarity for all metals. However, when EC50 values were expressed in terms of free ion activity they were not significantly different for Pb and Zn, supporting the free ion activity model. Conversely, EC50 values expressed as free Cu activity did vary significantly with K2SO4 molarity, possibly due to a greater degree of adsorption of Cu onto inactive sites on the cell surfaces than for Zn and Pb. Linear regression analysis of bioluminescence on free ion activity revealed significant correlations for each metal above the toxicity threshold. In conclusion, lux-marked E. coli is suitable for investigating the toxicity of metal ions and complexes in non saline systems although cell surface adsorption effects could be important for some metals, e.g. Cu.     

Optimisation of a microbial bioassay for contaminated soil monitoring: bacterial inoculum standardisation and comparison with Microtox assay

This work represents the first step to set up a toxicity testing procedure and to evaluate the sensitivity of the test microorganism to several classes of environmental pollutants. First, three different techniques were employed to standardise the microbial inoculum, then two different toxicity assessment protocols have been compared: Microtox and a dehydrogenase (DHase) activity inhibition test. The main goal was the optimisation of a microbial bioassay based on the dehydrogenase activity (DHase) inhibition in Pseudomonas fluorescens bacterial strain ATCC 13525. Triphenyl tetrazolium chloride (TTC) was used as electron acceptor and its reduction produces Triphenyl formazane (TPF). The P. fluorescens DHase inhibition bioassay was investigated for being a reliable and rapid method for assessing toxicity. The optimisation of the operating conditions resulted in a repeatable bioassay. Then, P. fluorescens and Vibrio fischeri sensitivity were firstly compared by testing Zn++, one of the reference compounds for Microtox test. In addition, other compounds (Ni++, Cd++, Cu++, phenol) were also tested with both bioassays. A high statistical significance of data was obtained with the logistic curve. The present work has demonstrated that P. fluorescens is as sensitive as Microtox culture (V. fischeri), for some of the metal ions. With reference to organic compounds, the lower sensitivity of P. fluorescens to phenol makes its use difficult in organic polluted samples.     

Influence of agricultural antibiotics and 17beta-estradiol on the microbial community of soil

Agricultural pharmaceuticals are a major environmental concern because of their hazardous effects on human and wildlife. This study analyzed phospholipid ester-linked fatty acids (PLFAs) and quinones to investigate the effects of a steroid (17beta-estradiol) and agricultural antibiotics (chlortetracycline and tylosin) on soil microbes in the laboratory. Two different types of soil were used: Sequatchie loam (0.8% organic matter) and LaDelle silt loam (9.2% organic matter). The soils were spiked with 17beta-estradiol and antibiotics, alone or in combination. In Sequatchie loam, 17beta-estradiol significantly increased the microbial biomass, especially the biomarkers for beta proteobacteria (16:1omega7c, 18:1omega7c, Cy17:0, and UQ-8). The coexistence of antibiotics decreased the stimulatory effect of 17beta-estradiol on the microbial community. In LaDelle silt loam, there were no significant differences in total microbial biomass and their microbial community structure among the treatments. Overall, 17beta-estradiol changed the microbial community of soil and the presence of antibiotics nullified the effect of 17beta-estradiol. However, the effects of 17beta-estradiol and antibiotics on soil microbes were sensitive to the soil properties, as seen in the LaDelle silt loam.     

Anaerobic microbial dechlorination: an approach to on-site treatment of toxaphene-contaminated soil

Enhanced microbial degradation of toxaphene by natural microorganisms occurred in soil and sediment amended with organic matter kept under anaerobic (flooded) conditions. Laboratory experiments yielded a dissipation half-life of approximately 3 and 1 week for soil and sediment, respectively, containing 10 ppm of technical toxaphene and a 1% alfalfa meal amendment. Dissipation was accompanied by an increase in early eluting gas chromatographic peaks and a decrease in later eluting peaks, indicating that dechlorination had occurred. Enhanced anaerobic dissipation also took place in soil containing 500 ppm of toxaphene, although at a lesser rate than at 10 ppm, and when cotton gin waste was used as amendment in place of alfalfa meal. Sediment in a toxaphene-contaminated pesticide waste disposal ditch was amended with 10% steer manure and flooded to ascertain field utility of the technique for on-site decontamination. Toxaphene residues were reduced from 63 to 23 ppm in 120 days, and some degradation activity still occurred up to 8 months after this single treatment.     

Temporal evolution of redox processes and free Cd dynamics in a metal-contaminated soil after rewetting

Soil testing procedures to address metals bioavailability currently use air-dried soil rewetted almost until saturation. Such practices may influence the redox state of soil and the related dynamics of metals. To assess this potential impact, a metal-contaminated soil was air-dried and rewetted to 90% water holding capacity. We monitored over a 21-day incubation period the temporal changes of soil redox potential and solution Cd concentration (either total or free). Other physico-chemical parameters were followed notably pH, ionic strength (I) and the concentrations of NO(3)(-), Mn, Fe and SO(4)(2-) in solution. Soil redox potential showed the progressive establishment of strong reducing conditions in soil, in agreement with the temporal changes of NO(3)(-), Mn, Fe and SO(4)(2-) concentrations. It decreased by 13 pe units over the culture period leading to sulphate-reducing conditions (pe<-3) within only 21days. Solution Cd concentration increased transitorily over the first 100-150h of incubation (2-fold increase) in relation with the parallel increase in the concentration of competing cations for adsorption (Ca, Mg). It steeply decreased over the last 300h of incubation (30-fold decrease) as a result of Cd precipitation as Cd sulphides. This biphasic evolution of Cd dynamics was related to the temporal changes of Cd resupply from the solid phase. Using the technique of DGT we described the kinetics of Cd resupply over time and needed to invoke the existence of two pools of Cd.     

Effects of biochar on nitrification and denitrification-mediated N2O emissions and the associated microbial community in an agricultural soil

Environmental Science and Pollution Research - Nitrous oxide (N2O) is a strong greenhouse gas, and it is of great significance for N2O reduction to study the effects of biochar on its production...     

Atrazine removal in agricultural infiltrate by bioaugmented polyvinyl alcohol immobilized and free Agrobacterium radiobacter J14a: a sand column study

Bench-scale sand column breakthrough experiments were conducted to examine atrazine removal in agricultural infiltrate by Agrobacterium radiobacter J14a (J14a) immobilized in phosphorylated-polyvinyl alcohol compared to free J14a cells. The effects of cell loading and infiltration rate on atrazine degradation and the loss of J14a were investigated. Four sets of experiments, (i) tracers, (ii) immobilized dead cells, (iii) immobilized cells, and (iv) free cells, were performed. The atrazine biodegradation at the cell loadings of 300, 600, and 900 mg dry cells L(-1) and the infiltration rates of 1, 3, and 6 cm d(-1) were tested for 5 column pore volumes (PV). The atrazine breakthrough results indicated that the immobilized dead cells significantly retarded atrazine transport. The atrazine removal efficiencies at the infiltration rates of 1, 3, and 6 cm d(-1) were 100%, 80-97%, and 50-70%, respectively. Atrazine degradation capacity for the immobilized cells was not significantly different from the free cells. Both infiltration rate and cell loading significantly affected atrazine removal for both cell systems. The bacterial loss from the immobilized cell system was 10-100 times less than that from the free cell system. For long-term tests at 50 PV, the immobilized cell system provided consistent atrazine removal efficiency while the atrazine removal by the free cells declined gradually because of the cell loss.     

黑土微生物群落对副球菌属(Paracoccus sp.)QD15-1修复邻苯二甲酸二甲酯污染土壤的响应

采用Illumina MiSeq高通量测序法探究黑土微生物群落对副球菌属(Paracoccus sp.)QD15-1修复邻苯二甲酸二甲酯(DMP)污染土壤的响应。结果表明,副球菌属QD15-1在7d内可快速有效降解土壤中的DMP,平均降解率为94.7%,并能保持稳定;Shannon、Sobs指数显示,土壤中微生物多样性、丰富性受到DMP的影响;土壤中的优势门为放线菌门(Actinobacteria),占总菌门数的50%以上;鞘氨醇单胞菌属(Sphingomonas)是第一优势菌属,第二优势菌属为类诺卡氏菌属(Nocardioides)。     

RAPD marker and substrate utilization pattern applied to study microbial community diversity in the soil affected by agricultural chemicals

Present analyses of random amplified polymorphic DNA (RAPD) and Biolog GN substrate utilization pattern are combined to further study the diversity of microbial communities in four soils affected by agricultural chemicals. The results showed that the four soil microbial communities were apparently distinguishable in the diversity at RAPD level in terms of the richness and modified richness in the summer, which supports our previous report using the same soils in winter. A significant difference for the average well color development (AWCD) at 72 h incubation was found among the soils in winter using Biolog GN substrate utilization pattern, but this difference was not found among the soils in summer. However, Shannon-Weaver indices for microbial communities in the summer soils polluted by agricultural chemicals were significantly higher than those in winter at metabolic level; in contrast, no significant difference existed between the two seasons for microbial communities in the soil without chemical pollution. Present results suggest that the combined approach using RAPD and substrate utilization pattern could be used to effectively quantify microbial community diversity and its changes among the seasons in the soils affected by agricultural chemicals, simultaneously at molecular and physiological levels.     

Enhanced uptake of Cd,Cr, and Cu in Catharanthus roseus (L.) G.Don by Bacillus cereus: application of moss and compost to reduce metal availability

Environmental Science and Pollution Research - Heavy metals (HMs) being the notorious and toxic are being introduced into the environment credited to natural and anthropogenic activities. The use...     

Dissipation of chlorothalonil, chlorpyrifos, and profenofos in a Malaysian agricultural soil: a comparison between the field experiment and simulation by the PERSIST model

A comparison of dissipation of chlorothalonil, chlorpyrifos, and profenofos in a Malaysian agricultural soil between the field experiment and simulation by the PERSIST model was studied. A plot of sweet pea (Pisum sativum) from a farm in the Cameron Highlands was selected for the field experiment. The plot was treated with chlorothalonil, chlorpyrifos, and profenofos. Core soil collection was conducted according to the sampling schedule. Residues of the three pesticides were analyzed in the laboratory. Simulations of the three pesticides' persistency were also conducted using a computer-run software PERSIST. Generally, predicted data obtained using PERSIST were found to be high for the three pesticides except for one field measurement of chlorpyrifos. The predicted data for profenofos, which is the most mobile of the three pesticides tested, was not well matched with the observed data compared to chlorothalonil and chlorpyrifos.     

Enhanced phytoextraction of an agricultural Cr- and Pb-contaminated soil by bioaugmentation with siderophore-producing bacteria

Bioaugmentation-assisted phytoextraction may enhance the phytoextraction efficiency thanks to larger metal mobilization by microbial metabolites. Green fluorescent protein-tagged cells of Pseudomonas aeruginosa, Pseudomonas fluorescens or Ralstonia metallidurans, able to produce siderophores, were inoculated in an agricultural soil containing Cr (488 mg kg(-1)) and Pb (382 mg kg(-1)) and maize was cultivated. Bacteria were inoculated as free or immobilized cells in Ca-alginate beads, with skim milk in the aim at improving both the bacterial survival and the in situ siderophore production. Skim milk addition increased inoculated Pseudomonads concentration in soil. Soil inoculation with free cells of R. metallidurans supplied with skim milk increased Cr accumulation in maize shoots by a factor of 5.2 and inoculation with immobilized P. aeruginosa cells supplied with skim milk increased Cr and Pb uptake by maize shoots by a factor of 5.4 and 3.8, respectively. However total metal taken up by the whole plant decreases almost always with bioaugmentation. Translocation factor also increased with P. aeruginosa or R. metallidurans by a factor of 6 up to 7. Inoculated bacteria concentration in soil was correlated with metals in the exchangeable fraction. Cr and Pb concentrations in the exchangeable fraction were correlated with metal contents in shoots or roots. Our results suggest that bioaugmentation-assisted phytoextraction is a relevant method in the aim at increasing the phytoextraction rate which usually limits the use of phytoremediation technologies.     

Saline irrigation and Zn amendment effect on Cd phytoavailability to Swiss chard (Beta vulgaris L.) grown on a long-term amended agricultural soil: a human risk assessment

Crops, particularly in the Northeast region of Mexico, have to cope with increasing soil salinization due to irrigation. Chloride (Cl^?) concentration has been strongly related to enhance cadmium (Cd) uptake by plants due to increased solubility in the soil solution. The effect of irrigation with slightly saline water from a local well was evaluated in this work on the accumulation and translocation of Cd in Swiss chard (Beta vulgaris L.) grown in soil historically amended with stabilized sewage sludge under a regime of phosphorus and zinc fertilization. A factorial pot experiment was conducted with two phosphate fertilizer levels (PF, 0 and 80 kg ha^?1 dry soil, respectively), two Zn levels (0 and 7 kg ha^?1 dry soil), and two sources of water for irrigation deionized water (DW) and slightly saline well water (WW) from an agricultural site. Additionally, a human risk assessment for Cd ingestion from plants was assessed. Results showed that Cl^? salinity in the WW effectively mobilized soil Cd and increased its phytoavailability. A higher level of Cd was found in roots (46.41 mg kg^?1) compared to shoots (10.75 mg kg^?1). Although the total content of Cd in the edible parts of the Swiss chard irrigated with WW exceeded permissible recommended consumption limit, bioavailable cadmium in the aboveground parts of the plant in relation to the total cadmium content was in the range from 8 to 32 %. Therefore, human health risks might be overestimated when the total concentration is taken into account.     

Impact of temperature on the dynamics of organic matter and on the soil-to-plant transfer of Cd,Zn and Pb in a contaminated agricultural soil

Predicting the soil-to-plant transfer of metals in the context of global warming has become a major issue for food safety. It requires a better understanding of how the temperature alters the bioavailability of metals in cultivated soils. This study focuses on one agricultural soil contaminated by Cd, Zn and Pb. DGT measurements were performed at 10, 20 and 30 °C to assess how the bioavailability of metals was affected by a rise in soil temperature. A lettuce crop was cultivated in the same conditions to determine if the soil-to-plant transfer of metals increased with a rise in soil temperature. A gradual decline in Cd and Zn bioavailability was observed from 10 to 30 °C, which was attributed to more intense complexation of metals in the pore water at higher temperatures. Together with its aromaticity, the affinity of dissolved organic matter (DOM) for metals was indeed suspected to increase with soil temperature. One main output of the present work is a model which satisfactorily explains the thermal-induced changes in the characteristics of DOM reported in Cornu et al. (Geoderma 162:65–70, 2011) by assuming that the mineralization of initial aliphatic compounds followed a first-order reaction, increased with soil temperature according to the Arrhenius law, and due to a priming effect, led to the appearance of aromatic molecules. The soil-to-plant transfer of Cd and Zn was promoted at higher soil temperatures despite a parallel decrease in Cd and Zn bioavailability. This suggests that plant processes affect the soil-to-plant transfer of Cd and Zn the most when the soil temperature rises.     

A field experiment with variable-suction multi-compartment samplers to measure the spatio-temporal distribution of solute leaching in an agricultural soil

Solutes spread out in time and space as they move downwards from the soil surface with infiltrating water. Solute monitoring in the field is often limited to observations of resident concentrations, while flux concentrations govern the movement of solutes in soils. A recently developed multi-compartment sampler is capable of measuring fluxes at a high spatial resolution with minimal disturbance of the local pressure head field. The objective of this paper is to use this sampler to quantify the spatial and temporal variation of solute leaching below the root zone in an agricultural field under natural rainfall in winter and spring. We placed two samplers at 31 and 25 cm depth in an agricultural field, leaving the soil above undisturbed. Each sampler contained 100 separate cells of 31 × 31 mm. Water fluxes were measured every 5 min for each cell. We monitored leaching of a chloride pulse under natural rainfall by frequently extracting the collected leachate while leaving the samplers buried in situ. This experiment was followed by a dye tracer experiment. This setting yielded information that widely surpassed the information that can be provided by separate anionic and dye tracer trials, and solute transport monitoring by coring or suction cups. The detailed information provided by the samplers showed that percolation at the sampling depth started much faster (approximately 3 h after the start of rainfall) in initially wet soil (pressure head above − 65 cm) than in drier soil (more than 14 h at pressure heads below − 80 cm). At any time, 25% of the drainage passed through 5–6% of the sampled area, reflecting the effect of heterogeneity on the flow paths. The amount of solute carried by individual cells varied over four orders of magnitude. The lateral concentration differences were limited though. This suggests a convective–dispersive regime despite the short vertical travel distance. On the other hand, the dilution index indicates a slight tendency towards stochastic–convective transport at this depth. There was no evidence in the observed drainage patterns and dye stained profiles of significant disturbance of the flow field by the samplers.