Mammalian cell line-based bioassays for toxicological evaluation of landfill leachate treated by Pseudomonas sp. ISTDF1

Landfill leachate has become a serious environmental concern because of the presence of many hazardous compounds which even at trace levels are a threat to human health and environment. Therefore, it is important to assess the toxicity of leachate generated and discharge it conforming to the safety standards. The present work examined the efficiency of an earlier reported Pseudomonas sp. strain ISTDF1 for detoxification of leachate collected from Okhla landfill site (New Delhi, India). GC-MS analysis performed after treatment showed the removal of compounds like alpha-limonene diepoxide, brominated dioxin-2-one, Bisphenol A, nitromusk, phthalate derivative, and nitrobenzene originally found in untreated leachate. ICP-AES analysis for heavy metals also showed reduction in concentrations of Zn, Cd, Cr, Fe, Ni, and Pb bringing them within the limit of safety discharge. Methyl tetrazolium (MTT) assay for cytotoxicity, alkaline comet assay for genotoxicity, and 7-ethoxyresorufin-O-deethylase (EROD) assay for dioxin-like behavior were carried out in human hepato-carcinoma cell line HepG2 to evaluate the toxic potential of treated and untreated leachates. The bacterium reduced toxicity as shown by 2.5-fold reduction of MTT EC₅₀ value, 7-fold reduction in Olive Tail Moment, and 2.8-fold reduction in EROD induction after 240 h of bacterial treatment.     

Fate of Bacillus thuringiensis (Bt) Cry3Bb1 protein in a soil microcosm

Transgenic crops expressing insecticidal, crystalline (Cry) Bacillus thuringiensis (Bt) proteins were commercialized in the US in 1996. There is little information in the peer-reviewed literature on the environmental fate of the coleopteran-active Bt Cry3Bb1 protein expressed in event MON863 corn. The exposure characterization of Bt proteins is unique in that the fate of the protein in soil and in crop residue must be considered. To date, the significance of macrodecomposing organisms, such as earthworms, isopods, and springtails, to the dissipation of Bt proteins present in crop residue has not been assessed. In addition, the input of Bt proteins into soil through leaching from post-harvest crop residue has not been examined. Two laboratory microcosm studies were conducted to determine the fate of Bt Cry3Bb1 in decomposing MON863 corn residue and to determine whether Bt proteins can enter soil by leaching from crop residue. In addition, the importance of macrodecomposing organisms to the degradation of Bt proteins in corn residue was assessed. Laboratory microcosms containing MON863 corn leaf, root, and stalk with and without macrodecomposers were used to examine the fate of Bt Cry3Bb1 in post-harvest MON863 corn residue. A half-life of less than five days was found for Bt Cry3Bb1 protein in decomposing MON863 corn residue. There was a trend of increasing half-life of Cry3Bb1 in microcosms containing macrodecomposers, however, this trend was only significant (p<0.05) for Bt Cry3Bb1 in MON863 leaf tissue and this increase is not likely relevant for non-target exposure. The recovery of Bt Cry3Bb1 protein from soil extracts was either below the limit of quantification (9 ng g(-1) soil) or below the limit of detection (0.7 ng mL(-1)) at all time points during the study. Based on the results from this study, Bt protein leaching from post-harvest crop residue is not a significant contributor to Bt protein input into soil.     

蔗渣碱法造纸制浆黑液资源化治理工业试验

通过工业性运行试验 ,验证“蔗渣碱法造纸制浆黑液资源化处理”项目工艺路线在工业过程中的可行性、工艺设备的可靠性、纸产品以及黑液处理回收产品的产量、质量及其工艺的稳定性 ;得出资源化回收产品产率、质量等工业实验数据 ,为项目产业化实施提供设计依据     

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)。     

Anaerobic biodegradation of 4-alkylphenols in a paddy soil microcosm supplemented with nitrate

Anaerobic degradation of phenol, p-cresol, 4-n-propylphenol (n-PP), 4-i-propylphenol (i-PP), 4-n-butylphenol (n-BP) and 4-sec-butylphenol (sec-BP) was observed in a paddy soil supplemented with nitrate. We detected the metabolites 4′-hydroxypropiophenone (HPP) from n-PP, 4-i-propenylphenol from i-PP, and 4-(1-butenyl)phenol and 4′-hydroxybutyrophenone (HBP) from n-BP. Compared with the original soils, Betaproteobacteria became predominant in the microcosm during the degradation of phenol and p-cresol whereas no remarkable change was observed in the community degrading propylphenols and butylphenols. The microcosm, however, did not degrade 4-t-butylphenol (t-BP), 4-t-octylphenol (t-OP) and 4-n-octylphenol (n-OP). Paddy soil supplemented with sulfate or iron (III) as electron acceptors did not degrade phenol and 4-alkylphenols with the exception of the degradation of p-cresol in sulfate-reducing conditions. It was demonstrated for the first time that anaerobic microbial degradation of alkylphenols, in a paddy soil supplemented with nitrate as an electron acceptor, occurred via oxidation of the alpha carbon in the alkyl chain.     

Effect of introduced phthalate-degrading bacteria on the diversity of indigenous bacterial communities during di-(2-ethylhexyl) phthalate (DEHP) degradation in a soil microcosm

Four previously isolated di-butyl-phthalate (DBP) degraders were tested for their abilities to degrade di-(2-ethylhexyl) phthalate (DEHP). In aqueous medium supplemented with 100mg/l of DEHP, both isolate G1 and Rhodococcus rhodochrous G2 showed excellent degradative activity; in three days they were able to degrade more than 97% of the added DEHP. Rhodococcus rhodochrous G7 degraded 32.5% of the added DEHP and Corynebacterium nitrilophilus G11 showed the least amount of DEHP degradation. The addition of surfactant Brij 30 at 0.1x critical micelle concentration (2mg/l) significantly improved DEHP degradation by Rhodococcus rhodochrous G2 (more than 90% of the added DEHP was degraded within 24 hours), but slightly inhibited the degradation of DEHP by the isolate G1 and Rhodococcus rhodochrous G7. Based on the 16S rDNA sequence data, isolate G1 was identified as Gordonia polyisoprenivorans. Soil inhibited DEHP degradation by G. polyisoprenivorans G1; fourteen days after a second addition of DEHP, 11.5% of the total added DEHP (i.e., 243.4 microg/g soil) remained detectable. Changes in the bacterial community were monitored using denaturing gradient gel electrophoresis (DGGE) and respective dendrogram analysis. It is clear that DEHP and DEHP plus G. polyisoprenivorans G1 substantially affected the bacterial community structure in the soils. However, as the population of indigenous DEHP degraders increased in the DEPH-treated soil, its bacterial communities resembled those in the DEHP plus G. polyisoprenivorans G1-inoculated soil by Day 17.     

Comparison of microbial indicators under two water regimes in a soil amended with combined paper mill sludge and decomposed cow manure

An incubation study was conducted under laboratory conditions to compare the effects of soil amendment of combined paper mill sludge (PS) and decomposed cow manure (DCM) on selected microbial indicators. A lateritic soil (Typic Haplustalf) was amended with 0 (control), 20 or 80tha(-1) (wet weight) of PS or DCM. The amended soils were then adjusted to 60% water holding capacity (WHC) or submerged conditions, and incubated at 27 degrees C in dark for up to 120days (d). The microbial biomass C (MBC), the basal soil respiration and the enzyme activities of the beta-glucosidase, acid phosphatase and sulphatase were analyzed at day 15, 30, 45, 60 and 120. Compared to the unamended soil (control), the MBC, the basal soil respiration and the enzyme activities increased with the rate of PS and DCM. At similar rate, the DCM treatment increased significantly the MBC, the soil respiration and the enzyme activities compared to the PS treatment. Also, the water regimes affected the microbial activities. At 60% WHC, the MBC and soil respiration increased during the first 30d and decreased thereafter. The enzyme activities showed similar trends, where they increased for the first 60d, and decreased thereafter. In contrast, under submerged condition, the MBC and enzymes activities declined during 120d, whereas the soil respiration increased. Compared to the control, the used of PS and DCM had no negative impact of the soil microbial parameters, even at the highest application rate. Long-term field experiments are required to confirm these laboratory results.     

Assessing the biological status of fish in a river receiving pulp and paper mill effluents

This study compared the use of sentinel species- and community-based field approaches for assessing the biological status of fish living in a river receiving pulp and paper mill effluents. Three approaches were compared. Two approaches used sentinel species. One of these involved an internal/external examination of the fish that leads to the calculation of a fish health assessment index (HAI) and the other involved biochemical measurements of hepatic mixed function oxidase (MFO) activity and plasma steroid levels. The third approach characterized the fish community structure according to an index of biotic integrity (IBI). The comparison focused on how the methods respond to the hypothesis that recent process modifications/effluent treatment changes, resulting in demonstrable improvements in effluent quality, have beneficial effects on fish. Neither of the approaches using sentinel fish indicated clear mill-related influences either before or after the process modifications/effluent treatment changes. There was no evidence of depressed plasma steroids and increased MFO activity in fish frequently associated with mill effluent exposure in previous studies. While the HAI was higher at stations downstream from two mills, this could not be linked to effluent exposure alone. In contrast, the study of community structure showed a substantial improvement in fish assemblages at all the mill sites.     

Bioaugmentation with a Bacillus sp. to reduce the phytoavailable Cd of an agricultural soil: comparison of free and immobilized microbial inocula

In order to reduce the cadmium potentially available for plants, soil bioaugmentation was performed by using a Bacillus sp. In a pot experimentation, sterilized and non-sterilized soils were inoculated using free or immobilized cells entrapped in alginate beads. This test was carried out with different inoculum sizes (2 x 10(10) and 2 x 10(11)CFU kg(-1) dw of soil) and alginate bead compositions (10 and 15 g of both alginate and CaCl(2) l(-1)). Then, the soil pots were incubated at 20 degrees C and the soil humidity was kept at a level of 20%. After 3 weeks of a batch incubation, the potentially phytoavailable Cd was reduced up to a factor of 14. The bioaugmentation resulted in the soil colonization by Bacillus sp. thanks to an increase of the cell concentration up to 1.8 log units. However, in comparison to the cells being inoculated in a free mode, the immobilization of the cells did not significantly improve the survival of the cells in the soil. Although the resulting effect not being highly pronounced, the potentially phytoavailable Cd correlated with the cell concentration in a surprisingly positive way. What is more, the Bacillus concentrations in the soil were positively correlated with the inoculum, too.     

Treatment of PAHs in contaminated soil by extraction with aqueous DNA followed by biodegradation with a pure culture of Sphingomonas sp

The biodegradation of polycyclic aromatic hydrocarbons (PAHs) in aqueous deoxyribonucleic acid (DNA) solution from contaminated soil washing was investigated. Initial data with a model effluent consisting of anthracene, phenanthrene, pyrene and benzo[a]pyrene that were individually dissolved in 1% aqueous DNA solution confirmed their positive degradation by Sphingomonas sp. at around 10(8)CFU mL(-1) initial cell loading. For anthracene and phenanthrene, complete removal was achieved within 1h treatment. Degradation of pyrene and benzo[a]pyrene took a relatively longer time of a few days and weeks, respectively. DNA-dissolved PAHs were also degraded relatively faster than PAH crystals in aqueous medium to suggest that the binding of the PAHs in the polymer does not pose serious constraint to bacterial uptake. The DNA was stable against the PAH-degrading bacteria. Parallel experiments with actual DNA solutions obtained during pyrene extraction from an artificially spiked soil also showed similar results. Close to 100% pyrene degradation was achieved after 1d treatment. With its chemical stability, the cell-treated DNA was re-used up to four cycles without a considerable decline in extraction performance.     

Microbial degradation characteristics and kinetics of novel pyrimidynyloxybenzoic herbicide ZJ0273 by a newly isolated Bacillus sp. CY

ZJ0273 (propyl 4-(2-(4,6-demethoxy pyrimidin-2-yloxy)benzylamino)benzoate) is a novel herbicide developed in China for oilseed crop. Sixteen bacteria capable of utilizing ZJ0273 as the sole carbon source were isolated from soils. One of the isolates was designated as Bacillus sp. CY based on its physiological and biochemical characteristics and phylogenetic analysis of 16S rDNA sequences. The present study aimed to investigate the ZJ0273 degradation characteristics and kinetics by Bacillus sp. CY which has the ability to utilize ZJ0273 as the sole source of carbon and energy under aerobic conditions. The optimum biodegradation temperature, pH, and ZJ0273 initial concentration were 20–40 °C, 5.0–9.0, and 50–400 mg/l, respectively. Strain CY degraded 65 % of ZJ0273 (initial concentration of 50 mg/l) during 30 days of incubation in basal mineral medium at pH 8.0 and 35 °C. DT50 (half-life value), k (degradation rate constant of ZJ0273), and R ² are 19.20 days, 0.0361 day^?1, and 0.9464, respectively.     

Enhanced remediation of dioxins-spiked soil by a plant-microbe system using a dibenzofuran-degrading Comamonas sp. and Trifolium repens L

In this study, rhizoremediation technology was applied to dioxins-spiked soil. A dibenzofuran-degrading bacterium Comamonas sp. strain KD7, reported in the previous paper, was used in combination with white clover (Trifolium repens L.). First, the effect of strain KD7 on clover seed germination and root elongation was examined in the presence of dioxins compounds. As a result, the white clover seeds inoculated with strain KD7 exhibited a higher germination efficiency and increased root elongation compared with uninoculated white clover. Next, the recovery efficiency of two extraction methods were considered for analyzing the dioxin concentration in soil samples, then, the potential of the plant-microbe combination was evaluated for the remediation of dioxins-spiked soil. After 12 week of growth, significant reductions in the soil were confirmed for most compounds. Our results demonstrated that clover can function as a carrier in order to increase the dioxin-degrading activity of strain KD7. The association of clover and strain KD7 is considered to be a potential tool in the remediation of dioxin-contaminated soil.     

Influence of linear alkylbenzene sulfonate (LAS) on the structure of Alphaproteobacteria,Actinobacteria, and Acidobacteria communities in a soil microcosm

Background, aim, and scope  

Linear alkylbenzene sulfonate (LAS) is the most used anionic surfactant in a worldwide scale and is considered a high-priority pollutant. LAS is regarded as a readily biodegradable product under aerobic conditions in aqueous media and is mostly removed in wastewater treatment plants, but an important fraction (20–25%) is immobilized in sewage sludge and persists under anoxic conditions. Due to the application of the sludge as a fertilizer, LAS reaches agricultural soil, and therefore, microbial toxicity tests have been widely used to evaluate the influence of LAS on soil microbial ecology. However, molecular-based community-level analyses have been seldom applied in studies regarding the effects of LAS on natural or engineered systems, and, to our knowledge, there are no reports of their use for such appraisals in agricultural soil. In this study, a microcosm system is used to evaluate the effects of a commercial mixture of LAS on the community structure of Alphaproteobacteria, Actinobacteria, and Acidobacteria in an agricultural soil.     

Interaction effects of insecticides on microbial populations and dehydrogenase activity in a black clay soil

Three insecticides, monocrotophos, quinalphos, and cypermethrin, were applied at 0, 5, 10, and 25 microg g(-1) either singly or in combination to a black clay soil to investigate their effects on the soil microflora and dehydrogenase activity. All three insecticides significantly enhanced the proliferation of bacteria and fungi and the soil dehydrogenase activity even at the highest level of 25 microg g(-1). Monocrotophos or quinalphos in combination with cypermethrin at tested levels interacted significantly to yield additive, synergistic, and antagonistic responses toward bacteria and fungi and dehydrogenase activity in soil. Antagonistic interactions were more pronounced toward soil microflora and dehydrogenase activity when the two (monocrotophos or quinalphos + cypermethrin) insecticides were present together in the soil at highest level (25 + 25 microg g(-1)), whereas synergistic or additive responses occurred at lower level with the same combination of insecticides in soil.     

Parameterization and evaluation of sulfate adsorption in a dynamic soil chemistry model

Sulfate adsorption was implemented in the dynamic, multi-layer soil chemistry model SAFE. The process is modeled by an isotherm in which sulfate adsorption is considered to be fully reversible and dependent on sulfate concentration as well as pH in soil solution. The isotherm was parameterized by a site-specific series of simple batch experiments at different pH (3.8-5.0) and sulfate concentration (10-260 micromol 1(-1)) levels. Application of the model to the Lake Gardsj6n roof covered site shows that including sulfate adsorption improves the dynamic behavior of the model and sulfate adsorption and desorption delay acidification and recovery of the soil. The modeled adsorbed pool of sulfate at the site reached a maximum level of 700 mmol/m(2) in the late 1980s, well in line with experimental data.     

Degradation characteristics of a dibenzofuran-degrader Terrabacter sp. strain DBF63 toward chlorinated dioxins in soil

Nonylphenols (NPs) are the primary stable metabolites of alkylphenol polyethoxylates (APEs), a family of compounds widely used in industry and in some domestic products. As NPs accumulate in sediments in aquatic environments, the risk to benthic organisms needs to be assessed. In this study 4NP-spiked sediments were tested on larvae of the dipteran Chironomus riparius. First instar larvae obtained from populations at three different sources were used. To spike the sediments, an equilibration procedure between water and sediment was adopted to avoid the use of solvents. Lower 10-d LC50 values were determined for two populations of C. riparius from clean environments (315-465 and 315-350 microg g(-1) d.w., respectively) than those of a strain deriving from a population collected in a polluted river (600-680 microg g(-1) d.w.). Larval growth always decreased with increasing 4NP concentration but without any defined trend. The results of this study suggest that tolerance to the toxicant can be developed in populations of polluted environments and that testing procedures should be standardised.     

A remediation strategy based on active phytoremediation followed by natural attenuation in a soil contaminated by pyrite waste

Phytoremediation of metal-polluted soils can be promoted by the proper use of soil amendments and agricultural practices. A 4-year phytoremediation programme was applied to a site affected by the toxic spill of pyrite residue at Aznalcóllar (Spain) in 1998, contaminated with heavy metals (Zn, Cu, Pb, Cd) and arsenic. This consisted of active phytoremediation, using organic amendments (cow manure and compost) and lime and growing two successive crops of Brassica juncea (L.) Czern., followed by natural attenuation without further intervention. Changes in soil pH, extractable metal and As concentrations, organic carbon content and microbial biomass was evaluated. The initial oxidation of metal sulphides from pyrite residues released soluble metals and reduced soil pH to extremely acidic values (mean 4.1, range 2.0-7.0). The addition of lime (up to 64 t ha(-1)) increased soil pH to adequate values for plant growth, resulting in a significant decrease in DTPA-extractable metal concentrations in all plots. The natural attenuation phase showed also a decrease in extractable metals. Organic treatments increased the soil total organic carbon, which led to higher values of microbial biomass (11.6, 15.2 and 14.9 g kg(-1) TOC and 123, 170 and 275 microg g(-1) biomass-C in control, compost and manure plots, respectively). Active phytoremediation followed by natural attenuation, was effective for remediation of this pyrite-polluted soil.     

Biotransformation of phenylurea herbicides by a soil bacterial strain, Arthrobacter sp. N2: structure, ecotoxicity and fate of diuron metabolite with soil fungi

In order to assess the influence of the aromatic substitution on the ability of a soil bacterial strain, Arthrobacter sp. N2, to degrade phenylurea herbicides, biotransformation assays were performed in mineral medium with resting cells of this soil bacterial strain on three phenylurea herbicides (diuron, chlorotoluron and isoproturon). Each herbicide considered, led to the formation of only one metabolite detected by HPLC analysis. After isolation, the metabolites were identified by NMR and MS, as the corresponding substituted anilines. According to the Microtox test (realized on the bacterium Vibrio fischeri), these metabolites presented non-target toxicity far more important (up to 600 times higher for 4-isopropylaniline) than the parent molecule. For isoproturon and chlorotoluron, the amount of substituted anilines obtained at the end of the biotransformation was very low, whereas the biotransformation of diuron into 3,4-dichloroaniline was almost quantitative. In this last case, the degradation product accumulated in the medium. In soil, other microorganisms are present that might degrade it. So the biotransformation of 3,4-dichloroaniline was then tested with four fungal strains: Aspergillus niger, Beauveria bassiana, Cunninghamella echinulata var. elegans and Mortierella isabellina. The aniline was further transformed with all the microorganisms tested. Only one metabolite was detected by HPLC analysis and after isolation, it was identified to be 3,4-dichloroacetanilide. This acetylated compound led to biological effects less important on V. fischeri than 3,4-dichloroaniline. These results stress the importance of identifying the degradation products to assess the impact of a polluting agent. Indeed, the pollutant may undergo transformation yielding compounds more toxic than the parent molecule.     

Hydrogeochemical evaluation of calcareous eolianite aquifer with saline soil in a semiarid area

The aim of this study is to evaluate the groundwater geochemistry in Burg Elarab area as an example of a calcareous eolianite aquifer that is covered with saline soil in a semiarid climatic condition. To conduct this study, 37 groundwater samples were taped from the production wells in addition to two surface water samples from Mallahet Mariut Lake and Bahig Canal. To elucidate the origin of dissolved ions and the geochemical processes influencing this groundwater, combinations of geomorphological, pedological, hydrogeological, hydrochemical, and statistical approaches were considered. Results suggest that the groundwater flows from both sides of the plain to the central area. Soil type and salinity and the intruded brackish lake water are the main factors controlling the groundwater chemistry. Chemically, the groundwater samples were classified into three groups. Group 1 samples have higher salinity range and characterize the area close to Mallahet Mariut and are influenced by cation exchange processes. Group 2 samples have an intermediate salinity range, occupy most of the plain area, and receive water from direct infiltrations and mixing between different recharge sources. Group 3 samples have low salinity range and limited areal extent and characterize the groundwater flowing from the Mariut Tableland. Reverse ion exchange is the predominant process in the latter group. Calcite precipitation is a general phenomenon characterizing all the groundwater types in the study area.     

Study of the biodegradation process of polychlorinated biphenyls in liquid medium and soil by a new isolated aerobic bacterium (Janibacter sp.)

We have isolated and characterised a novel aerobic bacterial strain, designated MS3-02, belonging to the genus Janibacter sp. The capability of this new strain to degrade polychlorinated biphenyls (PCBs) in a commercial mixture (Aroclor 1242) in liquid medium and in soil (sterile and non sterile soil), under laboratory scale, has been evaluated. MS3-02 was isolated from the soil around of an incinerator, located in the east of Madrid (Spain). Gas-chromatographic analysis showed that MS3-02 was able to reduce most peaks observed in the chromatogram between 70% and 100% after seven days of incubation in a culture mineral medium containing yeast extract, but without the addition of biphenyl. The presence of biphenyl in the culture medium decreased the rate of PCB degradation by this bacterium. Comparing the performance of the MS3-02 in liquid culture medium and in soil, degradation was less efficient in sterile soil and still less efficient in non sterile soil. Under the best conditions (sterile soil and 20 weeks of incubation) MS3-02 was able to reduce, between 50% and 100%, nine of the main gas-chromatographic peaks in Aroclor 1242.