Assessment of s-Triazine Catabolic Potential in Soil Bacterial Isolates Applying atz Genes as Functional Biomarkers

Fluorescence in situ hybridization (FISH) technique and qPCR analyses, targeting atz genes, were applied to detect the presence of simazine-degrading bacteria in an agricultural soil with a history of herbicide application. atzB-targeted bacteria detected by FISH represented 5% of total soil bacteria with potential capability to metabolize the herbicide. The soil natural attenuation capacity was confirmed in soil microcosms by measuring simazine degradation. Moreover, four bacterial strains were isolated from the soil and identified as Acinetobacter lwoffii, Pseudomonas putida, Rhizobium sp. and Pseudomonas sp. The isolates were able to grow using different s-triazine compounds and related metabolites as the sole carbon source. Growth parameters in presence of simazine were calculated using the Gompertz model. Rhizobium sp. showed the highest simazine degradation (71.2%) and mineralization (38.7%) rates, whereas the lowest values were found to A. lwoffii??50.4% of degradation and 22.4% of mineralization. Results from qPCR analyses of atzA, atzB and atzC genes revealed their presence in Rhizobium sp. and A. lwoffii, being atzB and atzC the most abundant functional genes. Rhizobium sp. showed a higher amount of the three biomarkers compared to A. lwoffii: the atzA, atzB and atzC gene copy number per microlitre were, respectively, 10¹, 10² and 10³-fold higher in the former. Therefore the proposed molecular approaches based on the use of atz genes as biomarkers can be considered as useful tools to evaluate the presence and potential capability of degrading-s-triazines soil microorganisms.     

Improvement of simazine degradation by inoculation of corn and soybean plants with rhizobacteria

We isolated the strains of aerobic bacteria, that were able to use the simazine (2-chloro4,6-bis(ethylamino)-s-triazine) herbicide as a sole source of nitrogen, from the roots of corn plants and the nodules of soybean plants. Partial base sequences of 16S rRNA genes and physiological characteristics of the representative isolates revealed that they belonged to Agrobacterium radiobacter and Bradyrhizobium japonicum, respectively. Inoculation of corn plants with the isolates of A. radiobacter resulted in a faster decrease in the amount of simazine in vermiculite soil. Neither bacteria nor corn plants alone were able to promote the decrease in the amount of simazine. Soybean plants modulated by the isolates of B. japonicum were more sensitive to simazine in a hydroponic medium compared to non-nodulated soybeans, though a significant decrease in the amount of simazine in the media was observed. The results obtained in the present study indicate that the rhizosphere of corn and soybean plants containing root nodules is an appropriate habitat for simazine-degrading bacteria to degrade the herbicide.     

Effects of solid olive-mill waste addition to soil on sorption, degradation and leaching of the herbicide simazine

Abstract. A laboratory study was conducted to investigate the effects of adding an intermediary byproduct of olive oil extraction ( alperujo or solid olive-mill waste, SOMW) on the sorption, degradation and leaching of the herbicide simazine in a sandy loam soil. The effect of SOMW addition on soil porosity was also assessed. The soil was amended in the laboratory with SOMW at two different rates (5% and 10% w/w). Simazine sorption isotherms showed a great increase in herbicide sorption after SOMW addition to soil; sorption increased with the amount of SOMW added. Incubation studies showed extended persistence by reduced biodegradation of simazine in the soil amended with SOMW compared with the unamended soil. Although the addition of SOMW to soil increased the total porosity, breakthrough curves of simazine in handpacked soil columns showed that SOMW addition retarded the vertical movement of the herbicide through the soil and reduced the total amount of herbicide leached. It appeared that the longer residence time of simazine in the amended soil columns (>20 days) compared with that in the unamended soil column (<20 days) allowed enhanced degradation and/or irreversible sorption under column leaching conditions. The results revealed important changes in herbicide behaviour upon SOMW addition, confirming the need to assess these changes in order to optimize the combined use of organic wastes and soil-applied pesticides.     

DNOC, a model pollutant, adversely affects the potential of soil microbial communities to mineralise the herbicide 2,4-D: an investigation using micro-sampling procedures

The effects of the herbicide, DNOC, 4,6-dinitro-o-cresol, a model pollutant, have been studied by comparing the potential of soil microbial communities present in individual soil aggregates or in larger soil microcosms as samples of soil aggregates to mineralise the herbicide 2,4-D. We have shown that 2-3 mm soil aggregates vary widely in their 2,4-D mineralisation potential and that ageing or exposure to DNOC considerably simplified the distribution patterns of this capacity. The main factors of variation have been quantified and classified using a quasi-likelihood method derived from the Generalised Linear Model approach. Besides DNOC concentration and duration of exposure, an additional ‘rank’ factor reflecting a desiccation gradient of the aggregates on the microtiter plates was found to have statistical significance. We concluded that it should be possible to derive an experimental approach, designated as ‘functional profiling’, with potential use to detect soil chemical contamination. Curves of 2,4-D mineralisation in individual soil aggregates could be classified according to three different types of kinetics, which were assumed to reflect heterogeneous spatial distribution, differences in microbial community composition and varying efficiency of the microbial consortia involved in 2,4-D degradation. Exposure to DNOC considerably simplified the distribution patterns of the different types of kinetics with one type, showing slow rate and low cumulative mineralisation, becoming predominant as ageing, concentration and duration of DNOC exposure increased. We argue on the possible use of ‘kinetic profiling’ as a sensitive bioindicator of soil quality. By comparison, in soil microcosms, 2,4-D mineralisation showed an extra mineralisation potential of 64% over individual aggregates in the control soil and exposure to DNOC was followed by concentration and time-dependent recovery of the 2,4-D mineralisation potential. It is likely that 2 g size soil microcosms gather a larger number of biochemical capacities which could complement each other to increase the potential of soil to mineralise xenobiotic compounds.     

Persistence of simazine and terbuthylazine in a semiarid soil after organic amendment with urban sewage sludge

The persistence of two herbicides, simazine and terbuthylazine, and appearance of their principal dealkylated chloro-s-triazine metabolites have been studied in agricultural soil after the addition of urban sewage sludge as organic amendment. Both herbicides and metabolites were monitored during long-term laboratory incubation (140 days) and analyzed by gas chromatography with a nitrogen-phosphorus detector (GC-NPD). Residues were confirmed by gas chromatography with a mass selective detector (GC-MSD). A sonication microextraction method was used to extract the compounds. The organic amendments used were urban sewage sludge and the humic fraction of this sludge, to increase the organic matter content of the soil from 1% to 2%. For both compounds, simazine and terbuthylazine, the degradation began earlier in the amended soils. Simazine showed a higher dissipation rate than terbuthylazine, the percentage of the former at the end of the experiment being lower than 2% in all cases, while for terbuthylazine the corresponding percentage ranged from 5% to 46%. Organic amendment, mainly its humic fraction, caused a certain stabilization of terbuthylazine in the soil, but did not greatly influence the residual amount of simazine at the end of the experiment. The periodic aeration of the soil caused a greater degradation in the case of terbuthylazine. Only mono-deethylsimazine and deethylterbuthylazine were isolated from the soil during the time the experiment lasted, while the di-deethylated metabolite of simazine was not found.     

Effects of Wood Amendments on the Degradation of Terbuthylazine and on Soil Microbial Community Activity in a Clay Loam Soil

The herbicide terbuthylazine is widely used within the EU; however, its frequent detection in surface and groundwater, together with its intrinsic toxicological properties, may pose a risk both for human and environmental health. Organic amendments have recently been proposed as a possible herbicide sorbent in soil, in order to limit herbicide movement from soil to water. The environmental fate of terbuthylazine depends not only in its mobility but also in its persistence. The latter is directly dependent on microbial degradation. For this reason, the effects of pine and oak residues on terbuthylazine soil microbial community functioning and on the potential of this community for terbuthylazine degradation were studied. For this purpose, degradation kinetics, soil dehydrogenase activity and the number of live bacteria were assessed in a clay loam soil treated with terbuthylazine and either amended with pine or oak wood or unamended (sterilised and non-sterilised). At day 65, 85?% of the herbicide applied still persisted in the sterile soil, 73?% in the pine-amended one and 63?% in the oak-amended and unamended ones. Pine residues increased the sorption of terbuthylazine to soil and hampered microbial degradation owing to its high terbuthylazine sorption capacity and a decrease in the bioavailability of the herbicide. On the contrary, in the presence of oak residues, the herbicide sorption did not increase significantly. The overall results confirm the active role of the soil microbial community in terbuthylazine degradation in amended and unamended soils and in a liquid enrichment culture performed using an aliquot of the same soil as the inoculum. In this clay loam soil, in the absence of amendments, the herbicide was found to be quite persistent (t _1/2?>?95?days), while in the enrichment culture, the same natural soil bacterial community was able to halve terbuthylazine in 24?days. The high terbuthylazine persistence in this soil was presumably ascribable to its texture and in particular to the mineralogy of the clay fraction.     

Screening of Microorganisms for Biodegradation of Simazine Pollution (Obsolete Pesticide Azotop 50 WP)

The capability of environmental microorganisms to biodegrade simazine—an active substance of 2-chloro-s-triazine herbicides (pesticide waste since 2007)—was assessed. An enormous metabolic potential of microorganisms impels to explore the possibilities of using them as an alternative way for thermal and chemical methods of utilization. First, the biotope rich in microorganisms resistant to simazine was examined. Only the higher dose of simazine (100 mg/l) had an actual influence on quantity of bacteria and environmental fungi incubated on substrate with simazine. Most simazine-resistant bacteria populated activated sludge and biohumus (vermicompost); the biggest strain of resistant fungi was found in floral soil and risosphere soil of maize. Compost and biohumus were the sources of microorganisms which biodegraded simazine, though either of them was the dominant considering the quantity of simazine-resistant microorganisms. In both cases of periodic culture (microorganisms from biohumus and compost), nearly 100% of simazine (50 mg/l) was degraded (within 8 days). After the repeated enrichment culture with simazine, the rate of its degradation highly accelerated, and just after 24 h, the significant decrease of simazine (20% in compost and 80% in biohumus) was noted. Although a dozen attempts of isolating various strains responsible for biodegradation of simazine from compost and biohumus were performed, only the strain identified as Arthrobacter urefaciens (NC) was obtained, and it biodegraded simazine with almost 100% efficiency (within 4 days).     

Previous exposure advances the degradation of an anthropogenic s-triazine regardless of soil origin

Purpose

Previous investigations—field samplings and laboratory experiments—support the hypothesis that the degradation of s-triazines is enhanced in previously exposed as compared to pristine soils in terrestrial environments. Despite this, bottlenecks of soil sampling and various soil modification practices in microcosm studies have made it difficult to guarantee that previous contamination history enhances contaminant degradation regardless of soil origin in terrestrial ecosystems. We test the hypothesis that the degradation of simazine (2-chloro-4,6-bis(ethylamino)-s-triazine) is enhanced in previously exposed soils as compared to pristine soils in 10 l buckets at the mesocosm scale.

Materials and methods

We collected soil at three separate sites consisting of a previously exposed and a pristine field. At every field, soil was collected at three separate plots and simazine degradation (days 0 and 65) and the response to atzB degrader gene primers (days 0 and 110) were followed. We analyzed the results using analysis of covariance (ANCOVA). Previous exposure and field site were assessed as fixed factors and initial simazine concentration and abiotic soil conditions as covariates.

Results and discussion

After the 65-day exposure, remaining simazine concentrations depended on previous exposure but not on collection site. The response to atzB gene primers was positive in all mesocosms where simazine degradation had been rapid. Soil moisture, pH, and organic matter content were insignificant. If soil moisture was not included in the ANCOVA model, previous exposure did not appear as a significant factor.

Conclusions

The results support the hypothesis that simazine is degraded more rapidly in previously exposed soils as compared to pristine environments, provided that degradation genes are available. Previously exposed soil might be used to enhance the degradation of simazine in recently contaminated terrestrial soils, supposing that the central requirements for microbial growth are adequate.     

Biochar mediated alterations in herbicide breakdown and leaching in soil

Biochar application to soil has been proposed as a mechanism for improving soil quality and the long term sequestration of carbon. The implications of biochar on pesticide behavior, particularly in the longer term, however, remains poorly understood. Here we evaluated the influence of biochar type, time after incorporation into soil, dose rate and particle size on the sorption, biodegradation and leaching of the herbicide simazine. We show that typical agronomic application rates of biochar (10-100 t ha⁻¹) led to alterations in soil water herbicide concentrations, availability, transport and spatial heterogeneity. Overall, biochar suppressed simazine biodegradation and reduced simazine leaching. These responses were induced by a rapid and strong sorption of simazine to the biochar which limits its availability to microbial communities. Spatial imaging of ¹⁴C-labeled simazine revealed concentrated hotpsots of herbicide co-localized with biochar in the soil profile. The rate of simazine mineralization, amount of sorption and leaching was inversely correlated with biochar particle size. Biochar aged in the field for 2 years had the same effect as fresh biochar on the sorption and mineralization of simazine, suggesting that the effects of biochar on herbicide behavior may be long lasting. We conclude that biochar application to soil will reduce the dissipation of foliar applied pesticides decreasing the risk of environmental contamination and human exposure via transfer in the food chain, but may affect the efficacy of soil-applied herbicides.     

Influence of soil moisture on sorption and degradation of hexazinone and simazine in soil.

Sorption and degradation rates of hexazinone and simazine on soil were determined in a sandy loam soil incubated, during 44 days, at 25 degrees C with moisture contents ranging from 4% to 18%. Herbicide levels in soil solution were also measured, after extraction of this solution by a centrifugation method. All experiments were conducted with treated soil in plastic columns, and the results showed that this method is suitable for the simultaneous study of pesticide sorption and degradation in soil at different environmental conditions. In general, sorption of both herbicides was higher for aged herbicide residues compared to recently applied herbicides, and soil subjected to drying and rewetting cycles had the highest sorption values. K(f) values ranged from 0.5 to 1.2 for simazine and from 0.2 to 0.4 for hexazinone. Degradation rates increased with soil moisture content for both herbicides, and drying-rewetting of soil yielded degradation rates slower than that obtained at 10% soil moisture content. Hexazinone concentration in soil solution decreased with incubation time faster than simazine.     

Lumbricid macrofauna alter atrazine mineralization and sorption in a silt loam soil

Atrazine is a widely used herbicide and is often a contaminant in terrestrial and freshwater ecosystems. It is uncertain, however, how the activity of soil macrofauna affects atrazine fate and transport. Therefore, we investigated whether earthworms enhance atrazine biodegradation by stimulating herbicide degrading soil microflora, or if they increase atrazine persistence by facilitating herbicide sorption. Short (43 d) and medium term (86 d) effects of the earthworms Lumbricus terrestris and Aporrectodea caliginosa on mineralization, distribution, and sorption of U-ring-¹⁴C atrazine and on soil C mineralization was quantified in packed-soil microcosms using silt loam soil. A priming effect (stimulation of soil C mineralization) caused by atrazine supply was shown that likely lowered the earthworm net effect on soil C mineralization in atrazine-treated soil microcosms. Although earthworms significantly increased soil microbial activity, they reduced atrazine mineralization to ¹⁴CO₂-C from15.2 to 11.7% at 86 d. Earthworms facilitated formation of non-extractable atrazine residues within C-rich soil microsites that they created by burrowing and ingesting soil and organic matter. Atrazine sorption was highest in their gut contents and higher in casts than in burrow linings. Also, gut contents exhibited the highest formation of bound atrazine residues (non-extractable atrazine). Earthworms also promoted a deeper and patchier distribution of atrazine in the soil. This contributed to greater leaching losses of atrazine in microcosms amended with earthworms (3%) than in earthworm-free microcosms (0.003%), although these differences were not significant due to high variability in transport from earthworm-amended microcosms. Our results indicated that earthworms, mainly by casting activity, facilitated atrazine sorption, which increased atrazine persistence. As a consequence, this effect overrode any increase in atrazine biodegradation due to stimulation of microbial activity by earthworms. It is concluded that the affect of earthworms of atrazine mineralization is time-dependent, mineralization being slightly enhanced in the short term and subsequently reduced in the medium term.     

Effects of nutrient concentration on the biodegradation of crude oil and associated microbial populations in the soil

The influence of nutrient amendments on the biodegradation of a crude oil and on associated microbial populations in an agricultural soil was studied in microcosms during a 150-day experiment. Concentration and chemical composition of residual hydrocarbons (HC) were periodically monitored in series of polluted soils unfertilized and fertilized with increasing concentrations of nutrients. The decrease in HC concentration was effective in all soils over time. The maximal biodegradation extent was 62%, meanwhile the natural attenuation contributed to 47% of degradation. A permanent inhibition of hydrocarbons assimilation was recorded with a high input of nutrients. The biodegradation of saturates, aromatics and polars was respectively, permanently, temporally and not reduced by excessive fertilization in soil. Accumulation of polar metabolic by-products was demonstrated. Enumerations of total heterotrophic bacteria and hydrocarbon-adapted bacteria showed a strong stimulation in both populations. Maximum stimulation was observed with the highest input of nutrients. However, the extents of biodegradation were not concurrently improved, thus indicating that the microbial degraders were selected depending on the nutrient supply. The permanent and/or temporally inhibition of the saturated and unsaturated HC assimilation revealed that different nutrient supplies were optimum for the degradation of aliphatic and aromatic HC.     

Enhanced microbial degradation of pentachlorophenol from soil in the presence of earthworms: Evidence of functional bacteria using DNA-stable isotope probing

This study investigated the effect of two earthworm species (Amynthas robustus Perrier and Eisenia fetida Savigny) on the soil microbial degradation of pentachlorophenol (PCP). PCP-degrading microbes were identified using DNA-stable isotope probing (SIP). The results showed that adsorption and fixing to soil particles and organic fractions dominated the fate of PCP in soil without any amendments. The inoculation of both earthworm species significantly enhanced soil PCP disappearance and basal respiration. The DNA-SIP results revealed that Klebsiella, Cupriavidus, Aeromonas, and Burkholderia spp. were present at higher relative abundances in [¹³C]-labeled-PCP-amended soil microcosms than [¹²C]-PCP-amended soil in the presence of A. robustus, indicating that these bacterial species were responsible for PCP assimilation. Cupriavidus and Aeromonas sp. were also detected in the earthworm gut before inoculation, and their relative abundance was affected by earthworms. These results demonstrated that earthworms can introduce functional bacteria into soils and increase the population of PCP-degrading bacteria, thereby accelerating soil PCP degradation.     

Structural and functional diversity of bacterial community in soil treated with the herbicide napropamide estimated by the DGGE,CLPP and r/K-strategy approaches

Napropamide is one of the most commonly used herbicide in agricultural practice and can exhibit toxic effect to soil microorganisms. Therefore, the main objective of this study was to examine the genetic and functional diversity of microbial communities in soil treated with napropamide at field rate (FR, 2.25 mg kg⁻¹ of soil) and 10 times the FR (10 × FR, 22.5 mg kg⁻¹ of soil) by the denaturing gradient gel electrophoresis (DGGE) and the community level physiological profile (CLPP) methods. In addition, the r/K-strategy approach was used to evaluate the effect of this herbicide on the community structure of the culturable soil bacteria. DGGE patterns revealed that napropamide affected the structure of microbial community; however, the richness (S) and genetic diversity (H) values indicated that the FR dosage of napropamide experienced non-significant changes. In turn, the 10 × FR dosage of herbicide caused significant changes in the S and H values of dominant soil bacteria. DGGE profiles suggest an evolution of bacteria capable of degrading napropamide among indigenous microflora. Analysis of the CLPPs indicated that the catabolic activity of microbial community expressed as AWCD (average well-color development) was temporary positively affected after napropamide application and resulted in an increase of the substrate richness (S_R) as well as functional biodiversity (H) values. Analysis of the bacterial growth strategy revealed that napropamide affected the r- or K-type bacterial classes (ecotypes). In treated-soil samples K-strategists dominated the population, as indicated by the decreased ecophysiological (EP) index. Napropamide significantly affected the physiological state of culturable bacteria and caused a reduction in the rate of colony formation as well as a prolonged time of growth rate. Obtained results indicate that application of napropamide may poses a potential risk for soil functioning.     

Degradation and Leaching of Simazine in Arctic Sandy Soils

Abstract

Degradation and leaching of ¹⁴C-labelled simazine in coarse sandy soils at 15 + 1°C were investigated using radiometric and mass-spectrometric methods. During 6 months incubation approx. 4–7% of the applied ¹⁴C-simazine was evolved as ¹⁴CO₂. 4–9% of the simazine still remained in the soil. Addition of hen manure or acidification by addition of peat did not clearly influence the rate of degradation of simazine, whereas mechanical treatment significantly increased its degradation. In a nitrogen atmosphere the rate of degradation of simazine was reduced.

9–15% of the simazine or its radioactive metabolites leached through a 33 cm sandy moraine soil column (diameter 6 cm) in ca. 1770 mm of precipitation over a 4 month period, and 2% was leached from a fine sand soil under the same conditions.     

Isoproturon mineralization in an agricultural soil

The impact of soil moisture content and temperature on isoproturon (3-(4-isopropylphenyl)-1,1-dimethyl-urea [IPU]) mineralization activity was assessed on an agricultural soil regularly exposed to this herbicide. Mineralization of ¹⁴C-IPU was monitored on soil microcosms incubated at different temperatures (10°C, 20°C, 28°C) and soil moisture contents (9%, 12%, 15, 18%, 21%, 24%). An increase in temperature and/or soil moisture significantly enhanced the maximum rate and percentage of IPU mineralization while it decreased the lag time before mineralization. The maximum rate and percentage of IPU mineralization respectively ranged from 0.18% day⁻¹ and 9% for the lowest temperature and soil moisture content pair (10°C–9%) to 1.51% day⁻¹ and 27.1% for the highest pair (28°C–24%). Statistics revealed a cross interaction of temperature and soil moisture content on the maximum rate of IPU mineralization. The optimum conditions for IPU mineralization, estimated from the double Gaussian model, were 25.8°C and 24% soil moisture content. The influence of fluctuations in soil moisture content on IPU-mineralization was investigated by subjecting the soil microcosms to drought stress. When IPU was added at the end of the drought stress, it had no statistical effect on IPU mineralization. However, when it was added before the drought stress, two mineralization phases were observed: (1) one corresponding to the drought stress for which mineralization was low and (2) another one observed after restoration of soil moisture content characterized by higher mineralization rate. It can be concluded that climatic fluctuations affect the activity of IPU mineralizing microbial community, and may lead to an increase in IPU persistence.     

Effects of carbon source amendment on the anaerobic degradation of 1,1,1-trichloroethane (TCA) in a contaminated aquifer

We conducted a 10-month anaerobic microcosm test and a 15-month field test to investigate the effects of carbon source addition on the in-situ 1,1,1-trichloroethane (TCA) degradation in a polluted aquifer at an industrial site near Antwerp, Belgium. In the microcosms TCA decreased from 15000 μ g/L to 1500 μ g/L. 1,1-Dichloroethane (DCA) end-concentrations were about the same as the concentrations at the start of the tests (1500 μ g/L). 1,1-Dichloroethene (DCE) decreased from 1200 μ g/L to 800 μ g/L. Carbon source unamended and amended microcosms showed equal concentration trends. Neither chloroethane (CA) nor vinyl chloride (VC) were produced in the microcosms. In the field test TCA dropped from 15000 μ g/L to below 2000 μ g/L. Upstream, TCA levels remained elevated proving that the C-source injection had stimulated degradation. Sulphate reducing bacteria were stimulated; FeS was produced in the microcosms and aquifer. Dehalococcoides species were stimulated in some microcosms and in the HRC-amended aquifer. Neither sulphate reducers nor Dehalococcoidesspecies are however believed to be responsible for the observed TCA degradation. The carbon source injection however did yield environmental conditions that increased TCA degradation.     

Harsh summer conditions caused structural and specific functional changes of microbial communities in an arable soil

The mineralization of the herbicide 3-(4-isopropylphenyl)-1,1-dimethylurea (isoproturon) was reduced after the dry and hot summer 2003 in a soil profile placed in a field lysimeter. A different isoproturon mineralization pattern remained in soil material taken at two different soil depths (0–5 cm and 15–20 cm), although soil material was re-equilibrated at adequate climatic conditions. Special soil microcosms were designed to determine if the changes in this special soil function 'isoproturon mineralization' were related to the climatic scenario of summer 2003. These microcosms were filled with lysimeter soil from the 15–20 cm depth and the temperature and dryness of summer 2003 were simulated. Afterwards, soil samples were taken from the microcosms and re-equilibrated under controlled conditions for 4 weeks. Subsequently, isoproturon mineralization was investigated. The soil microbial community reduced drastically its original capability of isoproturon mineralization in the course of the model experiments.
Analysis of 16S-rRNA by denaturing gel gradient electrophoresis (DGGE) revealed substantial differences in the band patterns of the bacterial communities from both depths of the field lysimeter soil and from the soil incubated in microcosms. The different soil microbial biomass determined by microcalorimetry reinforced these results. In conclusion, the factors higher temperature and smaller soil moisture content generated important and enduring changes in the microbial community structure and therefore in specific soil functions of the community, as shown here by the function of isoproturon degradation. Results are discussed in connection with environmental conditions and conservation tillage.     

Bioremedial potential of fenamiphos and chlorpyrifos degrading isolates: Influence of different environmental conditions

Previously isolated bacterial strains for chlorpyrifos and fenamiphos degradation were used to examine their potential as bioremedial agents in soils and water containing pesticide residues. Both, chlorpyrifos-degrading Enterobacter sp and fenamiphos-degrading consortium rapidly degraded pesticides when inoculated into natural and sterile water and soils. Degradation rate was slower in lower pH soils in comparison with natural and alkaline soils. Soil organic matter had no impact on pesticide degrading ability of isolates. Soil moisture <40% of maximum water-holding capacity slowed down degradation rate. The bacterial isolates were able to rapidly degrade fenamiphos and chlorpyrifos between 15 and 35 °C but their degradation ability was sharply reduced at 5 and 50 °C. Both groups of bacterial systems were also able to remove a range of pesticide degradation. An inoculum density of 10⁴ cells g⁻¹ of soil was required for initiating rapid growth and degradation. Ageing of pesticide in soils prior to inoculation produced contrasting results. Ageing of fenamiphos had no impact on subsequent degradation by the inoculated consortium. However, degradation of chlorpyrifos by Enterobacter sp after aging resulted in persistence of ∼10% of pesticide in soil matrix. Higher K_oc value of chlorpyrifos may have resulted in a lack of bioavailability of a smaller percentage of chlorpyrifos to degrading bacteria. Overall, this paper confirms bioremedial potential of a fenamiphos degrading consortium and a chlorpyrifos degrading bacterium under different soil and water characteristics.     

Degradation of alachlor in natural and sludge-amended soils, studied by gas and liquid chromatography coupled to mass spectrometry (GC-MS and HPLC-MS)

Alachlor [2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl)acetamide] is an herbicide used worldwide. The relative rates of disappearance of alachlor, the formation kinetics of alachlor ethane sulfonic acid (ESA), and the formation of other degradation products in two different soils (a soil with natural organic matter and a sludge-amended soil) has been studied. For such a purpose, soil samples were spiked with alachlor at 2.5 mg kg(-1), concentration generally applied in agricultural soils, and were submitted to sunlight, simulating natural field conditions. Extracts were analyzed by GC-MS and HPLC-MS in scan mode. A good correlation was observed between both techniques, and HPLC-MS allowed the determination of two eluting peaks corresponding to the two stereoisomeric forms of alachlor ESA. Degradation of alachlor in the two soils followed first-order kinetics. Half-life in the natural soil was 4.2 +/- 0.1 days, and half-life in the sludge-amended soil was 5.8 +/- 0.8 days. The higher half-life observed in the sludge-amended soil was attributed to the higher sorption of alachlor to this soil compared to the natural soil. The degradation of alachlor in both soils gave rise to the production of alachlor ESA. Its concentration increased during the incubation period, and after 27 days, its concentration was about 0.59 mg kg(-1) in the natural soil and 0.37 mg kg(-1) in the sludge-amended soil. The other two alachlor transformation products were identified using GC-MS, and the abundance of these degradation products increased while alachlor was degraded.