Acceleration of Anthraquinone-Type Dye Removal by White-Rot Fungus Under Optimized Environmental Conditions

The decolorization of the recalcitrant dye Remazol Brilliant Blue R (RBBR) by the culture filtrate of Polyporus sp. S133 and the effect of various environmental factors were investigated. Both biodegradation and biosorption were playing an important role in bioremoval mechanisms. The highest biosorption of RBBR in Polyporus sp. S133 was shown by all carbon sources such as sucrose, glucose, fructose, and starch. No biosorption was shown by the addition of aromatic compounds and metal ions; 97.1?% RBBR decolorization was achieved in 120-rpm culture for 96?h, as compared to 49.5?% decolorization in stationary culture. Increasing the shaking rotation of the culture to more than 120?rpm was proven to give a negative effect on decolorization. The highest production of laccase was shown at pH 4 and constantly decreases when the pH level increases. The addition of glucose, ammonium tartrate, Cu²⁺, and protocatechuic acid was the suitable environmental condition for RBBR decolorization. There was a positive relationship between all environmental conditions and laccase production in the decolorization of RBBR.     

Breakdown Products in the Metabolic Pathway of Anthracene Degradation by a Ligninolytic Fungus Polyporus sp. S133

Polyporus sp. S133 fungi were selected based on their ability to degrade anthracene in liquid media. The degradation efficiency of anthracene increased by adding 0.5% Tween 80 to reach 71%; agitation at 120 rev/min increased the degradation to 92% after 30?days of incubation. Enzymes such as manganese peroxidase (MnP), lignin peroxidase (LiP), laccase, 1,2-dioxygenase and 2,3-dioxgenase were produced by Polyporus sp. S133 during incubation, and the highest enzyme activity was 182.3 U l^?1 by 1,2-dioxygenase after 20?days of incubation. These results indicate that ligninolytic and dioxygenase enzymes secreted from Polyporus sp. S133 could play an important role in anthracene degradation efficiency. The metabolites detected through the degradation pathway were anthraquinone, phthalic acid, benzoic acid and catechol.     

Decolorization of Azo,Triphenylmethane and Anthraquinone Dyes by Laccase of a Newly Isolated <Emphasis Type="Italic">Armillaria</Emphasis> sp. F022

A newly isolated white-rot fungus, Armillaria sp. strain F022, was isolated from the decayed wood in a tropical rain forest. Strain F022 was capable of decolorizing a variety of synthetic dyes, including azo, triphenylmethane, and anthraquinone dyes, with an optimal efficiency of decolorization obtained when dyes added after 96 h of culture, with the exception of Brilliant Green. All of the tested dyes were decolorized by the purified laccase in the absence of any redox mediators, but only a few were completely removed, while others were not completely removed even when decolorization time was increased. The laccase, with possible contributions from unknown enzymes, played a role in the decolorization process carried out by Armillaria sp. F022 cultures, and this biosorption contributed a negligible part to the decolorization by cultures. The effect of dye to fungal growth was also investigated. When dyes were added at 0 h of culture, the maximum dry mycelium weight (DMW) values in the medium containing Brilliant Green were 1/6 of that achieved by the control group. For other dyes, the DMW was similar with control. The toxic tolerance of dye for the cell beads was excellent at least up to a concentration of 500 mg/l. The optimum conditions for decolorization of three synthetic dyes are at pH 4 and 40°C.     

Potential of Gonium spp. in Synthetic Reactive Dye Removal, Possible Role of Laccases and Stimulation by Triacontanol Hormone

In this study, Gonium sp. was investigated for possible usage in dye-containing wastewater treatment. Trials were performed in media including triacontanol hormone, Reactive Orange 14, Reactive Red 120, Reactive Black 5, Remazol Brilliant Blue R (RBBR), and also hormone against the controls. Algae could remove RBBR with the highest dye removal percentage (56%) among the tested dyes. The optimum pH was 9 in removing 50 mg L^?1 RBBR at a dye removal percentage of 47.1%. The role of laccase activity of Gonium sp. was also investigated. This first attempt in the literature showed the involvement of the enzyme in the algal growth and bioremoval process. In the presence of the plant growth hormone in the culture, the activity showed a steady and significant increase up to nearly sixfold between 5th and 14th days of incubation.     

The Production of Ligninolytic Enzymes by Marine-Derived Basidiomycetes and Their Biotechnological Potential in the Biodegradation of Recalcitrant Pollutants and the Treatment of Textile Effluents

Filamentous fungi derived from marine environments are well known as a potential genetic resource for various biotechnological applications. Although terrestrial fungi have been reported to be highly efficient in the remediation of xenobiotic pollutants, fungi isolated from the marine environment may possess biological advantages over terrestrial fungi because of their adaptations to high salinity and pH extremes. The present study describes the production of ligninolytic enzymes under saline and non-saline conditions and the decolorization of Remazol Brilliant Blue R (RBBR) dye by three basidiomycetes recovered from marine sponges (Tinctoporellus sp. CBMAI 1061, Marasmiellus sp. CBMAI 1062, and Peniophora sp. CBMAI 1063). Ligninolytic enzymes were primarily produced by these fungi in a salt-free malt extract and malt extract formulated with artificial seawater (saline condition). CuSO₄ and wheat bran were the best inducers of lignin peroxidase and manganese peroxidase activity. RBBR was decolorized up to 100% by the three fungi, and Tinctoporellus sp. CBMAI 1061 was the most efficient. Our results revealed the biotechnological potential of marine-derived basidiomycetes for dye decolorization and the treatment of colored effluent as well as for the degradation of other organopollutants by ligninolytic enzymes in non-saline and saline conditions that resemble the marine environment.     

Salt-tolerant rhizobacteria-mediated induced tolerance in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>) and chemical diversity in rhizosphere enhance plant growth

Salt-tolerant isolates Bacillus pumilus, Pseudomonas mendocina, Arthrobacter sp., Halomonas sp., and Nitrinicola lacisaponensis isolated from high saline habitats exhibited plant growth-promoting traits like P solubilization and indole acetic acid (IAA), siderophore, and ammonia production. These isolates were inoculated in wheat to assess microbe-mediated responses and plant growth promotion in salt affected soil. Maximum shoot and root length (33.8 and 13.6 cm) and shoot and root biomass (2.73 and 4.48 g dry weight) was recorded in plants inoculated with B. pumilus after 30 days. Total chlorophyll content was maximum in the leaves of the plants treated with Halomonas sp. (24.22 mg g⁻¹ dry weight) followed by B. pumilus (23.41 mg g⁻¹ dry weight) as compared to control (18.21 mg g⁻¹ dry weight) after 30 days. Total protein content was maximum in Arthrobacter sp. inoculated plant leaves (3.19 mg g⁻¹ dry weight) followed by B. pumilus (2.47 mg g⁻¹ dry weight) as compared to control (2.15 mg g⁻¹ dry weight) after 30 days. Total carotenoid content was maximum in plants inoculated with Halomonas sp. (1,075.45 and 1,113.29 μg g⁻¹ dry weight) in comparison to control (837.32 and 885.85 μg g⁻¹ dry weight) after 15 and 30 days. Inoculation of bacterial isolates increased presence of individual phenolics (gallic, caffeic, syringic, vanillic, ferulic, and cinnamic acids) and flavonoid quercetin in the rhizosphere soil. The concentration of IAA in rhizosphere soil and root exudates was also higher in all treatments than in control. Accumulation of phenolics and quercetin in the plants played a cumulative synergistic role that supported enhanced plant growth promotion of wheat in the stressed soil.     

Control of cotton <Emphasis Type="Italic">Verticillium</Emphasis> wilt and fungal diversity of rhizosphere soils by bio-organic fertilizer

Cotton Verticillium wilt is a destructive soil-borne disease affecting cotton production. In this study, application of bio-organic fertilizer (BIO) at the beginning of nursery growth and/or at the beginning of transplanting was evaluated for its ability to control Verticillium dahliae Kleb. The most efficient control of cotton Verticillium wilt was achieved when the nursery application of BIO was combined with a second application in transplanted soil, resulting in a wilt disease incidence of only 4.4%, compared with 90.0% in the control. Denaturing gradient gel electrophoresis patterns showed that the consecutive applications of BIO at nursery and transplanting stage resulted in the presence of a unique group of fungi not found in any other treatments. Humicola sp., Metarhizium anisopliae, and Chaetomium sp., which were considered to be beneficial fungi, were found in the BIO treatment, whereas some harmful fungi, such as Alternaria alternate, Coniochaeta velutina, and Chaetothyriales sp. were detected in the control. After the consecutive applications of BIO at nursery and transplanting stage, the V. dahliae population in the rhizosphere soil in the budding period, flowering and boll-forming stage, boll-opening stage, and at harvest time were 8.5 × 10², 3.1 × 10², 4.6 × 10², and 1.7 × 10² colony-forming units per gram of soil (cfu g⁻¹), respectively, which were significantly lower than in the control (6.1 × 10³, 3.4 × 10³, 5.2 × 10³, and 7.0 × 10³ cfu g⁻¹, respectively). These results indicate that the suggested application mode of BIO could effectively control cotton Verticillium wilt by significantly changing the fungal community structure and reducing the V. dahliae population in the rhizosphere soil.     

Use of Ozonization for the Treatment of Dye Wastewaters Containing Rhodamine B in the Agate Industry

The industrial processing of precious stones is a source of revenue for several Brazilian towns, especially in the state of Rio Grande do Sul. Given the growing number of small-sized companies that process precious stones, wastewater production is inevitable and is a cause for concern inasmuch as preservation of nature is considered. The present study investigates the detoxification of the wastewater produced by the process of rhodamine B dyeing using oxidation processes. Ozonization (O₃), ultraviolet irradiation (UV), and O₃/UV methods were assessed. Some of the parameters used to measure the efficiency of the analyzed treatments included COD, ecotoxicity (Daphnia magna), cytotoxicity, and genotoxicity assays (Allium cepa assays). Results show predominance of negative and local environmental impacts, which are reversible in more than 70% of cases. The major proposed reversibility measures were the change in the process layout and dye wastewater segregation. Among the analyzed methods, ozonization proved to be more efficient in decolorization, with 60 min of treatment, pH = 9 and dosage of 5.705 mg O₃/mg of rhodamine B. A pseudo first-order reaction, with a kinetic constant of 7.5 × 10⁻² min⁻¹, was observed. The cytotoxic and genotoxic effects were assessed for both raw and treated wastewaters. Despite complete decolorization, cytotoxicity and genotoxicity assays revealed an EC₅₀ of 28.6, in addition to chromosome aberrations in 40% of dividing cells for the treated wastewater.     

Ligninolytic fungi in bioremediation: extracellular enzyme production and degradation rate

Ligninolytic fungi can be used for remediation of pollutants in water and soil. Extracellular peroxidases and laccases have been shown to oxidize recalcitrant compounds in vitro but the likely significance of individual enzyme levels in vivo remains unclear. This study documents the amounts and activities of Mn-dependent peroxidase (MnP), lignin peroxidase and laccase (LAC) in various species of ligninolytic fungi grown in liquid medium and soil and their effect on degradation of polycyclic aromatic hydrocarbons (anthracene and pyrene), a polychlorinated biphenyl mixture (Delor 106) and a number of synthetic dyes. Stationary cultures of a highly degradative strain Irpex lacteus exhibited 380-fold and 2-fold increase in production of MnP and LAC, respectively, compared to submerged cultures. Addition of Tween 80 to the submerged culture increased MnP levels 260-fold. High levels of MnP correlated with efficient decolorization of Reactive Orange 16 azo dye but not of Remazol Brilliant Blue R anthraquinone dye. Degradation of anthracene and pyrene in spiked soil by straw-grown explorative mycelium of Phanerochaete chrysosporium, Trametes versicolor and Pleurotus ostreatus showed the importance of MnP and LAC levels secreted into the soil. The importance of high fungal enzyme levels for efficient degradation of recalcitrant compounds was better demonstrated in liquid media compared to the same strains growing in soil.     

Development of two culture media for mass cultivation of <Emphasis Type="Italic">Azospirillum</Emphasis> spp. and for production of inoculants to enhance plant growth

High yield culture medium is fundamental for production of inoculants for plant growth-promoting bacteria. Based on substitution of glucose in tryptone–yeast extract–glucose medium by Na-gluconate or glycerol, two new culture media were developed for mass cultivation of the commonly used plant growth-promoting bacterium Azospirillum sp. After 18 h of incubation, these modifications increased populations of different strains of Azospirillum (to ∼10¹¹ cells ml⁻¹ [single cell count] and ∼5 × 10⁹ CFU ml⁻¹ [plate count method]), significantly reduced generation time, and were also suitable for production of common synthetic inoculants.     

<Emphasis Type="Italic">Trichoderma</Emphasis> inoculation and trash management effects on soil microbial biomass,soil respiration,nutrient uptake and yield of ratoon sugarcane under subtropical conditions

A field experiment was conducted during 2003–2005 and 2004–2006 at the Indian Institute of Sugarcane Research, Lucknow, India to study the effect of Trichoderma viride inoculation in ratoon sugarcane with three trash management practices, i.e. trash mulching, trash burning and trash removal. Trichoderma inoculation with trash mulch increased soil organic carbon and phosphorus (P) content by 5.08 Mg ha⁻¹ and 11.7 kg ha⁻¹ over their initial contents of 15.75 Mg ha⁻¹ and 12.5 kg ha⁻¹, respectively. Soil compaction evaluated as bulk density in 0- to 15-cm soil layer, increased from 1.48 Mg m⁻³ at ratoon initiation (in April) to 1.53 Mg m⁻³ at harvest (in December) due to trash burning and from 1.42 Mg m⁻³ at ratoon initiation (in April) to 1.48 Mg m⁻³ at harvest (in December) due to trash mulching. The soil basal respiration was the highest during tillering phase and then decreased gradually, thereafter with the advancement of crop growth. On an average, at all the stages of crop growth, Trichoderma inoculation increased the soil basal respiration over no inoculation. Soil microbial biomass increased in all plots except in the plots of trash burning/removal without Trichoderma inoculation. The maximum increase (40 mg C kg⁻¹ soil) in soil microbial biomass C, however, was observed in the plots of trash mulch with Trichoderma inoculation treatment which also recorded the highest uptake of nutrient and cane yield. On an average, Trichoderma inoculation with trash mulch increased N, P and K uptake by 15.9, 4.68 and 23.6 kg ha⁻¹, respectively, over uninoculated condition. The cane yield was increased by 12.8 Mg ha⁻¹ with trash mulch + Trichoderma over trash removal without Trichoderma. Upon degradation, trash mulch served as a source of energy for enhanced multiplication of soil bacteria and fungi and provided suitable niche for plant–microbe interaction.     

Phytoremediation of Mixed Soil Contaminants

Tests were conducted to study the influence of non-ionic surfactants Triton X-100 and Tween 80 on the removal of mixed contaminants from a sandy soil using phytoremediation. Cd(II) and Pb(II) were used to form the inorganic contaminant, while used engine oil was selected to form the organic contaminant. The Indian mustard (Brassica juncea) plant was the plant chosen for phytoremediation of the sandy soil that contained the mixed contaminant. Thirty days after the plants were grown in the greenhouse, surfactants were applied to test pots in which the soil had been spiked with 50 mg kg⁻¹ of CdCl₂, 500 mg kg⁻¹ of PbCl₂ and 500 mg kg⁻¹ of used engine oil. Two control tests were conducted in this study. Planted and unplanted control tests were conducted using soil without surfactants. Following these tests, the tests were completed using the plants and surfactants at different concentrations. Test results showed that Triton X-100 and Tween 80 at concentrations higher than their critical micellar concentration enhanced Cd(II) and Pb(II) accumulation in the plant roots. Further, test data showed that translocation of contaminants to plant shoots occurred for Cd(II) but not for Pb(II). At the same concentrations, Tween 80 was more effective than Triton X-100 in facilitating rhizodegradation of used engine oil. This study demonstrates that simultaneous phytoremediation of Pb(II), Cd(II) and oil can be enhanced by using non-ionic surfactant Tween 80. Leaching test results indicated that the enhanced phytoremediation could remove the mixed contaminants safely from the point of view of limiting groundwater contamination.     

Promotion of mycorrhiza formation and growth of willows by the bacterial strain <Emphasis Type="Italic">Sphingomonas</Emphasis> sp. 23L on fly ash

Re-vegetation of fly ash, the principal by-product of coal fired power stations, is hampered by its unfavourable chemical and physical properties for plant growth. In the present study, we evaluated the use of inoculation with a mycorrhiza-associated bacterial strain (Sphingomonas sp. 23L) to promote mycorrhiza formation and plant growth of three willow clones (Salix spp.) on fly ash from an over-burdened dump in a pot experiment. The high pH_H2O (8.7) and low nitrogen content (N_t = 0.1 g kg⁻¹) in combination with hydrophobicity of the particle surfaces caused low plant growth. Inoculation of the willows with Sphingomonas sp. 23L improved the nitrogen uptake by plants, increased plant growth and stimulated formation of ectomycorrhizae with an autochthonous Geopora sp. strain on all three willow clones. The ectomycorrhiza formed by the Geopora sp. was morphologically and anatomically described. The inoculation significantly increased the shoot growth of two Salix viminalis clones and the root growth of a S. viminalis x caprea hybrid clone. We conclude that inoculation with mycorrhiza promoting bacterial strains might be a suitable approach to support mycorrhiza formation with autochtonous site-adapted ectomycorrhizal fungi in fly ash and thereby to improve re-vegetation of fly ash landfills with willows.     

Induced Degradation of Anthraquinone-Based Dye by Laccase Produced from <Emphasis Type="Italic">Pycnoporus sanguineus</Emphasis> (CS43)

In this study, in-house isolated laccase isoforms, i.e., Lac-I and Lac-II of the basidiomycete Pycnoporus sanguineus (CS43), were evaluated in relation to their Remazol Brilliant Blue R (RBBR) dye degradation capacity. A modified Dhouib medium additionally supplemented with 3% ethanol as a secondary inducer was used to propagate P. sanguineus CS43 for enhanced production of laccase under liquid state fermentation. The crude laccase extract was purified by passing through ion exchange diethylaminoethanol (DEAE)-Sepharose and gel filtration-based Sephadex G-200 column chromatography. The purified laccase fractions were subjected to the electrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed two laccase isoforms Lac-I and Lac-II with 66 and 68 kDa, respectively. To explore the industrial applicability, for RBBR dye, degradation efficiencies ranged from 82 to 88% after 3 h of incubation for both; Lac-I and Lac-II at both concentrations were recorded. However, with 8 U/mL, the degradation ranged between 70 to 80% during the first 5 min of incubation. Enhanced degradation of RBBR dye was obtained in the presence of violuric acid and N-hydroxypthalamide as laccase mediators. Finally, using RBBR as a substrate kinetic characterization of both Lac-I and Lac-II isoforms was performed that revealed K _m (0.243 and 0.117 mM for Lac-I and Lac-II) and V _max (1.233 and 1.012 mM/Sec for Lac-I and Lac-II) values, respectively.     

Correlation of Ligninolytic Enzymes from the Newly-Found Species Trametes versicolor U97 with RBBR Decolorization and DDT Degradation

Thirty strains of fungi collected from nature were investigated for their ability to grow on agar medium contaminated with Remazol Brilliant Blue R (RBBR) and 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT). The results showed that strain U97, later identified as Trametes versicolor, was the most active decomposer. This fungus decolorized 85?% of RBBR in 6?h and degraded 71?% of DDT in 30?days. In RBBR decolorization, high-performance liquid chromatography analysis revealed that two peaks were identified as metabolic products. Among inducers for ligninolytic enzymes, only veratryl alcohol improved RBBR decolorization and DDT degradation by 93?% and 77?%, respectively. A partial least squares method using Minitab 15 showed that lignin peroxidase exhibited a positive correlation to the abilities of T. versicolor U97 to decolorize RBBR and degrade DDT. A multivariate linear equation, with the same values of ligninolytic activity during RBBR decolorization and DDT degradation, revealed that 1?% RBBR decolorization represented 1.16?% DDT degradation. Screening with agar or liquid medium and improvement of the mathematical modeling could have practical importance in the exploitation of T. versicolor U97 for the removal of DDT on a commercial scale.     

Fungal degradation of chlorpyrifos by <Emphasis Type="Italic">Acremonium</Emphasis> sp. strain (GFRC-1) isolated from a laboratory-enriched red agricultural soil

The organophosphorus insecticide, chlorpyrifos, has been widely applied in agriculture; in veterinary, against household pests; and in subterranean termite control. Due to its slow rate of degradation in soil, it can persist for extended periods in soil with a significant threat to environment and public health. The mixed and pure fungi were isolated from three soils by enrichment technique. The enriched mixed fungal cultures were capable of biodegrading chlorpyrifos (300 mg L⁻¹) when cultivated in Czapek Dox medium. The identified pure fungal strain, Acremonium sp., utilized chlorpyrifos as a source of carbon and nitrogen. The highest chlorpyrifos degradation (83.9%) by Acremonium sp. strain GFRC-1 was found when cultivated in the nutrient medium with full nutrients. Desdiethyl chlorpyrifos was detected as a major biodegradation product of chlorpyrifos. The isolated fungal strain will be used for developing bioremediation strategy for chlorpyrifos-polluted soils.     

Perchlorate Depositional History as Recorded in North American Ice Cores from the Eclipse Icefield, Canada, and the Upper Fremont Glacier, USA

Temporal depositional rates are important in order to understand the production and occurrence of perchlorate (ClO₄⁻) as limited information exists regarding the impact of anthropogenic production or atmospheric pollution on ClO₄⁻ deposition. Perchlorate concentrations in discrete ice core samples from the Eclipse Icefield (Yukon Territory, Canada) and Upper Fremont Glacier (Wyoming, USA) were analyzed using ion chromatography tandem mass spectrometry to evaluate temporal changes in the deposition of ClO₄ ⁻ in North America. The ice core samples cover a time period from 1726 to 1993 and 1970 to 2002 for the Upper Fremont Glacier (UFG) and Eclipse ice cores, respectively. The average ClO₄ ⁻ concentration in the Eclipse ice core for the time period from 1970 to 1973 was 0.6 ± 0.3 ng L⁻¹, with higher values of 2.3 ± 1.7 and 2.2 ± 2.0 ng L⁻¹ for the periods 1982–1986 and 1999–2002, respectively. All pre-1980 ice core samples from the UFG had ClO₄ ⁻ concentrations <0.2 ng L⁻¹, and the post-1980 samples ranged from <0.2 ng L⁻¹ to a maximum of 2.6 ng L⁻¹ for the year 1992. A significant positive correlation (R = 0.75, N = 15, p < 0.001) of ClO₄⁻ with SO₄²⁻ was found for the annual UFG ice core layers and of ClO₄ ⁻ with SO₄²⁻ and NO₃⁻ in sub-annual Eclipse ice samples (R > 0.3, N = 121, p < 0.002). The estimated yearly ClO₄⁻ depositional flux for the Eclipse ice core ranged from 0.6 (1970) to 4.7 μg m⁻² year⁻¹ (1982) and the UFG from <0.1 (pre-1980) to 1.4 μg m⁻² year⁻¹ (1992). There was no consistent seasonal variation in the ClO₄⁻ depositional flux for the Eclipse ice core, in contrast to a previous study on the Arctic region. The presence of ClO₄⁻ in these ice cores might correspond to an intermittent source such as volcanic eruptions and/or any anthropogenic forcing that may directly or indirectly aid in atmospheric ClO₄⁻ formation.     

Crop residues and fertilizer nitrogen influence residue decomposition and nitrous oxide emission from a Vertisol

Crop residues with high C/N ratio immobilize N released during decomposition in soil, thus reducing N losses through leaching, denitrification, and nitrous oxide (N₂O) emission. A laboratory incubation experiment was conducted for 84 days under controlled conditions (24°C and moisture content 55% of water-holding capacity) to study the influence of sugarcane, maize, sorghum, cotton and lucerne residues, and mineral N addition, on N mineralization–immobilization and N₂O emission. Residues were added at the rate of 3 t C ha⁻¹ to soil with, and without, 150 kg urea N ha⁻¹. The addition of sugarcane, maize, and sorghum residues without N fertilizer resulted in a significant immobilization of soil N. Amended soil had significantly (P < 0.05) lower NO₃⁻–N, which reached minimum values of 2.8 mg N kg⁻¹ for sugarcane (at day 28), 10.3 mg N kg⁻¹ for maize (day 7), and 5.9 mg N kg⁻¹ for sorghum (day 7), compared to 22.7 mg N kg⁻¹ for the unamended soil (day 7). During 84 days of incubation, the total mineral N in the residues + N treatments were decreased by 45 mg N kg⁻¹ in sugarcane, 34 mg kg⁻¹ in maize, 29 mg kg⁻¹ in sorghum, and 16 mg kg⁻¹ in cotton amended soil compared to soil + N fertilizer, although soil NO₃⁻–N increased by 7 mg kg⁻¹ in lucerne amended soil. The addition of residues also significantly increased amended soil microbial biomass C and N. Maximum emissions of N₂O from crop residue amended soils occurred in the first 4–5 days of incubation. Overall, after 84 days of incubation, the cumulative N₂O emission was 25% lower with cotton + N fertilizer, compared to soil + N fertilizer. The cumulative N₂O emission was significantly and positively correlated with NO₃⁻–N (r = 0.92, P < 0.01) and total mineral N (r = 0.93, P < 0.01) after 84 days of incubation, and had a weak but significant positive correlation with cumulative CO₂ in the first 3 and 5 days of incubation (r = 0.59, P < 0.05).     

Effects of Dissolved Water Constituents on the Photodegradation of Fenitrothion and Diazinon

The photochemical degradation of two widely used organophosphorothioate insecticides, fenitrothion and diazinon, was investigated in aqueous solutions containing three separate dissolved constituents commonly found in natural waters (NO₃⁻, CO₃²⁻ and dissolved organic matter (DOC)). The effect of these constituents on pesticide photodegradation was compared to degradation in “constituent-free” pure water. Solutions were irradiated in an Atlas solar simulator fitted with a UV-filtered Xenon arc lamp with light irradiances (500 W m⁻²) measured using a spectral radiometer to allow derivation of quantum yields of degradation. Fenitrothion absorbs light within the solar UV range (λ, 295–400 nm) and underwent direct photolysis in pure water whereas diazinon (λ _max ∼250 nm) showed no observable loss over the experimental period. However, photodegradation conforming to pseudo-first-order kinetics was observed for both chemicals in the presence of the dissolved constituents (at concentrations typically observed in natural waters), with the rates of photodecay observed in the order of NO₃⁻ > CO₃²⁻ ≅ DOC, with the highest rates observed in the 3 mM NO₃⁻ solutions (k _Fen = 0.155 ± 0.041 h⁻¹; k _Dia = 0.084 ± 0.0007 h⁻¹). For diazinon this rate was comparable to fenitrothion photolysis in pure water (k _fen 0.072 ± 0.0078 h⁻¹), highlighting the importance of NO₃⁻ on a non-photolabile pesticide, with indirect photodegradation probably attributable to the light-induced release of aqueous hydroxyl radicals (^·OH) from NO₃⁻. Suwannee river fulvic acid (serving as DOC) did not statistically affect the rate of photodecay for fenitrothion relative to its photolysis in MilliQ water, although measured rates in DOC solutions were slightly lower. However, measurable rates of photodecay were apparent for diazinon in the DOC solutions, indicating that fulvic acid, possibly in the form of “excited” triplet-state-DOC plays a role in diazinon transformation. Hydrolysis was not apparent for fenitrothion (in buffered solutions of pH 5–9) but was notable for diazinon at the lower pHs of 5 and 3 (k _Dia-hyd 0.3414 h⁻¹ at pH 3 and 0.228 h⁻¹ at pH 5), resulting in the formation of the degradate, 2-isopropyl–6-methyl–4-pyrimidinol. This work highlights the importance of dissolved constituents on abiotic photodegradation of pesticides and it is recommended that these constituents be incorporated into laboratory-based fate-testing regimes.     

Effects of long-term mineral fertilization on microbial biomass,microbial activity,and the presence of <Emphasis Type="Italic">r</Emphasis>- and <Emphasis Type="Italic">K</Emphasis>-strategists in soil

Long-term effects of mineral fertilization on microbial biomass C (MBC), basal respiration (R _B), substrate-induced respiration (R _S), β-glucosidase activity, and the r–K-growth strategy of soil microflora were investigated using a field trial on grassland established in 1969. The experimental plots were fertilized at three rates of mineral N (0, 80, and 160 kg ha⁻¹ year⁻¹) with 32 kg P ha⁻¹ year⁻¹ and 100 kg K ha⁻¹ year⁻¹. No fertilizer was applied on the control plots (C). The application of a mineral fertilizer led to lower values of the MBC and R _B, probably as a result of fast mineralization of available substrate after an input of the mineral fertilizer. The application of mineral N decreased the content of C extracted by 0.5 M K₂SO₄ (C _ex). A positive correlation was found between pH and the proportion of active microflora (R _S/MBC). The specific growth rate (μ) of soil heterotrophs was higher in the fertilized than in unfertilized soils, suggesting the stimulation of r-strategists, probably as the result of the presence of available P and rhizodepositions. The cessation of fertilization with 320 kg N ha⁻¹ year⁻¹ (NF) in 1989 also stimulated r-strategists compared to C soil, probably as the result of the higher content of available P in the NF soil than in the C soil.