The influence of glucose on simazine decomposition.

Glucose added to soil and to the culture medium of Penicillium citrinum stimulated simazine decomposition. Differences in the rate of simazine decomposition depended on the dose of glucose. Both in soil and in the medium the rate of simazine decomposition was connected with the intensity of development of microorganisms.     

Effect of Herbicides on the Toxicity of Fungicides Against Rhizoctonia solani Causing Damping-off of Cotton

The effect of two herbicides (paraquat and simazine on the antifungal activity of two fungicides (captan and mounsrin) against Rhizoctonia solani was studied. when the herbicides paraquat and simazine were applied to soil they altered the effectiveness of both fungicides in controlling R. solani , thus causing damping-off of cotton. Both herbicides increased the toxicity of both fungicides against mycelial growth of the fungus. In pot tests, seeds or soil treated with captan or mounsrin, gave better control of R. solani damping-off disease when the soil was treated with paraquat or simazine compared to untreated soil. Captan was, however, found to be more effective in controlling the disease than mounsrin.     

Simazine application inhibits nitrification and changes the ammonia-oxidizing bacterial communities in a fertilized agricultural soil

s-Triazine herbicides are widely used for weed control, and are persistent in soils. Nitrification is an essential process in the global nitrogen cycle in soil, and involves ammonia-oxidizing Bacteria (AOB) and ammonia-oxidizing Archaea (AOA). In this study, we evaluated the effect of the s-triazine herbicide simazine on the nitrification and on the structure of ammonia-oxidizing microbial communities in a fertilized agricultural soil. The effect of simazine on AOB and AOA were studied by PCR-amplification of amoA genes of nitrifying Bacteria and Archaea in soil microcosms and denaturing gradient gel electrophoresis (DGGE) analyses. Simazine [50?μg g(-1) dry weight soil (d.w.s)] completely inhibited the nitrification processes in the fertilized agricultural soil. The inhibition by simazine of ammonia oxidation observed was similar to the reduction of ammonia oxidation by the nitrification inhibitor acetylene. The application of simazine-affected AOB community DGGE patterns in the agricultural soil amended with ammonium, whereas no significant changes in the AOA community were observed. The DGGE analyses strongly suggest that simazine inhibited Nitrosobacteria and specifically Nitrosospira species. In conclusion, our results suggest that the s-triazine herbicide not only inhibits the target susceptible plants but also inhibits the ammonia oxidation and the AOB in fertilized soils.     

Bioaugmentation with Pseudomonas sp. strain MHP41 promotes simazine attenuation and bacterial community changes in agricultural soils

Bioremediation is an important technology for the removal of persistent organic pollutants from the environment. Bioaugmentation with the encapsulated Pseudomonas sp. strain MHP41 of agricultural soils contaminated with the herbicide simazine was studied. The experiments were performed in microcosm trials using two soils: soil that had never been previously exposed to s -triazines (NS) and soil that had >20 years of s -triazine application (AS). The efficiency of the bioremediation process was assessed by monitoring simazine removal by HPLC. The simazine-degrading microbiota was estimated using an indicator for respiration combined with most-probable-number enumeration. The soil bacterial community structures and the effect of bioaugmentation on these communities were determined using 16S RNA gene clone libraries and FISH analysis. Bioaugmentation with MHP41 cells enhanced simazine degradation and increased the number of simazine-degrading microorganisms in the two soils. In highly contaminated NS soil, bioaugmentation with strain MHP41 was essential for simazine removal. Comparative analysis of 16S rRNA gene clone libraries from NS and AS soils revealed high bacterial diversity. Bioaugmentation with strain MHP41 promoted soil bacterial community shifts. FISH analysis revealed that bioaugmentation increased the relative abundances of two phylogenetic groups ( Acidobacteria and Planctomycetes ) in both soils. Although members of the Archaea were metabolically active in these soils, their relative abundance was not altered by bioaugmentation.     

Effect of simazine,lindane and ceresan on soil respiration and nitrification rates

Summary The effect of three pesticidesviz, simazine, lindane and ceresan upto 100-fold of field application was studied on organic matter mineralization as well as on nitrification rates in Delhi alluvial soil. The normal rate of simazine (2 ppm) did not influence CO₂ evolution but higher concentrations suppressed it. Lindane and ceresan inhibited CO₂ production from soil by normal concentrations but the same CO₂ production was enhanced in berseem roots treated soil upto 10 ppm of ceresan.The nitrification of ammonium sulphate in soil due to these pesticides was impaired significantly for varying periods. The inhibitory effect of higher concentrations was marked only upto 3 weeks, subsequently the rate of nitrification was restored. Among the nitrifiers, Nitrobacter were more susceptible as compared to Nitrosomonas.     

Combining supercritical fluid extraction of soil herbicides with enzyme immunoassay analysis

Supercritical fluid extraction (SFE) of soil herbicides followed by enzyme immunoassay analysis (EIA) is explained in a step-by-step process. Extracted herbicides, include 2,4-D, simazine, atrazine, and alachlor. The herbicide, trifluralin was not successfully analyzed by EIA because of crossreacting metabolites. Problems with SFE, including uneven packing of cells, leaks, uneven flow and clogging, can largely be eliminated as the method parameters are optimized. It was necessary to add modifiers including methanol or acetone to the SF CO₂ to increase the solubility of the analytes. Detection limits of 2.5 ng/g soil for atrazine and alachlor and 15 ng/g soil for simazine and 2,4-D without concentration of the sample were achieved. Recoveries above 80% and relative standard deviations (RSDs) less than 15% for 2,4-D simazine, atrazine and alachlor were achieved. Atrazine and alachlor recoveries were above 90% with RSDs below 10%. Forty soil samples could be extracted and analyzed in an 8-h day.     

Characterization of Arthrobacter nicotinovorans HIM, an atrazine-degrading bacterium, from agricultural soil New Zealand

Arthrobacter nicotinovorans HIM was isolated directly from an agricultural sandy dune soil 6 months after a single application of atrazine. It grew in minimal medium with atrazine as sole nitrogen source but was unable to mineralize 14C-ring-labelled atrazine. Atrazine was degraded to cyanuric acid. In addition to atrazine the bacterium degraded simazine, terbuthylazine, propazine, cyanazine and prometryn but was unable to grow on terbumeton. When added to soil, A. nicotinovorans HIM did enhance mineralization of 14C-ring-labelled atrazine and simazine, in combination with naturally occurring cyanuric acid degrading microbes resident in the soil. Using PCR, the atrazine-degradation genes atzABC were identified in A. nicotinovorans HIM. Cloning of the atzABC genes revealed significant homology (>99%) with the atrazine degradation genes of Pseudomonas sp. strain ADP. The atrazine degradation genes were held on a 96 kbp plasmid.     

Influence of temperature on phytotoxicity of triazine herbicides to pine seedlings

The effects of temperature, over a range of 5 to 30 C, on phytotoxicity of simazine, atrazine, propazine, prometryne, prometone, and ipazine to young Pinus resinosa seedlings were investigated in growth chambers. Herbicides were applied to the soil surface and then mixed into the soil before pine seeds were planted. Development of recently germinated seedlings was then studied for 7 weeks. High temperatures greatly accelerated herbicide toxicity, but the effects of temperature varied greatly among herbicides. Atrazine and simazine were more toxic than other herbicides tested at all temperatures. Toxicity of simazine and atrazine was apparent early, whereas effects of propazine, prometryne, prometone, and ipazine were somewhat delayed. After 7 weeks maximum dry-weight production of shoots under each herbicide treatment and control occurred at 20 C, with some decreases noted at lower temperatures and marked decreases at progressively higher ones. At 20 C final seedling dry weights following treatment with simazine or atrazine were only one-third as high as in control plants. Growth was also reduced in lesser amounts by propazine, prometryne, prometone, or ipazine. Variations in phytotoxicity of different triazine herbicides appeared to be related more to their structural differences than their solubilities. Under the constant environmental conditions of the experiments, toxicity symptoms in plants treated with triazine herbicides appeared more rapidly and decisively than in previous field experiments under fluctuating environments. The influence of high temperatures in enhancing triazine toxicity appeared to involve complex interactions of physiological activity of plants and temperature effects on herbicide uptake.     

Effect of simazine and atrazine on the mineralization of fertilizer and manure nitrogen

Summary Laboratory experiments were carried out with alluvial sandy loam soil to study the effect of simazine and atrazine herbicides at four levels (0.5, 1.0, 1.5 and 2.0 kg/ha) on the mineralization of nitrogen (ammoniacal and nitrate production) from fertilizer urea and sludge sources. The herbicides stimulated nitrate production. No specific trend in total mineralized nitrogen, ammoniacal and nitrate nitrogen was observed by varying the levels of herbicides. Mineralization of total nitrogen (ammoniacal and nitrate nitrogen) in presence of simazine and atrazine from the different sources in the descending order was:Urea > Sludge + Urea > Sludge > No Nitrogen.     

Herbicide usage and soil properties

Summary Nine annual applications at normal agricultural rates of MCPA, tri-allate, simazine and linuron produced no effect on the quantity of water-stable soil aggregates or the classical fractions of organic matter. re]19751007     

Effect of four common herbicides on the population of microorganisms in Red Sandy soil

Summary A study was conducted to know the effect of four common herbicidesviz, Lasso, Nitrofen, Propinol, and Simazine on soil microorganisms in Red Sandy soil. In general the application of different herbicides did not adversely affect the microbial population. The bacterial population was decreased by the application of propinol and nitrofen. Simazine and lasso stimulated the fungal population. The stimulation in Azotobacter population was observed in lasso, nitrofen and simazine treated soil.     

Analysis of herbicides: demonstration of the utility of enzyme immunoassay verification by HPLC

Evidence has accumulated that herbicides in the environment present a significant health hazard to the population. Therefore, the levels of heavily used substances such as atrazine and simazine and their metabolites need to be regularly assessed. The objective was to develop a rapid and simple tube ELISA procedure suitable for use in field studies and non-specialized laboratories. The antisera used were polyclonal antibodies raised in sheep against atrazine or simazine amido caproic acid conjugated to bovine serum albumin. The antibodies were first used to construct a two-step competitive ELISA procedure in 96-well microtitre plates. The 96-well format was then adapted to a coated-tube enzyme immunoassay, by immobilization of hapten-gelatine conjugates on polystyrene tubes. This enabled the colour to be read using a basic spectrophotometer. Soil samples were collected from agricultural and non-agricultural sites in Poland. Atrazine and simazine were extracted by liquid extraction from soil and assayed by tube ELISA. In addition, the samples were extracted by solid-phase extraction before analysis by HPLC. The immunoassays and chemical analysis were carried out by different individuals who were unaware of each other's results, which were then compared at the end of the study. Correlation of the two methods was excellent, with R=98.7 and 81.3 for atrazine and simazine, respectively. The immunoassay yielded the same order of results without having to perform solid-phase extraction before analysis. The study has demonstrated that the simple antigen-coated tube assay provides a cost-effective and valuable screening test. Comparison with the more elaborate, heavily labour-intensive HPLC analysis demonstrated that the results obtained by the simpler enzyme-immunoassay tests were within the same order.     

Isolation and characterization of a novel simazine-degrading bacterium from agricultural soil of central Chile, Pseudomonas sp. MHP41

s -Triazine herbicides are used extensively in South America in agriculture and forestry. In this study, a bacterium designated as strain MHP41, capable of degrading simazine and atrazine, was isolated from agricultural soil in the Quillota valley, central Chile. Strain MHP41 is able to grow in minimal medium, using simazine as the sole nitrogen source. In this medium, the bacterium exhibited a growth rate of μ=0.10 h⁻¹, yielding a high biomass of 4.2 × 10⁸ CFU mL⁻¹. Resting cells of strain MHP41 degrade more than 80% of simazine within 60 min. The atzA, atzB, atzC, atzD, atzE and atzF genes encoding the enzymes of the simazine upper and lower pathways were detected in strain MHP41. The motile Gram-negative bacterium was identified as a Pseudomonas sp., based on the Biolog microplate system and comparative sequence analyses of the 16S rRNA gene. Amplified ribosomal DNA restriction analysis allowed the differentiation of strain MHP41 from Pseudomonas sp. ADP. The comparative 16S rRNA gene sequence analyses suggested that strain MHP41 is closely related to Pseudomonas nitroreducens and Pseudomonas multiresinovorans . This is the first s -triazine-degrading bacterium isolated in South America. Strain MHP41 is a potential biocatalyst for the remediation of s -triazine-contaminated environments.     

Analysis of herbicides: demonstration of the utility of enzyme immunoassay verification by HPLC

Evidence has accumulated that herbicides in the environment present a significant health hazard to the population. Therefore, the levels of heavily used substances such as atrazine and simazine and their metabolites need to be regularly assessed. The objective was to develop a rapid and simple tube ELISA procedure suitable for use in field studies and non-specialized laboratories. The antisera used were polyclonal antibodies raised in sheep against atrazine or simazine amido caproic acid conjugated to bovine serum albumin. The antibodies were first used to construct a two-step competitive ELISA procedure in 96-well microtitre plates. The 96-well format was then adapted to a coated-tube enzyme immunoassay, by immobilization of hapten-gelatine conjugates on polystyrene tubes. This enabled the colour to be read using a basic spectrophotometer. Soil samples were collected from agricultural and non-agricultural sites in Poland. Atrazine and simazine were extracted by liquid extraction from soil and assayed by tube ELISA. In addition, the samples were extracted by solid-phase extraction before analysis by HPLC. The immunoassays and chemical analysis were carried out by different individuals who were unaware of each other's results, which were then compared at the end of the study. Correlation of the two methods was excellent, with R=98.7 and 81.3 for atrazine and simazine, respectively. The immunoassay yielded the same order of results without having to perform solid-phase extraction before analysis. The study has demonstrated that the simple antigen-coated tube assay provides a cost-effective and valuable screening test. Comparison with the more elaborate, heavily labour-intensive HPLC analysis demonstrated that the results obtained by the simpler enzyme-immunoassay tests were within the same order.     

Effect of nitrogen,Rhizobium inoculation and simazine on yield and quality of Bengal gram (Cicer arietinum L.)

Summary A field experiment was conducted to study the effect of Rhizobium inoculation, nitrogen and simazine application, individually and in combination, on yield and quality of Bengal gram. Application of nitrogen and simazine, and seed inoculation with Rhizobium increased the grain yield significantly. The combined treatment of Rhizobium, simazine and nitrogen increased the grain yield to the extent of 70 per cent over control. Application of simazine increased the methionine content. re]19760609     

Simazine treatment history determines a significant herbicide degradation potential in soils that is not improved by bioaugmentation with Pseudomonas sp. ADP

AIMS: To study biological removal of the herbicide simazine in soils with different history of herbicide treatment and to test bioaugmentation with a simazine-degrading bacterial strain. METHODS AND RESULTS: Simazine removal was studied in microcosms prepared with soils that had been differentially exposed to this herbicide. Simazine removal was much higher in previously exposed soils than in unexposed ones. Terminal restriction fragment length polymorphism analysis and multivariate analysis showed that soils previously exposed to simazine contained bacterial communities that were significantly impacted by simazine but also had an increased resilience. The biodegradation potential was also related to the presence of high levels of the atz-like gene sequences involved in simazine degradation. Bioaugmentation with Pseudomonas sp. ADP resulted in an increased initial rate of simazine removal, but this strain scarcely survived. After 28 days, residual simazine removals were the same in bioaugmented and not bioaugmented microcosms. CONCLUSIONS: In soils with a history of simazine treatment bacterial communities were able to overcome subsequent impacts with the herbicide. The success of bioaugmentation was limited by the low survival of the introduced strain. SIGNIFICANCE AND IMPACT OF THE STUDY: Conclusions from this work provided insights on simazine biodegradation potential of soils and the convenience of bioaugmentation.     

Chemostat selection of a bacterial community able to degrade s-triazinic compounds: continuous simazine biodegradation in a multi-stage packed bed biofilm reactor

Using a successive transfer method on mineral salt medium containing simazine, a microbial community enriched with microorganisms able to grow on simazine was obtained. Afterwards, using a continuous enrichment culture procedure, a bacterial community able to degrade simazine from an herbicide formulation was isolated from a chemostat. The continuous selector, fed with a mineral salt medium containing simazine and adjuvants present in the commercial herbicide formulation, was maintained in operation for 42 days. Following the lapse of this time, the cell count increased from 5 x 10(5) to 3 x 10(8) CFU mL(-1), and the simazine removal efficiency reached 96%. The chemostat's bacterial diversity was periodically evaluated by extracting the culture's bacterial DNA, amplifying their 16S rDNA fragments and analyzing them by thermal gradient gel electrophoresis. Finally, a stable bacterial consortium able to degrade simazine was selected. By PCR amplification, sequencing of bacterial 16S rDNA amplicons, and comparison with known sequences of 16S rDNA from the NCBI GenBank, eight bacterial strains were identified. The genera, Ochrobactrum, Mycobacterium, Cellulomonas, Arthrobacter, Microbacterium, Rhizobium and Pseudomonas have been reported as common degraders of triazinic herbicides. On the contrary, we were unable to find reports about the ability of the genus Pseudonocardia to degrade triazinic compounds. The selected bacterial community was attached to a porous support in a concurrently aerated four-stage packed-bed reactor fed with the herbicide. Highest overall simazine removal efficiencies eta (SZ) were obtained at overall dilution rates D below 0.284 h(-1). However, the multistage packed bed reactor could be operated at dilution rates as high as D = 3.58 h(-1) with overall simazine removal volumetric rates R (v,SZ) = 19.6 mg L(-1) h(-1), and overall simazine removal specific rates R (X,SZ) = 13.48 mg (mg cell protein)(-1) h(-1). Finally, the consortium's ability to degrade 2-chloro-4,6-diamino-1,3,5-triazine (CAAT), cyanuric acid and the herbicide atrazine, pure or mixed with simazine, was evaluated in fed batch processes.     

Contrasting effects of simazine on the photosynthetic physiology and leaf morphology of two Populus clones

Differential effects of simazine (2-chloro-4,6-bis(ethylamino)- s -triazine) on the physiology of two Populus clones were investigated in a greenhouse study. Additions of 5 mg/pot simazine to young plants had no deleterious morphological or physiological effects on clone NC 5328 ( P. x euramericana cv. I 45/51; Section Aigeiros), but reduced the rate of CO₂ fixation, increased CO₂ compensation concentrations and lowered the specific leaf weight of clone NE 388 ( P. maximowiczü x P. trichocarpa cv. Kingston; section Tacamahaca). Abaxial leaf conductance to water vapor was not affected in NE 388. Deleterious effects of simazine on NE 388 were detected ca 48 h after exposure of plants to simazine and generally became more pronounced thereafter. Visual symptoms of injury were evident at ca 2 weeks after simazine application.
Toxic responses to simazine in clone NE 388 varied in different portions of the crown. Inhibition of photosynthesis and increased CO₂ compensation concentrations were more pronounced in the region of recently matured leaves, but were somewhat less in the region of expanding leaves. Older mature leaves in the lower crown region showed no visual symptoms of injury and the rate of photosynthesis and CO₂ compensation concentrations were largely unaffected.     

Ongoing functional evolution of the bacterial atrazine chlorohydrolase AtzA

Triazine herbicides such as atrazine and simazine which were heavily used in the latter half of the twentieth century constituted a rich new source of nitrogen for soil microbes. An atzA dechlorinase active against both atrazine and simazine was isolated from various soil bacteria from diverse locations in the mid 1990s. We have surveyed the atzA genes from eight triazine-degrading Aminobacter aminovorans strains isolated from French agricultural soils recurrently exposed to triazines in 2000. Six amino acid differences from the original isolate were each found in more than one of the A. aminovorans strains. Three of these in particular (V92L, A170T and A296T) were recovered from a majority of the isolates and from locations separated by up to 900 km, so may reflect ongoing selection for the new function. Two of the latter (A170T and A296T) were indeed found to confer higher specificity for simazine, albeit not atrazine, and greater affinity for a metal ion required for activity, than did the original variant. In contrast, we found that ongoing maintenance of the original atzA-containing isolate in laboratory culture for 12 years in a medium containing high concentrations of atrazine has led to the fixation of another amino acid substitution that substantially reduces activity for the triazines. The high concentrations of atrazine in the medium may have relaxed the selection for a highly efficient triazine dechlorinase activity, and that there is some, as yet uncharacterised, counter selection against the activity of this enzyme under these conditions.     

Enzymatic activity, osmotic stress and degradation of pesticide mixtures in soil extract liquid broth inoculated with Phanerochaete chrysosporium and Trametes versicolor

In this study we examined the extracellular enzymatic activity of two white rot fungi (Phanerochaete chrysosporium and Trametes versicolor) in a soil extract broth in relation to differential degradation of a mixture of different concentrations (0-30 p.p.m.) of simazine, dieldrin and trifluralin under different osmotic stress (-0.7 and -2.8 MPa) and quantified enzyme production, relevant to P and N release (phosphomonoesterase, protease), carbon cycling (beta-glucosidase, cellulase) and laccase activity, involved in lignin degradation. Our results suggest that T. versicolor and P. chrysosporium have the ability to degrade different groups of pesticides, supported by the capacity for expression of a range of extracellular enzymes at both -0.7 and -2.8 MPa water potential. Phanerochaete chrysosporium was able to degrade this mixture of pesticides independently of laccase activity. In soil extract, T. versicolor was able to produce the same range of enzymes as P. chrysoporium plus laccase, even in the presence of 30 p.p.m. of the pesticide mixture. Complete degradation of dieldrin and trifluralin was observed, while about 80% of the simazine was degraded regardless of osmotic stress treatment in a nutritionally poor soil extract broth. The capacity of tolerance and degradation of high concentrations of mixtures of pesticides and production of a range of enzymes, even under osmotic stress, suggest potential bioremediation applications.