It has been proposed that levels of carbon and nitrogen in soil affect the rate at which triazine herbicides degrade. The present study was designed to investigate the effects of varying the levels of initial soil carbon and nitrogen as well as the effects of a later carbon addition on the mineralization of atrazine and its metabolites in soil. Atrazine degradation in soils amended with carbon as mannitol, and with nitrogen as urea, at levels of 10, 30, 50, or 80 mg kg−1 mannitol, urea, or mannitol and urea was similar to degradation in unamended soils. Only 39% of applied atrazine was mineralized after 326 days regardless of the initial carbon or nitrogen treatment. In contrast, a second mannitol amendment of 1 g kg−1 soil at day 140 increased [14C]carbon dioxide evolution by an additional 17% as a result of enhancing the mineralization rate of the atrazine metabolites. This conclusion is supported by our finding that periodic extraction of the soil with methanol followed by quantification on HPLC showed complete dissipation of the parent atrazine in 120 days. The metabolites hydroxyatrazine (HA), deisopropylatrazine (DIA), deethylatrazine (DEA), and diaminochlorotriazine (DACT) began to appear in the methanol extract 10 days following atrazine application. The greatest concentrations of HA, DEA, DIA. and DACT in the methanol extracts were 19, 12.4, 10.1, and 6.7% of applied atrazine, respectively. These concentrations were observed at day 95 except for DEA where the concentration continued to increase until day 142. A second soil extraction with hydrochloric acid (1m) + methanol (20 + 80 by volume) recovered additional HA, deethylhydroxyatrazine (DEHA), deisopropylhydroxyatrazine (DIHA), and DACT. When the extraction data were combined, 31.9, 12.4, 10.1. 10.2, 12.0 and 7.8% of applied atrazine was recovered as HA, DEA, DIA, DEHA. DIHA, and DACT, respectively. Combustion of the extracted soil showed 20% of the applied atrazine remained as soil‐bound residues after 326 days.
: Reported levels of atrazine in soils at pesticide mix-load sites can vary between 7É9 ] 10~5 mM and 1É9 mM. We report on a mixed microbial culture, capable of degrading concentrations of atrazine in excess of 1É9 mM. At initial concentrations of 0É046 M and 0É23 M, the mixed population degraded 78% and 21% of atrazine in soil (100 days), respectively. At the same initial concentrations in liquid cultures, 90% and 56% of the atrazine was degraded (80 days), respectively. Decreased degradation in soil samples may have resulted from atrazine sorption to soil surfaces or decreased contact between the population and the herbicide. In the 0É23 M system, we attribute incomplete degradation to phosphorous depletion. Data for carbon dioxide evolution was Ðtted to a three-halforder regression model, but we feel that there are limitations of the application of this model to atrazine degradation. The population uses the herbicide as a nitrogen source and little carbon is incorporated into biomass, as the energy status of carbons in the ring leads to their direct evolution as [14C]carbon dioxide. This situation contributes to an evolution pattern that, when Ðtted to the three-halforder model, results in underestimation of the biomass produced. Data from our study suggest that our mixed culture could be used for bioremediation of atrazine at concentrations up to and exceeding those currently reported for agrochemical mixing-loading facilities.
A new bioassay procedure was developed for the detection of erythromycin in aquaculture samples using a strain of a Stenotrophomonas as an indicator organism. Conventional diskplate and well-plate radial diffusion assay procedures were developed, as well as a third procedure using the same indicator organism in Luria-Bertani (LB) broth, supplemented with the indicator dye Brilliant Black (40 pg/mL) in a multi-well microtiter plate. For both the disk-plate and well-plate radial diffusion assays, the response reflected in the size (width) of the growth inhibition zone, which was linear over the tested concentration range of 0.05 to 2.0 pg erythromycidml. The limit of quantitation of the bioassay was at 0.05 pg erythromycidml. Among the three methods of assay tested with Stenotrophomonas sp., the semiquantitative dye reduction method is easy to read and is not diffusion dependent. This method allows for processing of more samples and more replication on a single titer plate. This new indicator organism is specific for erythromycin when tested in the presence of other antibacterial agents, i.e., oxytetracycline (Terramycin") and/or Romet-Ma. This new bioassay procedure is suitable for quantitation of low concentrations of erythromycin in aquaculture water and sediment samples.
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