“…To test these hypotheses, we measured diazotrophic activities of sediments, roots, and litter in laboratory incubation experiments with and without the addition of sodium molybdate, a 'specific' metabolic inhibitor for sulfur reducing bacteria (Oremland and Capone 1988). Nitrogenase activity was measured using the acetylene reduction assay, which was calibrated by 15 N 2 reduction assay.…”
We used established long-term experimental P-amended plots in freshwater marshes of northern Belize to determine the impact of P input on nitrogen (N) fixation. Marshes with different conductivities and sulfate concentrations were selected to elucidate the effect of salinity and the contribution of sulfur reducing bacteria to the overall N fixation. Rates of N fixation in sediment, roots, and cyanobacterial mats was measured in laboratory incubation experiments (acetylene reduction assay calibrated by 15 N 2 reduction assay) with and without the addition of sodium molybdate (sulfur reducing bacteria inhibitor). P has increased macrophyte primary production significantly, which led to the rapid elimination of cyanobacterial mats and the elimination of autotrophic N fixation. P addition enhanced heterotrophic N fixation in both the sediments and rhizosphere due primarily to increased C supply to the sediment. When expressed on a dry weight basis, root associated N fixation was higher than sediment N fixation, but the contribution of the root associated fixation to the total N fixation was small when expressed per square meter. Sulfur reducing bacteria were an important component of N fixation, contributing from 20 to 53% to the overall N fixation. A simple N budget was created to determine if N demands are met following P addition. The heterotrophic N fixation substituted in part for autotrophic cyanobacterial N fixation when P limitation was alleviated.
“…To test these hypotheses, we measured diazotrophic activities of sediments, roots, and litter in laboratory incubation experiments with and without the addition of sodium molybdate, a 'specific' metabolic inhibitor for sulfur reducing bacteria (Oremland and Capone 1988). Nitrogenase activity was measured using the acetylene reduction assay, which was calibrated by 15 N 2 reduction assay.…”
We used established long-term experimental P-amended plots in freshwater marshes of northern Belize to determine the impact of P input on nitrogen (N) fixation. Marshes with different conductivities and sulfate concentrations were selected to elucidate the effect of salinity and the contribution of sulfur reducing bacteria to the overall N fixation. Rates of N fixation in sediment, roots, and cyanobacterial mats was measured in laboratory incubation experiments (acetylene reduction assay calibrated by 15 N 2 reduction assay) with and without the addition of sodium molybdate (sulfur reducing bacteria inhibitor). P has increased macrophyte primary production significantly, which led to the rapid elimination of cyanobacterial mats and the elimination of autotrophic N fixation. P addition enhanced heterotrophic N fixation in both the sediments and rhizosphere due primarily to increased C supply to the sediment. When expressed on a dry weight basis, root associated N fixation was higher than sediment N fixation, but the contribution of the root associated fixation to the total N fixation was small when expressed per square meter. Sulfur reducing bacteria were an important component of N fixation, contributing from 20 to 53% to the overall N fixation. A simple N budget was created to determine if N demands are met following P addition. The heterotrophic N fixation substituted in part for autotrophic cyanobacterial N fixation when P limitation was alleviated.
“…On Day 291, 20 mM molybdate (added as Na 2 MoO•2H 2 0) was added to the influent as an inhibitor to sulfate reduction. 25 As seen in the figures, rapid loss of herbicide biotransformation was also observed. This further suggests that the sulfate-reducing organisms in the culture are likely responsible for the biotransformation of the herbicides, as has been observed by others.…”
Among the important factors affecting the biotransformation of xenobiotic chemicals upon their release into the environment are the dominant electron acceptor condition present and the presence of other, more readily degraded carbon sources. Here, glass-bead biofilm columns were used to investigate the effects of the presence of three different inorganic electron acceptor conditions (oxygen respiration, nitrate reduction, and sulfate reduction) on the biotransformation of the acetanilide herbicides alachlor and propachlor, and to determine the effects of two exogenous carbon sources (acetate and glucose) on their biotransformation under each of these conditions.Biotransformation of alachlor and propachlor occurred in the presence of both carbon sources and under each of the three electron acceptor conditions. Both were transformed most rapidly under sulfate-reducing conditions. Analysis by gas chromatography/mass spectrometry (GC/ MS) did not reveal any significant metabolic products. Both herbicides react abiotically with bisulfide, produced within the sulfate-reducing cultures, though most of the transformation was attributed to the microorganisms. The primary, readily degraded carbon source (acetate or glucose) was needed to establish each culture, and its continuous presence was required to sustain herbicide biotransformation in the sulfate-reducing reactors. Loss of either acetate or glucose from the column influent did not significantly affect herbicide biotransformation in the aerobic or nitratereducing reactors, at least for short periods. Temporary loss of the external electron acceptors (O 2 , NO 3 -, or SO 4 2-
“…Briefly, activated sludge samples were diluted to 1 g MLSS l 21 with nitrate-free effluent water and incubated with labelled glucose (3.7610 5 Bq) (D-[2-3 H]) and unlabelled glucose to a final concentration of 1.5 mM under anaerobic conditions for 6 h (labelled and unlabelled glucose were added at time 0, and again after 3 h). Addition of 1 mM molybdate and 10 mM BES (bromoethane sulfonate) was done 20 min prior to addition of tracer to inhibit sulfate reduction (Oremland & Capone, 1988) and methanogenic activity (Nollet et al, 1997). The incubations were terminated by addition of 50 % (v/v) ethanol (final concentration).…”
Microbiology in wastewater treatment has mainly been focused on problem-causing filamentous bacteria or bacteria directly involved in nitrogen and phosphorus removal, and to a lesser degree on flanking groups, such as hydrolysing and fermenting bacteria. However, these groups constitute important suppliers of readily degradable substrates for the overall processes in the plant. This study aimed to identify glucose-fermenting bacteria in a full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plant (WWTP), and to determine their abundance in similar WWTPs. Glucose-fermenting micro-organisms were identified by an in situ approach using RNA-based stable isotope probing. Activated sludge was incubated anaerobically with 13 C 6 -labelled glucose, and 13 C-enriched rRNA was subsequently reversetranscribed and used to construct a 16S rRNA gene clone library. Phylogenetic analysis of the library revealed the presence of two major phylogenetic groups of Gram-positive bacteria affiliating with the genera Tetrasphaera, Propionicimonas (Actinobacteria), and Lactococcus and Streptococcus (Firmicutes). Specific oligonucleotide probes were designed for fluorescence in situ hybridization (FISH) to specifically target the glucose-fermenting bacteria identified in this study. The combination of FISH with microautoradiography confirmed that Tetrasphaera, Propionicimonas and Streptococcus were the dominant glucose fermenters. The probe-defined fermenters were quantified in 10 full-scale EBPR plants and averaged 39 % of the total biovolume. Tetrasphaera and Propionicimonas were the most abundant glucose fermenters (average 33 and 4 %, respectively), while Streptococcus and Lactococcus were present only in some WWTPs (average 1 and 0.4 %, respectively). Thus the population of actively metabolizing glucose fermenters seems to occupy a relatively large component of the total biovolume.
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