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Situ Degradation (situ + degradation)
Selected AbstractsTranscription dynamics of the functional tfdA gene during MCPA herbicide degradation by Cupriavidus necator AEO106 (pRO101) in agricultural soilENVIRONMENTAL MICROBIOLOGY, Issue 3 2008Mette Haubjerg Nicolaisen Summary A modified protocol for simultaneous extraction of RNA and DNA, followed by real-time polymerase chain reaction quantification, was used to investigate tfdA gene expression during in situ degradation of the herbicide MCPA (4-chloro-2-methylphenoxy-acetic acid) in soil. tfdA encodes an ,-ketoglutarate-dependent dioxygenase catalysing the first step in the degradation pathway of MCPA and 2,4-D (2,4-dichlorophenoxy-acetic acid). A linear recovery of tfdA mRNA over three orders of magnitude was shown, and the tfdA mRNA level was normalized using the tfdA mRNA/DNA ratio. The density of active cells required for tfdA mRNA detection was 105 cells g,1 soil. Natural soil microcosms inoculated with Cupriavidus necator (formerly Ralstonia eutropha) AEO106 (pRO101) cells were amended with four different MCPA concentrations (2, 20, 50 and 150 mg kg,1). Mineralization rates were estimated by quantification of 14CO2 emission from degradation of 14C-MCPA. tfdA mRNA was detected 1 h after amendment at all four concentrations. In soils amended with 2 and 20 mg kg,1, the mRNA/DNA ratio for tfdA demonstrated a sharp transient maximum of tfdA expression from no to full expression within 3 and 6 h respectively, followed by a decline and complete loss of expression after 19 and 43 h. A more complex pattern of tfdA expression was observed for the higher 50 and 150 mg kg,1 amendments; this coincided with growth of C. necator AEO106 (pRO101) in the system. Repeated amendment with MCPA after 2 weeks in the 20 mg kg,1 scenario revealed a sharp increase of tfdA mRNA, and absence of a mineralization lag phase. For all amendments, tfdA mRNA was detectable only during active mineralization, and thus revealed a direct correlation between tfdA mRNA presence and microbial degrader activity. The present study demonstrates that direct analysis of functional gene expression dynamics by quantification of mRNA can indeed be made in natural soil. [source] atz gene expressions during atrazine degradation in the soil drilosphereMOLECULAR ECOLOGY, Issue 4 2010C. MONARD Abstract One of the various ecosystemic services sustained by soil is pollutant degradation mediated by adapted soil bacteria. The pathways of atrazine biodegradation have been elucidated but in situ expression of the genes involved in atrazine degradation has yet to be demonstrated in soil. Expression of the atzA and atzD genes involved in atrazine dechlorination and s -triazine ring cleavage, respectively, was investigated during in situ degradation of atrazine in the soil drilosphere and bulked samples from two agricultural soils that differed in their ability to mineralize atrazine. Interestingly, expression of the atzA gene, although present in both soils, was not detected. Atrazine mineralization was greatest in Epoisses soil, where a larger pool of atzD mRNA was consistently measured 7 days after atrazine treatment, compared with Vezin soil (146 vs. 49 mRNA per 10616S rRNA, respectively). Expression of the atzD gene varied along the degradation time course and was profoundly modified in soil bioturbated by earthworms. The atzD mRNA pool was the highest in the soil drilosphere (casts and burrow-linings) and it was significantly different in burrow-linings compared with bulk soil (e.g. 363 vs. 146 mRNA per 10616S rRNA, 7 days after atrazine treatment in Epoisses soil). Thus, consistent differences in atrazine mineralization were demonstrated between the soil drilosphere and bulk soil. However, the impact of bioturbation on atrazine mineralization depended on soil type. Mineralization was enhanced in casts, compared with bulk soil, from Epoisses soil but in burrow-linings from Vezin soil. This study is the first to report the effects of soil bioturbation by earthworms on s -triazine ring cleavage and its spatial variability in soil. [source] Investigating the in situ degradation of atrazine in groundwater,PEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 4 2006Robert Pearson Abstract This study focused on whether or not atrazine could be degraded by indigenous groundwater bacteria as part of an in situ remediation approach. Groundwater was taken from an unconfined middle upper chalk site where concentrations of atrazine and nitrate were typically in the ranges 0.02,0.2 µg litre,1 and 11.6,25.1 mg NO3 -N litre,1 respectively. Sacrificial batch studies were performed using this groundwater spiked with atrazine at a concentration of 10 µg litre,1 in conjunction with a minimal mineral salts liquid (Glu-MMSL) medium which contained glucose as the sole carbon source. Treatments comprised either the Glu-MMSL groundwater cultured bacteria or Pseudomonas sp. strain ADP. Results from sacrificial batches indicated the occurrence of bacterial growth and denitrification as monitored by optical density (absorbance at 600 nm) and NO3 -N content. Analysis of atrazine content by solid phase extraction coupled with high-performance liquid chromatography showed no degradation of atrazine over a period of 103 days in either treatment. These results indicated that no acclimatised bacterial community featuring positive degraders to the herbicide atrazine had become established within this chalk aquifer in response to the trace levels encountered. Copyright © 2006 Society of Chemical Industry [source] Effects of urea treatment of potato pulp and inclusion levels of potato pulp silage in supplements on digestibility and ruminal fermentation in beef steersANIMAL SCIENCE JOURNAL, Issue 6 2007Masahito SUGIMOTO ABSTRACT Six Wagyu (Japanese Black) steers fitted with a ruminal cannula were used in a split-plot design experiment comprising a 3 × 3 Latin square design (whole plot) and a randomized blocks design (subplot) to determine the effect of the treatment of potato pulp (PP) with urea and the effect of inclusion levels of PP silage in feed supplement on digestibility, ruminal in situ degradation and ruminal fermentation. The whole plot consisted of 20%, 50% and 80% PP silage (dry matter (DM) basis), with PP silage replacing formula feed. The subplot included untreated or 0.5% (on an as-fed basis) urea-treated PP. The treatment of PP with urea showed no effect on DM intake and digestibility. The percentage of the rapidly degradable DM fraction of the urea-treated PP silage was higher (P < 0.01) and the percentage of its slowly degradable DM fraction was lower (P < 0.01) than for the untreated PP silage. Ruminal ammonia concentration was greater (P < 0.01) for steers fed urea-treated PP silage than that for steers fed the untreated PP silage. The treatment of PP with urea caused a decrease in the molar proportion of acetate and an increase in the proportion of propionate in ruminal fluid after feeding. The rate of DM degradations in hay (linear, P < 0.01) and in PP silage decreased (linear, P < 0.01) as the inclusion level of PP silage increased. Increasing the inclusion level of PP silage in supplement decreased the effective degradability of DM in hay (linear, P < 0.05) and in PP silage (linear, P < 0.05). An increase in the amount of PP silage increased the molar proportion of acetate (linear, P < 0.01) and decreased the butyrate proportion (linear, P < 0.05) in ruminal fluid. The results suggest that urea treatment of PP facilitates microbial access to starch of PP silage in the rumen and that surplus level of PP silage in supplement have adverse effect on ruminal digestion. [source] |