Nitrification and Autotrophic Nitrifying Bacteria in a Hydrocarbon-Polluted Soil

In vitro ammonia-oxidizing bacteria are capable of oxidizing hydrocarbons incompletely. This transformation is accompanied by competitive inhibition of ammonia monooxygenase, the first key enzyme in nitrification. The effect of hydrocarbon pollution on soil nitrification was examined in situ. In a microcosm study, adding diesel fuel hydrocarbon to an uncontaminated soil (agricultural unfertilized soil) treated with ammonium sulfate dramatically reduced the amount of KCl-extractable nitrate but stimulated ammonium consumption. In a soil with long history of pollution that was treated with ammonium sulfate, 90% of the ammonium was transformed into nitrate after 3 weeks of incubation. Nitrate production was twofold higher in the contaminated soil than in the agricultural soil to which hydrocarbon was not added. To assess if ammonia-oxidizing bacteria acquired resistance to inhibition by hydrocarbon, the contaminated soil was reexposed to diesel fuel. Ammonium consumption was not affected, but nitrate production was 30% lower than nitrate production in the absence of hydrocarbon. The apparent reduction in nitrification resulted from immobilization of ammonium by hydrocarbon-stimulated microbial activity. These results indicated that the hydrocarbon inhibited nitrification in the noncontaminated soil (agricultural soil) and that ammonia-oxidizing bacteria in the polluted soil acquired resistance to inhibition by the hydrocarbon, possibly by increasing the affinity of nitrifying bacteria for ammonium in the soil.     

Anaerobic biodegradation of no. 2 diesel fuel in soil: a soil column study

Soil and sediments are contaminated with petroleum hydrocarbons in many parts of the world. Anaerobic degradation of petroleum hydrocarbon is very relevant in removing oil spills in the anaerobic zones of soil and sediments. This research investigates the possibility of degrading no. diesel fuel under anaerobic conditions. Anaerobic packed soil columns were used to simulate and study in situ bioremediation of soil contaminated with diesel fuel. Several anaerobic conditions were evaluated in soil columns, including sulfate reducing, nitrate reducing, methanogenic, and mixed electron acceptor conditions. The objectives were to determine the extent of diesel fuel degradation in soil columns under various anaerobic conditions and identify the best conditions for efficient removal of diesel fuel. Diesel fuels were degraded significantly under all conditions compared to no electron supplemented soil column (natural attenuation). However, the rate of diesel degradation was the highest under mixed electron acceptor conditions followed in order by sulfate reducing, nitrate reducing, and methanogenic conditions. Under mixed electron acceptor condition 81% of diesel fuel was degraded within 310 days. While under sulfate reducing condition 54.5% degradation of diesel fuel was observed for the same period. This study showed evidence for diesel fuel metabolism in a mixed microbial population system similar to any contaminated field sites, where heterogeneous microbial population exists.     

Influence of long-term diesel fuel pollution on nitrite-oxidising activity and population size of nitrobacter spp in soil

Previous investigations have shown that ammonia oxidation is not inhibited by diesel fuel in a soil with a long history of contamination contrary to a non-contaminated soil. As a consequence, ammonia oxidation does not constitute a limited step in nitrification process (Appl. Environ. Microbiol. 65 (1999) 4008). Moreover, this type of soil also has had the opportunity to develop an abundant microbial population able to metabolise the diesel hydrocarbons. Whether the properties of soil with a long history of diesel fuel contamination may affect the activity of nitrite-oxidising bacteria was investigated. It was observed that re-exposure of soil to diesel fuel apparently stimulated the proliferation of nitrite-oxidising bacteria, as determined by most probable number (MPN) culture technique and MPN-polymerase chain reaction technique. The potential of nitrite-oxidising activity in soil treated with diesel fuel was about 4 times higher than in the control without addition. In the presence of diesel fuel and ammonium, the potential nitrite-oxidising activity was 40% higher than in presence of ammonium only. However, in the presence of hydrocarbon only, low proliferation of Nitrobacter was observed, probably because the heterotrophic bacteria were strongly limited by lack of nitrogen and did not produce sufficient organic metabolites that could be used by the Nitrobacter cells.     

Biodegradation of Diesel Fuel in Soil Under Various Nitrogen Addition Regimes

Bioremediation is a growing technology for treating fuel-contaminated soils. Many biological, physical, and chemical parameters control the rate and efficiency of this process, including type and concentration of contaminants, temperature, oxygen content, and nutrient status. This study investigated the effect that nitrogen sources and concentrations had on the degradation rate of diesel fuel in nutrient limited soil at two carbon-to-nitrogen ratios. The different sources of nitrogen studied were ammonium nitrate, ammonium sulfate, potassium nitrate, urea, and urea oligomers (control release fertilizer). Laboratory experiments were conducted on field-contaminated soil using sealed bioreactors at a controlled temperature of 25°C. For both carbon-to-nitrogen ratios tested, hydrocarbon degradation rates were the highest for the ammonium sulfate (20:1 at 0.032?d^?1; 40:1 at 0.019d^?1) and urea treatments (20:1 at 0.025?d^?1; 40:1 at0.011?d^?1). A degradation rate correlation as a function of nitrate and ammonia concentrations was developed. The correlation suggests the occurrence of nitrate inhibition at elevated nitrate concentrations.     

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.     

Mechanistic Analysis of Ammonium Inhibition of Atmospheric Methane Consumption in Forest Soils

Methane consumption by forest soil was studied in situ and in vitro with respect to responses to nitrogen additions at atmospheric and elevated methane concentrations. Methane concentrations in intact soil decreased continuously from atmospheric levels at the surface to 0.5 ppm at a depth of 14 cm. The consumption rate of atmospheric methane in soils, however, was highest in the 4- to 8-cm depth interval (2.9 nmol per g of dry soil per day), with much lower activities below and above this zone. In contrast, extractable ammonium and nitrate concentrations were highest in the surface layer (0 to 2 cm; 22 and 1.6 μmol per g of dry soil, respectively), as was potential ammonium-oxidizing activity (19 nmol per g of dry soil per day). The difference in zonation between ammonium oxidation and methane consumption suggested that ammonia-oxidizing bacteria did not contribute significantly to atmospheric methane consumption. Exogenous ammonium inhibited methane consumption in situ and in vitro, but the pattern of inhibition did not conform to expectations based on simple competition between ammonia and methane for methane monooxygenase. The extent of ammonium inhibition increased with increasing methane concentration. Inhibition by a single ammonium addition remained constant over a period of 39 days. In addition, nitrite, the end product of methanotrophic ammonia oxidation, was a more effective inhibitor of methane consumption than ammonium. Factors that stimulated ammonium oxidation in soil, e.g., elevated methane concentrations and the availability of cosubstrates such as formate, methanol, or β-hydroxybutyrate, enhanced ammonium inhibition of methane oxidation, probably as a result of enhanced nitrite production.     

Effects of nitrobenzene contamination and of bioaugmentation on nitrification and ammonia-oxidizing bacteria in soil

Bioaugmentation of nitrobenzene-contaminated soil was performed by inoculation with Pseudomonas putida ZWL73, which can grow on nitrobenzene as carbon and nitrogen sources and release free ammonium from the aromatic ring via a partial-reductive pathway. Removal of nitrobenzene was effectively enhanced with concurrent accumulation of ammonium in the bioaugmented soil. Moreover, the negative impact of nitrobenzene contamination on culturable bacterial types and soil nitrification was reduced by strain ZWL73. Changes in the community structure of ammonia-oxidizing bacteria, determined by denaturing gradient gel electrophoresis, were associated with changes in environmental factors in nitrobenzene-contaminated soil, including concentrations of nitrobenzene, ammonium, nitrite and nitrate, but their influence was attenuated in the bioaugmented soil. Overall, P. putida ZWL73 shows promising abilities for effective removal of nitrobenzene and for attenuating the negative effects of nitrobenzene contamination on soil functioning.     

Abundance and community structure of ammonia-oxidizing microorganisms in reservoir sediment and adjacent soils

Ammonia oxidation is an important process for global nitrogen cycling. Both ammonia-oxidizing bacteria (AOB) and archaea (AOA) can be the important players in nitrification process. However, their relative contribution to nitrification remains controversial. This study investigated the abundance and community structure of AOA and AOB in sediment of Miyun Reservoir and adjacent soils. Quantitative PCR assays indicated that the highest AOA abundance occurred in unplanted riparian soil, followed by reservoir sediment, reed-planted riparian soil and agricultural soil. The AOB community size in agricultural soil was much larger than that in the other habitats. Large variations in the structures of AOA and AOB were also observed among the different habitats. The abundance of Nitrosospira-like AOB species were detected in the agricultural soil and reservoir sediment. Pearson’s correlation analysis showed the AOB diversity had positive significant correlations with pH and total nitrogen, while the AOA diversity might be negatively affected by nitrate nitrogen and ammonia nitrogen. This work could add new insights towards nitrification in aquatic and terrestrial ecosystems.     

Ammonia transformations and abundance of ammonia oxidizers in a clay soil underlying a manure pond

Unlined manure ponds are constructed on clay soil worldwide to manage farm waste. Seepage of ammonia-rich liquor into underlying soil layers contributes to groundwater contamination by nitrate. To identify the possible processes that lead to the production of nitrate from ammonia in this oxygen-limited environment, we studied the diversity and abundance of ammonia-transforming microorganisms under an unlined manure pond. The numbers of ammonia-oxidizing bacteria and anammox bacteria were most abundant in the top of the soil profile and decreased significantly with depth (0.5 m), correlating with soil pore-water ammonia concentrations and soil ammonia concentrations, respectively. On the other hand, the numbers of ammonia-oxidizing archaea were relatively constant throughout the soil profile (10(7) amoA copies per g(soil)). Nitrite-oxidizing bacteria were detected mainly in the top 0.2 m. The results suggest that nitrate accumulation in the vadose zone under the manure pond could be the result of complete aerobic nitrification (ammonia oxidation to nitrate) and could exist as a byproduct of anammox activity. While the majority of the nitrogen was removed within the 0.5-m soil section, possibly by combined anammox and heterotrophic denitrification, a fraction of the produced nitrate leached into the groundwater.     

Fertilization influences nitrogen dynamics in soils under Dalbergia sissoo

The influence of added ammonium, phosphorus, potassium, and gypsum on net nitrogen mineralization was studied in soil beneath a six-year-old plantation of the N₂-fixing tree Dalbergia sissoo in Pakistan. Soil with and without amendments was placed in polyethylene bags and incubated, buried in the soil, for 30 days. After that time the soil was analyzed and net ammonium and nitrate production and net nitrogen mineralization were calculated. The addition of ammonium stimulated nitrification indicating that the process was substrate limited. The inhibition of nitrification by Nitrapyrin showed that the process is autotrophic in these soils. Gypsum addition lowered soil pH from 8.0 to 7.2 and significantly stimulated ammonification, nitrification and net nitrogen mineralization. The addition of potassium more than tripled the soil K:Na ratio. Net ammonium and nitrate production and net nitrogen mineralization all increased in this treatment. The addition of phosphorus had no significant effect on soil nitrogen dynamics.     

Influence of Protozoa and Nutrient Availability on Nitrification Rates in Subsurface Sediments

Abstract Protozoan abundance, nitrification potential, and related factors in saturated subsurface sediments and the overlying soil were compared at a nonfertilized grassland and an agricultural cropland site. In a 6-week laboratory experiment, DOC, ammonium, and protozoan abundance were manipulated in flasks containing groundwater-sediment slurries. Microbial abundance (protozoa, actively respiring bacteria, and total bacteria) and nutrient concentrations (extractable ammonium and nitrate) were measured. Results from the soil profile analysis showed that protozoan abundance declined with depth at both sites, but significant numbers (392 cells g⁻¹dw) were found in groundwater sediments at the cropland site. Nitrification potential declined with depth at the grassland site and increased with depth at the cropland site. In the laboratory experiment, treatment responses generally were observed within 3 weeks, but had diminished by 6 weeks. Protozoa reduced bacterial populations through the first 3 weeks, but this effect was not significant by week 6. In the cropland sediments, increased net nitrate production occurred in the two reduced protozoa treatments that received ammonium, suggesting that nitrification was occurring and was limited by ammonium. High protozoan abundance in the cropland sediments increased the nitrate flux response, unless DOC was added; in this case, no response occurred. No such responses were recorded in the grassland sediments. Apparently, appreciable nitrification can occur in some groundwater sediments, if sufficient ammonium is present and DOC availability is low. Furthermore, nitrification can be enhanced when protozoan abundance is elevated. Finally, our results suggest that surface land use practices can alter subsurface nitrification rates and microbial community structure. Received: 12 July 1996; Accepted: 2 December 1996     

变性梯度凝胶电泳法研究我国不同土壤氨氧化细菌群落组成及活性

实验选用了我国3种不同土壤研究土壤硝化活性、硝化细菌数量,并使用变性梯度凝胶电泳(DGGE)的方法研究了不同土壤中氨氧化细菌(AOB)区系变化。通过28d的土壤培养实验研究发现,潮土具有最强的硝化势,几乎100%的铵态氮转化为硝态氮;而红壤中的硝化势最弱,只有4.9%的铵态氮转化为硝态氮。对这3种土壤硝化细菌进行计数发现,3种土壤氨氧化菌数量差异显著,而3种土壤亚硝酸氧化菌(NOB)处于一个数量级。采用氨氧化菌功能基因amoA(氨单加氧酶ammoniamonooxygenase)特异PCR结合DGGE的方法对土壤氨氧化菌区系进行分析。红壤有4个氨氧化菌种属,与潮土和黄泥土没有共同的氨氧化菌种属。4个种属中两个是与潮土和黄泥土亲源性比较远的,特有的氨氧化菌种属,这两个种属与已知的Nitrosospira属的cluster3bz97838和Nitrosospira属的cluster3aAF353263亲源性比较近。潮土有5个氨氧化菌种属,潮土与黄泥土有两个共同的氨氧化菌种属,这两个种属中的一个是潮土和黄泥土特有的,与其他氨氧化菌种属亲源性比较远的氨氧化菌种属,这个种属与已知的Nitrosospira属的cluster3bZ97849亲源性比较近。黄泥土有4个氨氧化菌种属,除了与潮土共有的一个种属是两种土壤特有的氨氧化菌种属外,黄泥土还有一个与其他氨氧化菌种属亲源性比较远的,黄泥土特有的种属,与Nitrosospira属的cluster3aAF353263亲源性很近。3种土壤中分离到的硝化细菌表现出不同的硝化能力。实验结果表明,以amoA基因为目标的PCR-DGGE是比以16SrDNA为目标的PCR-DGGE更有效的研究氨氧化菌种群的方法;3种土壤的氨氧化菌种群差异显著,尤其是红壤的氨氧化菌种群与另外两种土壤差异明显,这种差异可能与红壤的低pH条件对氨氧化菌种群的长期选择有关;3种土壤中的硝化活性与土壤中的硝化细菌数量没有显著相关,可能由于3种土壤差异显著的土壤环境对硝化活性的影响造成。因此在对不同土壤硝化细菌进行研究时不仅需要对硝化细菌数量进行研究,还需要研究不同土壤中硝化细菌的种属及不同土壤环境条件对硝化细菌硝化活性的影响。     

Autotrophic ammonia-oxidizing bacteria contribute minimally to nitrification in a nitrogen-impacted forested ecosystem

Deposition rates of atmospheric nitrogenous pollutants to forests in the San Bernardino Mountains range east of Los Angeles, California, are the highest reported in North America. Acidic soils from the west end of the range are N-saturated and have elevated rates of N-mineralization, nitrification, and nitrate leaching. We assessed the impact of this heavy nitrogen load on autotrophic ammonia-oxidizing communities by investigating their composition, abundance, and activity. Analysis of 177 cloned beta-Proteobacteria ammonia oxidizer 16S rRNA genes from highly to moderately N-impacted soils revealed similar levels of species composition; all of the soils supported the previously characterized Nitrosospira clusters 2, 3, and 4. Ammonia oxidizer abundance measured by quantitative PCR was also similar among the soils. However, rates of potential nitrification activity were greater for N-saturated soils than for soils collected from a less impacted site, but autotrophic (i.e., acetylene-sensitive) activity was low in all soils examined. N-saturated soils incubated for 30 days with ammonium accumulated additional soluble ammonium, whereas less-N-impacted soils had a net loss of ammonium. Lastly, nitrite production by cultivated Nitrosospira multiformis, an autotrophic ammonia-oxidizing bacterium adapted to relatively high ammonium concentrations, was significantly inhibited in pH-controlled slurries of sterilized soils amended with ammonium despite the maintenance of optimal ammonia-oxidizing conditions. Together, these results showed that factors other than autotrophic ammonia oxidizers contributed to high nitrification rates in these N-impacted forest soils and, unlike many other environments, differences in nitrogen content and soil pH did not favor particular autotrophic ammonia oxidizer groups.     

Nitrification exhibits Haldane kinetics in an agricultural soil treated with ammonium sulfate or dairy-waste compost

An agricultural soil was treated with dairy-waste compost, ammonium-sulfate fertilizer or no added nitrogen (control) and planted to silage corn for 6 years. The kinetics of nitrification were determined in laboratory-shaken slurry assays with a range of substrate concentrations (0-20 mM NH(4)(+)) over a 24-h period for soils from the three treatments. Determined concentrations of substrate and product were fit to Michaelis-Menten and Haldane models. For all the treatments, the Haldane model was a better fit, suggesting that significant nitrification inhibition may occur in soils under high ammonium conditions similar to those found immediately after fertilization or waste applications. The maximum rate of nitrification (V(max)) was significantly higher for the fertilized and compost-treated soils (1.74 and 1.50 mmol N kg(-1) soil day(-1)) vs. control soil (0.98 mmol kg(-1) soil day(-1)). The K(m) and K(i) values were not significantly different, with average values of 0.02 and 27 mM NH(4)(+), respectively. Our results suggest that both N sources increased nitrifier community size, but did not shift the nitrifier community structure in ways that influenced enzyme affinity or sensitivity to ammonium. The K(m) values are comparable to those determined directly in other soils, but are substantially lower than those from most pure cultures of ammonia-oxidizing bacteria.     

高效稳定性硫酸铵氮肥在黑土中的施用效果

为筛选高效稳定性氮肥,采用盆栽试验,通过监测施用不同处理的稳定性硫酸铵对黑土铵态氮和硝态氮含量、表观硝化率、硝化抑制率、玉米生长指标、产量和氮素效率等指标的影响,研究添加不同生化抑制剂配方的稳定性硫酸铵态氮肥在吉林黑土玉米栽培中的施用效果。本试验以不施氮肥(CK)和施硫酸铵(N)为对照,在硫酸铵中分别添加硝化抑制剂3,4-二甲基吡唑磷酸盐(DMPP)、2-氯-6-三甲基吡啶(CP),氮保护剂(N-GD)和肥料增效剂(HFJ)及其组合,制成9种稳定性硫酸铵氮肥。结果表明: 与单施硫酸铵氮肥处理相比,在黑土中添加DMPP和DMPP组合显著影响土壤中铵态氮和硝态氮含量及土壤表观硝化率,铵态氮含量提高1.4～2.0倍,硝态氮含量降低13.6%～17.9%,土壤表观硝化率降低55.3%～59.8%;添加DMPP、DMPP+HFJ和DMPP+N-GD组合硝化抑制率最高,达到16.5%以上;添加DMPP+HFJ+N-GD和HFJ的硫酸铵处理玉米叶片叶绿素含量增加最显著,增加4.5～5.3倍;硫酸铵添加硝化抑制剂和肥料增效剂对株高无显著影响;添加HFJ的硫酸铵处理玉米生物量、籽粒产量、经济系数、收获指数、氮肥农学利用率、氮素吸收利用率、肥料贡献率和氮肥偏生产力增加最显著,分别增加1.2、2.5、0.7、0.6、2.7、2.1、1.3和2.5倍。添加HFJ和DMPP、DMPP+HFJ、DMPP+N-GD处理的硫酸铵处理在黑土中施用效果最好,但是DMPP成本较高,因此,兼顾成本和氮肥利用率,建议稳定性硫酸铵态氮肥生化抑制剂首选氮肥增效剂HFJ,其次选择DMPP+HFJ或者DMPP+N-GD。     

On site bioremediation of hydrocarbon-contaminated Arctic tundra soils in inoculated biopiles

There is a need to develop technology to allow the remediation of soil in polar regions that have been contaminated by hydrocarbon fuel spills. Bioremediation is potentially useful for this purpose, but has not been well demonstrated in polar regions. We investigated biopiles for on-site bioremediation of soil contaminated with Arctic diesel fuel in two independent small-scale field experiments at different sites on the Arctic tundra. The results were highly consistent with one another. In biopiles at both sites, extensive hydrocarbon removal occurred after one summer. After 1 year in treatments with optimal conditions, total petroleum hydrocarbons were reduced from 196 to below 10 mg per kg of soil at one site, and from 2,109 to 195 mg per kg of soil at the other site. Addition of ammonium chloride and sodium phosphate greatly stimulated hydrocarbon removal and indicates that biodegradation was the primary mechanism by which this was achieved. Inoculation with cold-adapted, mixed microbial cultures further stimulated hydrocarbon removal during the summer immediately following inoculation. At one site, soil temperature was monitored during the summer season, and a clear plastic cover increased biopile soil temperature, measured as degree-day accumulation, by 30-49%. Our results show that on-site bioremediation of fuel-contaminated soil at Arctic tundra sites is feasible.     

Autotrophic growth of nitrifying community in an agricultural soil

The two-step nitrification process is an integral part of the global nitrogen cycle, and it is accomplished by distinctly different nitrifiers. By combining DNA-based stable isotope probing (SIP) and high-throughput pyrosequencing, we present the molecular evidence for autotrophic growth of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA) and nitrite-oxidizing bacteria (NOB) in agricultural soil upon ammonium fertilization. Time-course incubation of SIP microcosms indicated that the amoA genes of AOB was increasingly labeled by ¹³CO₂ after incubation for 3, 7 and 28 days during active nitrification, whereas labeling of the AOA amoA gene was detected to a much lesser extent only after a 28-day incubation. Phylogenetic analysis of the ¹³C-labeled amoA and 16S rRNA genes revealed that the Nitrosospira cluster 3-like sequences dominate the active AOB community and that active AOA is affiliated with the moderately thermophilic Nitrososphaera gargensis from a hot spring. The higher relative frequency of Nitrospira-like NOB in the ¹³C-labeled DNA suggests that it may be more actively involved in nitrite oxidation than Nitrobacter-like NOB. Furthermore, the acetylene inhibition technique showed that ¹³CO₂ assimilation by AOB, AOA and NOB occurs only when ammonia oxidation is not blocked, which provides strong hints for the chemolithoautotrophy of nitrifying community in complex soil environments. These results show that the microbial community of AOB and NOB dominates the nitrification process in the agricultural soil tested.     

Simultaneous nitrification and denitrification by controlling vertical and horizontal microenvironment in a membrane-aerated biofilm reactor

Nitrogen and carbon components in domestic modified wastewater were completely removed by simultaneous nitrification and denitrification using a membrane-aerated biofilm reactor where biofilm was fixed on a hollow-fiber membrane. To measure the spatial distribution of pH, ammonium and nitrate ions and to observe microbes inside the biofilm fixed on the membrane, microelectrodes and the fluorescence in situ hybridization (FISH) method were applied. Due to plug flow in the vertical direction (from the bottom to the top of the reactor), ammonium nitrogen was gradually removed and negligible nitrate nitrogen was detected throughout the reactor. FISH revealed that ammonia-oxidizing bacteria were mainly distributed inside the biofilm and other bacteria, which included denitrifying bacteria, were mainly distributed outside the biofilm and over the suspended sludge. In order to characterize bacterial activity in the vertical direction of the reactor, nitrification rates at lower, central and upper points were calculated using microelectrode data. The nitrification rate at the lower point was 7 and 125 times higher than those at the central and upper points, respectively. These results show that the removal of carbon and nitrogen compounds was accomplished efficiently by using various kinds of bacteria distributed vertically and horizontally in a single reactor.     

Effect of methyl 3-4-hydroxyphenyl propionate, a Sorghum root exudate, on N dynamic, potential nitrification activity and abundance of ammonia-oxidizing bacteria and archaea

Aims

It has been reported that root exudates of Sorghum bicolor can inhibit nitrification in a bioassay using Nitrosomonas, and methyl 3-(4-hydroxyphenyl) propionate (MHPP) was identified as one of the nitrification inhibiting compounds. Therefore, we have investigated the effects of this compound on nitrogen dynamic, potential nitrification activity and on soil microorganisms.

Methods

We conducted soil incubation experiments using synthetic MHPP to evaluate its effect on changes in inorganic soil nitrogen pools, on nitrification activity and on abundance of ammonia-oxidizing bacteria and archaea. Addition of MHPP at two concentrations equivalent to 70 and 350 μg C g^?1 soil was compared to glucose as a carbon source and to the commercially available nitrification inhibitor dicyandiamide (DCD).

Results

Soil amended with the high dose of MHPP and with DCD showed reduced nitrate content and low nitrification activity after 3 and 7 days of incubation. This was mirrored by a 70 % reduction in potential nitrification activity compared to a nitrogen-only control. None of the incubation treatments affected non-target microbial counts as estimated by 16S rRNA gene copy numbers, however, the high dose of MHPP significantly reduced the abundance of ammonia-oxidizing bacteria and archaea.

Conclusions

These findings suggest that MHPP is capable of suppressing nitrification in soil, possibly by reducing the population size and activity of ammonia-oxidizing microorganisms.     

The effect of the bioremediation of soil contaminated with petroleum derivatives on the occurrence of epigeic and edaphic fauna

This field study investigated the colonization process of soil contaminated with different petroleum products (petrol, diesel fuel, spent engine oil; dose: 6000 mg of fuel·kg^?1 dry mass [d.m.] of soil) by epigeic and edaphic invertebrates during the progress of natural bioremediation and bioremediation enhanced using selected microorganisms (ZB-01 biopreparation). Epigeic fauna was captured using pitfall traps. Occurrence of edaphic fauna in soil samples as well as total petroleum hydrocarbon contents (TPH) were also investigated. Results showed that inoculation with ZB-01 biocenosis allowed the degradation of petroleum derivatives in the soil contaminated with diesel fuel and engine oil, with 82.3% and 75.4% efficiency, respectively. Applying bioremediation to all contaminated soils accelerated the process of recolonization by edaphic invertebrates. However, the 28-month period was too short to observe full population recovery in soils contaminated with diesel fuel and engine oil. Microbe-enhanced bioremediation accelerated recolonization by epigeic invertebrates on soil contaminated with diesel fuel, whereas it exerted inhibitory effect on recolonization of soil contaminated with engine oil (especially by Collembola). The observed discrepancies in the rates of recolonization for soils contaminated with petrol and diesel fuel that were still noted at the stage of no longer different TPH levels justify the idea to include the survey of edaphic faunal density as one of the parameters in the ecological risk assessment of various bioremediation techniques.