The importance of autotrophic versus heterotrophic oxidation of atmospheric ammonium in forest ecosystems with acid soil

Abstract The role of autotrophic and heterotrophic nitrifying microorganisms in the oxidation of atmospheric ammonium in two acid and one calcareous location of a Dutch woodland area was investigated. In soil slurries nitrate formation was completely inhibited by acetylene, a specific inhibitor of autotrophic ammonium-oxidizing bacteria. A survey of nitrifiers in the forest soils showed that both autotrophic ammonium- and nitrite-oxidizing bacteria were present in high numbers and evidence was obtained that autotrophic bacteria are able to nitrify below pH 4. These results show that autotrophic nitrifying bacteria may account for most of the nitrification in the examined soils. To assess the contribution of heterotrophic nitrifiers, about 200 strains of heterotrophic bacteria and 21 morphologically distinct fungal strains were isolated from the acid soil locations and tested for their ability to nitrify. Only one Penicillium strain produced nitrate in test media, but its nitrate formation when added to acid soils was poor. These findings indicate that in the investigated soil heterotrophs are of minor importance in the oxidation of atmospheric ammonium.     

Fungi of a forest soil nitrifying at low pH values

Abstract No autotrophic nitrifying organisms were found in a podzolic brown earth forming nitrate. 350 fungi and aerobic heterotrophic bacteria were isolated from this soil and examined for their nitrifying abilities. About one quarter of the isolates produced 0.05–0.90 mg N·1⁻¹ nitrite or nitrate in peptone solution, soil extract mixture or sterilised soil. The nitrification rate of the most active fungus, Verticillium lecanii , was highest at pH 3.5 in defined media. The results support the significance of heterotrophic nitrification in acid soils.     

Nitrification at Low pH by Aggregated Chemolithotrophic Bacteria

A study was performed to gain insight into the mechanism of acid-tolerant, chemolithotrophic nitrification. Microorganisms that nitrified at pH 4 were enriched from two Dutch acid soils. Nitrate production in the enrichment cultures was indicated to be of a chemolithoautotrophic nature as it was (i) completely inhibited by acetylene at a concentration as low as 1 μmol/liter and (ii) strongly retarded under conditions of carbon dioxide limitation. Electron microscopy of the enrichment cultures showed the presence of bacteria that were morphologically similar to strains of known chemolithotrophic nitrifying genera. Many of the enriched bacteria, in particular those that were identified as ammonium oxidizers, were aggregated. Filtration experiments indicated that aggregated cells were able to nitrify at low pH, whereas single cells were not. It is hypothesized that cells inside the aggregates are protected against the toxicity of nitrous acid. Nitrification by aggregated chemolithoautotrophic bacteria may be the dominating process of nitrate formation in many acid soils as it does not appear to depend on the existence of microsites of high pH (acid-sensitive autotrophic nitrification) or on the availability of organic carbon (heterotrophic nitrification).     

High-rate nitrification at low pH in suspended- and attached-biomass reactors

This article reports on high-rate nitrification at low pH in biofilm and suspended-biomass reactors by known chemolithotrophic bacteria. In the biofilm reactor, at low pH (4.3 +/- 0.1) and low bulk ammonium concentrations (9.3 +/- 3.3 mg.liter(-1)), a very high nitrification rate of 5.6 g of N oxidized.liter(-1).day(-1) was achieved. The specific nitrification rate (0.55 g of N.g of biomass(-1).day(-1)) was similar to values reported for nitrifying reactors at optimal pH. In the suspended-biomass reactor, the average pH was significantly lower than that in the biofilm reactor (pH 3.8 +/- 0.3), and values as low as pH 3.2 were found. In addition, measurements in the suspended-biomass reactor, using isotope-labeled ammonium (15N), showed that in spite of the very low pH, biomass growth occurred with a yield of 0.1 g of biomass.g of N oxidized(-1). Fluorescence in situ hybridization using existing rRNA-targeted oligonucleotide probes showed that the nitrifying bacteria were from the monophyletic genus Nitrosomonas, suggesting that autotrophic nitrification at low pH is more widespread than previously thought. The results presented in this paper clearly show that autotrophic nitrifying bacteria have the ability to nitrify at a high rate at low pH and in the presence of only a negligible free ammonia concentration, suggesting the presence of an efficient ammonium uptake system and the means to cope with low pH.     

Nickel-hyperaccumulating Plants Provide a Niche for Nickel-resistant Bacteria

The occurrence and abundance of nickel-resistant bacteria was studied in various soils originating from serpentine and other ultramafic rocks from both temperate and tropical localities. Samples were taken from soil covered with herbs and shrubs not known to accumulate nickel and from areas inhabited by nickel-hyperaccumulating shrubs and trees. In samples taken from the humus and mineral soil layers underneath the canopy of nickel-hyperaccumulating trees, the number of bacteria able to grow at 3, 10 and 20 mM NiCl₂, was almost as high as the number of bacteria growing in the absence of nickel ions. In contrast, for soil samples taken outside these areas only moderately nickel-resistant (3 mM NiCl₂) bacteria and low numbers were encountered. It is concluded that, in humid climate zones, the predominance of nickel-resistant bacteria in the soil is not due to the nickel ions generated by the weathering of nickel-containing rocks but is due to nickel salts being continuously liberated by the decay of nickel-rich leaf litter. Nickel-hyperaccumulating plants can be considered to drive a nickel cycle resulting in the continuous percolation of the top soil by nickel ions.     

Nitrification activities and the changes in the populations of nitrifying bacteria in soil perfused at two different H-ion concentrations

Summary The rate of nitrification by soil aggregates at pH 5.5 and 7.5 was examined by a perfusion technique. Nitrification occurred at both levels of H-ion concentration but at the higher pH the rate of nitrification was greater. The population estimates of nitrifying bacteria were correspondingly greater at the high pH. Once the pH was lowered from 7.5 to 5.5, the nitrification rate decreased slowly with a corresponding decrease in the numbers of nitrifying bacteria. The distribution of nitrifying bacteria throughout soil aggregates was homogenous. The lower limit of pH for nitrification was 4.3.     

Nitrification of ammonium sulphate and urea in an acid red yellow podzolic tea soil in Sri Lanka in relation to soil fertility

Summary Two laboratory incubation experiments were carried out to study the rate of nitrification of ammonium sulphate and urea in an acid red yellow podzolic tea soil. One experiment was with a soil (A) collected from under a good cover of high yielding clonal tea and the other with a soil (B) collected from a bare plot adjoining the tea field. Soil pH was adjusted with calcium carbonate or calcium oxide. Nitrification was appreciable in soil A whereas it was very low in soil B. Elevation of pH substantially increased nitrification in soil A, slightly increased and even depressed nitrification, in certain treatments, in soil B. The difference in the rates of nitrification in the two soils is discussed in relation to soil fertility.     

A comparison of some populations of Eucalyptus viminalis Labill. growing on calcareous and acid soils in Victoria,Australia

A study of populations of Eucalyptus viminalis found on both acid and alkaline soils showed that seedlings differ in their tolerance of calcareous soils and their susceptibility to lime chlorosis. Seedlings from an open-forest population on calcareous dunes in the Otway Ranges, Victoria, averaged a significantly greater yield than seedlings of three other populations found on acid soils when all were grown on calcareous soil of pH 6.8. A tall ribbon gum form of E. viminalis, from Paradise gully in the Otways, was the least tolerant of alkaline conditions and showed severe signs of chlorosis and reddening of leaf margins. The latter appeared to be related to high uptake of phosphorus. This population showed rapid growth on a fertile acid loam. The data presented also emphasize the need to consider the variation in soils which can occur at any one site. Population differences were most marked when seedlings were grown on soils of pH near 7.0. Yield was reduced in the Otways calcareous population when seedlings were grown on the more leached soil from swales (pH 6.5) between dunes and on the highly calcareous soil from the dune crests (pH 7.8–8.0). The failure of any of the populations of E. viminalis studied to grow well on soils of high pH suggest that this species is not tolerant of highly calcareous soils. This may in part explain the absence of this species from the drier calcareous areas on the Mornington Peninsula and the Yanakie Isthmus, Victoria. Since seedlings had reduced root development on the soil collected from the Peninsula, E. viminalis may be restricted, in effect because of drought stress, to non-calcareous sites in the eastern part of Victoria.     

Urease gene‐containing Archaea dominate autotrophic ammonia oxidation in two acid soils

The metabolic traits of ammonia‐oxidizing archaea (AOA) and bacteria (AOB) interacting with their environment determine the nitrogen cycle at the global scale. Ureolytic metabolism has long been proposed as a mechanism for AOB to cope with substrate paucity in acid soil, but it remains unclear whether urea hydrolysis could afford AOA greater ecological advantages. By combining DNA‐based stable isotope probing (SIP) and high‐throughput pyrosequencing, here we show that autotrophic ammonia oxidation in two acid soils was predominately driven by AOA that contain ureC genes encoding the alpha subunit of a putative archaeal urease. In urea‐amended SIP microcosms of forest soil (pH 5.40) and tea orchard soil (pH 3.75), nitrification activity was stimulated significantly by urea fertilization when compared with water‐amended soils in which nitrification resulted solely from the oxidation of ammonia generated through mineralization of soil organic nitrogen. The stimulated activity was paralleled by changes in abundance and composition of archaeal amoA genes. Time‐course incubations indicated that archaeal amoA genes were increasingly labelled by ¹³CO₂ in both microcosms amended with water and urea. Pyrosequencing revealed that archaeal populations were labelled to a much greater extent in soils amended with urea than water. Furthermore, archaeal ureC genes were successfully amplified in the ¹³C‐DNA, and acetylene inhibition suggests that autotrophic growth of urease‐containing AOA depended on energy generation through ammonia oxidation. The sequences of AOB were not detected, and active AOA were affiliated with the marine Group 1.1a‐associated lineage. The results suggest that ureolytic N metabolism could afford AOA greater advantages for autotrophic ammonia oxidation in acid soil, but the mechanism of how urea activates AOA cells remains unclear.     

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。     

毛竹林和阔叶林凋落物互置对土壤氮矿化的影响及微生物贡献

凋落物输入方式的改变导致凋落物数量和质量发生变化,进而对森林土壤氮矿化产生影响。选择未被入侵的次生阔叶林和毛竹入侵后形成的毛竹纯林为对象,对地表凋落物进行保留、去除与置换处理,采用室内培养法同时添加抗生素(链霉素和放线菌酮)分析真菌和细菌在土壤氮素矿化中的贡献。结果表明：(1)去除凋落物处理使阔叶林土壤净氨化速率增加27.0%,净硝化速率降低11.4%;毛竹林土壤净氨化速率增加23.4%。(2)置换凋落物处理使阔叶林土壤净氨化速率增加43.8%,净硝化速率降低33.5%;毛竹林土壤净硝化速率增加73.1%。(3)添加抗生素后,凋落物置换处理与对照相比,置换凋落物后阔叶林土壤真菌和细菌在净氨化中发挥主要作用;真菌在两种林分土壤净硝化中发挥主要作用,细菌在阔叶林土壤净硝化中发挥主要作用。(4)结合测定的凋落物化学性质可知,置换凋落物后引起真菌和细菌在土壤氮素矿化中贡献发生变化,是由于输入凋落物中木质素和纤维素含量的变化。综上,凋落物去除和置换改变了土壤氮素矿化速率,置换凋落物后改变了真菌和细菌对土壤氮素矿化的贡献。解析凋落物质量在土壤氮素矿化中的作用及微生物群落的相对贡献,有助于阐明毛竹入...     

Influence of tree species and ground vegetation on nitrification in an acid forest soil

Soil N transformations were studied at Ironhill, near Liphook, UK as part of a forest fumigation experiment. Nitrification potential was measured in a humoferric podzol soil, of pH 3 (in 0.01 M CaCl₂). An initial experiment into nitrogen mineralisation potential indicated that nitrification was linked strongly to the species of coniferous tree growing in the soil. Transfer of soil solution between soils had no influence on mineralisation potential and allelopathic effects of the trees were not demonstrated. The initial finding was attributed subsequently to the type of ground vegetation and its management. Attempts to reproduce soil conditions, which promoted nitrification, were partially successful.Soil, from the Ironhill site, was incubated with various nitrogenous substrates and other nutrients and sources of carbon to test whether heterotrophs were responsible for nitrification. Organic N (which was ammonified) promoted nitrification, but the addition of ammonium was inhibitory unless supplied with a readily available carbon source such as acetate. Nitrification potential was unaffected when soils were incubated with an inhibitor of autotrophic nitrification. The results of these experiments supported strongly the hypothesis that heterotrophic organisms were responsible for nitrification in this soil.     

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.     

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 β-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.     

Model of ammonia volatilization from calcareous soils

A quantitative model of ammonia volatilization from the calcareous soil uppermost 1-cm layer was developed and tested. The model accounts for the following processes: ammonium-ammonia equilibration in the soil solution, cation exchange between calcium and ammonium which results in ammonium distribution between soil liquid and solid phases, nitrification of dissolved ammonium, distribution of ammonia between liquid and gaseous phases and diffusion of gaseous ammonia in the soil air. The combined effect of various characteristics such as soil pH, cation exchange capacity, water capacity and nitrification rate on ammonia losses from various soil types have been studied. The model was validated against experimental results of ammonia losses from different soils for its use as a predicting tool. The model shows that most of ammonia losses can be explained by the interactive effect of high soil pH and low cation exchange capacity. Computations show increased ammonia volatilization with decreasing soil water capacity. Increasing fertilizer application rate has a small effect on percentage of ammonia losses. Increased nitrification rate and shorter “lag” period of nitrification reduce ammonia losses considerably. Good agreement was obtained between model calculations and experimental results of ammonia volatilization from 13 soils.     

Nitrification and nitrous oxide production potentials in aerobic soil samples from the soil profile of a Finnish coniferous site receiving high ammonium deposition

Abstract Using aerobic soil slurry technique nitrification and nitrous oxide production were studied in samples from a pine site in Western Finland. The site received atmospheric ammonium deposition of 7–33 kg N ha⁻¹ a⁻¹ from a mink farm. The experiments with soil slurries showed that the nitrification potential in the litter layer was higher at pH 6 than at pH 4. However, the nitrification potentials in the samples from the organic and mineral horizons at pH 6 and 4 were almost equal. Also N₂O was produced at a higher rate at pH 6 than at pH 4 in slurries of the litter layer samples. The reverse was true for samples from the organic and mineral horizons. The highest N₂O production and nitrification rates were measured in the suspensions of litter layer samples. Nitrification activity in field-moist soil samples was lower than the activity in the slurries indicating that the availability of ammonium limited nitrification in these soils. Acetylene (2.5 kPa) retarded nitrification activity (70-–100%) and N₂O production (40 – 90%) in soil slurries. Acetylene inhibited the N₂O production by 40–60% during the first 3 days after its addition to field-moist samples incubated in aerobic atmosphere. After 3 days the inhibition became much lower (4–5%). The results indicate that, in soil profiles of boreal coniferous forests receiving ammonium deposition, chemolithotrophic nitrification may have importance in the N₂O production, and that changes in soil pH affect differently nitrification as well as N₂O production in litter and deeper soil layers.     

Low Nitrification Rates in Acid Scots Pine Forest Soils Are Due to pH-Related Factors

In a previous study, ammonia-oxidizing bacteria (AOB)-like sequences were detected in the fragmentation layer of acid Scots pine (Pinus sylvestris L.) forest soils (pH 2.9–3.4) with high nitrification rates (>11.0 μg g⁻¹ dry soil week⁻¹), but were not detected in soils with low nitrification rates (<0.5 μg g⁻¹ dry soil week⁻¹). In the present study, we investigated whether this low nitrification rate has a biotic cause (complete absence of AOB) or an abiotic cause (unfavorable environmental conditions). Therefore, two soils strongly differing in net nitrification were compared: one soil with a low nitrification rate (location Schoorl) and another soil with a high nitrification rate (location Wekerom) were subjected to liming and/or ammonium amendment treatments. Nitrification was assessed by analysis of dynamics in NH₄ ⁺-N and NO₃ ⁻-N concentrations, whereas the presence and composition of AOB communities were assessed by polymerase chain reaction–denaturing gradient gel electrophoresis and sequencing of the ammonia monooxygenase (amoA) gene. Liming, rather than ammonium amendment, stimulated the growth of AOB and their nitrifying activity in Schoorl soil. The retrieved amoA sequences from limed (without and with N amendment) Schoorl and Wekerom soils exclusively belong to Nitrosospira cluster 2. Our study suggests that low nitrification rates in acidic Scots pine forest soils are due to pH-related factors. Nitrosospira cluster 2 detected in these soils is presumably a urease-positive cluster type of AOB.     

High abundance of ammonia-oxidizing archaea in acidified subtropical forest soils in southern China after long-term N deposition

Nitrification has been believed to be performed only by autotrophic ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) until the recent discovery of ammonia-oxidizing archaea (AOA). Meanwhile, it has been questioned whether AOB are significantly responsible for NH(3) oxidation in acidic forest soils. Here, we investigated nitrifying communities and their activity in highly acidified soils of three subtropical forests in southern China that had received chronic high atmospheric N deposition. Nitrifying communities were analyzed using PCR- and culture (most probable number)-based approaches. Nitrification activity was analyzed by measuring gross soil nitrification rates using a (15) N isotope dilution technique. AOB were not detected in the three forest soils: neither via PCR of 16S rRNA and ammonia monooxygenase (amoA) genes nor via culture-based approaches. In contrast, an extraordinary abundance of the putative archaeal amoA was detected (3.2?×?10(8) -1.2?×?10(9) g?soil(-1) ). Moreover, this abundance was correlated with gross soil nitrification rates. This indicates that amoA-possessing archaea rather than bacteria were predominantly responsible for nitrification of the soils. Furthermore, sequences of the genus Nitrospira, a dominant group of soil NOB, were detected. Thus, nitrification of acidified subtropical forest soils in southern China could be performed by a combination of AOA and NOB.     

A Most Probable Number method (MPN) for the estimation of cell numbers of heterotrophic nitrifying bacteria in soil

A Most Probable Number (MPN) method was developed allowing for the first time estimation of populations of bacteria capable of heterotrophic nitrification. The method was applied to an acidic soil of a coniferous forest exhibiting nitrate production. In this soil nitrate production was unlikely to be catalyzed by autotrophic nitrifiers, since autotrophic ammonia oxidizers never could be detected, and autotrophic nitrite oxidizers were usually not found in appreciable cell numbers. The developed MPN method is based on the demonstration of the presence/absence of nitrite/nitrate produced by heterotrophic nitrifying bacteria during growth in a complex medium (peptone-meat-extract softagar medium) containing low concentrations of agar (0.1%). Both the supply of the growing cultures in MPN test tubes with sufficient oxygen and the presence of low agar concentrations in the medium were found to be favourable for sustainable nitrite/nitrate production. The results demonstrate that in the acidic forest soil the microbial population capable of heterotrophic nitrifcation represents a significant part of the total aerobic heterotrophic population. By applying the developed MPN method, several bacterial strains of different genera not previously described to perform heterotrophic nitrification have been isolated from the soil and have been identified by bacterio-diagnostic tests.