The effect of atmospheric SO2 pollution on the microflora of forest soils

The existing knowledge of the effects of industrial SO₂ immissions on forest soil microflora is reviewed. Most Czechoslovak data were obtained in heavily polluted spruce stands in the Ore Mountain and in the Slavkov Forest (NW Bohemia). The industrial SO₂ immissions soil. Pseudomonads yield to yellow-and red-pigmented microorganisms, among micromycetes there is a higher incidence of the generaRhizopus andMucor. The biochemical capabilities of the bacterial populations are reviewed: the immissions have a negative effect on the occurrence and efficiency of heterotrophic nitrifiers, increase the concentration of autotrophic as well as heterotrophic oxidizers of S⁰ and of sulfite-resistant bacteria. The soil activities of the C and N cycles are inhibited whereas the oxidation of S⁰ is stimulated. Changes in the soil of spruce stands are probably due to intoxication with sulfur dioxide (and possibly with sulfite) rather than to acidification. Replacement of withered spruce stands with mountain ash brings about a dramatic improvement of the soil microflora. Presented at the16th Congress of the Czechoslovak Microbiological Society, Banská Bystrica, October 21–23, 1983.     

Heterotrophic nitrification in add forest soils

Ninety cultures of heterotrophic organisms were isolated from soils of four acid Norwegian forest sites, which were active in nitrifying. The isolates were tested for ability to form nitrite in a glucose-ammonium-inorganic salts medium. Eleven cultures were found capable of oxidizing ammonium to nitrite. The nitrifying organisms consist of 4 bacteria and 7 fungi.     

Biochemical activities of soil microflora in SO2 polluted forest stands

The microbial colonization and soil biochemical activities were followed on the Ore Mountain tops in NW Bohemia for 1–2 years in a residual young spruce stand, grass-covered withered spruce stand and grass-covered mountain-ash and birch stands. In grass-covered stands the soil pH value spontaneously increased. Concentrations of heterotrophic bacteria were higher by one or two orders of magnitude than those in a spruce stand without ground vegetation; soil respiration, ammonification and nitrification increased. Bacterial communities of the fermentation horizon were better equipped with degradation and mineralization activities than communities of the spruce stand. These results are in contradiction to the assumption that SO₂ mmissions induce intoxication of the soil making microbial life impossible.     

Phosphorus composition of upland soils polluted by long-term atmospheric nitrogen deposition

Atmospheric N deposition can enhance biological P limitation in terrestrial ecosystems and increase the importance of organic P to plants and microorganisms. We used NaOH–EDTA extraction and solution ³¹P NMR spectroscopy to determine the P composition of soils in the Upper Teesdale National Nature Reserve, northern England, an upland region influenced by such deposition for at least 150 years. Three characteristic soil types were sampled on three occasions during an annual cycle: blanket peat (318 mg g^–1 total C, 607 g g^–1 total P, pH 3.9); acid organic soil under grassland (354 mg g^–1 total C, 1190 g g^–1 total P, pH 3.7); calcareous soil under grassland (140 mg g^–1 total C, 649 g g^–1 total P, pH 7.3). Between 58 and 99% of the total P in soil and litter layers was extracted by 0.25 M NaOH + 0.05 M EDTA. Extracts of all soils were dominated by organic P, mainly in the form of orthophosphate monoesters (43–69% extracted P). The two acidic soils also contained large proportions of orthophosphate diesters (6–19% extracted P) and phosphonates (7–16% extracted P), suggesting that these compounds become stabilised at low pH. However, a seasonal trend of increasing orthophosphate monoester-to-diester ratios, most evident in the calcareous grassland soil, indicated the preferential degradation of orthophosphate diesters during the growing season. Orthophosphate was the major inorganic P compound (17–34% extracted P), and all soils contained pyrophosphate (1–5% extracted P). However, orthophosphate determined in the NaOH–EDTA extracts by solution ³¹P NMR spectroscopy was substantially greater than that determined by molybdate colourimetry, suggesting that orthophosphate occurred in complexes with humic compounds that were not detected by conventional procedures. Our results suggest that organisms able to use recalcitrant soil organic P may have a competitive advantage in environments under enhanced atmospheric N deposition.     

Distribution of autotrophic nitrifying bacteria in a polluted river (the Passaic).

The abundance of nitrifying bacteria, determined by most-probable-number procedures, within habitats of the Passaic River was as follows: rooted aquatic plants greater than algae approximately equal to rocks greater than sediments greater than greater than water. On the average, NH4+ oxidizers were 540-fold more abundant in the topmost 1 cm of sediment than in the water, and NO2- oxidizers were 250-fold more abundant. The population densities in this surface sediment at two nearby stations, one with a predominantly mineral stream bed and the other an organic ooze, did not differ significantly. Large numbers of nitrifiers were present to a depth of about 5 cm in a mineral sediment core.     

Distribution of autotrophic nitrifying bacteria in a polluted river (the Passaic)

The abundance of nitrifying bacteria, determined by most-probable-number procedures, within habitats of the Passaic River was as follows: rooted aquatic plants greater than algae approximately equal to rocks greater than sediments greater than greater than water. On the average, NH4+ oxidizers were 540-fold more abundant in the topmost 1 cm of sediment than in the water, and NO2- oxidizers were 250-fold more abundant. The population densities in this surface sediment at two nearby stations, one with a predominantly mineral stream bed and the other an organic ooze, did not differ significantly. Large numbers of nitrifiers were present to a depth of about 5 cm in a mineral sediment core.     

Degradation of anthracene by bacteria isolated from oil polluted tropical soils

Four bacteria, identified as Pseudomonas aeruginosa, Alcaligenes eutrophus, Bacillus subtilis and Micrococcus luteus were isolated from crude oil polluted soils using anthracene as the sole carbon and energy source. All the organisms utilized n-hexadecane, n-tetradecane, diesel oil, engine oil and naphthalene as sole carbon sources. None could utilize hexane, cycloheptane, xylene, benzene, toluene, phenol, fluoranthene,and kerosene as carbon sources. Highest cell density obtained with 0.1% (w/v) anthracene were 4.5 x 10(7) (cfu/ml), 8.6 x 10(6) (cfu/ml), 5.4 x 10(6) and 2.4 x 10(6) (cfu/ml) respectively, for P. aeruginosa, A. eutrophus, B. subtilis and M. luteus after 30 days incubation. Growth of the organisms on a Nigerian crude oil resulted in a residual oil concentration of 22.2%, 33.3%, 39.3%, 44% and 91.7% respectively, for P. aeruginosa, A. eutrophus, B. subtilis, M. luteus and the noninoculated control on the 14 th day. Ring fission enzymes of the meta pathway were detected in induced cells of P. aeruginosa and A. eutrophus while ortho pathway enzymes were detected in B. subtilis and M. luteus. P. aeruginosa and A. eutrophus had specific catechol-2,3-dioxygenase activities of 3.8 +/- 0.183 and 0.64 +/- 0.032 micromol/min x mg protein respectively while catechol-1,2-dioxygenase activities of 1.95 +/- 0.029 and 1.89 +/- 0.026 micromol/min x mg protein were detected in B. subtilis and M. luteus respectively. This work, highlights the capability of these unreported tropical strains of A. eutrophus, B. subtilis and M. luteus as anthracene degraders.     

Ureolytic and nitrifying bacterial heterotrophs from acid soils

Nitrification following ureolysis in soil samples from tea growing soils (pH 4.5–5.5) was found to be chiefly due to the activity of heterotrophic bacteria belonging to generaBacillus, Arthrobacter, Sporosarcina, Micrococcus, Clostridium, Pseudomonas andProteus. A correlation between the intensity of ureolytic activity of organisms in a given soil sample and the yield levels of tea was observed. In culture media the increase in the quantity of NH ₄ ⁺ -N indicating ureolysis was not accompanied by proportional increase in biomass. Ureolysis and nitrification in sterile soil sample inoculated with the isolates improved through amendment of organic carbon to the soil.     

Attributes of atmospheric carbon monoxide oxidation by Maine forest soils

CO, one of the most important trace gases, regulates tropospheric methane, hydroxyl radical, and ozone contents. Ten to 25% of the estimated global CO flux may be consumed by soils annually. Depth profiles for (14)CO oxidation and CO concentration indicated that CO oxidation occurred primarily in surface soils and that photooxidation of soil organic matter did not necessarily contribute significantly to CO fluxes. Kinetic analyses revealed that the apparent K(m) was about 18 nM (17 ppm) and the V(max) was 6.9 micromol g (fresh weight)(-1) h(-1); the apparent K(m) was similar to the apparent K(m) for atmospheric methane consumption, but the V(max) was more than 100 times higher. Atmospheric CO oxidation responded sensitively to soil water regimes; decreases in water content in initially saturated soils resulted in increased uptake, and optimum uptake occurred at water contents of 30 to 60%. However, extended drying led to decreased uptake and net CO production. Rewetting could restore CO uptake, albeit with a pronounced hysteresis. The responses to changing temperatures indicated that the optimum temperature for net uptake was between 20 and 25 degrees C and that there was a transition to net production at temperatures above 30 degrees C. The responses to methyl fluoride and acetylene indicated that populations other than ammonia oxidizers and methanotrophs must be involved in forest soils. The response to acetylene was notable, since the strong initial inhibition was reversed after 12 h of incubation; in contrast, methyl fluoride did not have an inhibitory effect. Ammonium did not inhibit CO uptake; the level of nitrite inhibition was initially substantial, but nitrite inhibition was reversible over time. Nitrite inhibition appeared to occur through indirect effects based on abiological formation of NO.     

Nitrification and occurrence of Nitrobacter by MPN-PCR in low and high nitrifying coniferous forest soils

The nitrification of three coniferous forest soils was investigated: a podzol (Fontainebleau, France) with low N deposition and no nitrate accumulation, an acid mull (Haldon, UK) with low N deposition and moderate nitrate accumulation, and a podzol (Wekerom, The Netherlands) with high N deposition and high nitrate accumulation. Twenty-one months in situ lysimeter experiments and short-term and long-term in vitro incubations were performed to respectively establish the status of NO ₃ ^- accumulation and potential nitrification of these soils. These complementary approaches allowed to conclude that the absence of NO ₃ ^- accumulation in Fontainebleau soil was effectively due to a lack of nitrifing activity in the whole profile while the high NO ₃ ^- accumulation observed in the Wekerom soil essentially resulted from an active nitrifying activity in the Oh horizon of this soil. For the Haldon soil, the inadequacy between the lysimeter and the short-term in vitro experiment was discussed.Surprisingly, relatively high densities of the nitrite-oxidiser Nitrobacter (enumerated by PCR-MPN technique) were found in all the studied horizons of the three soils. Moreover, a long-term incubation experiment showed that inhibition of nitrification in the Oh Fontainebleau soil could be removed after submitting the soil samples to constant conditions during 82 days. This demonstrates that in these soils, ammonium-oxidisers (as well as nitrite-oxidisers) are present and that extrinsic factors were involved in the inhibition of nitrification.     

Nitrification and occurrence of salt-tolerant nitrifying bacteria in the Negev desert soils

Ammonia oxidation potential, major ammonia oxidizers and occurrence of salt-tolerant nitrifying bacteria were studied in soil samples collected from diverse ecosystems along the northern Negev desert. Great diversity in ammonia oxidation potential was observed among the soil samples, and ammonia oxidizers were the rate-limiting step of nitrification. Denaturing gradient gel electrophoresis and partial 16S rRNA gene sequences indicate that members of the genus Nitrosospira are the major ammonia oxidizers in the natural desert soil samples. Upon enrichment with different salt concentrations, salt-tolerant nitrifying enrichments were established from several soil samples. In two enrichments, nitrification was not inhibited by 400 mM NaCl. Electrophoretic analysis and partial 16S rRNA gene sequences indicate that Nitrosomonas species were dominant in the 400 mM salt enrichment. The results point towards the potential of the desert ecosystem as a source of stress-tolerant nitrifying bacteria or other microorganisms with important properties.     

Molecular analysis of the diversity of nitrifying bacteria in the soils of the forest and steppe zones of European Russia

Nitrifying bacteria play a key role in the global nitrogen cycle due to their ability to convert reduced nitrogen compounds (ammonium) to oxidized ones (nitrite and nitrate). Recent investigations based on the methods of molecular ecology revealed that bacteria are responsible for nitrification in natural ecosystems. At the same time, data on the species composition of the nitrifiers in soil microbial communities are scarce. Soil samples collected in the forest and steppe areas of European Russia and the enrichment cultures of nitrifying bacteria isolated from these samples were used for molecular studies of the diversity of the amoA gene encoding the synthesis of the key enzyme of autotrophic ammonium oxidation. The nitrifying bacteria of the genera Nitrosospira and Nitrosovibrio were found in all the studied soils from natural biocenoses and agrocenoses.     

Presence of Nitrosospira cluster 2 bacteria corresponds to N transformation rates in nine acid Scots pine forest soils

The relation between environmental factors and the presence of ammonia-oxidising bacteria (AOB), and its consequences for the N transformation rates were investigated in nine Scots pine (Pinus sylvestris L.) forest soils. In general, the diversity in AOB appears to be strikingly low compared to other ecosystems. Nitrosospira cluster 2, as determined by temporal temperature gradient electrophoresis and sequencing, was the only sequence cluster detected in the five soils with high nitrification rates. In the four soils with low nitrification rates, AOB-like sequences could not be detected. Differences in nitrification rates between the forest soils correlated to soil C/N ratio (or total N) and atmospheric N deposition.     

Seasonal dynamics of atmospheric methane oxidation in gray forest soils

Seasonal fluctuations in the methane flow in the soil-atmosphere system were determined for gray forest soils of Central Russia. Consumption of atmospheric methane was found to exceed methane emission in gray forest soils under forest and in agrocenosis. The average annual rates of atmospheric methane consumption by the soil under forest and in agrocenosis were 0.026 and 0.008 mg CH4-C/(m2 h), respectively. The annual rate of atmospheric methane oxidation in the gray forest soils of Moscow oblast was estimated to be 0.68 kton. Seasonal fluctuations in the methane oxidation activity were due to changes in the hydrothermal conditions and in the reserves of readily decomposable organic matter and mineral nitrogen, as well as to changes in the activity of methane oxidizers.     

Ecophysiological interaction between nitrifying bacteria and heterotrophic bacteria in autotrophic nitrifying biofilms as determined by microautoradiography-fluorescence in situ hybridization

Ecophysiological interactions between the community members (i.e., nitrifiers and heterotrophic bacteria) in a carbon-limited autotrophic nitrifying biofilm fed only NH(4)(+) as an energy source were investigated by using a full-cycle 16S rRNA approach followed by microautoradiography (MAR)-fluorescence in situ hybridization (FISH). Phylogenetic differentiation (identification) of heterotrophic bacteria was performed by 16S rRNA gene sequence analysis, and FISH probes were designed to determine the community structure and the spatial organization (i.e., niche differentiation) in the biofilm. FISH analysis showed that this autotrophic nitrifying biofilm was composed of 50% nitrifying bacteria (ammonia-oxidizing bacteria [AOB] and nitrite-oxidizing bacteria [NOB]) and 50% heterotrophic bacteria, and the distribution was as follows: members of the alpha subclass of the class Proteobacteria (alpha-Proteobacteria), 23%; gamma-Proteobacteria, 13%; green nonsulfur bacteria (GNSB), 9%; Cytophaga-Flavobacterium-Bacteroides (CFB) division, 2%; and unidentified (organisms that could not be hybridized with any probe except EUB338), 3%. These results indicated that a pair of nitrifiers (AOB and NOB) supported a heterotrophic bacterium via production of soluble microbial products (SMP). MAR-FISH revealed that the heterotrophic bacterial community was composed of bacteria that were phylogenetically and metabolically diverse and to some extent metabolically redundant, which ensured the stability of the ecosystem as a biofilm. alpha- and gamma-Proteobacteria dominated the utilization of [(14)C]acetic acid and (14)C-amino acids in this biofilm. Despite their low abundance (ca. 2%) in the biofilm community, members of the CFB cluster accounted for the largest fraction (ca. 64%) of the bacterial community consuming N-acetyl-D-[1-(14)C]glucosamine (NAG). The GNSB accounted for 9% of the (14)C-amino acid-consuming bacteria and 27% of the [(14)C]NAG-consuming bacteria but did not utilize [(14)C]acetic acid. Bacteria classified in the unidentified group accounted for 6% of the total heterotrophic bacteria and could utilize all organic substrates, including NAG. This showed that there was an efficient food web (carbon metabolism) in the autotrophic nitrifying biofilm community, which ensured maximum utilization of SMP produced by nitrifiers and prevented buildup of metabolites or waste materials of nitrifiers to significant levels.     

SO2 and NO2 effects on microbial activity in an acid forest soil

The rate of glucose decomposition and the pH fell in a forest soil (initial pH 4.06) exposed to 1.0 ppm SO₂. No such effect was noted if the soil was exposed to 1.0 ppm nitrogen dioxide (NO₂). Nitrite but not bisulfite (5g N or S/g of soil) inhibited O₂ consumption and CO₂ evolution in the glucose-amended forest soil, and nitrite and bisulfite acted synergistically in inhibiting these processes. Iron and manganese were solubilized when the soil was exposed to 10 ppm SO₂, but NO₂ caused no such change.     

Immobilization of nitrifying bacteria by polyethylene glycol prepolymer

The effects of immobilizing materials on the activity of nitrifying bacteria were investigated by using 11 kinds of prepolymers of polyethylene glycol. Relative respiratory activity of immobilized nitrifying bacteria with polyethylene glycol metacrylate prepolymer was higher than that of polyethylene glycol acrylate prepolymer, and there was a tendency for relative respiratory activity to be higher with a prepolymer of greater molecular weight. With the polyethylene glycol prepolymer, there was a drastic improvement over the conventional method of immobilization by acrylamide in the relative respiratory activity of the pellet. Inorganic synthetic wastewater was treated under a high loading rate of 1.14 kg-N/m³·d. Influent NH₄-N could be removed to 2 mg/l or less and the nitrogen removal was 90%.