Variation in Heterotrophic and Autotrophic Nitrifier Populations in Relation to Nitrification in Organic Soils

The occurrence of heterotrophic and autotrophic nitrifiers in Pahokee muck and the role of these organisms in the ecosystem were assessed by surveying their population densities under different field conditions and by observing the relationship of these populations with aerobic bacteria and soil moisture. Heterotrophic nitrifier populations varied from 2.0 × 10⁵ to 3.8 × 10⁶ bacteria per cm³ of muck in surface fallow (bare) Pahokee muck during the annual cycle. This population decreased 40-fold between the surface and the 60- to 70-cm depths of soil. Similar variations were noted with autotrophic nitrifier populations. Significant correlations were found between heterotrophic nitrifiers and both soil moisture and aerobic bacteria. These relationships did not exist for the autotrophic nitrifiers. In soil that had been heated to kill the autotrophic nitrifiers, while preserving a population of the heterotrophs, and then amended with sodium acetate or ammonium sulfate or both, no nitrate or nitrite accumulated, although significant increases in heterotrophic nitrifiers were detected. In unheated control soil, nitrate plus nitrite-N increased from 14.3 to 181 μg/g of wet soil, and 48 μg of nitrite-N per g was produced. These data suggest that the autotrophic nitrifiers were the sole population responsible for nitrification in Pahokee muck.     

Dynamics of carbon,nitrogen and phosphorus in soil amended with irradiated,pasteurized and limed biosolids

Sewage biosolids contain high concentrations of pathogens, which limits their use as soil amendment. This study investigated how application of lime (Ca(OH)2), irradiation, or pasteurization reduced pathogens in biosolids and how its application affected soil characteristics. A soil sampled outside the canopy of Mesquite trees (Prosopis laevigata) and from a pasture at Lerma (Mexico) was amended with treated or untreated biosolids, characterized and incubated aerobically while dynamics of carbon (C), nitrogen (N) and phosphorus (P) were monitored. Heavy metals concentrations in the biosolids were low, so it was of excellent quality (USEPA). The amount of pathogens in the biosolids made it a class "B" (USEPA) which can be used in forests. Only irradiation sufficiently reduced faecal coliforms to make it a class "A" biosolids without restrictions in application. C mineralization increased significantly when biosolids were added, but not concentrations of available P (P < 0.05). Ammonium (NH4+) concentrations in soil amended with biosolids were higher compared to unamended soil, but not the concentrations of nitrate (NO3-) except when biosolids treated with Ca(OH)2 was added to the Lerma soil.     

Ecologically significant effects of Pseudomonas putida PPO301(pRO103), genetically engineered to degrade 2,4-dichlorophenoxyacetate, on microbial populations and processes in soil

Pseudomonas putida PPO301 (pRO103), genetically engineered to degrade 2,4-dichlorophenoxyacetate, affected microbial populations and processes in a nonsterile xeric soil. In soil amended with 2,4-dichlorophenoxyacetate (500 micrograms/g soil) and inoculated with PPO301 (pRO103), the rate of evolution of carbon dioxide was retarded for approximately 35 days; there was a transient increase in dehydrogenase activity; and the number of fungal propagules decreased below detection after 18 days. In unamended soil inoculated with PPO301(pRO103), the rate of evolution of carbon dioxide and the dehydrogenase activity were unaffected, and the numbers of fungal propagules were reduced by about two orders of magnitude. The numbers of total, spore-forming, and chitin-utilizing bacteria were reduced transiently in soil either amended or unamended with 2,4-dichlorophenoxyacetate and inoculated with PPO301(pRO103). The activities of arylsulfatases and phosphatases in soil were not affected by the presence of PPO301(pRO103), either in the presence or absence of 2,4-dichlorophenoxyacetate. In soil amended with 2,4-dichlorophenoxyacetate and inoculated with the parental strain (PPO301) or not inoculated, the evolution of carbon dioxide, the numbers of fungal propagules and of total, spore-forming, and chitin-utilizing bacteria, and the dehydrogenase activity were not affected as in soil inoculated with PPO301(pRO103). These results demonstrated that a genetically engineered microorganism, in the presence of the substrate on which its novel genes can function, is capable of inducing measurable ecological effects in soil.     

Effects of agronomic application of olive mill wastewater in a field of olive trees on carbohydrate profiles, chlorophyll a fluorescence and mineral nutrient content

Olive mill wastewater (OMW) management is a serious environmental issue for the Mediterranean area where there is the most production of olive oil. OMW contains a high organic load, substantial amounts of plant nutrients but also several compounds with recognized toxicity towards living organisms. Moreover, OMW may represent a low cost source of water. We studied the influence of irrigation with OMW (amounts applied: 30, 60, 100 and 150 m³ h⁻¹) in a field of olive trees on root colonization, photosynthesis, chlorophyll fluorescence, leaf nutrient concentration and soluble carbohydrate. The soil fatty acid methyl ester (FAME) 16:1ω5 was used to quantify biomass of arbuscular mycorrhizal (AM) fungi and the root FAME 16:1ω5 analysis was used as index for the development of colonization in the roots. Agronomic application of OMW decreased significantly the abundance of the soil FAME 16:1ω5 and the root FAME 16:1ω5 in the soil amended with 60, 100 and 150 m³ ha⁻¹ OMW. Decreased root FAME 16:1ω5 due to OMW amendment was associated with a significant reduction of tissue nutrient concentrations in the olive trees. The highest application of OMW to the soil reduced significantly the olive trees uptake of N, P, K, Ca, Mg, Fe, Cu, Mn and Zn. Land spreading of OMW increased concentration of soluble carbohydrate in the olive leaves, mostly due to decreased sink demand for carbon by the root. In the olive trees amended with 150 m³ ha⁻¹ OMW, net CO₂ uptake rate (A), quantum yield of photosystem II electron transport (Φ_PSII), maximal photochemical efficiency of photosystem II (Fv/Fm), photochemical quenching (q_p) and the electron transport rate (ETR) were significantly depressed, whereas non-photochemical quenching (NPQ) was found to increase. Taken with data from experiments in field conditions, our results suggest that agronomic application of OMW alters the functioning of arbuscular mycorrhizas and can even disrupt the relationship between AM fungi and olive trees.     

Increased frequency of drug-resistant bacteria and fecal coliforms in an Indiana Creek adjacent to farmland amended with treated sludge

Many studies indicate the presence of human pathogens and drug-resistant bacteria in treated sewage sludge. Since one of the main methods of treated sewage disposal is by application to agricultural land, the presence of these organisms is of concern to human health. The goal of this study was to determine whether the frequency of drug-resistant and indicator bacteria in Sugar Creek, which is used for recreational purposes, was influenced by proximity to a farmland routinely amended with treated sludge (site E). Surface water from 3 sites along Sugar Creek (site E, 1 upstream site (site C) and 1 downstream site (site K)) were tested for the presence of ampicillin-resistant (Amp(R)) bacteria, fecal and total coliforms over a period of 40 d. Site E consistently had higher frequencies of Amp(R) bacteria and fecal coliforms compared with the other 2 sites. All of the tested Amp(R) isolates were resistant to at least 1 other antibiotic. However, no isolate was resistant to more than 4 classes of antimicrobials. These results suggest that surface runoff from the farmland is strongly correlated with higher incidence of Amp(R) and fecal coliforms at site E.     

Potential nitrification in alum-treated soil slurries amended with poultry manure

Alum is used to reduce environmental pollutants in poultry production. Alum decreases NH3 volatilization and increases total N and NH4+-N compared to untreated poultry manure. Nitrification in poultry wastes could therefore be stimulated due to higher NH4+ concentrations or could be inhibited because the soil environment is acidified. A 10-day laboratory study was conducted to study potential nitrification rates in soil slurries (20 g soil in 150 ml water) amended with 2.0 g alum-treated poultry manure. Fecal bacteria, NH4+, NO2-, NO3-, orthophosphate, pH, and NH3 were measured at 2-day intervals. Alum significantly reduced fecal bacteria concentrations through day 6. Water-soluble P was reduced 82% by day 10. Alum-treated manure had significantly increased NH4+ concentrations by day 8 and 10, and significantly decreased NO2- and NO3- concentrations by days 6-10. Alum's effect on potential nitrification was inhibitory in the soil environment. Slurries with alum-treated poultry manure had reduced nitrification rates, fecal bacteria, and soluble P. Therefore, in addition to reducing P loss, alum could temporarily reduce the risk for environmental pollution from land-applied manures in terms of both NO3- and fecal bacteria loss.     

Effects of olive mill wastewater on soil carbon and nitrogen cycling

This study investigated the cycling of C and N following application of olive mill wastewater (OMW) at various rates (0, 42, 84, and 168 m³/ha). OMW stimulated respiration rate throughout the study period, but an increase in soil organic matter was observed only at the highest rate. Soil phenol content decreased rapidly within 2 weeks following application but neither phenol oxidase and peroxidase activity nor laccase gene copies could explain this response. Soil NH₄ ⁺-N content increased in response to OMW application rate, while an opposite trend observed for NO₃ ^?-N, which attributed to immobilization. This decrease was in accordance with amoA gene copies of archaeal and bacterial ammonia oxidizers in the first days following OMW application. Afterwards, although amoA gene copies and potential nitrification rates recovered to values similar to or higher than those in the non-treated soils, NO₃ ^?-N content did not change among the treatments. A corresponding increase in denitrifying gene copies (nirK, nirS, nosZ) during that period indicates that denitrification, stimulated by OMW application rate, was responsible for this effect; a hypothesis consistent with the decrease in total Kjeldahl nitrogen content late in the season. The findings suggest that land application of OMW is a promising practice for OMW management, even at rates approaching the soil water holding capacity.     

黄瓜连作土壤酚酸类物质积累对土壤微生物和酶活性的影响

伴随连作年限的增加,日光温室黄瓜连作土壤中酚酸类物质(对羟基苯甲酸、阿魏酸、苯甲酸)明显积累,连作5～9年的土壤酚酸类物质含量显著高于连作1～3年的土壤.伴随外源酚酸类物质处理浓度的增加,黄瓜根区土壤中细菌、放线菌和微生物总量以及N生理群均呈先升后降趋势,在80 μg·g^-1处理浓度下细菌、放线菌数量最多,处理浓度在120μg·g^-1以下土壤真菌数量(包括尖孢镰刀菌、疫霉)急剧增长;多酚氧化酶、过氧化氢酶、蔗糖酶、脲酶和蛋白酶活性也同样呈先升后降趋势,但其峰值对应的浓度不同.     

Potential Regrowth and Recolonization of Salmonellae and Indicators in Biosolids and Biosolid-Amended Soil

This study evaluated the potential for conversion of Class B to Class A biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete lined drying beds. Anaerobically (8% solids) and aerobically (2% solids) digested Class B biosolids were pumped into field-scale drying beds, and microbial populations and environmental conditions were monitored. Numbers of fecal coliforms and salmonellae decreased as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A requirements were achieved in both biosolids for the pathogens and the indicators. However, following rainfall events, significant increase in numbers was observed for both fecal coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms was observed in both biosolids and biosolid-amended soil, but the regrowth of salmonellae observed in the concrete-lined drying beds did not occur. These laboratory studies demonstrated that pathogens decreased in numbers when soil was amended with biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete lined drying beds following rainfall events was most likely due to recolonization due to contamination from fecal matter introduced by animals and not from regrowth of salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined beds created a situation in which moisture added as rainfall accumulated in the beds, promoting the growth of fecal coliforms and salmonellae added from external sources.     

Potential regrowth and recolonization of salmonellae and indicators in biosolids and biosolid-amended soil

This study evaluated the potential for conversion of Class B to Class A biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete lined drying beds. Anaerobically (8% solids) and aerobically (2% solids) digested Class B biosolids were pumped into field-scale drying beds, and microbial populations and environmental conditions were monitored. Numbers of fecal coliforms and salmonellae decreased as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A requirements were achieved in both biosolids for the pathogens and the indicators. However, following rainfall events, significant increase in numbers was observed for both fecal coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms was observed in both biosolids and biosolid-amended soil, but the regrowth of salmonellae observed in the concrete-lined drying beds did not occur. These laboratory studies demonstrated that pathogens decreased in numbers when soil was amended with biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete lined drying beds following rainfall events was most likely due to recolonization due to contamination from fecal matter introduced by animals and not from regrowth of salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined beds created a situation in which moisture added as rainfall accumulated in the beds, promoting the growth of fecal coliforms and salmonellae added from external sources.     

Inorganic materials as ameliorants for soil remediation of metal toxicity to wild mustard (Sinapis arvensis L.)

The ameliorating effects of different inorganic materials were investigated on a soil originating from a zinc smelter dumping site contaminated by toxic metals. Wild mustard (Sinapis arvensis L.) was used as a test plant. The soil was amended with different doses of mining sludge, Perferric Red Latosol (LVj), steel shots, cyclonic ash, silifertil, and superphosphate. The most effective amendments improved plant growth with 45% and reduced metal uptake by over 70% in comparison to untreated soil. Reductions in availability as estimated by BaCl2-extractable metals reached up to 90% for Zn and 65% for Cd as compared to unamended soil. These reductions were associated with lower shoot and root metal contents. Shoot Zn content was reduced from 1,369 microg g(-1) in plants grown on untreated soil to 377 microg g(-1) when grown on cyclonic ash amended soil while Cd decreased from 267 to 44 microg g(-1) in steel shots amended soil. Superphosphate addition had no ameliorating effect. On the contrary, it increased BaCl2-extractable amounts of Zn. Considering all parameters we determined, steel shots, cyclonic ash and silifertil are the most promising for remediating metal contaminated soil in the tropics. Further studies evaluating impacts, cost-effectiveness and durability of effects will be conducted.     

Effect of continuous olive mill wastewater applications, in the presence and absence of nitrogen fertilization, on the structure of rhizosphere–soil fungal communities

Olive mill wastewater (OMW) is rich in potentially toxic organics precluding its disposal into water receptors. However, land application of diluted OMW may result in safe disposal and fertilization. In order to investigate the effects of OMW on the structure of soil fungal groups, OMW was applied daily to pepper plants growing in a loamy sand and a sandy loam at two doses for a period of 3 months (total OMW equivalents 900 and 1800 m³ ha⁻¹). Nitrogen (N) fertilization alleviated N scarcity and considerably enhanced plant biomass production; however, when applied in combination with the high OMW dose, it induced plant stress. OMW applications resulted in marked changes in the denaturing gradient gel electrophoresis patterns of soil basidiomycete communities, while concurrent N fertilization reduced these effects. In contrast, the ascomycete communities required N fertilization to respond to OMW addition. Cloning libraries for the basidiomycete communities showed that Cryptococcus yeasts and Ceratobasidium spp. dominated in the samples treated with OMW. In contrast, certain plant pathogenic basidiomycetes such as Thanatephorus cucumeris and Athelia rolfsii were suppressed. The observed changes may be reasonably explained by the capacity of OMW to enrich soils in organic substrates, to induce N immobilization and to directly introduce OMW-derived basidiomycetous yeasts.     

Comparative effect of different aerobic pretreatments on the kinetics and macroenergetic parameters of anaerobic digestion of olive mill wastewater in continuous mode

A comparative kinetic study was carried out on the anaerobic digestion of olive mill wastewater (OMW) and OMW that was previously fermented with Geotrichum candidum, Azotobacter chroococcum and Aspergillus terreus. The reactors used were continuously fed and contained sepiolite as support for the mediating bacteria. A kinetic model for multicomponent substrate removal by anaerobic digestion has been used. The model is based on the linear removal concept which is a special case of the broader Monod equation. The second-order kinetic constant, k _{2( s )}, was found to be influenced by the pretreatment carried out, and was 4.2, 4.0 and 2.5 times higher for Aspergillus, Azotobacter and Geotrichum-pretreated OMWs than that obtained in the anaerobic digestion of untreated OMW. This was significant at 95% confidence level. This behaviour is believed to be due to the lower levels of phenolic compounds and biotoxicity present in the pretreated OMWs. In fact, the kinetic constant increased when the phenolic compound content and biotoxicity of the pretreated OMWs decreased. In addition, the macroenergetic parameters of the anaerobic digestion of OMW, i.e. the specific rate of substrate uptake for cell maintenance, m, and the yield coefficient for the biomass, Y, decreased by a factor of 2.4, 3.6 and 5.1 and increased by a factor of 1.9, 2.2 and 2.4 respectively, for the OMWs previously treated with Geotrichum candidum, Azotobacter chroococcum and Aspergillus terreus in relation to the observed values for the untreated OMW.     

土壤中毒死蜱和微生物相互作用的研究

实验结果表明,50、500、5000g·kg^-1的毒死蜱在灭菌土中的降解速度十分缓慢,60d后,仅降解20％左右。在施过毒死蜱的土壤中的降解速度非常迅速,60d后,50mg·kg^-1的降解率接近100％,500mg·kg^-1的降解率达到90％,5000g·kg^-1的降解率在80％左右。在未灭菌但未施过毒死蜱的土壤中的降解速度则介于二者之间．在各处理的土壤,50和500mg·kg^-1的毒死蜱在处理前期对真菌和细菌数量有一定的刺激作用,60d后,基本恢复到正常水平,而5000mg·kg^-1的毒死蜱则对真菌和细菌的数量有较强的抑制作用。60d后仍不能恢复,在同样处理条件下,施过毒死蜱土壤中的真菌和细菌数量并不比未施过毒死蜱土壤中的占优势,表明毒死蜱在施过药的土壤中降解速度的加快并不是由细菌和真菌的数量决定的,而是由它们的降解能力决定的。     

Microbial populations, ammonification and nitrification in soil treated with urea and inorganic salts

Soil fertilization with urea at rates of 0.2 and 0.5 mg N/g soil was toxic for total counts of bacteria and fungi except with cellulolytic fungi where growth was promoted by addition of urea after 90-d incubation. Also, the population numbers of both bacteria and fungi were significantly decreased when soil was amended with CaCl₂ and K₂SO₄ applied at two levels (50 and 100 μmol/g soil). Some alleviation of the toxic effect of either urea or the inorganic salts was observed when they were applied in combination. Fungal species composition was found to be affected in the treated soil. The most tolerant fungi wereAlternaria alternata, Aspergillus flavus, A. fumigatus, A. niger, A. terreus, Eurotium amstelodami, E. chevalieri, Nectria hœmatococca, Pœcilomyces variotii andStachybotrys chartarum. Other species of fungi were either sensitive to all treatments or sensitive to some and tolerant to others. Ammonium nitrogen was found to be more accumulated in soil treated with urea mixed with inorganic salts compared with soil treated only with urea whereas NO ₃ ⁻ -N (and NO ₂ ⁻ -N) was decreased. The overall effect of the addition of inorganic salts on total mineralized nitrogen, however was promotive. Soil pH was increased in ureatreated soil but was depressed by application of CaCl₂ or K₂SO₄ to values lower than that of untreated soil. The results may be of agronomical interest in overcoming problems encountered with urea application as N fertilizer.     

Long-term fertilization regimes change soil nitrification potential by impacting active autotrophic ammonia oxidizers and nitrite oxidizers as assessed by DNA stable isotope probing

Chemoautotrophic ammonia-oxidizers and nitrite-oxidizers are responsible for a significant amount of soil nitrate production. The identity and composition of these active nitrifiers in soils under different long-term fertilization regimes remain largely under-investigated. Based on that soil nitrification potential significantly decreased in soils with chemical fertilization (CF) and increased in soils with organic fertilization (OF), a microcosm experiment with DNA stable isotope probing was further conducted to clarify the active nitrifiers. Both ammonia-oxidizing archaea (AOA) and bacteria (AOB) were found to actively respond to urea addition in soils with OF and no fertilizer (CK), whereas only AOB were detected in soils with CF. Around 98% of active AOB were Nitrosospira cluster 3a.1 in all tested soils, and more than 90% of active AOA were Nitrososphaera subcluster 1.1 in unfertilized and organically fertilized soils. Nitrite oxidation was performed only by Nitrospira-like bacteria in all soils. The relative abundances of Nitrospira lineage I and VI were 32% and 61%, respectively, in unfertilized soils, and that of Nitrospira lineage II was 97% in fertilized soils, indicating long-term fertilization shifted the composition of active Nitrospira-like bacteria in response to urea. This finding indicates that different fertilizer regimes impact the composition of active nitrifiers, thus, impacting soil nitrification potential.     

β-Glucan production by Botryosphaeria rhodina on undiluted olive-mill wastewaters

Botryosphaeria rhodina produced beta-glucan when grown on undiluted olive-mill wastewaters (OMW). The production of exopolysaccharide increased with the COD up to 17.2 g l(-1) on the most loaded OMW (151 and 66 g l(-1) of COD and total sugar, respectively). The total phenol content of OMW was reduced from 8 to 4.1 g l(-1).     

Phytoremediation potentials of cowpea (Vigina unguiculata) and maize (Zea mays) for hydrocarbon degradation in organic and inorganic manure-amended tropical typic paleustults

A field study on phytoremediation of hydrocarbon contaminated soil was designed to assess the effects of organic manures (poultry droppings and cassava peels) and NPK fertilization on the potentials of cowpea (Vigina unguiculata) and maize (Zea mays) to stimulate hydrocarbon degradation in soil. Cowpea and maize crops were established on the hydrocarbon contaminated soil amended with three rates (0, 4, and 8 ton/ha) of the soil amendments, and arranged in 3 x 3 x 3 factorial in Randomized Complete Block Design. Hydrocarbon was significantly (P < 0.05) reduced in plots treated with the combined forms of the soil amendments. While the treatment combinations of 8 t/ha Poultry Droppings (PD) + 8 t/ha Cassava Peels (CP) + 4 t/ha NPK fertilizer was optimal for hydrocarbon degradation in the cowpea plots, 4 t/ha PD + 8 t/ha CP + 8 t/ha NPK fertilizer was the most preferred in the maize plot. Cowpea showed greater potential for hydrocarbon degradation at the first year. The mean values of hydrocarbon concentrations at the cowpea and maize plots indicated no significant difference at the second year. Grain yield of cowpea increased by 87% at the second year, while maize was unable to grow to maturity in the first year.     

Persistence and distribution of pollution indicator bacteria on land used for disposal of piggery effluent.

Numbers of pollution indicator bacteria (fecal coliforms and fecal streptococci) were assessed on land to which effluent from intensively housed pigs had been applied. Topsoil (to a 30-mm depth) was found to provide a more favorable environment for fecal coliform persistence than was pasture or subsoil. Times required for a 90% reduction in number (T90) in topsoil (calculated by linear regression of log counts obtained in a 6-week period after effluent application) ranged from 7 to 20 days (mean T90, 11 days). T90 values for fecal coliforms fell within this range irrespective of the season of application and for a number of soil types and climatic conditions. The range in die-off times was encountered irrespective of the fecal coliform count in the applied effluent or the application regimen (125 to 1,000 kg of elemental nitrogen in the form of effluent per ha; return periods, 3 to 12 months). Autumn and winter conditions were conducive to the persistence of a survivor tail of these bacteria at 10(1) to 10(3) cells per g of topsoil. Fecal streptococci survived similarly on soil and pasture (T90, ca. 14 days) and appeared slightly more suited to survival in the environment than did fecal coliforms. Contamination of subsoils after effluent applications occurred at a rate well in excess of the infiltration capacity of the soil, presumably by percolation of the effluent through soil cracks. Contamination levels of subsoils in the experimental area generally remained low.     

Effects of Management Practices on Nematode and Fungus Populations and Cucumber Yield

Three crops of cucumber were grown in succession in beds by use of trickle irrigation, plastic film mulch, and soil chemical treatments over a 17-month period, including a fallow winter season. Total yield for the three crops was highest (1208 quintals/ha) in film-mulched plots treated with MBR-CP, and next-highest in film-mulched plots treated with DD-MENCS (1094 quintals/ha); total yield was only 456 quintals/ha in film-mulched control (untreated) plots. Yield in untreated film-mulched plots was 256% of that in untreated unmulched plots (178 quintals/ha). Plant growth and yields were greatest when populations of nematodes and soil-borne fungi were suppressed to very low levels. The residual control by soil treatments lasted longest on Meloidogyne incognita and Fusarium solani.