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.     

Biostimulation and bioaugmentation for on-site treatment of weathered diesel fuel in Arctic soil

Bioremediation of weathered diesel fuel in Arctic soil at low temperature was studied both on-site in small-scale biopiles and in laboratory microcosms. The field study site was on Ellesmere Island (82°30'N, 62°20'W). Biostimulation was by fertilization with phosphorous and nitrogen. Bioaugmentation was with an enrichment culture originating from the field site. In biopiles, total petroleum hydrocarbons (TPH) were reduced from 2.9 to 0.5 mg/g of dry soil over a period of 65 days. In microcosms at 7 °C, TPH were reduced from 2.4 to 0.5 mg/g of dry soil over a period of 90 days. Inoculation had no effect on hydrocarbon removal in biopiles or in microcosms. Maximum TPH removal rates in the biopiles were approximately 90 μg of TPH g^–1 of soil day^–1, occurring during the first 14 days when ambient temperature ranged from 0 to 10 °C. The fate of three phylotypes present in the inoculum was monitored using most-probable-number PCR, targeting 16S rRNA genes. Populations of all three phylotypes increased more than 100-fold during incubation of both uninoculated and inoculated biopiles. The inoculum increased the initial populations of the phylotypes but did not significantly affect their final populations. Thus, biostimulation on site enriched populations that were also selected in laboratory enrichment cultures. Electronic Publication     

Improving the biotreatment of hydrocarbons-contaminated soils by addition of activated sludge taken from the wastewater treatment facilities of an oil refinery

Addition of activated sludge taken from the wastewater treatment facilities ofan oil refinery to a soil contaminated with oily sludge stimulated hydrocarbonbiodegradation in microcosms, bioreactors and biopile. Microcosms containing50 g of soil to which 0.07 % (w/w) of activated sludge was added presented ahigher degradation of alkanes (80 % vs 24 %) and polycyclic aromatic hydrocarbons(PAHs) (77 % vs 49 %) as compared to the one receiving only water, after 30days of incubation at room temperature. Addition of ammonium nitrate or sterilesludge filtrate instead of activated sludge resulted in a similar removal of PAHsbut not of alkanes suggesting that the nitrogen contained in the activated sludgeplays a major role in the degradation of PAHs while microorganisms of thesludge are active against alkanes. Addition of sludge also stimulated hydrocarbonbiodegradation in 10-kg bioreactors operated during 60 days and in a 50-m³ biopile operated during 126 days. This biopile treatment allowed the use of the soil for industrial purpose based on provincial regulation (``C' criteria). In contrast, the soil of the control biopile that received only water still exceeded C criteria for C₁₀–C₅₀ hydrocarbons, total PAHs, chrysene and benzo[a]anthracene.The stimulation effect of sludge was stronger on the 4-rings than on 2-rings PAHs.The soil of the biopile that received sludge was 4–5 times less toxic than the control. These results suggest that this particular type of activated sludge could be used to increase the efficiency of the treatment of hydrocarbon-contaminated soils in a biopile.     

Temperature Dependence of Methane Production in Tropical Rice Soils

Although CH 4 production is sensitive to temperature, it is not clear how temperature controls CH 4 production directly versus the production of organic substrates that methanogens convert into CH 4 . Therefore, this study was done to better understand how CH 4 production in rice paddy soil responded to temperature when the process was not limited by the availability of substrates. In a laboratory-incubation study using three Indian rice soils under flooded conditions, the effect of temperature on CH 4 production was examined. CH 4 production in acid sulphate, laterite, and alluvial soil samples under flooded conditions distinctly increased with increase in temperature from 15°C to 35°C. Laterite and acid sulphate soils produced distinctly less CH 4 than alluvial soils. CO 2 production increased with increase in temperature in all the soils. The readily mineralizable carbon C and Fe 2+ contents in soils were least at 15°C and highest at 35°C, irrespective of soil type. Likewise, a significant correlation existed between microbial population (methanogens and sulphate reducers) and CH 4 production. Comparing the temperature coefficients ( Q 10 ) for methane production within each soil type at low (15°C-25°C) and medium (25°C-35°C) temperature intervals revealed that these values were not uniform for both alluvial and laterite soils. But acid sulphate soil had Q 10 values that were near 2 at both temperature intervals. When these soil samples were amended with substrates (acetate, H 2 -CO 2 , and rice straw), there were stimulatory effects on methane production rates and consequently on the Q 10 values. The pattern of temperature coefficients was characteristic of the soil type and the nature of substrates used for amendment.     

Temperature response and flowering of white clover (Trifolium repens) varieties in controlled environments and the field

Floral initiation of 10 white clover varieties growing in three controlled day/night temperature regimes, 22“/10°C, 20°/10°C, 17°/10°C, was recorded. Effects of artificial soil heating on floral initiation of the same plants subsequently transferred to the field were also examined. In the controlled environments only a slight increase in day temperature (2–5°C) was necessary to significantly increase flowering. Defoliation at the three temperatures had contrasting effects on subsequent flower production. Results from the soil heating experiment suggested that increased temperature might compensate for short daylengths, by bringing forward reproductive bud initiation by 1 month. Soil heating increased the total number of inflorescences produced.     

Changes in starch structure during manufacturing of starch microspheres for use in parenteral drug formulations: Effects of temperature treatment

Starch microspheres were produced by emulsification of a starch dispersion in an aqueous polyethylene glycol (PEG) solution. Crystalline/ordered structure was formed within these starch droplets during incubation at 6 °C for 25 h followed by incubation at 37 °C for 28 h. After incubation at 37 °C the crystalline structure in the samples was of type B. The crystallization process of microspheres was compared with crystallization in a model system. The crystalline structure of the microspheres melted at temperatures almost 20 °C lower than in the model system incubated under the same conditions, as determined by differential scanning calorimetry. It was thus concluded that the crystallization process within microspheres was different than that of bulk starch and the ability of the starch molecules to reorganize themselves within the dispersed starch phase of an aqueous two-phase system at the higher incubation temperature was limited. It was also observed that the presence of PEG or carbonate buffer protected the molecular order formed by the starch molecules during incubation from breakdown during freeze-drying.     

Photoperiodic and thermal regulation of antifreeze protein levels in the beetle Dendroides canadensis

The importance of photoperiod, temperature and their interaction in controlling the seasonal pattern of haemolymph antifreeze protein levels in larvae of the beetle Dendroides canadensis was investigated. A complete photoperiodic response curve for antifreeze protein production was generated at 20°C with larvae collected in early fall. Individuals exposed to a 10-h photoperiod or less, including constant darkness, had significantly elevated antifreeze levels over those maintained in an 11-h photoperiod or more, including constant light. The critical daylength resulting in 50% population response lies between LD 11:13 and LD 10:14. This photoperiodic response was masked at sufficiently low (threshold between 15 and 10°C) and high (threshold between 25 and 30°C) temperatures. Partial photoperiodic response curves (at 17 and 25°C) obtained within this specified temperature range indicate that the position of the critical photoperiod (between 10 and 11 h) is stable while the amplitude of the response curve is temperature dependent.Experiments investigating the mechanisms controlling the spring depletion of protein antifreeze levels suggest that both photoperiod and temperature are important.The dominant response of photoperiod in the fall along with the modifying effects of temperature are considered to provide the necessary precision to assure adequate cold tolerance early in the fall and the flexibility to protect the species from yearly variation in weather conditions.     

Effect of Mulch Surface Color on Root-knot of Tomato Grown in Simulated Planting Beds

The effect of different-colored polyethylene mulches on quantity and spectra of reflected light, plant morphology, and root-knot disease was studied in tomato (Lycopersicon esculentum) grown in simulated planting beds. Tomato plants were inoculated with Meloidogyne incognita at initial populations (Pi) of 0, 1,000, 10,000, or 50,000 eggs/plant, and grown in a greenhouse for 50 days over white, red, or black mulch. Soil temperature was kept constant among the mulch treatments by placing an insulation barrier between the colored mulch and the soil surface. Soil temperature varied less than 0.5 °C between soil chambers at solar noon. Tomatoes grown over white mulch received more reflected photosynthetic light and had greater shoot weights (27%), root weights (32%), and leaf area (20%) than plants grown over black mulch. Plants grown over red mulch received a higher far-red-to-red ratio in the reflected light. Mulch color altered the plant''s response to root-knot nematode infection by changing the distribution of mass in axillary shoots. At high Pi, axillary leaf area and leaf weight were greater in tomato grown over white mulch than when grown over red mulch. The root-gall index was lower for plants grown over white mulch than similar plants grown over red mulch.     

Decomposition of soil organic matter from boreal black spruce forest: environmental and chemical controls

Black spruce forests are a dominant covertype in the boreal forest region, and they inhabit landscapes that span a wide range of hydrologic and thermal conditions. These forests often have large stores of soil organic carbon. Recent increases in temperature at northern latitudes may be stimulating decomposition rates of this soil carbon. It is unclear, however, how changes in environmental conditions influence decomposition in these systems, and if substrate controls of decomposition vary with hydrologic and thermal regime. We addressed these issues by investigating the effects of temperature, moisture, and organic matter chemical characteristics on decomposition of fibric soil horizons from three black spruce forest sites. The sites varied in drainage and permafrost, and included a “Well Drained” site where permafrost was absent, and “Moderately well Drained” and “Poorly Drained” sites where permafrost was present at about 0.5 m depth. Samples collected from each site were incubated at five different moisture contents (2, 25, 50, 75, and 100% saturation) and two different temperatures (10°C and 20°C) in a full factorial design for two months. Organic matter chemistry was analyzed using pyrolysis gas chromatography-mass spectrometry prior to incubation, and after incubation on soils held at 20°C, 50% saturation. Mean cumulative mineralization, normalized to initial carbon content, ranged from 0.2% to 4.7%, and was dependent on temperature, moisture, and site. The effect of temperature on mineralization was significantly influenced by moisture content, as mineralization was greatest at 20°C and 50–75% saturation. While the relative effects of temperature and moisture were similar for all soils, mineralization rates were significantly greater for samples from the “Well Drained” site compared to the other sites. Variations in the relative abundances of polysaccharide-derivatives and compounds of undetermined source (such as toluene, phenol, 4-methyl phenol, and several unidentifiable compounds) could account for approximately 44% of the variation in mineralization across all sites under ideal temperature and moisture conditions. Based on our results, changes in temperature and moisture likely have similar, additive effects on in situ soil organic matter (SOM) decomposition across a wide range of black spruce forest systems, while variations in SOM chemistry can lead to significant differences in decomposition rates within and among forest sites.     

Passive heat treatment of sweet basil crops suppresses Peronospora belbahrii downy mildew

Sweet basil (Ocimum basilicum) is an annual herb crop grown in polyethylene‐covered structures in Israel. It is Israel's leading herb crop, grown in warm regions of the country. Downy mildew (caused by Peronospora belbahrii) is a severe disease in Israel and in many other crop‐growing regions worldwide. Experiments were carried out to identify potential climate‐management techniques for suppression of this disease on basil in non‐heated greenhouses. Disease severity was evaluated under commercial‐like conditions in three experiments, with 8–10 walk‐in tunnels at each location. Pathogen inoculum was introduced into all walk‐in tunnels. Regression analysis was performed between the disease values and air temperature, relative humidity (RH) and soil temperature. Downy mildew severity was negatively related to high (>25°C) air temperature, RH in the range of 65–85% and high (>21°C) soil temperature. The increase in air temperature did not result in a significant increase in leaf temperature; canopy surface median temperatures only reached 30°C. Symptomless plants from relatively warmer tunnels (peak temperatures of 45–48°C) that were transferred to conditions that promote downy mildew (22 ± 2°C, RH > 95%) became severely diseased, showing sporulation of P. belbahrii, suggesting that infection occurred but at the high temperatures symptom expression/tissue colonisation was suppressed. Pot experiments in which aerial and subterranean plant organs were differentially heated revealed that treating the roots with a high temperature (26–31°C), similar to the soil temperatures in the warmer greenhouses, while maintaining the upper plant parts at ambient temperature (20°C), suppresses canopy downy mildew. The effect lasted for 1–2 weeks after the plants were removed from the heated soil treatments and maintained under optimal conditions for pathogen development. Furthermore, oospores were found in the symptomatic leaves. Oospores are minimally affected by high temperature, and therefore the high temperature presumably did not affect pathogen survival. In conclusion, the effect of high greenhouse temperature on basil downy mildew may not result from a direct negative effect of high temperature on the pathogen but from an indirect high‐temperature effect on the host, rendering it less susceptible to pathogen development.     

Analysis of spatial and yearly variation in winter survival of winter wheat

 Four years of winter survival data for winter wheat (Triticum aestivum L.) were collected on a loam soil located on the Central Experimental Farm at Ottawa, Ontario (45° 23′N, 75° 43′W). The site was low-lying and subject to frequent winter flooding and ice sheet formation. It appeared level although there was microtopographic variation with a range in elevation of approximately 0.15 m. The objective of the study was to gain insight into factors which might affect winter survival. Two varieties, a soft white and a hard red winter wheat, were planted in September. Crop establishment was measured in late fall and the percentage survival was measured in April of the following year. We assumed the large spatial differences in survival were not totally random, but rather were affected by spatial variation in environmental factors such as snow and ice depth, soil moisture and temperature. Hourly measurements of soil temperature at a depth of 0.05 m were recorded throughout the fall, winter and spring. Fall and spring soil moisture at the same depth were measured on the plot, as well as snow and ice depth at selected times throughout the winter. Measurements were taken on a grid covering the plot to help explain spatial variation in survival. In addition, detailed measurements of elevation were taken on a grid. Meteorological data were available from the nearby weather site. While soil temperatures were never low enough to kill plants according to the CERES model, the spatial variation in winter survival was associated with differences in elevation and the resulting surface drainage patterns. Received: 23 March 1998/Accepted: 21 October 1998     

Zur Dynamik der Bodenmikroflora nach Anwendung der Fungizide Olpisan (Trichlordinitrobenzol), Captan und Thiuram

It has been established that the survival time of black rot organism in unsterilized soil depends on soil temperature. At 20 °C the theoretical survival time was calculated 20 days and at 5 °C, it was 47 days. Similarly in stem debris, lying on soil surface and at depth of 20 cm, the survival time of pathogen was calculated 493 and 551 days respectively.     

Protein-bound SH groups in frozen-thawed egg cells of the sea urchin

Unfertilized egg cells of the sea urchin St. intermedius could survive slow freezing to ?15 °C for a short period of time, but at the same freezing temperature extracellular freezing became fatal within a few hours. Such freezing injury resulted in “black” or “white” cytolysis in frozen-thawed cells. “Black” cytolysis took place in the process of both freezing and thawing, while “white” cytolysis occurred only on thawing. Rapid rewarming consistently produced “white” cytolysis in extracellularly frozen cells. The observed behavior of the injured cells during freeze-thawing appeared favorable for the explanation of freezing injury by the SH-SS hypothesis. Protein-bound SH groups were quantitatively determined in both whole cell and cortex with plasma membrane before and after freeze-thawing. However, no significant change in the SH value was observed between freeze-thaw cytolysed materials and unfrozen ones.     

A field enclosure apparatus for measuring crop photosynthesis

The field enclosure is a transparent box covering a soil area of 1.5 m². It is a semi-closed system in which concentrations of water vapour and carbon dioxide are maintained constant: the required rate of input of carbon dioxide being a measure of the photosynthesis rate, and the rate of condensation of water, on cooling coils, a measure of transpiration. The air within the enclosure is circulated rapidly by fans to decrease concentration gradients, and under steady radiation inputs the air temperature is controlled to ±0.5 °C. Both photosynthesis and transpiration rates are corrected for air exchange with the surroundings, as measured through the injection of the inert gas, nitrous oxide.     

Genipa americana L.: A New Phytochemical for White Hair Coloring

This work describes a new hair dyeing methodology using a chemical reaction between geniposide, an iridoid glycoside extracted from the fruit of Genipa americana (geniposide extract, GE) and the amine group of hair keratin. The influence of reaction conditions (pH, temperature, and extract concentration) on the staining of hair fibers, color development, fiber morphology, and mechanical hair properties of black and white human hair samples, was evaluated before and after GE dyeing treatment. Eye contact safety of GE was also studied using HET-CAM. The treatment of white hair fibers using GE at 20 mg mL⁻¹, temperature of 80 °C and pH 5.5 presented the greatest color change (ΔE=54.0). The higher pH influence was observed at pH 10.0 on white hair tresses (ΔE=6.8), using an GE concentration of 20 mg mL⁻¹ and room temperature (25 °C). Treated samples showed marked changes on mechanical and morphological properties. The HET-CAM did not show any change, thus demonstrating that using GE is safe. In conclusion, the temperature and concentration of the extract were the variables that mostly influenced the color and hair damage. A new approach for hair dyeing was established where iridoids may potentially be useful as a natural hair dyeing.     

The effect of acclimation temperature on the activation energies of state III respiration and on the unsaturation of membrane lipids of goldfish mitochondria

The influence of the acclimation temperature on the thermotropic behaviour of mitochondrial respiration and on the degree of unsaturation of mitochondrial membrane lipids has been studied. The mitochondria were isolated from red muscle, white muscle and liver of goldfish acclimated to 5, 20 and 30°C. ADP-activated succinate oxidation was measured at different temperatures and resulted in non-linear Arrhenius-plots with breaks between 10 and 23°C. As for the break-temperatures, there was found a shift downwards in preparations of decreased acclimation temperatures. This could be caused by a changed composition of membrane lipids and a simultaneous shift of the membrane phase transition temperature. Therefore, the fatty acid composition of all membrane preparations was analyzed. However, no consistent change of the degree of unsaturation due to a changed acclimation temperature could be found.     

Out on a limb: Thermal microenvironments in the tropical forest canopy and their relevance to ants

Small, cursorial ectotherms like ants often are immersed in the superheated air layers that develop millimeters above exposed, insolated surfaces (i.e., the thermal boundary layer). We quantified the thermal microenvironments around tree branches in the tropical rainforest canopy, and explored the effects of substrate color on the internal body temperature and species composition of arboreal ants. Branch temperatures during the day (09:00–16:00) were hottest (often > 50 °C) and most variable on the upper surface, while the lowest and least variable temperatures occurred on the underside. Temperatures on black substrates declined with increasing distance above the surface in both the field and the laboratory. By contrast, a micro-scale temperature inversion occurred above white substrates. Wind events (ca. 2 m s⁻¹) eliminated these patterns. Internal temperatures of bodies of Cephalotes atratus workers experimentally heated in the laboratory were 6 °C warmer on white vs. black substrates, and 6 °C cooler than ambient in windy conditions. The composition of ant species foraging at baits differed between black-painted and unpainted tree branches, with a tendency for smaller ants to avoid the significantly hotter black surfaces. Collectively, these outcomes show that ants traversing canopy branches experience very heterogeneous thermal microenvironments that are partly influenced in predictable ways by branch surface coloration and breezy conditions.     

Soil solarisation, amendments and bio-control agents for the control of Macrophomina phaseolina and Fusarium oxysporum f.sp. cumini in aridisols

The effects of soil solarisation, residue incorporation, summer irrigation and biocontrol agents singly or in combination on survival of Macrophomina phaseolina and Fusarium oxysporum f.sp. cumini were ascertained in the 2000 and 2001 summer seasons. In amended plots, temperature increased by 2.5°C over non‐amended plots (42–51°C) at various soil depths. Combining amendments and soil solarisation elevated the soil temperatures by 0.5–5°C and 2.5–13.0°C compared to non‐amended solarised and non‐solarised plots, respectively. These treatment combinations significantly reduced M. phaseolina and Fusarium propagules compared to control. Of these, combining mustard pod residues with soil solarisation almost eliminated viable propagules of both the pathogens at 0–30 cm soil depth. However, a combination of mustard pod residue and oil‐cake (2.5 + 0.5 ton ha^?1) with only one summer irrigation also caused pronounced reduction in pathogenic propagules, which was equal to that recorded in non‐amended solarised plots. The effect of surviving propagules of M. phaseolina and Fusarium on incidence of dry root rot on clusterbean and wilt on cumin was studied in subsequent rainy and winter seasons, respectively. Significant reductions in both diseases were recorded in residue and biocontrol amended plots with or without polyethylene mulching compared to non‐amended control. The lowest plant mortality in both the crops was recorded in mustard residue amended solarised plots in a two year field experiment. However, the disease indices in the plots having a combination of mustard residues and oil‐cake amendment with one summer irrigation was equal to that achieved in the treatment having polyethylene mulching. These results suggest that in hot arid regions use of Brassica residues can be a practical and feasible substitute for polyethylene mulching in managing soil‐borne diseases.     

The promotive effect on subsequent germination of treating imbibed celery seeds with high temperature before or during drying

Abstract High temperature (32°C) prevented germination of celery seeds even if given after 4 d of germination induction at 17°C in white light, but germination occurred if the seeds were then returned to 17°C. Celery seeds incubated for 3 d at 17°C in white light and then air-dried at 20°C germinated slowly when re-sown at 17°C in the light, achieving only 24% germination after 21 d. Exposure of such seeds to 32°C prior to and during drying resulted in 50% germination after 3.6 d at 17°C in white light, with no loss in viability, compared to 5.7 d for seeds not given a germination induction treatment. If celery seeds were dried rapidly germination was poor, an effect which could be overcome by high temperature treatment. It is suggested that the mechanism which imposes dormancy at 32°C also conditions the seed to withstand desiccation damage.     

Prediction model for cabbage stem weevil Ceutorhynchus pallidactylus Mrsh. occurrence on winter rape based on an artificial neural network

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