Alpine microorganisms: useful tools for low-temperature bioremediation

Cold environments, including polar and alpine regions, are colonized by a wide diversity of microorganisms able to thrive at low temperatures. There is evidence of a wide range of metabolic activities in alpine cold ecosystems. Like polar microorganisms, alpine microorganisms play a key ecological role in their natural habitats for nutrient cycling, litter degradation, and many other processes. A number of studies have demonstrated the capacity of alpine microorganisms to degrade efficiently a wide range of hydrocarbons, including phenol, phenol-related compounds and petroleum hydrocarbons, and the feasibility of low-temperature bioremediation of European alpine soils by stimulating the degradation capacity of indigenous microorganisms has also been shown.     

Biodegradation and bioremediation of hydrocarbons in extreme environments

Many hydrocarbon-contaminated environments are characterized by low or elevated temperatures, acidic or alkaline pH, high salt concentrations, or high pressure, Hydrocarbon-degrading microorganisms, adapted to grow and thrive in these environments, play an important role in the biological treatment of polluted extreme habitats. The biodegradation (transformation or mineralization) of a wide range of hydrocarbons, including aliphatic, aromatic, halogenated and nitrated compounds, has been shown to occur in various extreme habitats. The biodegradation of many components of petroleum hydrocarbons has been reported in a variety of terrestrial and marine cold ecosystems. Cold-adapted hydrocarbon degraders are also useful for wastewater treatment. The use of thermophiles for biodegradation of hydrocarbons with low water solubility is of interest, as solubility and thus bioavailability, are enhanced at elevated temperatures. Thermophiles, predominantly bacilli, possess a substantial potential for the degradation of environmental pollutants, including all major classes. Indigenous thermophilic hydrocarbon degraders are of special significance for the bioremediation of oil-polluted desert soil. Some studies have investigated composting as a bioremediation process. Hydrocarbon biodegradation in the presence of high salt concentrations is of interest for the bioremediation of oil-polluted salt marshes and industrial wastewaters, contaminated with aromatic hydrocarbons or with chlorinated hydrocarbons. Our knowledge of the biodegradation potential of acidophilic, alkaliphilic, or barophilic microorganisms is limited.     

Hydrocarbon phytoremediation in the family Fabaceae--a review

Currently, studies often focus on the use of Poaceae species (grasses) for phytoremediation of hydrocarbon-contaminated soils. Research into the use of Fabaceae species (legumes) to remediate hydrocarbons in soils has been conducted, but these plants are commonly overlooked due to slower recorded rates of degradation compared with many grass species. Evidence in the literature suggests that in some cases Fabaceae species may increase total degradation of hydrocarbons and stimulate degradative capacity of the soil microbial community, particularly for contaminants which are normally more recalcitrant to degradation. As many recalcitrant hydrocarbons have negative impacts on human and ecosystem health, development of remediation options is crucial. Reconsideration of Fabaceae species for removal of such contaminants may lead to environmentally and economically sustainable technologies for remediation of contaminated sites.     

Oil degradation by basidiomycetes in soil and peat at low temperatures

A total of 17 basidiomycete strains causing white rot and growing on oil-contaminated substrates have been screened. Three strains with high (Steccherinum murashkinskyi), average (Trametes maxima), and low (Pleurotus ostreatus) capacities for the colonization of oil-contaminated substrates have been selected. The potential for degrading crude oil hydrocarbons has been assessed with the use of fungi grown on nonsterile soil and peat at low temperatures. Candida sp. and Rhodococcus sp. commercial strains have been used as reference organisms with oil-degrading ability. All microorganisms introduced in oil-contaminated soil have proved to be ineffective, whereas the inoculation of peat with basidiomycetes and oil-degrading microorganisms accelerated the destruction of oil hydrocarbons. The greatest degradation potential of oil-aliphatic hydrocarbons has been found in S. murashlinskyi. T. maxima turned out to be the most successful in degrading aromatic hydrocarbons. It has been suggested that aboriginal microflora contributes importantly to the effectiveness of oil-destructing microorganisms. T. maxima and S. murashkinskyi strains are promising for further study as oil-oxidizing agents during bioremediation of oil-contaminated peat soil under conditions of low temperatures.     

Microbial degradation of hydrocarbons in the environment.

The ecology of hydrocarbon degradation by microbial populations in the natural environment is reviewed, emphasizing the physical, chemical, and biological factors that contribute to the biodegradation of petroleum and individual hydrocarbons. Rates of biodegradation depend greatly on the composition, state, and concentration of the oil or hydrocarbons, with dispersion and emulsification enhancing rates in aquatic systems and absorption by soil particulates being the key feature of terrestrial ecosystems. Temperature and oxygen and nutrient concentrations are important variables in both types of environments. Salinity and pressure may also affect biodegradation rates in some aquatic environments, and moisture and pH may limit biodegradation in soils. Hydrocarbons are degraded primarily by bacteria and fungi. Adaptation by prior exposure of microbial communities to hydrocarbons increases hydrocarbon degradation rates. Adaptation is brought about by selective enrichment of hydrocarbon-utilizing microorganisms and amplification of the pool of hydrocarbon-catabolizing genes. The latter phenomenon can now be monitored through the use of DNA probes. Increases in plasmid frequency may also be associated with genetic adaptation. Seeding to accelerate rates of biodegradation has been shown to be effective in some cases, particularly when used under controlled conditions, such as in fermentors or chemostats.     

Effects of oxygen,nitrogen, and temperature on gasoline biodegradation in soil

Biodegradation was considered to be a feasible approach to remediate petroleum hydrocarbon-contaminated soil from a site at the University of Idaho. Before a full-scale treatment process was designed, the biodegradative capacity of the soil's indigenous microorganisms was tested. Gas chromatography was used to measure gasoline vapor components in the headspace above the contaminated soils held in closed containers. In a study of biodegradation kinetics, gasoline degradation rates under various conditions (different soil cores, temperatures, oxygen concentrations, and nutrient concentrations) were tested. It was found that gasoline hydrocarbons could be biodegraded at relatively high rates after appropriate nutrient additions. An unexpected observation was that the optimal concentration of oxygen for the gasoline-degrading microorganisms in these soils was only 10%.Publication No. 94505 of the Idaho Agricultural Experiment Station.     

Specificity of the chitinolytic microbial complex of soils incubated at different temperatures

The structural and functional specificity of the chitinolytic microbial complex changes dramatically depending on the incubation temperature of soil microcosms. It was shown that the highest rates of chitin degradation occurred in desert soils at high temperatures (50°C); in the moderate and northern zones, these rates peaked at lower temperatures (5°C). The role of prokaryotes as the main chitin degraders in soils incubated at high temperatures, with fungi more actively participating in chitin decomposition at low temperatures, was shown for the first time. Fluorescent in situ hybridization (FISH) revealed the predominance of actinomycetes in the metabolically active chitinolytic prokaryotic complex of desert soils (high temperatures); in the soils of the northern latitudes (low temperatures), proteobacteria prevailed. The relationship between the taxonomic position of the dominant members of the chitinolytic complex of soil microorganisms, isolated in pure cultures with the dominant phylogenetic groups and the sequence types obtained by using molecular biological techniques (FISH) was revealed.     

Microbial degradation of model petroleum at low temperatures

Two areas of Chesapeake Bay, Colgate Creek in Baltimore Harbor and Eastern Bay, are presently under study, with routine sampling of water and sediment for petroleum-degrading microorganisms (bacteria, yeasts, and fungi) by direct plating and enrichment culture. Selected physical and chemical parameters are recorded for each sampling site, and water and sediment samples are extracted for hydrocarbons. Numbers of petroleum-degrading microorganisms enumerated by direct plating were found to correlate with the concentration of benzene-extractable material and were higher for the Colgate Creek than for the Eastern Bay site. Petroleum-degrading microorganisms were isolated from water and sediment samples at environmental temperatures of 0°, 5°, and 10°C. A salts medium supplemented with nitrate and phosphate was used to provide optimum conditions for petroleum degradation, whereas Chesapeake Bay water was used to simulate natural environmental conditions. Use of a model petroleum permitted quantitative measurement of utilization of individual hydrocarbons ranging in complexity from simple alkanes to polynuclear aromatic hydrocarbons. Higher growth yields and maximum hydrocarbon degradation was observed for microorganisms in the salts medium at 0°, 5°, and 10°C, although significant quantities of hydrocarbons were utilized in some samples grown in a medium for which Chesapeake Bay water was the diluent. Bacterial hydrocarbon degradation accounted for most of the model petroleum utilization at 0° and 5°C. However, oscillations of bacterial populations, with significant growth of yeasts, was observed at 10°C. Photomicroscopy and scanning electron microscopy revealed aggregates of bacteria, yeasts, and fungi associated with oil globules. From preliminary identification and classification of the hydrocarbon-utilizing bacteria, members of the generaVibrio, Aeromonas, Pseudomonas, andAcinetobacter were present in the enrichment cultures. From results of this study, it is concluded that utilization of model petroleum at low temperatures is a function of the types and numbers of microorganisms present in an original inoculum taken from the natural environment.     

Temperature impacts on anaerobic biotransformation of LNAPL and concurrent shifts in microbial community structure

Thermally-enhanced bioremediation is a promising treatment approach for petroleum contamination; however, studies examining temperature effects on anaerobic biodegradation in zones containing light non-aqueous phase liquids (LNAPLs) are lacking. Herein, laboratory microcosm studies were conducted for a former refinery to evaluate LNAPL transformation, sulfate reduction, and methane generation over a one-year period for temperatures ranging from 4 to 40 °C, and microbial community shifts were characterized. Temperatures of 22 and 30 °C significantly increased total biogas generation compared to lower (4 and 9 °C) and higher temperatures (35 and 40 °C; p < 0.1). Additionally, at 22 and 30 °C methane generation commenced ~6 months earlier than for 35 and 40 °C. Statistically significant biodegradation of benzene, toluene and xylenes was observed at elevated temperatures but not at lower temperatures (p < 0.1). Additionally, a novel differential chromatogram approach was developed to overcome challenges associated with resolving losses in complex mixtures of hydrocarbons, and application of this method revealed greater losses of hydrocarbons at 22 and 30 °C as compared to lower and higher temperatures. Finally, molecular biology assays revealed that the composition and activity of microbial communities shifted in a temperature-dependent manner. Collectively, results demonstrated that anaerobic biodegradation processes can be enhanced by increasing the temperature of LNAPL-containing soils, but biodegradation does not simply increase as temperature increases likely due to a lack of microorganisms that thrive at temperatures well above the historical high temperatures for a site. Rather, optimal degradation is achieved by holding soils at the high end of, or slightly higher than, their natural range.     

Cold-adapted enzymes produced by fungi from terrestrial and marine Antarctic environments

Antarctica is the coldest, windiest, and driest continent on Earth. In this sense, microorganisms that inhabit Antarctica environments have to be adapted to harsh conditions. Fungal strains affiliated with Ascomycota and Basidiomycota phyla have been recovered from terrestrial and marine Antarctic samples. They have been used for the bioprospecting of molecules, such as enzymes. Many reports have shown that these microorganisms produce cold-adapted enzymes at low or mild temperatures, including hydrolases (e.g. α-amylase, cellulase, chitinase, glucosidase, invertase, lipase, pectinase, phytase, protease, subtilase, tannase, and xylanase) and oxidoreductases (laccase and superoxide dismutase). Most of these enzymes are extracellular and their production in the laboratory has been carried out mainly under submerged culture conditions. Several studies showed that the cold-adapted enzymes exhibit a wide range in optimal pH (1.0–9.0) and temperature (10.0–70.0?°C). A myriad of methods have been applied for cold-adapted enzyme purification, resulting in purification factors and yields ranging from 1.70 to 1568.00-fold and 0.60 to 86.20%, respectively. Additionally, some fungal cold-adapted enzymes have been cloned and expressed in host organisms. Considering the enzyme-producing ability of microorganisms and the properties of cold-adapted enzymes, fungi recovered from Antarctic environments could be a prolific genetic resource for biotechnological processes (industrial and environmental) carried out at low or mild temperatures.     

低温微生物修复石油烃类污染土壤研究进展

耐冷菌、嗜冷菌等低温微生物广泛存在于极地、高山以及高纬度等土壤环境中,是石油烃类污染物在低温条件下降解与转化的重要微生物资源.利用低温微生物的独特优势,石油污染土壤的低温生物修复技术的研究成为当前热点领域.本文系统综述了低温石油烃降解菌的分类及冷适机制,低温微生物对不同类型石油烃组分的降解特征和降解机理,低温环境中接种降解菌、添加营养物质和表面活性剂等强化技术在石油污染土壤中生物修复的应用.以及微生物分子生物学技术在低温微生物降解石油烃的研究现状,为拓展我国石油污染土壤生物修复技术提供参考.     

Microbial degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in soils.

The microbial degradation of tensile test pieces made of poly(3-hydroxybutyrate) [P(3HB)] or a copolymer of 90% 3-hydroxybutyric acid and 10% 3-hydroxyvaleric acid was studied in soils incubated at a constant temperature of 15, 28, or 40 degrees C for up to 200 days. In addition, hydrolytic degradation in sterile buffer at temperatures ranging from 4 to 55 degrees C was monitored for 98 days. Degradation was measured through loss of weight (surface erosion), molecular weight, and mechanical strength. While no weight loss was recorded in sterile buffer, samples incubated in soils were degraded at an erosion rate of 0.03 to 0.64% weight loss per day, depending on the polymer, the soil, and the incubation temperature. The erosion rate was enhanced by incubation at higher temperatures, and in most cases the copolymer lost weight at a higher rate than the homopolymer. The molecular weights of samples incubated at 40 degrees C in soils and those incubated at 40 degrees C in sterile buffer decreased at similar rates, while the molecular weights of samples incubated at lower temperatures remained almost unaffected, indicating that molecular weight decrease is due to simple hydrolysis and not to the action of biodegrading microorganisms. The degradation resulted in loss of mechanical properties. From the samples used in the biodegradation studies, 295 dominant microbial strains capable of degrading P (3HB) and the poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer in vitro were isolated and identified.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mechanisms of salt tolerance in transgenic Arabidopsis thaliana constitutively overexpressing the tomato 14-3-3 protein TFT7

A hydroponics experiment was conducted to investigate the effects of iron plaque on root surfaces with respect to selenite uptake and translocation within the seedlings of two cultivars of rice (Oryza sativa L. cv Xiushui48 and Bing9652). Different amounts of iron plaque were formed by adding 0, 10, 30, 50, 70 mg Fe l⁻¹ in the nutrient solution. After 24 h of growth, the amount of iron plaque was positively correlated with the Fe²⁺ addition to the nutrient solution. These concentrations of Fe, inducing plaque, had no significant effect on the shoot and root growth of rice plants in 50 μg Se l⁻¹ nutrient solution. The amount of Se accumulated in iron plaque was positively correlated to the amount of iron plaque. Increasing iron plaque decreased the selenium concentration in shoots and in roots. At the same time, the translocation of Se from roots to shoots was reduced with increasing amounts of iron plaque. At both the shorter and longer exposure times, the ratio of root- to-shoot selenium was higher than in the controls. More Se stayed in the roots at the longer exposure time than at the shorter time. The concentration of selenium in the xylem sap was sharply decreased with increasing amount of iron plaque on the rice roots. The DCB (dithionite-citrate-bicarbonate)-extracted Se was up to 89.9–91.1% of the total Se when roots with iron plaque (Fe 70) were incubated in 50 μg Se l⁻¹ solution for 30 min. This DCB-extracted Se, however, accounted for only 21.9–28.7% of total Se when roots with iron plaque were incubated in the same solution for 3 days. Se adsorbed in iron plaque can be desorbed by low-molecular-weight organic acids, similar to the desorption of Se from ferrihydrite. These results suggest that iron plaque might act as a ‘buffer’ for Se in the rhizosphere.     

Low temperature bioremediation of oil-contaminated soil using biostimulation and bioaugmentation with a Pseudomonas sp. from maritime Antarctica

AIMS: To identify native Antarctic bacteria capable of oil degradation at low temperatures. METHODS AND RESULTS: Oil contaminated and pristine soils from Signy Island (South Orkney Islands, Antarctica) were examined for bacteria capable of oil degradation at low temperatures. Of the 300 isolates cultured, Pseudomonas strain ST41 grew on the widest range of hydrocarbons at 4 degrees C. ST41 was used in microcosm studies of low temperature bioremediation of oil-contaminated soils. Microcosm experiments showed that at 4 degrees C the levels of oil degradation increased, relative to the controls, with (i) the addition of ST41 to the existing soil microbial population (bioaugmentation), (ii) the addition of nutrients (biostimulation) and to the greatest extent with (iii) a combination of both treatments (bioaugmentation and biostimulation). Addition of water to oil contaminated soil (hydration) also enhanced oil degradation, although less than the other treatments. Analysis of the dominant species in the microcosms after 12 weeks, using temporal temperature gradient gel electrophoresis, showed Pseudomonas species to be the dominant soil bacteria in both bioaugmented and biostimulated microcosms. CONCLUSIONS: Addition of water and nutrients may enhance oil degradation through the biostimulation of indigenous oil-degrading microbial populations within the soil. However, bioaugmentation with Antarctic bacteria capable of efficient low temperature hydrocarbon degradation may enhance the rate of bioremediation if applied soon after the spill. SIGNIFICANCE AND IMPACT OF THE STUDY: In the future, native soil bacteria could be of use in bioremediation technologies in Antarctica.     

Mobility and Persistence of Petroleum Hydrocarbons in Peat Soils of Southeastern Mexico

Spilled crude petroleum from oil wells contains numerous hydrocarbons, some of which are toxic and threaten life. We have studied the mobility and persistence of hydrocarbons in waterlogged soils that contain large proportions of fermented organic matter (Histosols) and large concentrations of dissolved organic carbon (DOC) in the State of Tabasco, Mexico. We sampled soil and phreatic water at sites polluted by oil spills for several decades, as well as at sites that had only recently (few weeks) been polluted, and compared their hydrocarbon contents with those of unaffected sites in the same area. Samples were analyzed for 16 non-alkylated polyaromatic hydrocarbons (PAHs) and n-alkanes from nC9 to nC34. The spilled hydrocarbons had remained predominantly in the organic surface horizons of the soil where spillage occurred; there was little evidence of movement within the soil. The fraction of low molecular weight compounds was larger at sites of recent spills than where spills happened several decades ago. Nevertheless, sites of old spills still contained large concentrations of hydrocarbons, among which those of low molecular weight represented from 30 to 49% of total PAHs and from 50 to 84% of total n-alkanes, indicating that volatilization or microbial degradation is slow in these soils. In the peat horizons the measured organic carbon partition coefficients (K _oc ) for the higher molecular weight PAHs were consistently smaller than those estimated by empirical equations by up to two orders of magnitude. The dissolved organic carbon of these peat soils seems to influence this behavior. At sites of old spills, partition coefficients for the PAHs were larger than at sites of recent spills.     

Degradation of BTEX by anaerobic bacteria: physiology and application

Pollution of the environment with aromatic hydrocarbons, such as benzene, toluene, ethylbenzene and xylene (so-called BTEX) is often observed. The cleanup of these toxic compounds has gained much attention in the last decades. In situ bioremediation of aromatic hydrocarbons contaminated soils and groundwater by naturally occurring microorganisms or microorganisms that are introduced is possible. Anaerobic bioremediation is an attractive technology as these compounds are often present in the anoxic zones of the environment. The bottleneck in the application of anaerobic techniques is the lack of knowledge about the anaerobic biodegradation of benzene and the bacteria involved in anaerobic benzene degradation. Here, we review the existing knowledge on the degradation of benzene and other aromatic hydrocarbons by anaerobic bacteria, in particular the physiology and application, including results on the (per)chlorate stimulated degradation of these compounds, which is an interesting new alternative option for bioremediation.     

Bioremediation and phytoremediation of total petroleum hydrocarbons (TPH) under various conditions

Remediation of contaminated soils is often studied using fine-textured soils rather than low-fertility sandy soils, and few studies focus on recontamination events. This study compared aerobic and anaerobic treatments for remediation of freshly introduced used motor oil on a sandy soil previously phytoremediated and bioacclimated (microorganisms already adapted in the soil environment) with some residual total petroleum hydrocarbon (TPH) contamination. Vegetated and unvegetated conditions to remediate anthropogenic fill containing residual TPH that was spiked with nonaqueous phase liquids (NAPLs) were evaluated in a 90-day greenhouse pot study. Vegetated treatments used switchgrass (Panicum virgatum). The concentration of aerobic bacteria were orders of magnitude higher in vegetated treatments compared to unvegetated. Nevertheless, final TPH concentrations were low in all saturated soil treatments, and high in the presence of switchgrass. Concentrations were also low in unvegetated pots with fertilizer. Acclimated indigenous microbial communities were shown to be more effective in breaking down hydrocarbons than introducing microbes from the addition of plant treatments in sandy soils. Remediation of fresh introduced NAPLs on pre-phytoremediated and bioacclimated soil was most efficient in saturated, anaerobic environments, probably due to the already pre-established microbial associations, easily bioavailable contaminants, and optimized soil conditions for microbial establishment and survival.     

Effects of Rapeseed Oil on the Rhizodegradation of Polyaromatic Hydrocarbons in Contaminated Soil

Plants have the ability to promote degradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated soil by supporting PAH degrading microorganisms in the rhizosphere (rhizodegradation). The aim of this study was to evaluate if rapeseed oil increases rhizodegradation because various studies have shown that vegetable oils are able to act as extractants for PAHs in contaminated soils and therefore might increase bioavailability of PAHs for microbial degradation. In this study different leguminous and grass species were tested. The results suggested a significant impact of vegetable oil (1 and 3% w/w) on plant growth (decrease of plant height and biomass). The results of the pot experiment showed a decrease in the PAH content of the soil without amendment of rapeseed oil after six months. In soil amended with 1% and 3% of oil, there was no decrease in PAH content within this period. Although no enhancement of PAH degradation by plants could be measured in the bulk soil of the pot experiments, a rhizobox experiment showed a significant reduction of PAH content in the rhizosphere of alfalfa (Medicago sativa cv. Europe). Our investigations also showed significant differences in the degradation behaviour of the 16 individually analysed PAHs.     

Microbial degradation of hydrochlorofluorocarbons (CHCl2F and CHCl2CF3) in soils and sediments.

The ability of microorganisms to degrade trace levels of the hydrochlorofluorocarbons HCFC-21 and HCFC-123 was investigated. Methanotroph-linked oxidation of HCFC-21 was observed in aerobic soils, and anaerobic degradation of HCFC-21 occurred in freshwater and salt marsh sediments. Microbial degradation of HCFC-123 was observed in anoxic freshwater and salt marsh sediments, and the recovery of 1,1,1-trifluoro-2-chloroethane indicated the involvement of reductive dechlorination. No degradation of HCFC-123 was observed in aerobic soils. In some experiments, HCFCs were degraded at low (parts per billion) concentrations, raising the possibility that bacteria in nature remove HCFCs from the atmosphere.