Translocation and turnover of rhizodeposit carbon within soil microbial communities of an extensive grassland ecosystem

Background and Aims

A substantial amount of photosynthesized plant-C is allocated belowground in grassland ecosystems where it influences the structure and function of the soil microbial community with potential implications for C cycling and storage. We applied stable isotope probing of microbial PLFAs and repeated soil sampling in a grassland over a period of 1 year to assess the role of microbial communities in the cycling of rhizodeposit-C.

Methods

Pulse-labeling with ¹³CO₂ was performed in a grassland site near Gent (Belgium). Soil samples were taken 24 h, 1 week, 1 month, 4 months, 9 months and 1 year following labeling and analyzed for ¹³C in soil, roots and microbial PLFAs.

Results

C enrichment of PLFAs occurred rapidly (within 24 h) but temporally varied across microbial groups. PLFAs indicative for fungi and gram-negative bacteria showed a faster ¹³C uptake compared to gram-positive bacteria and actinomycetes. However, the relative ¹³C concentrations of the latter communities increased after 1 week, while those of fungi decreased and those of gram-negative bacteria remained constant. PLFA ¹³C mean residence times were much shorter for fungi compared to bacteria and actinomycetes.

Conclusions

Our results indicate temporally varying rhizodeposit-C uptake by different microbial groups, and faster turnover rates of mycorrhizal versus saprotrophic fungi and fungi versus bacteria. Fungi appeared to play a major role in the initial processing and possible rapid channeling of rhizodeposit-C into the soil microbial community. Actinomycetes and gram-positive bacteria appeared to have a delayed utilization of rhizodeposit-C or to prefer other C sources upon rhizodeposition.     

Intensification of the aerobic bioremediation of an actual site soil historically contaminated by polychlorinated biphenyls (PCBs) through bioaugmentation with a non acclimated, complex source of microorganisms

Background  

The biotreatability of actual-site polychlorinated biphenyl (PCB)-contaminated soils is often limited by their poor content of autochthonous pollutant-degrading microorganisms. In such cases, inoculation might be the solution for a successful bioremediation. Some pure and mixed cultures of characterized PCB degrading bacteria have been tested to this purpose. However, several failures have been recorded mostly due to the inability of inoculated microbes to compete with autochthonous microflora and to face the toxicity and the scarcity of nutrients occurring in the contaminated biotope. Complex microbial systems, such as compost or sludge, normally consisting of a large variety of robust microorganisms and essential nutrients, would have better chances to succeed in colonizing degraded contaminated soils. However, such sources of microorganisms have been poorly applied in soil bioremediation and in particular in the biotreatment of soil with PCBs. Thus, in this study the effects of Enzyveba, i.e. a consortium of non-adapted microorganisms developed from composted material, on the slurry- and solid-phase aerobic bioremediation of an actual-site, aged PCB-contaminated soil were studied.     

Contrasts between subsurface microbial communities and their metabolic adaptation to polycyclic aromatic hydrocarbons at a forested and an urban coal-tar disposal site

The abundance and distribution of microorganisms and their potential for mineralizing polycyclic aromatic hydrocarbons (PAHs) were measured in subsurface sediment samples at two geographically separate buried coal-tar sites. At a relatively undisturbed forested site in the northeastern United States, metabolic adaptation to the PAHs was evident: Radiolabeled naphthalene and phenanthrene were converted to ¹⁴CO₂ in core material from inside but not outside a plume of groundwater contamination. However, at the urban site in the midwestern United States these PAHs were mineralized in sediments from both contaminated and uncontaminated boreholes. Thus, clear qualitative evidence showing an adaptational response by the subsurface microbial community was not obtained at the urban site. Instead, subtler clues suggesting metabolic adaptation by subsurface microorganisms from the urban site were discerned by comparing lag periods and extents of ¹⁴CO₂ production from radiolabeled PAHs added to samples from contaminated and uncontaminated boreholes. Despite slightly higher PAH mineralization activity in contaminated borehole samples, p-hydroxybenzoate was mineralized equally in all samples from the urban site regardless of location. No striking trends in the abundances of actinomycetes, fungi, and either viable or total bacteria were encountered. However, colonies of the soil bacterium, Bacillus mycoides, were detected on enumeration plates of several samples from unsaturated and saturated zones in both urban boreholes. Furthermore, other common soil bacteria, Myxococcus xanthus and Chromobacterium violaceum, were identified in samples from the uncontaminated urban borehole. The occurrence of bacteria usually restricted to surface soil, combined with the observation of fragments of building materials in many of the core samples, suggested that past excavation and backfilling operations may have caused mixing of surface soil with subsurface materials at the urban site. We speculate that this mixing, as well as non-coal-tar-derived sources of PAHs, contributed to the PAH-mineralizing activity present in the sediment samples from the uncontaminated urban borehole.     

A computational approach to discovering the functions of bacterial phytochromes by analysis of homolog distributions

Background  

Phytochromes are photoreceptors, discovered in plants, that control a wide variety of developmental processes. They have also been found in bacteria and fungi, but for many species their biological role remains obscure. This work concentrates on the phytochrome system of Agrobacterium tumefaciens, a non-photosynthetic soil bacterium with two phytochromes. To identify proteins that might share common functions with phytochromes, a co-distribution analysis was performed on the basis of protein sequences from 138 bacteria.     

Fate of Escherichia coli O145 present naturally in bovine slurry applied to vegetables before harvest,after washing and simulated wholesale and retail distribution

Aims

To determine the fate of Escherichia coli on vegetables that were processed through commercial wash treatments and stored under simulated retail conditions at 4°C or wholesale at fluctuating ambient temperatures (0–25°C, dependent on season).

Methods and Results

Bovine slurry that was naturally contaminated with E. coli O145 was applied without dilution or diluted 1:10 using borehole water to growing potatoes, leeks or carrots. Manure was applied 1 week prior to harvest to simulate a near‐harvest contamination event by manure deposition or an application of contaminated water to simulate a flooding event or irrigation from a contaminated water source. At harvest, crops were contaminated at up to 2 log cfu g^?1. Washing transferred E. coli into the water of a flotation tank used for potato washing and did not completely remove all traces of contamination from the crop. Manure‐contaminated potatoes were observed to contain 0·72 cfu E. coli O145 g^?1 after processing and retail storage. Manure‐contaminated leeks harboured 0·73–1·55 cfu E. coli O145 g^?1 after washing and storage. There was no cross‐contamination when leeks were spray washed. Washing in an abrasive drum resulted in less than perfect decontamination for manure‐contaminated carrots. There were five post‐distribution isolations from carrots irrigated with contaminated water 24 h prior to harvest.

Conclusions

Standard commercial washing and distribution conditions may be insufficient to reliably control human pathogenic E. coli on fresh produce.

Significance and Impact

Previous speculation that the cause of a UK foodborne disease outbreak was soil from imperfectly cleaned vegetables is plausible.     

Changes in microbial activities and biomasses over a forest floor gradient in C-to-N ratio

Background and aims

Under chronically elevated N deposition, N retention mainly occur at high soil C-to-N ratio. This may be mediated through soil microbes, such as ectomycorrhizal (EM) fungi, saprotrophic fungi and bacteria, and the aim of this study was to evaluate the relationship between soil microbes and forest floor C-to-N ratios.

Methods

Soil samples from 33 Norway spruce (Picea abies (L.) H. Karst) forests in Denmark and southern Sweden in a forest floor C-to-N ratio gradient (ranging from 14 to 35) were analysed regarding the content of phospholipid fatty acids (PLFAs) to estimate their soil microbial community composition and the relative biomasses of different microbial groups. The relation of EM biomass to total fungal biomass was estimated as the loss of the fungal PLFA 18:2ω6,9 during incubation of soils and the production of EM mycelia was estimated using fungal in-growth mesh bags. The soil microbial variables were correlated to forest floor C-to-N ratio, NO ₃ ^- leaching, soil pH and stand age.

Results

Fungal proportions of microbial biomass, EM to total fungi and EM mycelial production were all positively related to C-to-N ratio, while NO ₃ ^- leaching was negatively related to C-to-N ratio.

Conclusions

Both EM and saprotrophic fungi change with forest floor C-to-N ratios and appear to play a central role in N retention in forest soil. A better understanding of the mechanisms behind this process may be revealed if the role of recalcitrant fungal metabolites for N retention (and soil C sequestration) can be identified. Research along this line deserves further studies.     

As,Pb, Sb,and Zn transfer from soil to root of wild rosemary: do native symbionts matter?

Background and aims

This is an in natura study aimed to determine the potential of Rosmarinus officinalis for phytostabilization of trace metal and metalloid (TMM)-contaminated soils in the Calanques National Park (Marseille, southeast of France). The link between rosemary tolerance/accumulation of As, Pb, Sb, and Zn and root symbioses with arbuscular mycorrhizal (AM) fungi and/or dark septate endophytes (DSE) was examined.

Methods

Eight sites along a gradient of contamination were selected for soil and root collections. TMM concentrations were analyzed in all the samples and root symbioses were observed. Moreover, in the roots of various diameters collected in the most contaminated site, X-ray microfluorescence methods were used to determine TMM localization in tissues.

Results

Rosemary accumulated, in its roots, the most labile TMM fraction in the soil. The positive linear correlation between TMM concentrations in soil and endophyte root colonization rates suggests the involvement of AM fungi and DSE in rosemary tolerance to TMM. Moreover, a typical TMM localization in root peripheral tissues of thin roots containing endophytes forming AM and DSE development was observed using X-ray microfluorescence.

Conclusions

Rosemary and its root symbioses appeared as a potential candidate for a phytostabilization process of metal-contaminated soils in Mediterranean area.     

Effects of oil spills on microbial heterotrophs in Antarctic soils

Oil spillage on the moist coastal soils of the Ross Sea region of Antarctica can impact on populations of microbial heterotrophs in these soils, as determined by viable plate counts and a most probable number technique. Elevated numbers of culturable hydrocarbon degraders, bacteria and fungi were detected in surface and subsurface soils from oil-contaminated sites, compared with nearby control sites. Culturable yeasts were not detected in soil from coastal control sites, yet reached >10⁵ organisms g^-1 dry weight in contaminated soils. The presence of hydrocarbons in soils resulted in a shift in the genera of culturable filamentous fungi. Chrysosporium dominated control soils, yet Phialophora was more abundant in oil-contaminated soils. Hydrocarbon degraders are most likely bacteria; however, fungi could play a role in degradation of hydrocarbons or their metabolites. Depleted levels of nitrate detected in some contaminated soils and decreased pH may be the result of growth of hydrocarbon degraders. Numbers and diversity of culturable microbes from Antarctic soil varied depending on whether a pristine site or a human-impacted (in this case, by fuel spills) site is studied.     

Expressions and clinical significances of c-MET, p-MET and E2f-1 in human gastric carcinoma

Background

Studies in different parts of the world have implicated money as a vehicle for transmission of pathogens. Such information which is necessary to facilitate infection control strategies is lacking in many sub-Saharan countries including Cameroon. This study analyzed the Franc de la Communauté Financiere d’Afrique (Franc CFA), the currency used in Cameroon and other countries in the Central African sub-region, as a potential vehicle for transmission of pathogenic bacteria and fungi, particularly drug-resistant strains, to generate findings which could create awareness on currency contamination and serve as a guide when formulating health policies on currency.

Methods

Two hundred and thirteen currency samples representing various denominations of notes and coins randomly collected from diverse sources in Buea, Cameroon were analyzed for bacteria and fungi. The sensitivity of bacterial isolates to antibiotics was tested using the disc diffusion method. The relationship between contamination and physical state, source or denomination of currency was assessed using the χ² test. All statistics were discussed at 0.05 significance level.

Results

Two hundred (93.9%) samples were contaminated with notes (96.6%) showing higher contamination than coins (88.2%). Uncirculated (mint) samples showed no contamination. There was a significant difference (P?0.05) in contamination with respect to currency denomination, physical state and source. All samples from butchers and patients/personnel in hospitals were contaminated. Lower denominations showed significantly higher (P = 0.008) levels of contamination than higher denominations. Dirty currency was more contaminated than clean currency. Nine bacterial species were isolated. Coagulase-negative Staphylococcus (CoNS) (54.9%) and Staphylococcus aureus (20.1%) predominated. Among the fungi detected, Aspergillus sp (17.3%) and Penicillium sp (15.9%) showed higher frequency of occurrence. Bacteria were susceptible (100%) to ceftriaxone, gentamicin, norfloxacin and ofloxacin. Susceptibility to amoxicillin, penicillin, ampicillin, vancomycin and cotrimoxazole was low. Staphylococci were resistant (100%) to vancomycin, penicillin G, and amoxicillin. CoNS in addition showed resistance (100%) to cotrimoxazole.

Conclusions

The CFA franc circulating in Buea could serve as a vehicle for transmission of drug resistant pathogenic or potential organisms and contamination could be due to currency usage and handling as mint notes were not contaminated. Hygiene practices during or after handling currency is greatly encouraged to prevent infection.     

Biphenyl-Metabolizing Bacteria in the Rhizosphere of Horseradish and Bulk Soil Contaminated by Polychlorinated Biphenyls as Revealed by Stable Isotope Probing

DNA-based stable isotope probing in combination with terminal restriction fragment length polymorphism was used in order to identify members of the microbial community that metabolize biphenyl in the rhizosphere of horseradish (Armoracia rusticana) cultivated in soil contaminated with polychlorinated biphenyls (PCBs) compared to members of the microbial community in initial, uncultivated bulk soil. On the basis of early and recurrent detection of their 16S rRNA genes in clone libraries constructed from [¹³C]DNA, Hydrogenophaga spp. appeared to dominate biphenyl catabolism in the horseradish rhizosphere soil, whereas Paenibacillus spp. were the predominant biphenyl-utilizing bacteria in the initial bulk soil. Other bacteria found to derive carbon from biphenyl in this nutrient-amended microcosm-based study belonged mostly to the class Betaproteobacteria and were identified as Achromobacter spp., Variovorax spp., Methylovorus spp., or Methylophilus spp. Some bacteria that were unclassified at the genus level were also detected, and these bacteria may be members of undescribed genera. The deduced amino acid sequences of the biphenyl dioxygenase α subunits (BphA) from bacteria that incorporated [¹³C]into DNA in 3-day incubations of the soils with [¹³C]biphenyl are almost identical to that of Pseudomonas alcaligenes B-357. This suggests that the spectrum of the PCB congeners that can be degraded by these enzymes may be similar to that of strain B-357. These results demonstrate that altering the soil environment can result in the participation of different bacteria in the metabolism of biphenyl.Polychlorinated biphenyls (PCBs) are very stable chloroorganic compounds with the general formula C₁₂H_10-xCl_x. Mixtures of PCBs have been used as coolants and lubricants in transformers, capacitors, and other electrical equipment as they do not burn easily and are good insulators. It is estimated that some 1.5 million tons of PCBs were produced up to 1988 worldwide (11; http://www.atsdr.cdc.gov/cercla; http://www.epa.gov/epawaste/hazard/tsd/pcbs/pubs/about.htm). Although production of these compounds was stopped, due to their long-term persistence, many sites all over the world are still contaminated with PCBs. Moreover, not only do PCBs threaten human health in the vicinity of the contaminated area, but lower PCB congeners volatilize and migrate to places far from where they were originally released (2, 3, 16). Also, their metabolic products have environmental significance; activities of both plants and microorganisms result in formation of different intermediates and final products whose toxicity can in some cases be even higher than that of the original toxicant (24, 26; http://www.atsdr.cdc.gov/cercla).Physical-chemical methods used for the removal of PCBs often cause further natural disturbance and pollution; in contrast, biological methods of removal (i.e., bioremediation) are less expensive and more environmentally sound and thus have aroused much interest (7). These methods include the use of microorganisms and also exploitation of plants (i.e., phytoremediation) (19) and the cooperation of plants with microorganisms in the rhizosphere (i.e., rhizoremediation) (21). These bioremediation options also include the use of genetically modified bacteria (6) and/or plants (18, 23). PCBs were only recently introduced into the environment, and no completely efficient pathways for the aerobic bacterial degradation of all of these compounds have evolved (34); however, lower chlorinated PCB congeners can be degraded via the pathway that is used by aerobic bacteria to degrade biphenyl (35). Therefore, metabolism of biphenyl as a potential cometabolite of PCBs was the subject of this study.The biphenyl degradation pathway is the same in all aerobic bacteria, and enzymes of this pathway degrade biphenyl in four steps into benzoate and 2-hydroxypenta-2,4-dienoate (21). The first enzyme of the pathway, biphenyl dioxygenase, has broad substrate specificity and thus permits degradation of biphenyl-related compounds (9). Substrates for biphenyl dioxygenase comprise, in addition to biphenyl itself, other diphenyl or benzene skeletons with several substituents, including halogens and bicyclic or tricyclic fused heterocyclic aromatics (35). These substrates also include certain natural compounds, including some plant flavonoids, phenols, or terpenes (10). Bacteria capable of metabolizing biphenyl are thus pervasive members of many microbial communities in vegetated soil.As reported previously (20), there are two main problems with introduction of a new population of degrading or genetically modified microorganisms to enhance the biodegradation of PCBs in a contaminated environment: legislative barriers and the inability of strains added to the soil to survive. Therefore, the use of microorganisms for bioremediation of contaminated sites is not likely to be successful. Hence, understanding the biodegradative processes in the natural communities is necessary for planning remediation strategies. Identification of members of the community potentially responsible for the degradative process has recently been enabled by DNA-based stable isotope probing (SIP), as reviewed previously; therefore, this technique has become an efficient tool in microbial ecology (33). In this study, by tracking the transfer of ¹³C from [¹³C]biphenyl into bacterial DNA, it was possible to identify biphenyl-metabolizing bacteria in PCB-contaminated soil. To analyze how the bacterial diversity can be changed by introduction of a plant and subsequent cultivation in a greenhouse, bacteria in the rhizosphere of horseradish (Armoracia rusticana) cultivated in a contaminated soil were studied.     

Analysis of phenanthrene degradation by Ascomycota fungi isolated from contaminated soil from Reynosa,Mexico

Polycyclic aromatic hydrocarbons (PAHs) are organic compounds generated mainly by anthropogenic sources. They are considered toxic to mammals, since they have carcinogenic, mutagenic and genotoxic properties, among others. Although mycoremediation is an efficient, economical and eco-friendly technique for degrading PAHs, the fungal degradation potential of the phylum Ascomycota has not been widely studied. In this work, we evaluated different fungal strains from the polluted soil of ‘La Escondida’ lagoon in Reynosa, Mexico to know their potential to degrade phenanthrene (PHE). Forty-three soil isolates with the capacity to grow in the presence of PHE (0·1% w/v) were obtained. The fungi Aspergillus oryzae MF13 and Aspergillus flavipes QCS12 had the best potential to degrade PHE. Both fungi germinated and grew at PHE concentrations of up to 5000 mg l⁻¹ and degraded 235 mg l⁻¹ of PHE in 28 days, with and without an additional carbon source. These characteristics indicate that A. oryzae MF13 and A. flavipes QCS12 could be promising organisms for the remediation of sites contaminated with PAHs and detoxification of recalcitrant xenobiotics.     

Soil nitrogen balance resulting from N fixation and rhizodeposition by the symbiotic association Anthyllis vulneraria/Mesorhizobium metallidurans grown in highly polluted Zn,Pb and Cd mine tailings

Background and aims

The association of the legume Anthyllis vulneraria and the grass Festuca arvernensis, was found to be very efficient for the phytostabilisation of highly multi-metal contaminated mine tailings. Our objective was to quantify the contribution of Anthyllis inoculated with its symbiotic bacteria Mesorhizobium metallidurans to the soil N pool and to test whether a starter nitrogen fertilization may improve symbiotic nitrogen fixation and the growth of Festuca.

Methods

Plants of Festuca and of Anthyllis inoculated with M. metallidurans were grown separately during eight months in pots filled with mine contaminated soil. Estimation of the N fluxes was realized using ¹⁵?N isotopic methods.

Results

Starter N fertilization (28 kg N ha^?1) improved symbiotic N₂ fixation and the growth of both species. Belowground N balance (N rhizodeposition – soil N uptake) of the non-fertilized Anthyllis at maturity was negative (?30.6 kg N ha^?1). However, the amount of N derived from fixation, including above- and belowground parts, was 78.6 kg N ha^?1, demonstrating the ability of this symbiotic association to improve soil N content after senescence.

Conclusions

i) soil N enrichment by the N₂-fixing symbiotic association occurs after plant senescence, when decaying leaves and shoots are incorporated into the soil; ii) application of a starter fertilization is an efficient solution to improve phytostabilisation of highly contaminated sites.     

Airborne concentrations of bacteria and fungi in Thailand border market

Thailand border market is where the local Thais, Cambodians, Laotians, and Burmeses exchange their goods and culture at the border checkpoints. It is considered to be the source of aerial disease transmission especially for foreigners because it is always very crowded with people from all walks of life. Unhealthy air quality makes this area high risk of spread of airborne diseases. This study assessed airborne concentrations of bacteria and fungi in a border market to improve exposure estimates and develop efficient control strategies to reduce health risk. The density and distribution of airborne bacteria and fungi were investigated in the Chong Chom border market in Surin Province, Thailand. Eighteen air sampling sites were taken from outdoors and various work environments including indoor footpaths, wooden handicraft shops, electronic shops, the secondhand clothing shops, and fruit market areas. Exposed Petri plate method and liquid impinger sampler were used for sampling at the breathing zone, 1.5 m above the floor level, during weekend and holiday. Meteorological factors such as relative humidity, temperature, and light intensity were collected by portable data logger. The relative humidity was 67–73%, and temperature 29–33°C, and light varied between 18 and 270 Lux m⁻². Gram-positive and Gram-negative bacteria were found at a mean value of 10⁴ CFU m⁻³, and airborne fungi of 10³ CFU m⁻³ were recorded. The highest concentration of culturable airborne microorganisms was found along the indoor footpath (9.62 × 10⁴ CFU m⁻³ and 750.00 CFU/plate/h for impingement and sedimentation methods, respectively), the fruit market area (7.86 × 10⁴ CFU m⁻³ and 592.42 CFU/plate/h for impingement and sedimentation methods, respectively), and the secondhand clothing shop (4.59 × 10³ CFU m⁻³ and 335.42 CFU/plate/h for impingement and sedimentation methods, respectively) for Gram-positive bacteria, Gram-negative bacteria, and fungi, respectively. The lowest concentration of Gram-positive bacteria, Gram-negative bacteria, and fungi was found only at the outdoor area at 1.53 × 10⁴ CFU m⁻³, 0.93 × 10⁴ CFU m⁻³ and 0.80 × 10³ CFU m⁻³ by means of impingement method and 136.67 CFU/plate/h, 69.25 CFU/plate/h, and 62.00 CFU/plate/h by means of sedimentation methods for Gram-positive bacteria, Gram-negative bacteria, and fungi, respectively. The most frequently present airborne bacteria were identified as Bacillus, Corynebacteria, Diplococcus, Micrococcus, Acinetobacter, Alcaligenes, Enterobacter, and spore former rods. Acremonium, Aspergillus, Cladosporium, Penicillium, and Sporotrichum were the most frequently found aerosol fungi genera. The distribution of airborne microorganisms correlated with relative humidity and light factors based on principal component analysis. In conclusion, the border market is a potential source of aerial disease transmission and a various hazards of bioaerosols for workers, consumers, sellers, and tourists. The bioaerosol concentration exceeded the standard of occupational exposure limit. Many major indicators of allergenic and toxigenic airborne bacteria and fungi, Acinetobacter, Enterobacter, Pseudomonas, Cladosporium, Alternaria, Aspergillus, and Penicillium, were found in the various market environments.     

Screening of white-rot fungi for their ability to mineralize polycyclic aromatic hydrocarbons in soil

Soil samples from an agricultural field contaminated with 10 ppm¹⁴C-benz(a)anthracene in glass tubes were brought into contact with cultures of wood-rotting fungi, precultivated on wheat straw substrate. Forty-five strains of white-rot fungi and four brown-rot fungi were tested for their ability to colonize the soil and to mineralize¹⁴C-benz(a)anthracene to¹⁴CO₂ within a 20-week incubation time. Twenty-two white-rot fungi and all brown-rot fungi were unable to colonize the soil. Twenty-three strains of white-rot fungi, all belonging to the genusPleurotus, colonized the soil. During the experiment the noncolonizing fungi and their substrate disintegrated more and more to a nonstructured pulp from which water diffused into the soil. The same phenomenon was observed in the control which contained only straw without fungus and contaminated soil. In samples with colonizing fungi the substrate as well as the mycelia in the soil remained visibly unchanged during the entire experiment. Surprisingly, most samples with fungi not colonizing the soil and the control without fungus liberated between 40 and 58 % of the applied radioactivity as¹⁴CO₂ whereas the samples with the colonizing fungi respired only 15–25 % as¹⁴CO₂. This was 3–5 times more¹⁴CO₂ than that liberated from the control (4.9 %) which contained only contaminated soil without straw and fungus. A similar result was obtained with selected colonizing and noncolonizing fungi and soil contaminated with 10 ppm¹⁴C-pyrene. However, in pure culture studies in which¹⁴C-pyrene was added to the straw substrate,Pleurotus sp. (P2), as a representative of the colonizing fungi, mineralized 40.3 % of the added radioactivity to¹⁴CO₂. The noncolonizing fungiDichomitus squalens andFlammulina velutipes liberated only 17.2 or 1.7 %, respectively, as¹⁴CO₂. These results lead to the hypothesis that the native soil microflora stimulated by the formed products of straw lysis is responsible for high degradation rates found with noncolonizing fungi.     

Bioaerosols and Particle Release During Composting of Contaminated Sawmill Soil

Compost windrows for bioremediation of soil were built at a wood-preserving site contaminated with chlorophenols, polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). Sampling of airborne particles during the mixing of the compost windrows found concentrations of PCDDs and PCDFs in different particle sizes. The congener distribution of PCDDs and PCDFs in the collected air particle fractions was similar to that in the compost windrows, and the level of PCDDs and PCDFs was 1000-fold higher than the atmospheric background values reported previously. Viable particle-sizing samplers and several selective growth media were used to enumerate bacteria and fungi in the airborne particles. From the collected air samples, 40 bacteria were isolated and identified. Among the isolated bacteria, 80% were Gram-positive and spore-forming. Two of the identified airborne bacteria, Pseudomonas aeruginosa and Bacillus cereus, may cause human disease and are classified in biological agent hazard group 2. The amounts of airborne fungi, molds, and yeasts were 1000 to 2000 colony-forming units (CFUs) per m³. The number of actinomycetes was up to 6-fold, and the number of bacteria was 2- to 20-fold compared to background values. The overall level of airborne bacteria (200 to 3500 CFUs per m³) was low compared to the level of bacteria (10⁵ to 10⁸ CFUs per m³) found when composting municipal waste.     

Bioremediation of DDT contaminated soil using brown-rot fungi

The ability of brown-rot fungi (BRF) to eliminate DDT in artificially and historically contaminated soil was investigated to determine whether the BRF would be suitable for the bioremediation of DDT in soil. Gloeophyllum trabeum, Fomitopsis pinicola and Daedalea dickinsii showed an ability to eliminate DDT in artificially contaminated sterilized (SL) and un-sterilized (USL) soils. The addition of Fe²⁺ to the soil system enhanced the ability of some BRF to eliminate DDT. In the contaminated SL soil, the DDT was eliminated by approximately 41%, 9% and 15% by G. trabeum, F. pinicola and D. dickinsii, respectively. Compared with the controls, in the USL soil approximately 43%, 29% and 32% of DDT was eliminated and approximately 20%, 9% and 26% of DDD (1,1-dichloro-2,2-bis (4-chlorophenyl) ethane) was detected as a metabolic product with G. trabeum, F. pinicola and D. dickinsii, respectively. Of the BRF, G. trabeum demonstrated the greatest ability to eliminate DDT both in the SL and USL soils. G. trabeum was applied to a historically contaminated soil which had a DDT concentration more than three times the artificially contaminated soil. G. trabeum remediated about 64% of the initial DDT with the addition of Fe²⁺. There were no significant differences in the results with or without the addition of Fe²⁺, indicating that G. trabeum can be used directly for the degradation of DDT in soil without any other additional treatment. This study identified that G. trabeum is the most promising BRF for use in the bioremediation of DDT contaminated soil.     

Biodegradation of petroleum hydrocarbons by filamentous fungi (Aspergillus ustus and Purpureocillium lilacinum) isolated from used engine oil contaminated soil

The use of microorganisms for remediation and restoration of hydrocarbons contaminated soils is an effective and economic solution. The current study aims to find out efficient telluric filamentous fungi to degrade petroleum hydrocarbons pollutants. Six fungal strains were isolated from used engine (UE) oil contaminated soil. Fungi were screened for their ability to degrade crude oil, diesel and UE oil using 2.6-dichlorophenol indophenol (DCPIP). Two isolates were selected, identified and registered at NCBI as Aspergillus ustus HM3.aaa and Purpureocillium lilacinum HM4.aaa. Fungi were tested for their tolerance to different concentration of petroleum oils using radial growth diameter assay. Hydrocarbons removal percentage was evaluated gravimetrically. The degradation kinetic of crude oil was studied at a time interval of 10 days. A.ustus was the most tolerant fungi to high concentration of petroleum oils in solid medium. Quantitative analysis showed that crude oil was the most degraded oil by both isolate; P. lilacinium and A. ustus removed 44.55% and 30.43% of crude oil, respectively. The two fungi were able to degrade, respectively, 27.66 and 21.27% of diesel and 14.39 and 16.00% of UE oil. As compared to the controls, these fungi accumulated high biomass in liquid medium with all petroleum oils. Likewise, crude oil removal rate constant (K) and half-lives (t_1/2) were 0.02 day⁻¹, 34.66 day and 0.015 day⁻¹, 46.21 day for P. lilacinium and A. ustus, respectively. The selected fungi appear interesting for petroleum oils biodegradation and their application for soil bioremediation require scale-up studies.     

Accumulation of polychlorinated biphenyls in adipocytes: selective targeting to lipid droplets and role of caveolin-1

Background

Polychlorinated biphenyls (PCBs) are persistent environmental pollutants that preferentially accumulate in lipid-rich tissues of contaminated organisms. Although the adipose tissue constitutes a major intern reservoir of PCBs and recent epidemiological studies associate PCBs to the development of obesity and its related disorders, little is known about the mechanisms involved in their uptake by the adipose tissue and their intracellular localization in fat cells.

Methodology/Principal Findings

We have examined the intracellular distribution of PCBs in mouse cultured adipocytes and tested the potential involvement of caveolin-1, an abundant adipocyte membrane protein, in the uptake of these compounds by fat cells. We show that 2,4,4′-trichlorobiphenyl (PCB-28), 2,3′,4,4′,5-pentachlorobiphenyl (PCB-118) and 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB-153) congeners rapidly and extensively accumulate in 3T3-L1 or mouse embryonic fibroblast (MEF) derived cultured adipocytes. The dynamics of accumulation differed between the 3 congeners tested. By subcellular fractionation of primary adipocytes, we demonstrate that these pollutants were almost exclusively recovered within the lipid droplet fraction and practically not associated to cell membranes. The absence of caveolin-1 expression in primary adipocytes from cav-1 deficient mice did not modify lipid droplet selective targeting of PCBs. In cav-1 KO MEF differentiated adipocytes, PCB accumulation was decreased, which correlated with reduced cell triglyceride content. Conversely, adenoviral mediated cav-1 overexpressing in 3T3-L1 cells, which had no impact on total cell lipid content, did not change PCB accumulation.

Conclusion/Significance

Our data indicate that caveolin-1 per se is not required for selective PCB accumulation, but rather point out a primary dependence on adipocyte triglyceride content. If the crucial role of lipid droplets in energy homeostasis is considered, the almost exclusive accumulation of PCBs in these organelles warrants future attention as the impairment of their function could be linked to the worldwide obesity epidemic.     

Nitrogen deposition and soil carbon content affect nitrogen mineralization during primary succession in acid inland drift sand vegetation

Background and aims

Two inland dunes in the Netherlands receiving low (24) and high (41 kg N ha^?1 yr^?1) nitrogen (N) deposition were compared for N dynamics and microbial activity to investigate the potential effect of N on succession rate of the vegetation and loss of pioneer habitats.

Methods

Primary succession stages were sampled, including bare sand, and vegetation dominated by Polytrichum piliferum, Campylopus introflexus, lichens and grasses respectively, representing a series of vegetation types in undisturbed drift sand sites with succession starting on bare sand containing virtually no organic matter. Microbial characteristics and potential N mineralization were analysed in a laboratory experiment.

Results

Organic matter accumulated during succession, resulting in a lower pH and in higher microbial biomass (bacteria and fungi), respiration and net N mineralization. The increase in respiration and N mineralization was largely due to the development of an ectorganic layer in the middle stages of succession. The observed effects of N deposition were (1) decrease of microbial biomass, (2) higher net N mineralization per m², (3) higher levels of free nitrogen in the soil, and (4) a higher microbial N:P ratio.

Conclusions

Elevated N deposition leads to higher N availability which may cause accelerated succession.     

Soil microorganisms respond to five years of climate change manipulations and elevated atmospheric CO2 in a temperate heath ecosystem

Background and aims

Soil microbial responses to global change can affect organic matter turnover and nutrient cycling thereby altering the overall ecosystem functioning. In a large-scale experiment, we investigated the impact of 5 years of climate change and elevated atmospheric CO₂ on soil microorganisms and nutrient availability in a temperate heathland.

Methods

The future climate was simulated by increased soil temperature (+0.3 °C), extended pre-summer drought (excluding 5–8 % of the annual precipitation) and elevated CO₂ (+130 ppm) in a factorial design. Soil organic matter and nutrient pools were analysed and linked to microbial measures by quantitative PCR of bacteria and fungi, chloroform fumigation extraction, and substrate-induced respiration to assess their impact of climate change on nutrient availability.

Results

Warming resulted in higher measures of fungi and bacteria, of microbial biomass and of microbial growth potential, however, this did not reduce the availability of nitrogen or phosphorus in the soil. Elevated CO₂ did not directly affect the microbial measures or nutrient pools, whereas drought shifted the microbial community towards a higher fungal dominance.

Conclusions

Although we were not able to show strong interactive effects of the global change factors, warming and drought changed both nutrient availability and microbial community composition in the heathland soil, which could alter the ecosystem carbon and nutrient flow in the long-term.