Effect of surfactant solubilization on biodegradation of polychlorinated biphenyl congeners by Pseudomonas LB400

A variety of commercial surfactants were tested to determine their effect on polychlorinated biphenyl (PCB) transformation by Pseudomonas LB400. Initial tests determined that most surfactants were fully or partially able to solubilize the PCB congeners 2,5,2′-chlorobiphenyl (CBP), 2,4,2′,4′-CBP, 2,3,5,2′,5′-CBP and 2,4,5,2′,4′,5′-CBP, at concentrations above the surfactants' critical micelle concentration (CMC). Surfactants were also found to have no negative effect on bacterial survival, as cell numbers were the same or higher after incubation in the presence of surfactants than after incubation without surfactants. A comparison of the extent of biotransformation of single PCB congeners by the bacterium revealed that, at surfactant concentrations above the CMC, the presence of an anionic surfactant promoted while nonionic surfactants inhibited PCB transformation, compared to a control with no surfactant. The rates of transformation of PCB congeners were also higher in the presence of the anionic surfactant compared to the control. The inhibitory effects of a nonionic surfactant, Igepal CO-630 at a concentration above its CMC, on transformation of 2,4,5,2′,5′-CBP could be eliminated by diluting the surfactant/PCB solution to a concentration close to the surfactant CMC. Received: 26 October 1998 / Received revision: 5 March 1999 / Accepted: 14 March 1999     

The effects on operation conditions of sludge retention time and carbon/nitrogen ratio in an intermittently aerated membrane bioreactor (IAMBR)

An intermittently aerated membrane bioreactor (IAMBR) system has been developed to improve the efficiency of nutrient removal, and for the stable treatment of organic matter which is contained as suspended solid (SS) in the influent. The important operating factors of an intermittently aerated bioreactor (IABR) are sludge retention times (SRTs) and carbon/nitrogen (C/N) ratios. Because research on IAMBR is young, this paper explores the effect of SRTs and C/N ratios on these systems. For SRTs of 20, 25, 30, and 40 days, there was little difference in the removal of COD, T–N, and T–P. In comparing C/N ratios of 4.5, 7, and 10, the COD concentration in permeate with a C/N ratio of 10 was most stable, although the concentration of organic matter in the influent was high. The removal efficiencies of T–N and T–P in permeate with a C/N ratio of 10 were the highest at 92.9% and 88.9%, respectively. This implies that a C/N ratio above 10 should be maintained for a nutrient removal efficiency of approximately 90%.     

Psychrotolerant Bacteria Isolated from Arctic Soil That Degrade Polychlorinated Biphenyls at Low Temperatures

Psychrotolerant polychlorinated biphenyl (PCB)-degrading bacteria were isolated at 7°C from PCB-contaminated Arctic soil by using biphenyl as the sole organic carbon source. These isolates were distinguished from each other by differences in substrates that supported growth and substrates that were oxidized. 16S ribosomal DNA sequences suggest that these isolates are most closely related to the genus Pseudomonas. Total removal of Aroclor 1242, and rates of removal of selected PCB congeners, by cell suspensions of Arctic soil isolates and the mesophile Burkholderia cepacia LB400 were determined at 7, 37, and 50°C. Total removal values of Aroclor 1242 at 7°C by LB400 and most Arctic soil isolates were similar (between 2 and 3.5 μg of PCBs per mg of cell protein). However the rates of removal of some individual PCB congeners by Arctic isolates were up to 10 times higher than corresponding rates of removal by LB400. Total removal of Aroclor 1242 and the rates of removal of individual congeners by the Arctic soil bacteria were higher at 37°C than at 7°C but as much as 90% lower at 50°C than at 37°C. In contrast, rates of PCB removal by LB400 were higher at 50°C than at 37°C. In all cases, temperature did not affect the congener specificity of the bacteria. These observations suggest that the PCB-degrading enzyme systems of the bacteria isolated from Arctic soil are cold adapted.     

Bioremediation of polychlorinated biphenyl-contaminated soil using carvone and surfactant-grown bacteria

Partial bioremediation of polychlorinated biphenyl (PCB)-contaminated soil was achieved by repeated applications of PCB-degrading bacteria and a surfactant applied 34 times over an 18-week period. Two bacterial species, Arthrobacter sp. strain B1B and Ralstonia eutrophus H850, were induced for PCB degradation by carvone and salicylic acid, respectively, and were complementary for the removal of different PCB congeners. A variety of application strategies was examined utilizing a surfactant, sorbitan trioleate, which served both as a carbon substrate for the inoculum and as a detergent for the mobilization of PCBs. In soil containing 100 μg Aroclor 1242 g⁻¹ soil, bioaugmentation resulted in 55–59% PCB removal after 34 applications. However, most PCB removal occurred within the first 9 weeks. In contrast, repeated addition of surfactant and carvone to non-inoculated soil resulted in 30–36% PCB removal by the indigenous soil bacteria. The results suggest that bioaugmentation with surfactant-grown, carvone-induced, PCB-degrading bacteria may provide an effective treatment for partial decontamination of PCB-contaminated soils. Received: 9 March 2000 / Received revision: 27 June 2000 / Accepted: 16 July 2000     

Optimization of Influencing Parameters on Phenanthrene Removal Efficiency in Soil Washing Process by Using Response Surface Methodology

Response surface methodology (RSM) under Box–Behnken design (BBD) was applied to evaluate the effect of the influencing parameters including surfactant concentration, liquid/soil ratio, Humic Acid concentration, and washing time on phenanthrene removal efficiency in soil washing process by using the nonionic surfactant Tween 80 and find an optimal operational conditions to achieve the highest removal efficiency. A polynomial quadratic model was used to correlate phenanthrene removal efficiency and four independent variables (R² = 0.9719). Based on the obtained results the most influential parameter on phenanthrene removal efficiency was surfactant concentration with an impact value of 69.519%. Liquid/soil ratio was also another factor that significantly influenced on removal efficiency with an impact value of 25.014%. The interaction between surfactant concentration and liquid/soil ratio was also shown to have a positive significant effect on removal efficiency (p_value = 0.0027). However, the other independent variables Humic Acid concentration and time were not significant in the ranges selected in this study. Based on the optimization results maximum removal efficiency of 70.692 ± 3.647% was achieved under the conditions of surfactant concentration 5000 mg L^?1, liquid/soil ratio 30 v/w, HA concentration 9.88 mg L^?1, and washing time 2 h, which was in good agreement with predicted value (66.643%).     

Methyl-beta-cyclodextrin-enhanced solubilization and aerobic biodegradation of polychlorinated biphenyls in two aged-contaminated soils

The bioremediation of aged polychlorinated biphenyl (PCB)-contaminated soils is adversely affected by the low bioavailability of the pollutants. Randomly methylated-beta-cyclodextrins (RAMEB) were tested as a potential PCB-bioavailability-enhancing agent in the aerobic treatment of two aged-contaminated soils. The soils, contaminated by about 890 and 8500 mg/kg of Aroclor 1260 PCBs, were amended with biphenyl (4 g/kg), inorganic nutrients (to adjust their C:N ratio to 20:1), and variable amounts of RAMEB (0%, 0.5%, or 1.0% [w/w]) and treated in both aerobic 3-L solid-phase reactors and 1.5-L packed-bed loop reactors for 6 months. Notably, significant enhancement of the PCB biodegradation and dechlorination, along with a detectable depletion of the initial soil ecotoxicity, were generally observed in the RAMEB-treated reactors of both soils. RAMEB effects were different in the two soils, depending upon the treatment conditions employed, and generally increased proportionally with the concentration at which RAMEB was applied. RAMEB, which was slowly metabolized by the soil's aerobic microorganisms, was found to markedly enhance the occurrence of the indigenous aerobic, cultivable biphenyl-growing bacteria harboring genes homologous to those of two highly specialized PCB degraders (i.e., bphABC genes of Pseudomonas pseudoalcaligenes KF707 and bphA1A2A3A4BC1 genes of Rhodococcus globerulus P6) and chlorobenzoic acid-degrading bacteria as well as the occurrence of PCBs in the water phase of the soil reactors. These findings indicate that RAMEB enhanced the aerobic bioremediation of the two soils by increasing the bioavailability of PCBs and the occurrence of specialized bacteria in the soil reactors.     

Carbon and nitrogen stocks in physical fractions of a subtropical Acrisol as influenced by long-term no-till cropping systems and N fertilisation

The long-term soil management effects on C and N stocks of soil physical fractions are still poorly understood for South American subtropical soils. This study aimed (i) to evaluate the influence of cereal- and legume-based cropping systems and N fertilisation on C and N stocks of the sand-, silt- and clay-size fractions of a no-tilled subtropical Acrisol in southern Brazil, (ii) to compute the Carbon Management Index (CMI) for those cropping systems using physical fractionation data, and (iii) to investigate the possible existence of finite capacity of those soil physical fractions to store C and N. Soil samples of a long-term experiment were collected from the 0–2.5 and 2.5–7.5 cm layers of three no-till cropping systems [fallow bare soil, oat/maize (O/M) and pigeon pea+maize (P+M)] under two N fertilisation levels (0 and 180 kg N ha^–1). However, for fallow bare soil, only the non-fertilised sub-plot was sampled. An adjacent native grassland soil was sampled as a reference. The C and N stocks of the three soil physical fractions were higher in the legume-based cropping system (P+M) than in O/M and bare soil, because of the higher residue input in the former. The P+M cropping system restored the C and N stocks in sand- and silt-size fractions to the same levels found in grassland soil. Higher C and N stocks in all physical fractions were also obtained with N fertilisation. The C and N stocks and the C:N ratio were most affected by cropping systems in the sand- and least in the clay-size fraction. Particulate organic matter was found in the silt-size fraction, showing this fraction is not only constituted by mineral-associated organic mater, as commonly believed. Taking grassland soil as reference (CMI = 100), the CMI ranged from 46, in O/M no N, to 517, in P+M with N, pointing to a better soil management in the latter. The clay-size fraction tended to show a finite capacity to store C and N (48.8 g C kg^–1 and 4.9 g N kg^–1 of clay), which was not verified in sand- and silt-size fractions. The adoption of no-tillage and legume-based cropping systems with high residue input are adequate soil management strategies to improve soil quality and make the agricultural production systems more sustainable in subtropical regions.     

Development of field application vectors for bioremediation of soils contaminated with polychlorinated biphenyls.

Field application vectors (FAVs), which are a combination of a selective substrate, a host, and a cloning vector, have been developed for the purpose of expressing foreign genes in nonsterile, competitive environments in which the gene products provide no advantage to the host. Such gene products are exemplified by the enzymes for the cometabolism of polychlorinated biphenyls (PCBs) through the biphenyl degradation pathway. Attempts to use highly competent PCB-cometabolizing strains in the environment in the absence of biphenyl have not been successful, while the addition of biphenyl is limited by its human toxicity and low water solubility. Broad-substrate-specificity PCB-degradative genes (bphABC) were cloned from a naturally occurring isolate. Pseudomonas sp. strain ENV307, into broad-host-range plasmid pRK293. The resulting PCB-degrading plasmids were transferred to the FAV host Pseudomonas paucimobilis 1IGP4, which utilizes the nontoxic, water-soluble, nonionic surfactant Igepal CO-720 as a selective growth substrate. Plasmid stability in the recombinant strains was determined in the absence of antibiotic selection. PCB-degrading activity was determined by resting cell assays. Treatment of contaminated soil (10, 100, or 1,000 ppm of Aroclor 1242) by surfactant amendment (1.0% [wt/wt]Igepal CO-720 in wet soil) and inoculation with recombinant isolates of strain 1IGP4 (approximately 4 x 10(6) cells per g of soil) resulted in degradation of many of the individual PCB congeners in the absence of biphenyl.(ABSTRACT TRUNCATED AT 250 WORDS)     

Aerobic biodegradation of biphenyl and polychlorinated biphenyls by Arctic soil microorganisms.

We examined the degradation of biphenyl and the commercial polychlorinated biphenyl (PCB) mixture Aroclor 1221 by indigenous Arctic soil microorganisms to assess both the response of the soil microflora to PCB pollution and the potential of the microflora for bioremediation. In soil slurries, Arctic soil microflora and temperate-soil microflora had similar potentials to mineralize [14C]biphenyl. Mineralization began sooner and was more extensive in slurries of PCB-contaminated Arctic soils than in slurries of uncontaminated Arctic soils. The maximum mineralization rates at 30 and 7 degrees C were typically 1.2 to 1.4 and 0.52 to 1.0 mg of biphenyl g of dry soil-1 day-1, respectively. Slurries of PCB-contaminated Arctic soils degraded Aroclor 1221 more extensively at 30 degrees C (71 to 76% removal) than at 7 degrees C (14 to 40% removal). We isolated from Arctic soils organisms that were capable of psychrotolerant (growing at 7 to 30 degrees C) or psychrophilic (growing at 7 to 15 degrees C) growth on biphenyl. Two psychrotolerant isolates extensively degraded Aroclor 1221 at 7 degrees C (54 to 60% removal). The soil microflora and psychrotolerant isolates degraded all mono-, most di-, and some trichlorobiphenyl congeners. The results suggest that PCB pollution selected for biphenyl-mineralizing microorganisms in Arctic soils. While low temperatures severely limited Aroclor 1221 removal in slurries of Arctic soils, results with pure cultures suggest that more effective PCB biodegradation is possible under appropriate conditions.     

Patterns of restoration of soil physciochemical properties and microbial biomass in different landslide sites in the sal forest ecosystem of Nepal Himalaya

The pattern of natural restoration in soil components and processes was documented in five landslide-damaged (1–58-year-old) sites in the moist tropical sal (Shorea robusta) forest ecosystem of Nepal Himalaya. Comparisons were made with an undisturbed forest site in the same region. Concentrations of soil organic C, total N, total P and extractable nutrients (Ca, Mg and K) increased with the age of sites. The 58-year-old site showed concentrations of soil organic C, total N and total P that were 75–89% of concentrations in the undisturbed sal forest. The soil microbial biomass, the active fraction of soil organic matter, showed similar seasonal variations at all sites. The amount of mean microbial biomass (expressed as C, N and P contents) increased 4–5 times at the 58-year-old site relative to the 1-year-old site, and the bulk increase occurred within the initial 15 year. The increase in the C/N ratio of soil microbial biomass with age (9.4–11.6 years) reflected change in its composition. Although the net N-mineralization rate increased consistently until 58 years of age, the proportion of nitrification rate relative to ammonification rate distinctly decreased beyond 40 years. On the other hand, the soil available-N (both NO₃⁻ and NH₄⁺) concentrations increased from 1 to 40 year and then declined; with age the proportion of NH₄⁺ increased, however. Rates of restoration in soil properties were faster in the early successional stages (1–15 year) than late stages. Among different soil properties the restoration of soil microbial biomass (C and N) was faster than soil organic C and total N. Best fit power function models showed that the estimated times for the 58-year-old site to reach the level of the undisturbed, mature sal forest would be about 30–35 year for microbial biomass (C and N) and about 100–150 year for organic C and total N. Higher accumulation of soil microbial biomass and high N-mineralization rate at late successional stages indicated the re-establishment of enriched soil and restitution of nutrient cycling during the course of ecosystem restoration.     

Enhancing plant uptake of polychlorinated biphenyls and cadmium using tea saponin

The potential of natural surfactant tea saponin to enhance uptake of polychlorinated biphenyls (PCBs) and cadmium (Cd) by Zea Mays L. and Saccharum officinarum L. was investigated. With addition of tea saponin at 0.01% in solution culture, the concentrations of PCB 14, PCB 18, PCB 77 and PCB 156 in root of corn seedling were 2.72, 2.68, 1.94 and 2.40 times as those of treatments without adding any surfactant, respectively. Application of tea saponin to the soil significantly elevated PCB 5 accumulation in shoots and roots (p < 0.05) by sugarcanes. With addition of 0.3% tea saponin, Cd concentration was increased by 96.9% in roots, 156.8% in stems and 30.1% in leaves compared with the treatment without addition of surfactant in sugarcane grown in soil. Tea saponin had potential of assisting the uptake of PCBs and Cd by plants from water solution and soil.     

In Situ emulsification and mobilization of gasoline range hydrocarbons using surfactants

In this article the conditions that govern surfactant‐enhanced emulsification and mobilization of petroleum hydrocarbons in soil are reviewed. The effect of soil properties, groundwater constituents, and differing surfactant solutions on the emulsification process is discussed. A constant head soil flushing apparatus used to characterize surfactant‐enhanced mobilization of m‐xylene is described. Data showing the effect of surfactant‐enhanced mobilization on m‐xylene removal efficiency in washed sand is presented. Flushing solutions were used at concentrations from below to well above the critical micelle concentration (CMC) of the surfactants used. Removal efficiencies are shown to vary with surfactant concentration and with surfactant type. Flushing solutions of anionic, nonionic, and anionic/nonionic surfactant mixtures were evaluated.     

Application of Biosurfactants Produced by Pseudomonas aeruginosa SP4 for Bioremediation of Soils Contaminated by Pyrene

Biosurfactants are considered to facilitate PAHs dissolution in soil slurries for bioremediation applications. In this work, the carbon and nitrogen sources, pH, C/N ratio, and salinity, were considered for optimization of biosurfactant production by Pseudomonas aeruginosa SP4 isolate to enhance pyrene removal from the contaminated soil. Analysis of ANOVA indicated that the carbon source was the most effective factor, followed by pH, nitrogen source, C/N ratio, and salinity. Taguchi experimental design proposed the optimum operating conditions of olive oil, NH₄NO₃, C/N ratio of 5, salinity of 0.5%, and pH 7. Applying the conditions determined by Taguchi design led to a production yield of 452 mg L^?1 (13% improvement) at the optimum conditions. The main characteristics of produced biosurfactant included the critical micelle concentration (CMC) of 60 mg L^?1 and liquid medium surface tension of 29.5 mN m^?1. Produced biosurfactant was used for bioremediation of soil artificially contaminated with 500 mg kg^?1 of pyrene. Following the addition of 250 mg L^?1 biosurfactant, the pyrene removal of 84.6% was obtained compared to 59.8% for control sample without any surfactant.     

Soya lecithin effects on the aerobic biodegradation of polychlorinated biphenyls in an artificially contaminated soil

The effects of the phytogenic surfactant soya lecithin (SL) on the aerobic biodegradation of polychlorinated biphenyls (PCBs) spiked into a synthetic soil were studied. Soil was spiked with both biphenyl (4 g/kg) and Fenclor 42 (1,000 mg/kg) and treated in aerobic batch slurry-phase microcosms (17.5% w/v). Microcosms were prepared either with or without the exogenous aerobic PCB-dechlorinating bacterial co-culture ECO3 (inoculum:10(8) CFU/mL). In some inoculated microcosms, SL was added at 15 or 30 g/kg. Indigenous bacteria having the capability of metabolizing biphenyl and 2-chlorobenzoic acid were found to develop in the microcosms during the experiment, and were responsible for the significant PCB biodegradation and dechlorination observed in the uninoculated controls. The addition of ECO3 bacteria resulted in only a slight PCB biodegradation increase. In the presence of SL, a higher availability of biphenyl- and chlorobenzoic acid-degrading bacteria and higher PCB biodegradation and dechlorination yields were observed; the effects increased proportionally with the concentration of the applied SL. A significant decrease of soil ecotoxicity was also revealed in SL-supplemented microcosms. At both concentrations, SL was found to be a good carbon source for both the indigenous and ECO3 bacteria, as well as a product capable of enhancing the PCB bioavailability in the microcosms.     

Degradation of nonionic surfactants and polychlorinated biphenyls by recombinant field application vectors

Degradation of polychlorinated biphenyls (PCBs) in the environment is limited by their aqueous solubility and the degradative competence of indigenous populations. Field application vectors (FAVs) have been developed in which surfactants are used to both increase the solubility of the PCBs and support the growth of surfactant-degrading strains engineered for PCB degradation. Surfactant and PCB degradation by two recombinant strains were investigated. Pseudomonas putida IPL5 utilizes both alkylethoxylate [polyoxyethylene 10 lauryl ether (POL)] and alkylphenolethoxylate [Igepal CO-720 (IGP)] surfactants as growth substrates, but only degrades the ethoxylate moiety. The resulting degradation products from the alkyl- and alkylphenolethoxylate surfactants were 2-(dodecyloxy)ethanol and nonylphenoldiethoxylates, respectively. Ralstonia eutropha B30P4 grows on alkylethoxylate surfactants without the appearance of solvent-extractable degradation products. It also degrades the 2-(dodecyloxy)ethanol produced by strain IPL5 from the alkylethoxylate surfactants. The extent of degradation of the alkylethoxylate surfactant (POL) was greater for strain IPL5 (90%) than for B30P4 (60%) as determined by the cobaltothiocyanate active substances method (CTAS). The recombinant strain B30P4::TnPCB grew on biphenyl. In contrast, the recombinant strain IPL5::TnPCB could not grow on biphenyl, and PCB degradation was inhibited in the presence of biphenyl. The most extensive surfactant and PCB degradation was achieved by the use of both recombinant strains together in the absence of biphenyl. PCB (Aroclor 1242) and surfactant (POL) concentrations were reduced from 25 ppm and 2000 ppm, respectively, to 6.5 ppm and 225 ppm, without the accumulation of surfactant degradation products. Given the inherent complexity of commercial surfactant preparations, the use of recombinant consortia to achieve extensive surfactant and PCB degradation appears to be an environmentally acceptable and effective PCB remediation option. Received 04 October 1996/ Accepted in revised form 04 August 1997     

Biomagnification of Aroclor 1242 in Decomposing Spartina Litter

The accumulation of a polychlorinated biphenyl (PCB) mixture (Aroclor 1242) in the process of detritus formation by a shredded marshgrass (Spartina alterniflora) under aerobic conditions was monitored in percolators for 4 months at 20°C. Dissolved PCB in the influent solution was 14 to 16 μg/liter. Parameters monitored in addition to PCB accumulation were CO₂ evolution, NH₄⁺ uptake, NO₃⁻ production, and total organic nitrogen and carbon in the effluent. An NaN₃₋ poisoned control served to assess nonbiological PCB absorption. Up to 90% of the PCB dissolved in the influent water was removed by the biologically active detritus. Biomagnification resulted in three to four times higher PCB concentrations in the active detritus than in the abiotic control. Evolution of CO₂ was slightly depressed by PCB, but the overall quality (C:N ratio) of the detritus was not affected. The results indicate that bioaccumulation of PCB in detritus is an important means of entry for this pollutant into estuarine food webs.     

Effects of nonionic surfactants on the solubilization and mineralization of phenanthrene in soil-water systems

The solubilization and mineralization of (14)C-phenanthrene in soil-water systems was examined with several commercially available surface-active agents, viz., an alkyl ethoxylate C(12)E(4); two alkylphenol ethoxylate surfactants: C(8)PE(9.5) and C(9)PE(10.5); two sorbitan ethoxylate surfactants: the sorbitan monolaurate (Tween 20) and the sorbitan monooleate (Tween 80); two pairs of nonionic ethoxylate surfactant mixtures: C(12)E(4)/C(12)E(23) at a 1:1 ratio, and C(12-15)E(3)/C(12-15)E(9) at a 1:3 ratio; and two surfactants possessing relatively high critical micelle concentration (CMC) values and low aggregation numbers: CHAPS and octyglucoside. Surface tension experiments were performed to evaluate surfactant sorption onto soil and the surfactant doses required to attain the CMC in the soil-water systems. Surfactant solubilization of (14)C-phenanthrene commenced with the onset of micellization. The addition of surface-active agents was observed not to be beneficial to the microbial mineralization of phenanthrene in the soil-water systems and, for supra-CMC surfactant doses, phenanthrene mineralization was completely inhibited for all the surfactants tested. A comparison of solubilization, surface tension, and mineralization data confirms that the inhibitory effect on microbial degradation of phenanthrene is related to the CMC of the surfactant in the presence of soil. Additional tests demonstrated the recovery of mineralization upon dilution of surfactant concentration to sub-CMC levels, and a relatively high exit rate for phenanthrene from micelles. These tests suggest that the inhibitory effect is probably related to a reversible physiological surfactant micelle-bacteria interaction, possibly through partial complexing or release of membrane material with disrupting membrane lamellar structure. This study indicates that nonionic surfactant solubilization of sorbed hydrophobic organic compounds from soil may not be beneficial for the concomitant enhancement of soil bioremediation. Additional work is needed to address physicochemical processes for bioavailability enhancement, and effects of solubilizing agents on microorganisms for remediation and treatment of hydrophobic organic compounds and nonaqueous phase liquids. (c) 1992 John Wiley & Sons Inc.     

Phenanthrene Removal in a Selected Mexican Soil by the Fungus Penicillium frequentans: Role of C:N Ratio and Water Content

The purpose of this study was to determine the ability of the nonbasidiomycete, filamentous fungi Penicillium frequentans, isolated and grown on sugar cane bagasse pith, to remove phenanthrene in a solidstate culture. Additionally, the study investigated whether phenanthrene removal could be enhanced by manipulating the carbon-to-nitrogen ratio (C:N) and moisture content over a relatively short-term period (29 days). To evaluate the combined effect of moisture content and the C:N ratio, a combined experimental design, composed with a 2² factorial and both central and axial points, was used. It was shown that the moisture content (p < 0.0003) and the combined effect of moisture content and C:N ratio have a significant (p < 0.002) positive effect on the phenanthrene removal. It was also found that heterotrophic activity was not correlated to phenanthrene removal. An optimum phenanthrene removal efficiency of 74% was realized at a moisture content of 40% and a C:N ratio of 60. This suggests that Penicillium frequentans was able to effectively remove phenanthrene in a solidstate culture and that the combination of nutrient addition and moisture adjustment could enhance the phenanthrene removal activity.     

Effects of Triton X-100 and Quillaya Saponin on the ex situ bioremediation of a chronically polychlorobiphenyl-contaminated soil

The possibility of enhancing the ex situ bioremediation of a chronically polychlorinated biphenyl (PCB)-contaminated soil by using Triton X-100 or Quillaya Saponin, a synthetic and a biogenic surfactant, respectively, was studied. The soil, which contained about 350 mg/kg of PCBs and indigenous aerobic bacteria capable of growing on biphenyl or on monochlorobenzoic acids, was amended with inorganic nutrients and biphenyl, saturated with water and treated in aerobic batch slurry- and fixed-phase reactors. Triton X-100 and Quillaya Saponin were added to the reactors at a final concentration of 10 g/l at the 42nd day of treatment, and at the 43rd and 100th day, respectively. Triton X-100 was not metabolised by the soil microflora and it exerted inhibitory effects on the indigenous bacteria. Quillaya Saponin, on the contrary, was readily metabolised by the soil microflora. Under slurry-phase conditions, Triton X-100 negatively influenced the soil bioremediation process by affecting the availability of the chlorobenzoic acid degrading indigenous bacteria, whereas Quillaya Saponin slightly enhanced the biological degradation and dechlorination of the soil PCBs. In the fixed-phase reactors, where both the surfactant availability and the mixing of the soil were lower, Triton X-100 did not exert inhibitory effects on the soil biomass and enhanced significantly the soil PCB depletion, whereas Quillaya Saponin did not influence the bioremediation process. Received: 28 April 1998 / Received last revision: 15 July 1998 / Accepted: 29 July 1998     

Effects of long-term experimental warming on plants and soil microbes in a cool temperate semi-natural grassland in Japan

To clarify the effects of long-term warming on ecosystem matter cycling, we conducted an in situ 7-year experimental warming (2009–2015) using infrared heaters in a cool temperate semi-natural grassland in Japan. We measured plant aboveground biomass, soil total C and N, soil inorganic N (NH₄ ⁺-N and NO₃ ^?-N), and soil microbial biomass for 7 years (2009–2015). We also measured heterotrophic respiration for 2 years (2013–2014) and assessed net N mineralization and nitrification in 2015. We found that warming immediately increased plant aboveground biomass, but this effect ceased in 2013. However, the soil microbial biomass was continuously depressed by warming. Soil inorganic N concentrations in warmed plots substantially increased in the later years of the experiment (2013–2015) and the potential net N mineralization rate was also higher than in the earlier years. In contrast, heterotrophic respiration decreased with warming in 2013–2014. Our observations indicate that long-term warming has a contrasting effect on plants and soil microbes. In addition, the warming could have different effects on subterranean C and N cycling. To enhance the accuracy of estimation of future climate change, it is essential to continuously observe the warming effects on ecosystems and to focus on the change in subterranean C and N cycling.