The relationship between a leaf-rolling moth (Dactylioglypha tonica) and fungi covering the cocoon.

To discover the relationship between a leaf-rolling moth and the fungi densely covering its cocoons, the rolled nest leaves were collected in two districts in Japan and antibacterial properties of the fungi were examined. Cocoons and fungi isolated from the nest were classified into 5 categories by the growth stages of the insects, and 7 categories based on taxonomic properties and pigment productivity, respectively. The dominant genus was Penicillium in each location. However, the composition of the fungal categories was different and seemed to depend on their circumstances. From all cocoons with larvae, the strains that belonged to the same fungal category and produced the same antibiotic (deoxyherqueinone) were isolated. From these results, the species-specific relationship between the insect and fungi or fungal products was considered to be not extremely tight, and it was suggested the period of the larval spinning of the cocoon is a key stage of this unique relationship.     

Survival of mature larvae of codling moth (Cydia pomonella) on apple trees and ground

The effects of predation by carabid and staphylinid beetles and birds (blue- and great-tits) on mature larvae of the codling moth Cydia pomonella were investigated in a field experiment. The number of predatory beetles on the ground beneath eight of 16 apple trees was reduced by an insecticide spray, and tits were excluded from the trunks and lower branches of eight trees by means of Terylene netting. Beetles had no significant effect on survival of larvae on trees, but only 8% of larvae survived to adulthood on trees exposed to blue- and great-tits, whereas on trees protected from them 48% survived. Larvae were taken rapidly by the tits from the time larvae first built cocoons in summer and, by the time the majority of larvae had emerged from apples in late summer or autumn, tits had removed most from cocoons beneath flakes of bark glued to trees. Thus tits fed on larvae mainly in summer and autumn. On trees protected from tits, 8% of mature larvae were killed by fungi, of which Verticillium lecanii was most important. It was calculated that, of the larvae maturing in apples, 44% failed to build cocoons on the trees, 47% were taken by tits, then 1% were killed by fungi. Larvae failing to build cocoons on trees may build them on the ground. Of larvae in cocoons planted on or in the ground in 1975, only one (0.3%) persisted through the winter; 98.6% disappeared, and 1.1% were found dead, covered by fungi; in 1976, all larvae disappeared by December. Larvae disappeared even in the absence of predators, and are thought to have moved in response to damp ground conditions.     

Phenolic removal of olive-mill dry residues by laccase activity of white-rot fungi and its impact on tomato plant growth

We studied the influence of the laccase activity of two white-rot fungi on the toxic effect of water-soluble substances from dry residues of olives (ADOR) on tomato plants. Pycnoporus cinnabarinus and Coriolopsis rigida decreased the phenol content of ADOR to 73% after 15 days. P. cinnabarinus and C. rigida produced laccase activity after 5 and 15 days, respectively, and the highest activity in both fungi was detected at 20 days. The treatment of ADOR with these white-rot fungi decreased the phytotoxicity of this residue on tomato plants. A close relationship was found between the amount of laccase produced, the decrease in phenol content of ADOR by the saprobic fungi, decrease of phytotoxicity of ADOR, and the increase in dry weight of tomato plants. These results show that phenol removal by the laccase activity of white-rot fungi can be important in the elimination of phytotoxic substances present in olive-mill dry residues.     

Illness associated with contamination of drinking water supplies with phenol.

In January 1984 the River Dee in north Wales was contaminated with phenol, with subsequent contamination of the tap water received by about two million consumers. A retrospective postal survey of 594 households was undertaken to determine whether consumption of this contaminated water was associated with illness. Subjects in areas that received contaminated water reported significantly more gastrointestinal illness than those in a nearby unexposed area (32.6% v 8.7%, p less than 0.00001) as well as reporting a higher incidence of any symptoms (43.6% v 18.4%, p less than 0.00001). Symptoms were consistent with phenol poisoning and bore a strong temporal relation to the pollution of the supply, but they developed at concentrations of phenols previously considered to be safe by the water authorities concerned. Chlorophenols produced during the treatment of water may have aggravated the problem.     

Biodegradation of phenol, <Emphasis Type="Italic">o-</Emphasis>cresol, <Emphasis Type="Italic">m-</Emphasis>cresol and <Emphasis Type="Italic">p-</Emphasis>cresol by indigenous soil fungi in soil contaminated with creosote

Seven non-basidiomycetic fungi, Aspergillus, Candida, Cladosporium, Fusarium, Monicillium, Trichoderma and Penicillium, and two basidiomycetic fungi, Pleurotus and Phanerochaete were isolated from a creosote-contaminated soil by using mineral salts medium and soil extract broth containing antibiotics. Soil contaminated with phenol, o-cresol, m-cresol and p-cresol was collected from the yard of a wood treatment plant in South Africa and inoculated with the strains of Aspergillus, Cladosporium, Fusarium, Monicillium, Penicillium and Phanerochaete, selected from the isolate. The soil in some of the treatment reactors was amended with nutrient supplements to give a C:N:P ratio of 25:5:1. A total of 18 duplicate treatments were established and incubated in the dark at 25°C for 70days. The soil in all the reactors was tilled weekly and moisture was maintained at 70% field capacity. Soil samples were collected every 2weeks for analysis of residual concentrations of the phenols tested, pH measurement and moisture content determination. The nutrient-supplemented treatments were more effective in degrading the phenols (between 84 and 100%) than those that were not supplemented. Barley, which was used as bulking agent enhanced the growth of the fungi and subsequently the degradation of the phenols. Inoculation with a mixture of the six fungal isolates promoted more phenol degradation than with single isolates.     

Suspending cocoons to evade ant predation in Meteorus pulchricornis, a braconid parasitoid of exposed-living lepidopteran larvae

We tested the hypothesis that cocoon suspension by a thread in hymenopteran parasitoids is a defense tactic against predators, by comparing predation against suspended and non-suspended cocoons of the braconid wasp Meteorus pulchricornis on a Quercus phillyraeoides hedge on which workers of the common small ant Crematogaster matsumurai were foraging. The lost proportion of non-suspended cocoons, which were artificially attached to leaves of Q. phillyraeoides , markedly decreased with cocoon age, indicating a critical phase of predation on young cocoons. No suspended cocoons at age 1–12 h at the beginning of exposure were lost within 12 h, whereas more than 75% of same-aged non-suspended cocoons were lost in the same period. Predation against such young cocoons would be a strong force driving the evolution of cocoon suspension in parasitoids of exposed-living host insects.     

Presence of Culturable Bacteria in Cocoons of the Earthworm Eisenia fetida

Viable bacteria were found to coexist with developing embryos in egg capsules (cocoons) of the earthworm Eisenia fetida. Earthworms were reared under standardized conditions, and bacterial densities were measured in distinct batches of cocoons collected weekly for 10 weeks. Cocoons weighing 12 mg contained a mean viable bacterial population of approximately 10⁸ CFU/g of cocoons. No difference was found in viable counts obtained from cocoons incubated at 15°C and cocoons incubated at 24°C. Viable bacterial numbers increased with cocoon age, while acridine orange direct counts of microbial cells were stable at approximately 10⁹ cells per g of cocoons. Bacteria isolated from cocoons were used to develop antisera in rabbits for the production of strain-specific fluorescent antibodies. Fluorescent antibody and selective plating techniques were used to monitor populations of these bacteria in earthworm bedding and to determine whether cocoons acquire bacteria from the environment in which they are formed. Cocoon isolates were readily recovered from cocoons formed in inoculated bedding at densities of 10⁸ CFU/g of cocoons. Bradyrhizobium japonicum USDA 110 and UMR 161 added to bedding were also recovered from cocoons, but at lower densities than cocoon isolates. Escherichia coli K-12(pJP4) inoculum was recovered from bedding but not from cocoons. The bacterial complement of Eisenia fetida cocoons is affected by inoculation of selected bacterial isolates in the worm growth environment.     

一株高效苯酚降解真菌的分离鉴定及其菌剂的制备

【背景】含酚废水是普遍存在的有毒、难降解的有机污染物之一,生物法处理含酚废水因成本低、无二次污染而具有广阔的应用前景。可降解苯酚的微生物中,真菌比细菌对恶劣环境的适应性更好。针对液态菌液保存时间较短和运输困难的瓶颈,制备固体菌剂可以提高菌体存活率和储藏稳定性。【目的】筛选一株能够高效降解苯酚的真菌,优化其降酚性能并选择合适的载体制备菌剂。【方法】通过逐级驯化和纯化分离降酚菌,筛选得到降酚性能较强的真菌并通过ITS r DNA基因测序进行种属鉴定,通过参数优化进一步提高菌株降解苯酚的性能;以不同材料为载体制备菌剂,通过稀释平板计数法和苯酚降解实验探究菌剂在不同温度下的保存效果。【结果】分离筛选得到一株高效降解苯酚真菌QWD1,通过鉴定证明其属于Magnusiomyces capitatus,其最适降解条件:(NH_4)_2SO_4为氮源,接种量为15%,pH为7.0,温度为35°C,氮源浓度为14 mmol/L。在此条件下,28 d内对1 600 mg/L苯酚去除率可以达到97.15%;制备菌剂最合适载体为谷糠,适宜保存温度为4°C,保存时间可达到90 d甚至更长,活菌数高达2.5×10~8 CFU/g左右,降解苯酚效果良好。【结论】筛选得到了一株高效降解苯酚真菌,优化其降解性能并将其制备成菌剂,为处理含酚废水提供了新菌种和理论支持。     

Selective Degradation of Wood Components by White-Rot Fungi

In order to find naturally occurring white-rot fungi which preferentially degrade lignin. 25 different species of such fungi were cultivated on pine wood blocks and on kraft lignin agar plates with and without cellulose. Due to differences in phenol oxidase reactions on the kraft lignin agar plates, the 25 fungi could be divided into two groups, 1 and 2, which also differed in other properties. The three Group I fungi Sporotrichum pulverulentum, Phanerochaete sp. L1 and Polyporus dichrous produced high levels of endo-l,4-β-glucanase and cellobiose:quinone oxidoreductase in shaking cellulose flasks and a low level of phenol oxidase in standing wood meal flasks, The four fungi Merulius tremellosus, Phlebia radiata, Pycuoporus cinnabarinus and Pleurotus ostreatus from Group 2, on the other hand, produced low levels of endo-1,4-β-glucanase and cellobiose:.quinone oxidoreductase in the cellulose. flasks and a high level of phenol oxidase in the wood meal flasks. Analyses of pine wood blocks degraded by the above-mentioned fungi in the presence of either malt extract, asparagine or NH₄H₂PO₄ revealed that malt extract gave good lignin degradation. In the presence of this nutrient source. P. cinnabarinus, at 3.4% weight loss, even degraded 12.5% lignin without loss of cellulose or mannan. No common degradation pattern was, however, obtained using mall extract, asparagine or NH₄H₂PO₄, It is suggested that while-rot fungi, which preferentially degrade lignin, may be found among Group 2 fungi producing large amounts of phenol oxidases.     

Fish mucous cocoons: the 'mosquito nets' of the sea

Mucus performs numerous protective functions in vertebrates, and in fishes may defend them against harmful organisms, although often the evidence is contradictory. The function of the mucous cocoons that many parrotfishes and wrasses sleep in, while long used as a classical example of antipredator behaviour, remains unresolved. Ectoparasitic gnathiid isopods (Gnathiidae), which feed on the blood of fish, are removed by cleaner fish during the day; however, it is unclear how parrotfish and wrasse avoid gnathiid attacks at night. To test the novel hypothesis that mucous cocoons protect against gnathiids, we exposed the coral reef parrotfish Chlorurus sordidus (Scaridae) with and without cocoons to gnathiids overnight and measured the energetic content of cocoons. Fish without mucous cocoons were attacked more by gnathiids than fish with cocoons. The energetic content of mucous cocoons was estimated as 2.5 per cent of the fish's daily energy budget fish. Therefore, mucous cocoons protected against attacks by gnathiids, acting like mosquito nets in humans, a function of cocoons and an efficient physiological adaptation for preventing parasite infestation that is not used by any other animal.     

Addition of arthropod cocoons to house wren nests is correlated with delayed pairing

Males in the cavity-nesting house wren (Troglodytes aedon) frequentlyadd arthropod cocoons to their nests during building, possiblyas an ornamental cue for female choice. We tested this hypothesisby comparing the time to pairing for males that did and didnot add cocoons to their nests and for males in whose nestswe manipulated the number of cocoons prior to pairing. We alsotested the hypothesis that females acquire fitness-related benefitsby selecting mates based on their use of cocoons. The use ofcocoons by males was not consistently related to habitat, butthe number of cocoons added per nest increased during the courseof the breeding season. Contrary to prediction, the time topairing for males adding cocoons was significantly longer thanthat for males without cocoons in their nests at both unmanipulatedand experimental nests. There was also no consistent fitness-relatedbenefit for females related to the use of cocoons by their mates.Therefore, we conclude that females did not prefer males thatadded cocoons to their nests, and that the increased time topairing for males that add cocoons likely results in fitness-relatedcosts brought about by delayed breeding. Nonetheless, male housewrens routinely use cocoons, and why they do so remains unknown.     

根际圈在污染土壤修复中的作用与机理分析

根际圈以植物根系为中心聚集了大量的生命物质及其分泌物,构成了极为独特的“生态修复单元”。本文叙述了根在根际圈污染土壤修复中的生理生态作用,富集、固定重金属,吸收、降解有机污染物等功能;菌根真菌对根际圈内重金属的吸收、屏障及螯合作用,对有机污染物的降解作用;根际圈内细菌对重金属的吸附与固定,对有机污染物的降解作用以及根际圈真菌和细菌的联合修复作用等,同时对可能存在的机理进行了分析,认为根际圈对污染土壤的修复作用是植物修复的重要组成部分和主要理论基础之一,并指出利用重金属超富集植物修复重金属污染土壤具有广阔的应用前景;筛选对水溶性有机污染物高吸收富集及其根 发泌能力强的特异植物,同时接种利于有机污染物降解的专性或非专性真菌和细菌可能会成为有机污染土壤植物修复研究的重要方向之一。     

Disinfection of Aerosolized Pathogenic Fungi on Laboratory Surfaces: I. Tissue Phase

The effect of several fungicides on laboratory surfaces contaminated with the tissue phase of aerosolized Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, and Histoplasma capsulatum was ascertained. A statistical analysis of the data shows the correlation between fungi, surfaces, time, and concentration of disinfectant. All fungicides were effective at established times and concentrations, and the type of contaminated surface affected the fungicidal efficacy. By interpolating plotted graphs, laboratory personnel may determine, with a given fungicide, the concentration and time required to disinfect instruments, pipettes, gloves, bench tops, and floors contaminated with a tissue phase of pathogenic fungi.     

Gaseous TCE and PCE removal by an activated carbon biofilter

Gaseous trichloroethylene (TCE) and tetrachloroethylene (PCE) are emitted in the treatment of contaminated groundwaters with air stripping and/or the remediation of contaminated soils using vapor extraction techniques. This study investigated the application of biofiltration using cometabolic process to remediate gaseous TCE and PCE that are highly recalcitrant to biodegradation. The investigation was conducted using two specially built stainless steel columns, one for TCE and the other for PCE, packed with granular activated carbon (GAC) coated with phenol-oxidizing microorganisms at residence times of 1.5–7 min. Two activated carbon biofilters were fed with phenol at a specific concentration along with a nutrient solution to optimize the various catalyzed biochemical reactions. The removal efficiency of gaseous TCE was 100% at a residence time of 7 min and average inlet concentration of 85 ppm. For gaseous PCE, 100% removal efficiency was obtained at residence times of 4–7 min and average concentrations of 47–84 ppm. It was found that phenol fed to the biofilters was completely utilized by the phenol-oxidizing microorganisms, as an indirect indicator of the microorganisms growth in the biofilters, throughout the period of the biofilter operation. Transformation yields of gaseous TCE and PCE were about 8–48 g of TCE/g of phenol and 6–25 g of PCE/g of phenol, depending on different residence times. It was found that adsorption by GAC and absorption by the influent nutrient solution were a minor negligible mechanism for TCE and PCE removal in the activated carbon biofilters.     

Back to basics: an evaluation of NaOH and alternative rapid DNA extraction protocols for DNA barcoding,genotyping, and disease diagnostics from fungal and oomycete samples

The ubiquity, high diversity and often‐cryptic manifestations of fungi and oomycetes frequently necessitate molecular tools for detecting and identifying them in the environment. In applications including DNA barcoding, pathogen detection from plant samples, and genotyping for population genetics and epidemiology, rapid and dependable DNA extraction methods scalable from one to hundreds of samples are desirable. We evaluated several rapid extraction methods (NaOH, Rapid one‐step extraction (ROSE), Chelex 100, proteinase K) for their ability to obtain DNA of quantity and quality suitable for the following applications: PCR amplification of the multicopy barcoding locus ITS1/5.8S/ITS2 from various fungal cultures and sporocarps; single‐copy microsatellite amplification from cultures of the phytopathogenic oomycete Phytophthora ramorum; probe‐based P. ramorum detection from leaves. Several methods were effective for most of the applications, with NaOH extraction favored in terms of success rate, cost, speed and simplicity. Frozen dilutions of ROSE and NaOH extracts maintained PCR viability for over 32 months. DNA from rapid extractions performed poorly compared to CTAB/phenol‐chloroform extracts for TaqMan diagnostics from tanoak leaves, suggesting that incomplete removal of PCR inhibitors is an issue for sensitive diagnostic procedures, especially from plants with recalcitrant leaf chemistry. NaOH extracts exhibited lower yield and size than CTAB/phenol‐chloroform extracts; however, NaOH extraction facilitated obtaining clean sequence data from sporocarps contaminated by other fungi, perhaps due to dilution resulting from low DNA yield. We conclude that conventional extractions are often unnecessary for routine DNA sequencing or genotyping of fungi and oomycetes, and recommend simpler strategies where source materials and intended applications warrant such use.     

A hierarchical Bayesian model to estimate the unobservable predation rate on sawfly cocoons by small mammals

Predation by small mammals has been reported as an important mortality factor for the cocoons of sawfly species. However, it is difficult to provide an accurate estimate of newly spun cocoons and subsequent predation rates by small mammals for several reasons. First, all larvae do not spin cocoons at the same time. Second, cocoons are exposed to small mammal predation immediately after being spun. Third, the cocoons of the current generation are indistinguishable from those of the previous generation. We developed a hierarchical Bayesian model to estimate these values from annual one‐time soil sampling datasets. To apply this model to an actual data set, field surveys were conducted in eight stands of larch plantations in central Hokkaido (Japan) from 2009 to 2012. Ten 0.04‐m² soil samples were annually collected from each site in mid‐October. The abundance of unopened cocoons (I), cocoons emptied by small‐mammal predation (M), and empty cocoons caused by something other than small‐mammal predation (H) were determined. The abundance of newly spun cocoons, the predation rate by small mammals before and after cocoon sampling, and the annual rate of empty cocoons that remained were estimated. A posterior predictive check yielded Bayesian P‐values of 0.54, 0.48, and 0.07 for I, M, and H, respectively. Estimated predation rates showed a significant positive correlation with the number of trap captures of small mammals. Estimates of the number of newly spun cocoons had a significant positive correlation with defoliation intensity. These results indicate that our model showed an acceptable fit, with reasonable estimates. Our model is expected to be widely applicable to all hymenopteran and lepidopteran insects that spin cocoons in soil.     

Growth of phenol-mineralizing microorganisms in fresh water.

A method was developed to enumerate the procaryotic and eucaryotic phenol-mineralizing microorganisms present in samples of fresh water. Sixty-five percent or greater mineralization of [U-14C]phenol was considered a positive tube (contained phenol-mineralizing microorganisms) in the most-probable-number technique. Replicate most-probable-number tubes contained no microbial inhibitors, streptomycin and tetracycline, or cyclohexamide and nystatin plus 200 pg to 100 micrograms of phenol per ml. Phenol mineralization rates were obtained by measuring the amount of exogenous phenol that disappeared from solution over time in the presence or absence of the microbial inhibitors. Initially, less than 100 phenol-mineralizing bacteria per ml and 1 phenol-mineralizing fungus per ml were present at both 200 pg and 100 micrograms of phenol per ml. Phenol mineralization rates were 6.3 times greater for the mineralizing bacteria than for the fungi at 200 pg of phenol per ml. Phenol concentrations above 10 micrograms/ml were inhibitory to the microorganisms capable of mineralizing phenol. The phenol mineralizers grew in the water samples in the absence of phenol, indicating that there were sufficient indigenous nutrients in the lake water to support growth. There was no difference in the growth rate of these microorganisms in the presence or absence of 1 ng of phenol per ml, whereas the growth rate was more rapid at 1 microgram of phenol per ml than in its absence. There was a correlation between microbial growth and the amount of phenol mineralized at 1 microgram but not at 1 ng of phenol per ml.(ABSTRACT TRUNCATED AT 250 WORDS)     

Growth of phenol-mineralizing microorganisms in fresh water

A method was developed to enumerate the procaryotic and eucaryotic phenol-mineralizing microorganisms present in samples of fresh water. Sixty-five percent or greater mineralization of [U-14C]phenol was considered a positive tube (contained phenol-mineralizing microorganisms) in the most-probable-number technique. Replicate most-probable-number tubes contained no microbial inhibitors, streptomycin and tetracycline, or cyclohexamide and nystatin plus 200 pg to 100 micrograms of phenol per ml. Phenol mineralization rates were obtained by measuring the amount of exogenous phenol that disappeared from solution over time in the presence or absence of the microbial inhibitors. Initially, less than 100 phenol-mineralizing bacteria per ml and 1 phenol-mineralizing fungus per ml were present at both 200 pg and 100 micrograms of phenol per ml. Phenol mineralization rates were 6.3 times greater for the mineralizing bacteria than for the fungi at 200 pg of phenol per ml. Phenol concentrations above 10 micrograms/ml were inhibitory to the microorganisms capable of mineralizing phenol. The phenol mineralizers grew in the water samples in the absence of phenol, indicating that there were sufficient indigenous nutrients in the lake water to support growth. There was no difference in the growth rate of these microorganisms in the presence or absence of 1 ng of phenol per ml, whereas the growth rate was more rapid at 1 microgram of phenol per ml than in its absence. There was a correlation between microbial growth and the amount of phenol mineralized at 1 microgram but not at 1 ng of phenol per ml.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of Phenol Feeding Pattern on Microbial Community Structure and Cometabolism of Trichloroethylene

Cometabolism of trichloroethylene (TCE) by phenol-fed enrichments was evaluated in four reactors with distinct phenol feeding patterns. The reactors were inoculated from the same source, operated at the same average dilution rate, and received the same mass of phenol over time. Only the timing of phenol addition differed. Reactor C received phenol continuously; reactor SC5 received phenol semicontinuously--alternating between 5 h of feed and 3 h without feed; reactor SC2 alternated between 2 h of feed and 6 h without feed; and reactor P received a single pulse every 24 h. The structure of the enrichments and their capacity for TCE transformation were analyzed. In long-term operation, reactors C and SC5 were dominated by fungi, had higher levels of predators, were more susceptible to biomass fluctuations, and exhibited reduced capacity for TCE transformation. Reactors P and SC2 were characterized by lower levels of fungi, higher bacterial biomass, higher concentrations of TCE-degrading organisms, and higher rates of TCE transformation. After 200 days of operation, rates of TCE transformation increased 10-fold in reactor P, resulting in TCE transformation rates that were 20 to 100 times higher than the rates of the other reactor communities. The cause of this shift is unknown. Isolates capable of the highest rates of TCE transformation were obtained from reactor P. We conclude that cometabolic activity depends upon microbial community structure and that the community structure can be manipulated by altering the growth substrate feeding pattern.     

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by native microflora and combinations of white-rot fungi in a coal-tar contaminated soil

Four white-rot fungi (Phanerochaete chrysosporium IMI 232175, Pleurotus ostreatus from the University of Alberta Microfungus Collection IMI 341687, Coriolus versicolor IMI 210866 and Wye isolate #7) and all possible combinations of two or more of these fungi, were incubated in microcosms containing wheat straw and non-sterile coal-tar contaminated soil to determine their potential to degrade polycyclic aromatic hydrocarbons (PAHs). Biotic and abiotic controls were prepared similarly and PAH concentrations remaining in each microcosm were determined after 8, 16 and 32 weeks by GC-MS following extraction with dichloromethane. The greatest PAH losses were in the biotic control, compared to small or negligible differences in microcosms inoculated with one or more fungi. These results suggest that in the biotic control native microorganisms colonised the straw added as organic substrate and degraded PAH as an indirect consequence of their metabolism. By contrast, in other microcosms, colonisation of straw by the natural microflora was inhibited because the straw was previously inoculated with fungi. Soil cultures prepared at the end of the experiment showed that though introduced fungi were still alive, they were unable to thrive and degrade PAH in such a highly contaminated soil and remained in a metabolically inactive form.