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Soil microorganisms play a crucial role in the bioremediation of pentachlorophenol (PCP)-contaminated soils. However, whether and how soil bacterial networks with keystone taxa affect PCP dechlorination is not well understood. The present study investigated the effects of citrate on soil bacterial networks mediating PCP dechlorination by direct and indirect transformation in iron-rich upland and paddy soils. The rates of PCP dechlorination and Fe(II) generation were accelerated by citrate addition, particularly in the paddy soils. Network analysis revealed that the topological properties of bacterial networks were changed by citrate addition; more modules and keystone taxa were significantly correlated with PCP dechlorination and Fe(II) generation in the networks. Random forest modeling indicated that Clostridiales was the most important bacterial order; it was significantly involved in both the direct and indirect pathways of PCP dechlorination. Citrate addition had less influence on the balance between the direct and indirect pathways of PCP dechlorination in the upland soils, whereas it enhanced biological PCP dechlorination more directly and efficiently in the paddy soils. Our results suggested that land-use type and citrate addition play a critical role in controlling the biogeochemical mechanisms of PCP dechlorination., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Compost-derived humic acids (HAs) as cheap soil conditioners have potential to facilitate pentachlorophenol (PCP) bio-dechlorination but lack proof and studies. To clarify this, PCP bio-dechlorination mediated by compost-derived HAs under Fe(III) reduction conditions was investigated. Reverse phase high-performance liquid chromatography and high-performance size exclusion chromatography were employed to identify the functional components within compost-derived HAs. Our results showed that compost-derived HAs facilitated the bio-dechlorination of PCP under Fe(III) reduction conditions, and four kinds of byproducts were detected during the process. The relatively hydrophilic and high molecular weight (MW) components within compost-derived HAs presented significant associations with the concentration of byproducts from bio-dechlorination of PCP in Fe 2 O 3 reduction conditions. In contrast, the hydrophobic and low MW components were the main functional components for PCP bio-dechlorination in Fe 3 O 4 reduction environment. These findings clarified the effects of polarity and MW of compost-derived HAs on PCP bio-dechlorination, giving clues to optimize composting technology to utilize compost products for in situ contamination remediation of paddy soil.

An anaerobic incubation was launched with varying nitrate (1, 5, 10 and 20 mM exogenous NaNO 3 ) and molybdate (20 mM Na 2 MoO 4 , a sulfate-reducing inhibitor) additions to investigate the characteristics of PCP dechlorination, as well as the reduction of natural co-occurring electron acceptors, including NO 3 - , Fe(III) and SO 4 2- , and the responses of microbial community structures under a unique reductive mangrove soil. Regardless of exogenous addition, nitrate was rapidly eliminated in the first 12 days. The reduction process of Fe(III) was inhibited, while that of SO 4 2- reduction depended on addition concentration as compared to the control. PCP was mainly degraded from orth-position, forming the only intermediate 2,3,4,5-TeCP by anaerobic microbes, with the highest PCP removal rate of average 21.9% achieved in 1 and 5 mM NaNO 3 as well as 20 mM Na 2 MoO 4 treatments and the lowest of 7.5% in 20 mM NaNO 3 treatment. The effects of nitrate on PCP dechlorination depended on addition concentration, while molybdate promoted PCP attenuation significantly. Analyses of the Illumina sequencing data and the relative abundance of dominant microorganisms indicated that the core functional groups regulated PCP removal at genera level likely included Bacillus, Pesudomonas, Dethiobacter, Desulfoporosinus and Desulfovbrio in the nitrate treatments; while that was likely Sedimentibacter and Geosporobacter_Thermotalea in the molybdate treatment. Nitrate supplement but not over supplement, or addition of molybdate are suggested as alternative strategies for better remediation in the nitrate-deficient and sulfur-accumulated soil ecosystem contaminated by PCP, through regulating the growth of core functional groups and thereby coordinating the interaction between dechlorination and its coupled soil redox processes due to shifts of more available electrons to dechlorination. Our results broadened the knowledge regarding microbial PCP degradation and their interactions with natural soil redox processes under anaerobic soil ecosystems., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Pentachlorophenol (PCP) has been widely used as a pesticide in paddy fields and has imposed negative ecological effect on agricultural soil systems, which are in typically anaerobic conditions. In this study, we investigated the effect of repeated additions of PCP to paddy soil on the microbial communities under anoxic conditions. Acetate was added as the carbon source to induce and accelerate cycles of the PCP degradation. A maximum degradation rate occurred at the 11th cycle, which completely transformed 32.3 μM (8.6 mg L -1 ) PCP in 5 days. Illumina high throughput sequencing of 16S rRNA gene was used to profile the diversity and abundance of microbial communities at each interval and the results showed that the phyla of Bacteroidates, Firmicutes, Proteobacteria, and Euryarchaeota had a dominant presence in the PCP-dechlorinating cultures. Methanosarcina, Syntrophobotulus, Anaeromusa, Zoogloea, Treponema, W22 (family of Cloacamonaceae), and unclassified Cloacamonales were found to be the dominant genera during PCP dechlorination with acetate. The microbial community structure became relatively stable as cycles increased. Treponema, W22, and unclassified Cloacamonales were firstly observed to be associated with PCP dechlorination in the present study. Methanosarcina that have been isolated or identified in PCP dechlorination cultures previously was apparently enriched in the PCP dechlorination cultures. Additionally, the iron-cycling bacteria Syntrophobotulus, Anaeromusa, and Zoogloea were enriched in the PCP dechlorination cultures indicated they were likely to play an important role in PCP dechlorination. These findings increase our understanding for the microbial and geochemical interactions inherent in the transformation of organic contaminants from iron rich soil, and further extend our knowledge of the PCP-transforming microbial communities in anaerobic soil conditions.

Pentachlorophenol (PCP) is a toxic and persistent wood and cellulose preservative extensively used in the past decades. The production process of PCP generates polychlorinated dibenzo- p -dioxins and polychlorinated dibenzofurans (PCDD/Fs) as micropollutants. PCDD/Fs are also known to be very persistent and dangerous for human health and ecosystem functioning. Several physico-chemical and biological technologies have been used to remove PCP and PCDD/Fs from the environment. Bacterial degradation appears to be a cost-effective way of removing these contaminants from soil while causing little impact on the environment. Several bacteria that cometabolize or use these pollutants as their sole source of carbon have been isolated and characterized. This review summarizes current knowledge on the metabolic pathways of bacterial degradation of PCP and PCDD/Fs. PCP can be successfully degraded aerobically or anaerobically by bacteria. Highly chlorinated PCDD/Fs are more likely to be reductively dechlorinated, while less chlorinated PCDD/Fs are more prone to aerobic degradation. The biochemical and genetic basis of these pollutants' degradation is also described. There are several documented studies of effective applications of bioremediation techniques for the removal of PCP and PCDD/Fs from soil and sediments. These findings suggest that biodegradation can occur and be applied to treat these contaminants., Competing Interests: The authors declare no conflict of interest.

Vertical variations of pentachlorophenol (PCP) dissipation and microbial community were investigated in a paddy soil with the addition of electron acceptors (NO3(-), SO4(2-)) and donors (crop residues). Crop residues enhanced PCP dissipation by supplying dissolved organic carbon (DOC) as an electron donor, whereas NO3(-) and SO4(2-) inhibited it. The dissipation of PCP in electron donor treatments resulted in the accumulation of 3,4,5-trichlorophenol (3,4,5-TCP) except for wheat residues. The abundance and diversity of phospholipid fatty acids (PLFAs) decreased with increasing soil depth. The succession of predominant PLFAs shifted from aerobic bacteria to anaerobic bacteria when electron acceptors were changed to electron donors. The saturated/monounsaturated fatty acids (S/M) ratio increased with soil depth, which probably implied that nutrient turnover rate declined after the accumulation of 3,4,5-TCP. The results showed that the addition of electron donors and acceptors modified the microbial communities, which then further influenced the degradation pathway of PCP.

Immobilized solid-phase humin on a graphite electrode set at -500 mV (vs. standard hydrogen electrode) significantly enhanced the microbial reductive dechlorination of pentachlorophenol as a stable solid-phase redox mediator in bioelectrochemical systems (BESs). Compared with the suspended system, the immobilized system dechlorinated PCP at a much higher efficiency, achieving 116 μmol Cl(-)g(-1) humin d(-1). Fluorescence microscopy showed a conspicuous growth of bacteria on the negatively poised immobilized humin. Electron balance analyses suggested that the electrons required for microbial dechlorination were supplied primarily from the humin-immobilized electrode. Microbial community analyses based on 16S rRNA genes showed that Dehalobacter and Desulfovibrio grew on the immobilized humin as potential dechlorinators. These findings extend the potential of BESs using immobilized solid-phase humin as the redox mediator for in situ bioremediation, given the wide distribution of humin and its efficiency and stability as a mediator., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Combined effects of enrichment procedure and non-fermentable acetate or fermentable glucose on system performance and bacterial community for pentachlorophenol (PCP) degradation in microbial fuel cells (MFCs) were determined in this study. Co-substrate and PCP were added into MFCs either simultaneously or sequentially. Simultaneous addition with glucose (simultaneous-glucose) achieved the shortest acclimation time and the most endurance to heavy PCP shock loads. Species of Alphaproteobacteria (simultaneous-acetate, 33.9%; sequential-acetate, 31.3%), Gammaproteobacteria (simultaneous-glucose, 44.1%) and Firmicutes (sequential-glucose, 31.8%) dominated the complex systems. The genus Sedimentibacter was found to exist in all the cases whereas Spirochaetes were merely developed in simultaneous-acetate and simultaneous-glucose. While Epsilonproteobacteria were only absent from sequential-acetate, simultaneous-glucose benefited to the evolution of Lentisphaerae. These results demonstrate simultaneous-glucose is a strategy for efficient system performance and the microbiological evidence can contribute to improving understanding of and optimizing PCP degradation in MFCs., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

This study explored biostimulation mechanisms with an electron donor and a shuttle for accelerating pentachlorophenol (PCP) transformation in iron-rich soils. The results indicated that indigenous microbial communities are important for PCP transformation in soils. Biostimulation of indigenous microbial communities by the addition of lactate and anthraquinone-2,6-disulfonate (AQDS) led to the enhanced rates of PCP dechlorination by the dechlorinating- and iron-reducing bacteria in soils. The electrochemical studies using cyclic voltammograms and microbial current measurements confirmed the high reduction potential and the large amount of electrons generated under biostimulation conditions, which were responsible for the higher rates of PCP transformation. After biostimulation treatments by the additions of lactate and/or AQDS during PCP dechlorination processes, microbial community analysis by the terminal restriction fragment length polymorphism (T-RFLP) method showed the abundance terminal restricted fragments (T-RFs), an indicator of bacterial abundance, which represents the dechlorinating- and iron-reducing bacteria, suggesting their critical roles in PCP dechlorination in soils.

Pentachlorophenol (PCP) was more rapidly degraded in acetate and glucose-fed microbial fuel cells (MFCs) than in open circuit controls, with removal rates of 0.12 ± 0.01 mg/Lh (14.8 ± 1.0 mg/g-VSS-h) in acetate-fed, and 0.08 ± 0.01 mg/L h (6.9 ± 0.8 mg/g-VSS-h) in glucose-fed MFCs, at an initial PCP concentration of 15 mg/L. A PCP of 15 mg/L had no effect on power generation from acetate but power production was decreased with glucose. Coulombic balances indicate the predominant product was electricity (16.1 ± 0.3%) in PCP-acetate MFCs, and lactate (19.8 ± 3.3%) in PCP-glucose MFCs. Current generation accelerated the removal of PCP and co-substrates, as well as the degradation products in both PCP-acetate and PCP-glucose reactors. While 2,3,4,5-tetrachlorophenol was present in both reactors, tetrachlorohydroquinone was only found in PCP-acetate MFCs. These results demonstrate PCP degradation and power production were affected by current generation and the type of electron donor provided., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Bioaugmentation is a promising technology to clean up sites contaminated with recalcitrant chemicals. White-rot fungi have proven to be effective in the degradation of pentachlorophenol. Here, we report the bioremediation of soil contaminated with pentachlorophenol (PCP) by Anthracophyllum discolor and its impact on the soil microbial community. In this study three types of microcosms were established: fresh soil (C(0)), fresh soil plus wheat straw (WS(0)) and, fresh soil plus wheat straw inoculated with A. discolor (WSAD(0)). Additionally, similar treatments and a control of sterile soil spiked with PCP (C(250), WS(250) and WSAD(250)) were used to evaluate the remediation and adsorption of PCP. The PCP removal, total microbial activity, and enzymatic activities were evaluated. This study also investigated the structure of soil microbial community by denaturing gradient gel electrophoresis (DGGE), identifying some of the dominant bacterial and fungal species. The results showed that PCP was effectively degraded in soils by A. discolor and by indigenous soil microorganisms. The addition of wheat straw increased the PCP degradation and enzymatic activities. Only laccase activity was negatively affected by PCP contamination. The PCP degradation was associated with changes in microbial communities, mainly stimulation of members of bacterial phylum Proteobacteria (Xanthomonadaceae, Burkholderiaceae and Enterobacteriaceae), and fungal phylum Ascomycota and Basidiomycota. This study shows the ability of A. discolor to degrade PCP from contaminated soil, and demonstrates that agricultural residues, such as wheat straw, can be used as growth substrate by microorganisms in PCP-contaminated soil, demonstrating a great potential of autochthonous microorganisms for soil remediation., (Copyright 2010 Elsevier B.V. All rights reserved.)

The anaerobic biological treatment of pentachlorophenol (PCP) and methanol as the main carbon source was investigated in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor at 30+/-1 degrees C, during a 220-day trial period. The reactor biomass was developed as an attached biofilm on polyurethane foam particles, with 24h of hydraulic retention time. The PCP concentrations, which ranged from 2.0 to 13.0 mg/L, were controlled by adding synthetic substrate. The HAIB reactor reduced 97% of COD and removed 99% of PCP. The microbial biofilm communities of the HAIB reactor amended with PCP, without previous acclimatization, were characterized by polymerase chain reaction (PCR) and amplified ribosomal DNA restriction analysis (ARDRA) with specific Archaea oligonucleotide primers. The ARDRA technique provided an adequate analysis of the community, revealing the profile of the selected population along the reactor. The biomass activities in the HAIB reactor at the end of the experiments indicated the development of PCP degraders and the maintenance of the population of methanogenic Archaea, ensuring the high efficiency of the system treating PCP with added methanol as the cosubstrate. The use of the simplified ARDRA method enabled us to monitor the microbial population with the addition of high concentrations of toxic compounds and highlighting a selection of microorganisms in the biofilm.

A microbial consortium was developed by continuous enrichment of bacterial population isolated from sediment core of pulp and paper mill effluent in mineral salts medium (MSM) supplemented with pentachlorophenol (PCP) as sole source of carbon and energy in the chemostat. The enriched consortium contained three bacterial strains identified as Escherichia coli (PCP1), Pseudomonas aeruginosa (PCP2) and Acinetobacter sp. (PCP3) by morphological and biochemical tests, further confirmation was done using 16S rDNA sequence analysis. The potency of bacterial isolates in degradation of PCP was monitored in terms of growth and utilization of PCP as substrate with spectrophotometer and gas chromatograph-mass spectrometer (GC-MS) analysis. The strains were tested for their utilization of various organic compounds. The strain PCP3, showed higher potency to utilize PCP as sole source of carbon and energy than PCP1 and PCP2. The bacterial strain were able to utilize PCP through an oxidative and reductive route as indicated with the formation of tetrachloro-p-hydroquinone (TeCH), 2-chloro-1,4-benzenediol and 2,3,4,6-tetrachlorophenol, respectively.

Bacterial biodegradation (bioremediation) is the use of microorganisms to break down organic materials into simpler compounds; it plays a pivotal role in the clean-up of hazardous wastes in the environment. Following the completion of genome sequencing in bacteria capable of biodegradation, functional genomic studies have played a major role in obtaining information on bacterial biodegradation pathways. Novel proteomics technologies have recently been developed to make it possible to analyze global protein expression. Proteomics can also provide important information on the life cycle, regulation, and post-translational modification of proteins induced under specific conditions. Proteomics technologies have been applied to the comprehensive study of bacterial biodegradation. In this paper, we introduce the proteomics technologies applicable to bacterial biodegradation studies, review the results of the proteomics analysis of representative biodegrading bacteria, and discuss the potential use of proteomics technologies in future biodegradation studies.

Bio-surfactant is a new type of surfactant that is produced in microbial metabolism. Adding bio-surfactant during composting process, especially to those contain some toxic substances, has been proved to be a promising way. In this study, Strains III (2), a bacterial with high activity to produce bio-surfactant, were isolated firstly. Following comparison experiments with and without adding Strains III (2), namely Run 1 and Run R, were conducted, respectively. The experimental results showed that, by adding Strains III (2), the surface tension could reduce from 46.5 mN/m to 39.8 mN/m and the corresponding time to maintain the surface tension under 50 mN/ m could prolong from 60 h to 90 h. The oxygen uptake rate and total accumulated oxygen consumption with Stains III (2) were both higher than those without Strains III (2), while the accumulation of H2S in outlet gas was reduced to around 50% of Run R. Moreover, two additional experiments were also carried out to examine the effects of strains coming from different systems. One is adding Strains III (2) with a dose of 0.4% (Run 2), and the other is seedling commercial Strains at the same conditions, the composting experiments showed that: Run 2 was more effective than Run 3, because the commercial Strains can be suppressed significantly in a complex composting system with different pH, high temperature and some of metals. The bio-surfactant was also added into the solid waste, which contained some toxic substances, the corresponding results showed that the remove rate of Hg and sodium pentachlorophenolate (PCP-Na) could be improved highly. Thus, the microenvironment, reactionrate and composting quality could be enhanced effectively by adding bio-surfactant to the composting process.

Biodegradation of organic contaminants in multi-phase systems, such as soils and aquifers, is often limited by the extent and the rate of contaminant sorption onto the solid matrix. However, information about biodegradation in complex matrices is largely limited to studies of closed systems under batch conditions in which potential bioavailability is characterised by the aqueous-phase concentration of a contaminant. There is little knowledge regarding the influence of flow-through conditions on the availability of contaminants to microbes (contaminant bioavailability). Thus, the aim of this study was to assess and compare contaminant bioavailability, in the presence of a sorptive medium, under both batch and flow-through conditions. Accordingly, experiments were designed in which pentachlorophenol (PCP) was introduced into a mixture of inoculated silica sand and a PCP-retaining resin, under either batch or flow-through (columns) conditions. The results indicated that an increase in the amount of resin (0.1-0.2 g) clearly lowered PCP availability to microbes after 170 h under batch conditions (30 and 45% respectively); whereas, the initial decrease in bioavailability observed under flow-through conditions (45 and 70% respectively) was reversed and no longer observable after 170 h. This increase in PCP availability was linked to an improvement in the contaminant biodegradation capacity from 0.03 to 0.13 mg x l(-1) x h(-1) over 200 h.

Competition for molecular hydrogen exists among hydrogen-utilizing microorganisms in anoxic environments, and evidence suggests that lower hydrogen concentrations are observed with more energetically favorable electron-accepting processes. The transfer of electrons to organochlorines via reductive dehalogenation reactions plays an important role in hydrogen dynamics in impacted systems. We studied the flux of aqueous hydrogen concentrations in methanogenic sediment microcosms prior to and during reductive dehalogenation of a variety of substituted chlorophenols (CP) and tetrachloroethene (perchloroethylene, PCE). Mean hydrogen concentrations during reductive dehalogenation of 2,4-CP, 2,3,4-CP, and PCP were 3.6 nM, 4.1 nM, and 0.34 nM, respectively. Sediment microcosms that were not dosed with chlorophenols yet were actively methanogenic maintained a significantly higher mean hydrogen concentration of 9.8 nM. During active PCE dehalogenation, sediment microcosms maintained a mean hydrogen concentration of 0.82 nM. These data indicate that during limiting hydrogen production, the threshold ecosystem hydrogen concentration is controlled by microbial populations that couple hydrogen oxidation to thermodynamically favorable electron accepting reactions, including reductive dehalogenation of chloroaromatic and chloroaliphatic compounds. We also present revised estimates for the Gibbs free energy available from the reductive dehalogenation of a variety of substituted chlorophenols based on recently published values of vapor pressure, solubility, and pKa for these compounds.

Aims: To determine whether composting with animal manure can be used to effectively remediate soil from a pentachlorophenol (PCP)-contaminated site, and to establish the fate of the degraded xenobiotic., Methods and Results: Contaminated soil from a sawmill site was mixed with farm animal manure and composted in a 0.5 m3 silo under fully aerobic conditions. The disappearance and fate of PCP was monitored by gas chromatography (GC-ECD) and extensive mineralization confirmed in experiments with 14C-radiolabelled PCP. The disappearance of PCP was rapid and virtually complete within 6 days, prior to the onset of thermophilic conditions. Dechlorination of the PCP was found to be both reductive and sequential., Conclusions: PCP removal from contaminated soil by aerobic composting with animal manure is efficient and proceeds via reductive dechlorination to virtually complete mineralization. This contrasts with other chlorophenol composting regimes in which mineralization is achieved but dechlorination intermediates do not accumulate to detectable levels., Significance and Impact of the Study: The results of this study demonstrate that anaerobic reductive dechlorination can proceed in an aerobic composting environment and contribute to efficient pentachlorophenol removal. Farmyard manure composts may represent a rapid, low-cost, low-technology option for treatment of chlorophenol-contaminated soils.

The composition and chlorophenol-degrading potential of groundwater bacterial community in a permanently cold, oxygen-deficient chlorophenol contaminated aquifer at Kärkölä, Finland was studied with the aim of evaluating in situ bioremediation potential. The groundwater contained from 10(4) to 10(7) microscopically counted cells/ml and up to 10(5) CFU/ml heterotrophic bacteria cultivable at 8 and 20 degrees C. Of the 102 pure cultures, of which 86% Gram-negative, from the plume area (10,000 microg of chlorophenols/l), 57% degraded 2, 3, 4, 6-tetrachlorophenol (TeCP), the main component of the wood preservative which was the source of contamination: 17% also degraded pentachlorophenol (PCP). The degraders were scattered among 16 different clusters of Gram-negatives mainly proteobacteria and members of Cytophaga/Flexibacter/Bacteroides phylum judged by the composition of whole-cell fatty acids. Only one Gram-positive degrading cluster was found containing seven actinobacteria closest to Nocardioides. Of the 88 pure cultures isolated from outside the plume (< 10 microg of chlorophenols/l) 67% were Gram-negative. Seven percent of the isolates degraded 2, 3. 4, 6-TeCP and/or PCP. Five of the Gram-positive isolates from outside the plume were Mycobacterium/Rhodococcus-related actinobacteria and O-methylated 2, 3, 4, 6-TeCP and PCP. The results show that chlorophenol degrading bacterial flora had been enriched as a result of contamination of the aquifer. This suggests significant in situ bioremediation potential of the site.

A rapid enrichment approach based on a pentachlorophenol (PCP) feeding strategy which linked the PCP loading rate to methane production was applied to an upflow anaerobic sludge bed reactor inoculated with anaerobic sludge. Due to this strategy, over a 140-day experimental period the PCP volumetric load increased from 2 to 65 mg L(R)(-1) day(-1) with a near zero effluent concentration of PCP. Dechlorination dynamics featured sequential appearance of 3,4,5-chlorophenol, 3,5-chloro- phenol, and 3-chlorophenol in the reactor effluent. Profiling of the reactor population by denaturing gradient gel electrophoresis (DGGE) revealed a correlation between the appearance of dechlorination intermediates and bands on the DGGE profile. Nucleotide sequencing of newly detected 16S rDNA fragments suggested the proliferation of Clostridium and Syntrophobacter/Syntrophomonas spp. in the reactor during PCP degradation. Published by John Wiley & Sons, Inc.

A new most probable number (MPN) method for the determination of pentachlorophenol (PCP) degraders in soil using the change in pH due to PCP degradation is compared with a well documented MPN method using radiolabeled PCP. The results of all MPN counts were similar within a 95% confidence limit. The results obtained in MPN per gram of dry soil using pH measurements were 1.8 (+3.1, -1.03) x10 (4) compared to 0.64 (+1.34, -0.42) x 10(4) when using production of [(14)C]CO(2).

Chlorophenol-degrading bacteria from a long-term polluted groundwater aquifer were characterized. All isolates degraded 2,4,6-trichlorophenol and 2,3,4,6-tetrachlorophenol at concentrations detected in the contaminated groundwater (< 10 mg 1(-1)). Pentachlorophenol was degraded by three isolates when present alone. In two gram-positive isolates, 2,3,4,6-tetrachlorophenol was required as an inducer for the degradation of pentachlorophenol. The gram-positive isolates were sensitive to pentachlorophenol, with an IC50 value of 5 mg/l. Isolates belonging to the Cytophaga/Flexibacter/Bacteroides phylum had IC50 values of 25 and 63 mg/l. Isolates belonging to alpha-, beta- and gamma-Proteobacteria generally tolerated the highest pentachlorophenol concentrations (> 100 mg/l). Polychlorophenol-degrading capacity was found in strains of Nocardioides, Pseudomonas, Ralstonia, Flavobacterium, and Caulobacter previously not known to degrade polychlorophenols. In addition, six polychlorophenol-degrading sphingomonads were found.

Air-lift percolator experiments simulated in situ subsurface degradation of 2,4,6-trichlorophenol, 2,3,4,6-tetrachlorophenol and pentachlorophenol, in mixtures and individually, by indigenous microorganisms from a chlorophenol-contaminated aquifer. Inoculation with a chlorophenol(CP)-degrading gram-positive isolate from the CP-contaminated groundwater did not significantly increase CP degradation rates. Feed concentrations of up to 55 mg CP l-1 were degraded. Stable CP degradation was maintained for over 6 months. CP degradation rates up to 54.3 mg l-1 day-1 and effluent concentrations below 40 micrograms CP l-1 were achieved. CP were mineralized as shown by CP reduction, dissolved organic carbon removal and release of inorganic chloride.

With a new enrichment protocol, pentachlorophenol (PCP)-degrading bacteria were isolated from soil, water, and sewage. When characterized, all isolates were related and shared characteristics of the genus Arthrobacter. Growth rates for strain NC were determined for a number of substrates, including PCP and 2,4,6-trichlorophenol. Changes in PCP concentration affected growth rate and length of the lag phase but not cell yield. Increasing the pH from 6.8 to 7.8 decreased the length of the lag phase for growth on PCP. Cessation of growth, upon incremental addition of PCP, was found to be related to a decrease in pH rather than to a buildup of a toxic metabolite. Degradation of PCP by strain NC was shown to be complete.

The biodegradation, photolysis, and adsorption of pentachlorophenol (PCP) in outdoor, aquatic environments were examined with man-made channels built by the U.S. Environmental Protection Agency at a field station on the Mississippi River near Monticello, Minn. Four channels were used, each channel being approximately 520 m long and receiving river water that flowed through the channels for about 10 h before reentering the river. The channels were dosed continuously during the summer of 1982 with various concentrations of PCP (approximately 0, 48, 144, and 432 micrograms/liter). We monitored the biotic and abiotic degradation of PCP in these channels for approximately 16 weeks. Photolysis of PCP was rapid at the water surface, but greatly attenuated with depth. Depending on sunlight conditions, photolysis accounted for a 5 to 28% decline in initial PCP concentration. Adsorption of PCP by sediment and uptake by biota accounted for less than 15% and probably less than 5% in unacclimated water. Microbial degradation of PCP became significant about 3 weeks after the initiation of dosing and eventually became the primary mechanism of PCP removal, accounting for a 26 to 46% (dose-dependent) decline in initial PCP. Most of the PCP-mineralizing microorganisms that developed in the channels were either attached to surfaces (e.g., rocks and macrophytes) or associated with surface sediments. Total bacterial numbers (direct microscopic counts) in the various channels were not affected significantly by PCP concentrations of micrograms per liter. Numerous strains of bacteria able to grow at the expense of PCP were isolated from the adapted channels. The experiments reported here will help predict the responses of flowing aquatic ecosystems to contamination by biocides such as pentachlorophenol.

Batch and fed-batch experiments were conducted to examine the kinetics of pentachlorophenol utilization by an enrichment culture of pentachlorophenol-degrading bacteria. The Haldane modification of the Monod equation was found to describe the relationship between the specific growth rate and substrate concentration. Analysis of the kinetic parameters indicated that the maximum specific growth rate and yield coefficients are low, with values of 0.074 h-1 and 0.136 g/g, respectively. The Monod constant (Ks) was estimated to be 60 micrograms/liter, indicating a high affinity of the microorganisms for the substrate. However, high concentrations (KI = 1,375 micrograms/liter) were shown to be inhibitory for metabolism and growth. These kinetic parameters can be used to define the optimal conditions for the removal of pentachlorophenol in biological treatment systems.

The antibacterial activity of pentachlorophenol and 35 of its known or possible metabolites against 30 different species of bacteria was tested. In comparison with pentachlorophenol, no increase of inhibitory activity was found for any of the chlorinated anisoles tested (except for pentachloroanisole against Streptomyces spp.), 2-chlorophenol, 2,6-dichlorophenol, 2,3,6- and 2,4,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, tetrachloro-1,4- and -1,3-benzenediol (except for the 1,3-isomer against Streptomyces spp.), tetrachloro-1,3-dimethoxybenzene, and tetrachloro-1,3-benzenediol diacetate. Two chlorophenols, five dichlorophenols, four trichlorophenols, two tetrachlorophenols, and tetrachloro-1,2-benzenediol were more active than pentachlorophenol against some, but not all, of the strains tested.

A bacterial isolate obtained from a continuous-flow enrichment culture has been shown to metabolize pentachlorophenol as a sole source of organic carbon and energy.