Impact of four pesticides on the growth and metabolic activities of two photosynthetic algae

The acute toxicity was determined for soil algae Chlorella kesslerei and Anabaena inaequalis, exposed to pesticides lindane, pentachlorophenol (PCP), isoproturon (IPU), and methyl parathion (MP). Toxicity markers included growth inhibition, chlorophyll biosynthesis, and total carbohydrate content, as a function of dose and time. Concentration response functions (EC50) were estimated by probit data transformation and weighted linear regression analyses. Lindane's toxicity to Chlorella increased sharply with time (EC50 = 7490, 10.3, 0.09 mg L(-1); 24, 48, 72 h), but remained nearly constant through 72 h with Anabaena (8.7-6.7 mg L(-1); 24-72 h). PCP at low concentrations stimulated algal growth and chlorophyll a production, an effect reversed at higher doses. Anabaena was less tolerant of PCP and MP than was Chlorella. The 96-h static EC50 values for Chlorella were: 0.003, 34, 0.05, and 291 mg L(-1) for lindane, PCP, isoproturon, and MP, respectively; for Anabaena, these were 4.2, 0.13, 0.21, and 19 mg L(-1). Carbohydrate production responses were similar to those of cell density (growth) and chlorophyll biosynthesis, with MP having the lowest adverse impact. The overall relative toxicity among the four tested pesticides was: for Chlorella, lindane > IPU > PCP > MP; and for Anabaena, PCP > IPU > lindane > MP. The results confirm that toxicants such as these pesticides may affect individual (though related) species to significantly different degrees.     

Pentachlorophenol induced physiological-biochemical changes in Chlorella pyrenoidosa culture

Present study investigated physiological and biochemical changes in a green alga culture (Chlorella pyrenoidosa) upon exposure to varied concentrations of pentachlorophenol (PCP) (0, 0.4 mgL(-1), 2.5 mgL(-1) and 10 mgL(-1)). The results revealed that with the increase of PCP concentration, a decrease of the algal biomass, levels of photosynthetic pigments (chlorophyll a, b and carotenoid), soluble protein and an increase of nitrate reductase (NR) activity were observed. This suggests that the PCP may serve as an uncoupler, causing low ATP level within the algal cells and led most of the NR molecules to be in the dephosphoration state (i.e. active from). Moreover, it shows that photosynthetic pigments especially carotenoid were more sensitive indicators to indicate PCP toxicity as compared to the other parameters.     

IMPACT OF FOUR PESTICIDES ON THE GROWTH AND METABOLIC ACTIVITIES OF TWO PHOTOSYNTHETIC ALGAE

The acute toxicity was determined for soil algae Chlorella kesslerei and Anabaena inaequalis, exposed to pesticides lindane, pentachlorophenol (PCP), isoproturon (IPU), and methyl parathion (MP). Toxicity markers included growth inhibition, chlorophyll biosynthesis, and total carbohydrate content, as a function of dose and time. Concentration response functions (EC₅₀) were estimated by probit data transformation and weighted linear regression analyses. Lindane's toxicity to Chlorella increased sharply with time (EC₅₀=7490, 10.3, 0.09 mg L^?1; 24, 48, 72 h), but remained nearly constant through 72 h with Anabaena (8.7?6.7 mg L^?1; 24–72 h). PCP at low concentrations stimulated algal growth and chlorophyll a production, an effect reversed at higher doses. Anabaena was less tolerant of PCP and MP than was Chlorella. The 96-h static EC₅₀ values for Chlorella were: 0.003, 34, 0.05, and 291 mg L^?1 for lindane, PCP, isoproturon, and MP, respectively; for Anabaena, these were 4.2, 0.13, 0.21, and 19 mg L^?1. Carbohydrate production responses were similar to those of cell density (growth) and chlorophyll biosynthesis, with MP having the lowest adverse impact. The overall relative toxicity among the four tested pesticides was: for Chlorella, lindane>IPU?PCP?MP; and for Anabaena, PCP>IPU>lindane>MP. The results confirm that toxicants such as these pesticides may affect individual (though related) species to significantly different degrees.     

Simultaneous removal of nitrate and pentachlorophenol from simulated groundwater using a biodenitrification reactor packed with corncob

Both nitrate and pentachlorophenol (PCP) are familiar pollutants in aqueous environment. This research is focused on the simultaneous removal of nitrate and PCP from simulated contaminated groundwater using a laboratory-scale denitrification reactor packed with corncob as both carbon source and biofilm support. The reactor could be started up readily, and the removal efficiencies of nitrate and PCP reached up to approximately 98 % and 40–45 % when their initial concentrations were 50 mg N/L and 5 mg/L, respectively, after 15-day continuous operation at 10 h of hydraulic retention time (HRT) and 25 °C. Approximately 91 % of PCP removal efficiency was achieved, with 2.47 mg/L of chloride ion release at 24 h of HRT. Eighty-two percent of chlorine in PCP removed was ionized. The productions of 3-chlorophenol and phenol and chloride ion release indicate that the reductive dechlorination reaction is a major degradation pathway of PCP under the experimental conditions.     

Chromium (VI) accumulation reduces chlorophyll biosynthesis, nitrate reductase activity and protein content in Nymphaea alba L

Plants of Nymphaea alba L. grown at various levels of chromium (VI) ranging from 1 to 200 microM accumulated chromium in concentration and duration-dependent manner. At all Cr levels, chromium accumulation by various plant tissues followed the order roots > leaves > rhizomes. Approximately 93% of total chromium present in the medium was accumulated by plants at lowest conentration (1 microM) used in the experiment. Chromium-induced toxicity appears at 1 microM chromium resulting in the build-up of delta-aminolevulinic acid (ALA) and reduced activities of delta-aminolevulinic acid dehydratase (ALAD) and nitrate reductase (NR), total chlorophyll (Chl) and protein contents. Ch1a was more sensitive than Ch1b to chromium toxicity. It could be inferred that chromium toxicity is not located at the level of ALA synthesis, but, probably at the ALAD activity which was more severely affected during chlorophyll biosynthesis. Finally, impaired chlorophyll biosynthesis resulted in reduced total chlorophyll content.     

Bioconcentration factor of relatively low concentrations of chlorophenols in Japanese medaka

Bioconcentration factors (BCF) for pentachlorophenol (PCP) and 2,4-dichlorophenol (2,4-DCP) in Japanese medaka (Oryzias latipes) were determined at five different concentrations of the chemicals, between 0.1 and 10 microg/l (PCP), 0.3 and 30 microg/l (2,4-DCP), in the ambient water. Medaka were exposed to each chemicals in a continuous-flow system during the embryonic development period and 60 days after hatching from eggs collected in the laboratory. Both the exposure time and the aqueous concentrations are much more realistic and closer to natural aquatic environments than those used in conventional BCF studies. The BCF values of PCP were from (4.9+/-2.8)x10(3) at the aqueous concentration of 0.074+/-0.028 microg/l to (2.1+/-1.4)x10(3) at 9.70+/-0.56 microg/l. The BCF value of 2,4-DCP were from (3.4+/-3.0)x10(2) at 0.235+/-0.060 microg/l to 92+/-27 at 27.3+/-1.6 microg/l. Generally, BCF values increased as the aqueous concentrations of PCP or 2,4-DCP decreased. This finding suggests that a relatively low and realistic aqueous concentration of these compounds is necessary to more accurately determine their BCF values in natural aquatic environments. Conventional BCF experiments at higher aqueous concentrations may underestimate the BCF values.     

Phytotoxicity of diuron alone and in combination with copper or folpet on duckweed (Lemna minor)

The photosystem II-herbicide diuron is widely used for weed control in Champagne's vineyards. Its important use and its relative persistence make it of particular interest for ecotoxicological studies. Toxicity of diuron was assessed on Lemna minor L., a representative aquatic macrophyte regularly used for toxicological studies. Toxicity assessments were based on inhibition of growth and total chlorophyll content of L. minor cultures after 7 days. Growth was inhibited and IC(50) and IC(90) were, respectively, 25 and 60 microg l(-1), but chlorophyll content of L. minor increased in response to the herbicide. When diuron was combined with copper, growth inhibition of L. minor depended on the concentrations of both chemicals. For some concentrations, combination of these chemicals resulted in a slight (but non-significant) antagonism. Additivity was observed for all other mixtures. When diuron was combined with folpet, growth and chlorophyll content of L. minor only depended on the concentration of the herbicide. Diuron was also found to prevent the copper-induced decrease of chlorophyll content when it was combined with this metal. A multifactorial model was found more appropriate to characterize interactions between pesticides than Abott's model.     

Nitrate reductase activity as an indicator of ponderosa pine response to atmospheric nitrogen deposition in the San Bernardino Mountains

Determinations of nitrate reductase (NR) activity in ponderosa pine (Pinus ponderosa Dougl. ex. Laws.) needles were performed during summer 1994 in two areas (consisting of six different sites) with different nitrogen (N) deposition levels in the San Bernardino Mountains, southern California. Nitrate reductase activity was used as an integrative indicator of atmospheric nitrogen deposition to pine trees (direct uptake of N species from the atmosphere and N transported from the soil). Deposition of nitrate (NO3-) to pine branches was measured in order to determine dry atmospheric inputs of the oxidized N species to tree foliage. High NR activity was detected in all of the experimental sites. Activity of the enzyme was significantly higher at the locations characterized by higher NO3- deposition to branches--slight positive correlation between branch deposited NO3- and NR activity was found. However, high variability of NR in time and between the experimental sites discredit the NR assay as a reliable indicator of N deposition for ponderosa pine in the field conditions. This could be caused by substantial interference from other abiotic and biotic factors with tropospheric ozone as probably the most important one.     

The changes of nitric oxide production during the growth of Microcystis aerugrinosa

This study characterized the changes of nitric oxide (NO) production during the growth of Microcystis aerugrinosa, a cyanobacterium which usually cause cyanobacterial blooms. Results showed a drastic NO release accompanying with cell density and Chl-a content sharp rises when M. aerugrinosa grew from fifth day to sixth day. Moreover, high N:P ratio accelerated the cyanobacterial growth and NO burst. Sodium nitroprusside, an exogenous NO donor, promoted M. aerugrinosa growth with the optimal concentration of 0.1 mg/L. Experiments by supplementing with sodium nitrite and l-arginine demonstrated NO production in M. aerugrinosa cells was mainly through nitrate reductase (NR) pathway while minorly through NO synthase pathway. All these data suggested M. aerugrinosa produced increasing NO during its growth mainly by NR pathway, during which NO positively regulated the growth of M. aerugrinosa.     

Comparative study of the effects of MCPA, butylate, atrazine, and cyanazine on Selenastrum capricornutum

The herbicides MCPA, butylate, atrazine and cyanazine are extensively used in Canadian agriculture and information regarding their effects on indigenous biota is scarce. Phytotoxicity assessments were conducted in the laboratory on the common green alga Selenastrum capricornutum using both the active ingredient of the herbicides and their formulated products (for MCPA and butylate). Endpoints determined after the 96 h exposure included algal population growth inhibition (IC50-cell counts), percent lethality (LC50-flow cytometry derived) and photosynthetic electron transport inhibition (EC50-fluorescence induction). Pesticide formulations had greater toxic effects than the active ingredient alone. The 96 h IC50 (50% Inhibition Concentration) and LOEC (Lowest Observable Effects Concentration) using cell counts of S. capricornutum were 18.4 and 8.9 mg l(-1) respectively for MCPA-active ingredient and for MCPA-formulated, these were 0.62 and 0.0062 mg l(-1) respectively. Those for butylateactive ingredient were 61.0 and 8.3 mg l(-1) and for butylate-formulated 1.46 and 0.17 mg l(-1), respectively. The triazines active ingredient, which are photosynthetic inhibitors, had greater effects than either the MCPA or butylate. The IC50 for cyanazine and atrazine were 0.059 and 0.026 mg l(-1), respectively. By comparing the IC50 and LC50 values for the tested active ingredients, it was found that the effects of atrazine were algicidal, whereas those of cyanazine, butylate and MCPA were algistatic.     

Metallothionein induction, antioxidative responses, glycogen and growth changes in Tubifex tubifex (Oligochaete) exposed to the fungicide, fenhexamid

Laboratory studies were conducted to determine the effects of different concentrations of fenhexamid (0.1, 1, and 10 mg L(-1)) on growth, oxidative stress, protein, glycogen, and metallothionein (MT) contents in Tubifex tubifex after an exposure of 2, 4, and 7 days. In addition, residues of the fungicide were followed in water and in the worms. In water, fenhexamid concentration decreased slowly (maximum -2 +/- 0.03% after 2 days for 1 mg L(-1)). In the worms, it increased after 4 days and decreased thereafter, confirming that the worms were exposed to the fungicide and not to a degradation product. LC50 values were between 95.22 +/- 5.36 and 32.11 +/- 1.8 mg L(-1) depending on exposure time. Exposure to fenhexamid had a negative effect on T. tubifex growth (maximum effect -12.2 +/- 0.8% after 7 days with 10 mg L(-1)) demonstrating the toxic effect of the pesticide. This growth rate decrease was accompanied by a reduction in protein and glycogen contents. The activity of catalase (CAT), and glutathione reductase (GR) increased in response to the fungicide demonstrating an oxidative stress in the worms. In contrast glutathion-S-transferase activity (GST) decreased. Exposure to fenhexamid also induced synthesis of MT (maximum +78 +/- 8% after 2 days for 10 mg L(-1)). The specificity of MT concentration increase in response to metals is discussed.     

Studies on nitric oxide (NO) formation by the green alga Scenedesmus obliquus and the diazotrophic cyanobacterium Anabaena doliolum.

This study provides preliminary evidence that NO production could be a general attribute of algae. Anabaena doliolum was found to be a better NO producer than Scenedesmus and Synechoccocus. Experiments conducted with inhibitors of photosynthesis (DCMU), ATP synthesis (DCCD), and the uncoupler (2,4-DNP) and its analog arsenate clearly revealed that inhibition of nitrite assimilation through the blockage of nitrite reductase (NiR) is primarily responsible for NO emission. A linear relationship between nitrite concentration in the culture medium and NO in the exhaust gas supports the view that accumulation of nitrite is responsible for NO formation. A failure of Scenedesmus, grown in the medium substituted with W for Mo, to produce either NO/NO-2 in light or a 'light-off' peak, and a resumption of these activities upon the addition of Mo proved beyond doubt that a functional nitrate reductase (NR) is necessary for the production of nitrite and NO by algae grown on nitrate as the nitrogen source. Moreover, the appearance of a NO peak immediately after nitrite supplementation under dark conditions in W-substituted cultures with or without glucose ruled out an enzymatic role of NR in NO emission.     

The effects of SO2 fumigation on the nitrogen metabolism of aseptically grown spruce seedlings

Aseptically grown spruce seedlings were cultivated in a hydroponic system, where the roots were separated from the shoots by a gastight, silicone material. The plants were fumigated with four SO(2) concentrations (93, 190, 270 and 530 microg m(-3)) for nine weeks. Up to 270 microg m(-3) of SO(2), an inhibition of nitrogen metabolism (enzyme activities of nitrate reductase (NR) and glutamine sythetase (GS) and nitrate content) in the shoot was compensated by a stimulation in the root, while nitrogen uptake was unaffected. Only the treatment with 530 microg m(-3) of SO(3) decreased enzyme activities, nitrate content in both roots and shoots as well as nitrate uptake, and inhibited the growth of plants. Increases in the content of thiols and superoxidismutase activity are discussed in terms of SO(2) detoxification.     

Toxicity and tissue distribution of pentachlorophenol and permethrin in pipistrelle bats experimentally exposed to treated timber

The dependence of bats in Britain on houses as roosts may result in them being exposed to pesticides used in remedial timber treatments. Pentachlorophenol (PCP) and permethrin are used as a fungicide and an insecticide for timber treatment, respectively. The present study investigated toxicity and distribution in body tissues of these two pesticides in pipistrelle bats. Four groups of nine to ten bats were kept in separate outdoor flight enclosures and were provided with roost boxes treated with either PCP only, permethrin, PCP/permethrin mixture or solvent only (control). At the start of the experiment, mean (+/-SE) PCP and permethrin concentrations on the surface of wooden blocks that had been treated in the same way as roost boxes were 69.32+/-6.76 mg g(-1) (n=6) and 3.3+/-1.6 mg g(-1) (n=3), respectively. All bats exposed to PCP and PCP/permethrin treated boxes died within 24 and 120 h, respectively; nine out of the ten controls survived the 32 day experimental period (P<0.001; both groups compared with control). Bats exposed to permethrin treated boxes survived as well as controls. Mean (+/-SE) carcass PCP concentration (excluding deposits on fur) of bats exposed to PCP and PCP/permethrin treated boxes was 13.11+/-2.52 microg g(-1)BW (n=20). PCP burdens on fur were positively correlated with total weight of PCP in the carcass (P<0.001). PCP was present in fat depots, liver, kidney and the remainder of the body which, despite containing low PCP concentrations, was the main PCP reservoir (66.4+/-5.0% of carcass PCP load; n=20). Total PCP in the carcass was significantly correlated with lipid weight (P<0.005). Permethrin was not detectable in body washes and tissues of bats exposed to PCP/permethrin mixture or permethrin.     

Antioxidant enzyme activities as affected by trivalent and hexavalent chromium species in Fontinalis antipyretica Hedw

The detoxification mechanisms of the aquatic moss, Fontinalis antipyretica Hedw., exposed to Cr was analyzed. In addition, the influence of Cr salts (as Cr nitrate, chloride and potassium bichromate) on these mechanisms has also been studied. The activity of antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1.), catalase (EC 1.11.1.6.), ascorbate peroxidase (APX, EC 1.11.1.11.), guaiacol peroxidase (GPX, EC 1.11.1.7.) and glutathione reductase (GR, EC 1.6.4.2.) increased in plants treated with Cr concentrations ranging from 6.25x10(-5) to 6.25mM when given as Cr(NO(3))(3). Antioxidant enzymes responded to the other two salts CrCl(3) and K(2)Cr(2)O(7) only with Cr concentrations higher than 6.25x10(-2)mM. Glutathione level and GSSG/GSH ratio also responded to Cr exposure but no dose-effect relationship could be observed. Moreover, two unknown thiol compounds were observed in mosses exposed to the highest Cr concentrations. Effects on chlorophyll contents and chlorophyll a/b ratios were also shown even at low Cr concentrations. Our results indicated that environmentally realistic concentrations of Cr could lead to impairment of the cellular activity towards F. antipyretica and that Cr(III), when present as a nitrate salt, was as harmful as Cr(VI).     

Phytotoxicity and antioxidative enzymes of green microalga (Desmodesmus subspicatus) and duckweed (Lemna minor) exposed to herbicides MCPA, chloridazon and their mixtures

In this study, we evaluate the toxicity of MCPA (auxin-like growth inhibitor), chloridazon (CHD) (PSII-inhibitor) and their mixtures to floating plants and planktonic algae. Toxicity of MCPA (4-chloro-2-methylphenoxyacetic acid) and CHD (5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone) was first assessed in two growth inhibition tests with Lemna minor (ISO/DIS 20079) and Desmodesmus subspicatus (ISO 8692). Next, herbicide mixtures at concentrations corresponding to the EC values were used to assess their interactive effects, and the biomarkers were: for duckweed fresh weight, frond area, chlorophyll content and number of fronds, and for algae cell count and cell volume. The 3d EC?? and EC?? values using cell counts of D. subspicatus were 142.7 and 529.1 mg/L for MCPA and 1.7 and 5.1 mg/L for CHD. The 7d EC?? and EC?? values using frond number of L. minor amounted to 0.8 and 5.4 mg/L for MCPA and 0.7 and 10.4 mg/L for CHD. Higher sensitivity of reproductive (number of cells/fronds) than growth processes (cell volume/frond area) to herbicides applied individually and in mixtures was especially pronounced in the responses of Desmodesmus. Herbicide interactions were assessed by the two-way ANOVA and Abbott's formula. Generally, an antagonistic interaction with Lemna was revealed by MCPA and chloridazon, whereas additive effect of both herbicides was observed for Desmodesmus. A significant stimulation of SOD and APX activity by binary mixtures was noted in algal cells mainly after 24 and 48 hours of exposure. The extremely high stimulation of the activity of both enzymes was induced by the combination EC??CHD + EC??MCPA (48 h). Presumably due to oxidative stress, the treatment with CHD at concentration EC?? after 72 h was lethal for algae grown in aerated cultures, in contrast to standardized test conditions. Taking into account the consequences of risk assessment for herbicide mixtures we can state that a relatively low toxicity, as well as the lack of significant synergy between MCPA and CHD to non-target plants appears to be the most important result.     

Assessment of silver nanoparticle-induced physiological and molecular changes in Arabidopsis thaliana

In this study, the effect of silver nanoparticles and silver ions on Arabidopsis thaliana was investigated at physiological and molecular levels. The seedlings were grown in sublethal concentrations of silver nanoparticles and silver ions (0.2, 0.5, and 1 mg/L) in 1/4 Hoagland’s medium for 14 days under submerged hydroponic conditions. Significantly higher reduction in the total chlorophyll and increase in anthocyanin content were observed after exposure to 0.5 and 1 mg/L silver nanoparticles as compared to similar concentrations of silver ions. Lipid peroxidation increased significantly after exposure to 0.2, 0.5, and 1 mg/L of silver nanoparticles and 0.5 and 1 mg/L of silver ions. Qualitative analysis with dichloro-dihydro-fluorescein diacetate and rhodamine 123 fluorescence showed a dose-dependent increase in reactive oxygen species production and changes in mitochondrial membrane potential in the roots of seedlings exposed to different concentrations of silver nanoparticles. Real-time PCR analysis showed significant upregulation in the expression of sulfur assimilation, glutathione biosynthesis, glutathione S-transferase, and glutathione reductase genes upon exposure to silver nanoparticles as compared with silver ions. Overall, based on the physiological and molecular level responses, it was observed that exposure to silver nanoparticles exerted more toxic response than silver ions in A. thaliana.     

Removal of biocide pentachlorophenol in water system by the spent mushroom compost of Pleurotus pulmonarius

Pentachlorophenol (PCP) has been widely used as a wood preservative since 1980s. Although it has been banned worldwide, residues of PCP are still commonly found. The spent compost of oyster mushroom Pleurotus pulmonarius (SMC) which was a degraded paddy straw-based substrate, contained 25% chitin. Five percentage of the SMC could remove 89.0 +/- 0.4% of 100 mg PCPl(-1) within 2 days at room temperature predominantly by biodegradation. The maximum removal capacity was 15.5 +/- 1.0 mg g(-1) SMC. The sorption kinetics of PCP by SMC can be described by the Freundlich monolayer model with a theoretical sorption capacity similar to that found for chitin. A PCP-degradative bacterium was isolated from the SMC. Yet, biodegradation was predominantly contributed by the immobilized ligninolytic enzymes secreted by the mushroom to the SMC. Degradation of PCP involves dechlorination, methylation, carboxylation and ring cleavage as verified by GC-MSD and ion chromatography. Thus, the SMC has a potential for treating PCP-contaminated water.     

Two-phase partitioning bioreactor for the biodegradation of high concentrations of pentachlorophenol using Sphingobium chlorophenolicum DSM 8671

An organic-aqueous two-liquid-phase partitioning system was developed to degrade high concentrations of pentachlorophenol (PCP). Dioctyl sebacate was selected among 12 non-aqueous phases as the most suitable solvent to control the delivery of PCP to the aqueous phase for being non-biodegradable and biocompatible. In shake-flask experiments, the two-phase system was able to support the removal of 1g PCP l(-1) of total liquid phase. The performance of the two-liquid phase partitioning system (TLPPS) in shake-flask was evaluated under different conditions. At the initial biomass concentrations of 7, 25, and 58 mg dry weight l(-1), the volumetric removal rates of PCP obtained were 25.7+/-0.5, 32.1+/-0.1, and 39.3+/-2.9 mg PCP l(-1)h(-1), respectively. Higher performance was observed at lower organic-aqueous phase ratios (16% and 28%) than higher ones (37% and 44%). In a 2-l TLPPS, the degradation of 10 g PCP was completed in less than 100 h at a total volumetric rate of 142 mg l(-1) h(-1). Kinetics study using Monod model showed that compared to monophasic systems, the biphasic system significantly enhanced the maximum specific growth rate and PCP removal rate. Results of this biphasic system showed no accumulation of unknown by-product(s) which has been reported for physical-pretreatment or high-performance biphasic systems of PCP degradation.     

Biodegradation of low aqueous concentration pentachlorophenol (PCP) contaminated groundwater

Bioremedial treatment to remove low level organic contamination to regulatory standards has met with limited success. In this study source water from a contaminated surficial aquifer at a former wood treatment facility was used to evaluate the potential for indigenous microorganisms to degrade low level (< 1.0 mg) pentachlorophenol (PCP) to a regulatory drinking water standard of 0.001 mg/L. PCP degradation was evaluated in series of batch reactors in a two phase study to (a) determine the rate and extent of PCP removal and (b) evaluate the impact of nutrient amendment (N and P) on removal rate. All reactors with the exception of the abiotic control demonstrated PCP removal to a level < 0.002 mg/L within a maximum period of 32 d with and without nutrient amendment. A regression analysis of reactive phosphate (ortho-P) concentration versus removal rate produced an R2 of 0.94 (p = 0.006) indicating a significant correlation between the level of available phosphate and PCP degradation rate. Selective bacterial enumeration (for PCP degrading bacteria) revealed PCP-degrading bacteria increased in abundance prior to and in conjunction with the degradation phase to a density of between 10(3) to 10(4) CFU/ml. Isolates were also analyzed for total fatty acids using Fatty Acid Methyl Ester (FAME) methodology and the results indicated that PCP degrading bacteria were present in the aquifer and consisted of predominately fluorescent, oxidase positive Pseudomonas species. Overall, data indicate that autochthonous microbes are capable of removing low level PCP (< 1.0 mg/L) to approach if not reach the regulatory standard of 0.001 mg/L with the addition of oxygen, with or without nutrient amendment. Results of this research can be applied to full-scale implementation of in-situ or ex-situ bioremediation of groundwater at former wood treatment facilities.