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PubMed2001

In situ polychlorophenol bioremediation potential of the indigenous bacterial community of boreal groundwater.

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.

Bacteria↗
PubMed2002

Desorption kinetics of PCP-contaminated soil: effect of temperature.

A thermally enhanced pump-and-treatment method for pentachlorophenol (PCP)-contaminated soil and groundwater has shown potential advantages over the traditional pump-and-treatment method. Studies on the desorption kinetics of PCP from aquifer soil are the first step toward quantitative assessment of the newly proposed method. Five series of desorption batch tests were conducted at different temperatures, and the test results were analyzed by linear regression to determine the best-fit kinetic model for PCP desorbed from the contaminated field soil under all temperature conditions. The first-order kinetic model, parabolic diffusion model, and modified Freundlich model were discussed in linear regression work. A modified Freundlich model was found to describe the PCP desorption kinetics from soil in consideration of a temperature effect.

Absorption↗
PubMed2002

Migration of dissolved heavy metal compounds and PCP in the presence of colloids through a heterogeneous calcareous gravel and a homogeneous quartz sand-pilot scale experiments.

In preparation to field experiments with in situ mobilized colloids, a set of pilot scale aquifer tank experiments was performed to gain an insight into the mass transfer of several heavy metal ions and pentachlorophenol (PCP) in a heterogeneous gravel aquifer and a homogeneous sandy aquifer. The experiments simulate a spill of dissolved contaminants and their subsequent transport with in situ mobilized colloids that are not yet in equilibrium with the contaminant. Contaminants with a weak tendency to adsorb to the stationary sediment matrix, e.g., PCP, establish a dynamic equilibrium with the mobile colloidal phase. The fraction of contaminants sorbed to colloids remains constant along the observed flow path. In contrast, for contaminants that strongly sorb to the sediment matrix, the fraction associated with colloids increases with increasing flow distance. In either case, the filtration of colloids is the limiting factor for the colloidal mass transfer. It is also shown that the recovery rates decrease rapidly within both sediments, and that the transport parameters (flow velocity, dispersivity) are only representative for the mobile fraction of the contaminant. On the given experimental scale, there was no evidence of a preferred colloidal transport, or even a transport of colloids ahead of the conservative tracer.

Adsorption↗
PubMed2003

Toxicokinetics, toxicity and lethal body residues of two chlorophenols in the oligochaete worm, Lumbriculus variegatus, in different sediments.

Bioavailability, toxicokinetics and toxicity (LC(50)) of water- and sediment-associated 2,4,5-trichlorophenol (2,4,5-TCP) and pentachlorophenol (PCP) were measured in Lumbriculus variegatus Müller in a set of experiments. The critical body residue approach was applied by measuring also the lethal body residues (LBR(50)). Freshwater and three different sediments with various sediment organic carbon (SOC) concentrations were used as exposure media. SOC decreased the bioavailability of both chlorophenols, and the uptake rates decreased by 81% and 91% for 2,4,5-TCP and PCP, respectively, in the sediment with a SOC of 6.9% compared to those in sediment with a SOC of 0.5%. SOC appeared to be an important factor controlling the bioavailability as after the carbon normalisation the difference between the sediments was much smaller. The 96-h LC(50) values for instance for PCP were 145.3 microg/l in freshwater, and 6.8 and 8.1 microg/g dry weight in sediments with SOC concentrations of 0.5% and 2.4%, respectively. The LBR(50) values, were practically the same in freshwater and sediments: between 1.0 and 1.6 and from 0.4 to 0.9 micromol/g wet weight for 2,4,5-TCP and PCP, respectively, demonstrating the usefulness of this method for accurate, and more comparable, measurement of toxicity of chemicals with the same mode of toxic action in varying conditions. L. variegatus expressed a dose-response sediment avoidance behaviour but the PCP tissue concentrations were not affected by this behaviour.

Animals↗
PubMed2003

An episode of dioxin contamination in feedingstuff: the choline chloride case.

In 2000, as part of a survey programme, the German authorities detected high levels of dioxins in a choline chloride (CC) premix used as animal food component. The contaminated additive consisted of different products of mineral and vegetable origin acting as a carrier. The CC was manufactured in Belgium. The final product was produced in and distributed from a plant in Spain. The German authorities informed all European Community members of the incident. The Spanish Ministry of Agriculture, Fisheries and Food and the Regional Authorities immediately conducted a survey in collaboration with the Spanish Council for Scientific Research (CSIC) to determine and isolate the source of the contamination. Analysis of a large number of samples of pure CC, pine sawdust, almond shell and other substances currently used in the preparation of the premix confirmed the presence of a significant amount of polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans in mixtures in which pine sawdust was present. An analysis of the congener profiles revealed similarities with those found in technical pentachlorophenol (PCP) formulations. The conclusion was that PCP-contaminated sawdust as carrier for CC was source of dioxin contamination in feedingstuff.

Animal Feed↗
PubMed2003

Biodegradation capability of Absidia fusca Linnemann towards environmental pollutants.

The purpose of this work was to study the bioremediation capability of Absidia fusca Linnemann (Zygomycete) towards different classes of xenobiotics (lignin-derived compounds, chloroaromatic compounds, polycyclic aromatic hydrocarbons) the presence of which in contaminated soils, water and sediments poses a significant risk to the environment and human health. Two strains from different origins were compared. One was from an official collection and grown in non-inducing conditions, while the other was isolated during the course of the survey of fungal flora in a polluted soil from Annaba (Algeria). All data were analyzed and results validated via a statistical treatment. We showed the effect of the factors studied (origin of the strain, xenobiotic) but also the interactions between these factors. The strain of A. fusca isolated from a polluted soil was able to efficiently degrade most of the xenobiotics tested, particularly: pentachlorophenol, phenol, catechol, guaiacol and ferulic acid. This property also existed in the other strain but at a very low level.

Absidia↗
PubMed2000

Can adsorption isotherms predict sediment bioavailability?

The adsorption and desorption of 2,4-dichlorophenol (DCP) and pentachlorophenol (PCP) were studied for a range of synthetic particles, a dimethylditallowammonium exchanged clay and a natural sediment. The synthetic particles were Dowex 1X8400, Toyopearl Phenyl 650M and Toyopearl SP 650M. The bioaccumulation of the DCP and PCP from these particles was then studied using the oligochaete, Lumbriculus variegatus. There is a correlation between contaminant-particle interactions, as determined from adsorption and desorption isotherms, and bioaccumulation. Bioaccumulation by L. variegatus was found to be highest from the systems where differences in the classification of adsorption and desorption isotherms were observed.

Adsorption↗
PubMed2002

Investigation of the potential release of polychlorinated dioxins and furans from PCP-treated utility poles.

The United States (US) Environmental Protection Agency (EPA) estimated that the use of technical grade pentachlorophenol (PCP) between 1970 and 1995 to treat wood was approximately 400,000 metric tons in the US, and that between 4800 and 36,000 g of 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) were incorporated annually in treated wood. The EPA has been unable, however, to estimate the rate of release of polychlorinated dibenzo-p-dioxins and dibenzofurans (CDD/Fs) from treated utility poles into the environment. There is some evidence that CDD/Fs leach from treated poles into the surrounding soils, but these studies do not allow for the calculation of a rate of release from this mechanism. Another possible release mechanism is the volatilization of dioxins into the atmosphere, but there are no data to demonstrate, much less quantify, this release. While not directly measuring the release of dioxins from treated utility poles into the environment, this study was designed to examine the potential for such release. The general approach taken was to collect PCP-treated poles of varying ages, to remove and analyze multiple samples from each pole cross-section, and to compare the spatial distribution of CDD/F congeners among poles of different ages. Evidence of concentration-depth profile changes over time may provide insight into the potential for dioxins to migrate through and then out of PCP-treated utility poles. It was found that the CDD/F concentrations were consistently higher in the outer portions of the poles than the center. This trend tends to be most marked in older poles and for the lower chlorinated congeners. The trend for dioxins to concentrate in the outer portions of the pole over time suggest migration within the poles, and this migration may result in some environmental release. Other possible explanations were also offered.

Benzofurans↗
PubMed1999

PCP in the freshwater and marine environment of the European Union.

This study collates monitoring data principally from 1991 to 1996, relating levels of pentachlorophenol (PCP) in the freshwater and marine environments of the European Union (EU) Member States of Belgium, France, Germany, the Netherlands and the UK. In general, PCP levels in the open freshwater and marine environments displayed a downward trend in these countries, and where increased PCP concentrations were observed, these were tentatively linked to specific point discharges. The monitoring data were compared against the relevant predicted no-effect concentration (PNEC) for PCP in a particular environmental medium. On this basis, the study suggests that PCP does not pose a risk to the coastal, estuarine, marine waters or marine sediments of the above countries, and that any risks posed by PCP to the freshwater environment can largely be attributed to sediments contaminated with PCP from historic usage, or due to the continued use of small quantities of sodium pentachlorophenyl laurate in northern France. Further analysis of the data was not possible owing to the lack of systematic reporting by Member States, the lack of information on usage quantities and patterns, and inconsistent environmental surveillance and reporting of environmental data. This is particularly relevant for south-west France, Portugal and north-east Spain, the region which accounts for almost 90% of the EU's consumption of NaPCP. These issues need to be addressed at a pan-European level to better inform the selection of risk management measures and to improve the effectiveness of such measures.

Animals↗
PubMed2003

Evaluation of genotoxicity of PCP and 2,4-D by micronucleus test in freshwater fish Channa punctatus.

An in vivo study on the genotoxic effects of pentachlorophenol (PCP) and 2,4-dichlorophenoxyacetic acid (2,4-D) was carried out with freshwater air-breathing fish Channa punctatus. The fish were exposed to three sublethal doses of PCP (0.2, 0.4, and 0.6 ppm) and 2,4-D (25, 50, and 75 ppm) by medium treatment. Micronucleated erythrocytes were sampled at intervals of 48, 72, and 96 h post-treatment. Student's t test revealed significant increases in micronucleus (MN) frequency. Maximum incidence of MN was recorded at highest concentrations and durations for both chemicals. A time- and dose-dependent response of the MN frequency for both chemicals was confirmed. PCP-treated groups had 8-10 MN per cell, specifically at higher concentrations and duration. Echinocytes were recorded at higher concentrations of 2,4-D. The study confirmed the genotoxicity of PCP and 2,4-D in this organism. PCP was found to be more toxic than 2,4-D in terms of MN induction. This assay has further potential for detecting genotoxic compounds in aquatic ecosystems.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1996

The possible role of acetyltransferase in the induction of cytogenetic effects by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in cultured Chinese hamster cells.

When metabolically activated, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a heterocyclic amine isolated from cooked food, is clastogenic in cultured Chinese hamster and human cells. Secondary metabolites of PhIP are formed via acetyltransferase (AT) and sulfotransferase (ST) activity; however, which is responsible for its clastogenic effect is unknown. We addressed this question. We used a parental Chinese hamster lung cell line and three sublines transfected with different AT genes to test the clastogenic (i.e., micronucleus-inducing) effects of metabolically activated PhIP and 7,12-dimethylbenz[a]anthracene (DMBA) in the presence and absence of pentachlorophenol (PCP), a ST inhibitor. PhIP was significantly more clastogenic in the three AT-enriched sublines than in the parental line (p < 0.001). DMBA (a ST-activated mutagen), on the other hand, equally induced MNs in all the cell lines. When PCP was added to the test system, the MN-induction ability of DMBA, but not of PhIP, decreased significantly (p < 0.001). These findings strongly suggest that PhIP clastogenicity is due to AT activity and not to ST activity.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed2001

The effects of fluid motion on toxicant sensitivity of the rotifer Brachionus calyciflorus.

Standardized methods for estimating the toxicity of anthropogenic compounds to aquatic organisms frequently fail to consider key elements of the test organisms' environment. Aquatic organisms exist in a fluid environment, and fluid dynamics may have an important influence on the response to toxicants. Rotifers are one of the three major groups of zooplankton and have been increasingly utilized in standardized toxicity testing. However, like other toxicity tests, assays with the species Brachionus calyciflorus are performed under static conditions in the absence of fluid motion. We investigated how fluid motion modifies pentachlorophenol (PCP) toxicity to B. calyciflorus using 24 h acute and 48 h reproductive toxicity tests. Estimates of PCP LC50s and reproduction EC50s in static conditions decreased from 738 and 1082 microg l(-1), respectively, to as low as 262 and 136 microg l(-1), respectively, in fluid motion. Flow analysis indicated that increased toxicant sensitivity can occur at ecologically relevant levels of fluid motion. Mechanistic studies indicated that fluid motion/toxicant interactions may result from the ability of fluid motion to cause shear stress, alter toxicant uptake, and/or alter the bioavailability of food. As fluid motion may have an important effect on the life histories of a wide variety of aquatic organisms, fluid motion/toxicant interactions may be an important consideration in other standard test organisms. These results raise questions about the accuracy of ecological risk assessments based on toxicity data from static conditions.

Animals↗
PubMed2002

Evidence of differences in the biotransformation of organic contaminants in three species of freshwater invertebrates.

Acute static bioassays were performed using three freshwater invertebrate species (the oligochaete Lumbriculus variegatus, the fingernail clam Sphaerium corneum and the larvae Chironomus riparius) exposed separately to a variety of 14C radiolabelled contaminants. The aim of this work was to investigate if the chemicals remained as parent compounds after the treatments. Chemicals used were 2,4-dichlorophenol; 2,4,5-trichlorophenol; pentachlorophenol; pyrene; Fenpropidin, and Trifluralin. Homogenates of the whole body tissue of each organism were prepared and total radioactivity was measured. Contaminants were then extracted into organic solvents and analysed by high-pressure liquid chromatography techniques. Chromatograms showed that most of the substances extracted were present as parent compounds in S. corneum and in L. variegatus. In contrast, for C. riparius a low proportion of the chemicals was recovered as parent compounds. These results suggest that different metabolic processes could take place in the different species.

Animals↗
PubMedDate not supplied

Influence of body size and developmental stage of coho salmon (Oncorhynchus kisutch) on lethality of several toxicants.

Acute 96 h bioassays were conducted on coho salmon (Oncorhynchus kisutch) to determine their sensitivity to each of four toxicants during development. Alevins, swim-up fry and juveniles (up to 175 days post-hatch) were exposed to lethal concentrations of copper, zinc, nickel, or pentachlorophenol each week to monitor changes in susceptibility as the fish grew. Fish exhibited a high degree of susceptibility to all of the toxicants during the alevin stage. Swim-up fry showed drastic changes in sensitivity followed by more sustained patterns of change in the subsequent juvenile stage. Lethality could be related to both concentration of toxicant (C) and some exponential function of body weight (W) by the use of the multiple regression equation, Y = a + b(lnC) + g(lnW) where Y is the percent mortality expressed in probits. Through the use of this weighted expression of toxicity it was observed that coho alevins were consistently more sensitive to the toxicants than the juveniles. An exponential relationship between toxicity and weight could be calculated as -g/b. This exponent was found to be specific for each toxicant tested and changed in magnitude as the fish progressed through the various stages of development. Results indicated that lethality (LC50's) varied from one stage of development to the next, but within any defined stage, the relationship between toxicity and body weight was relatively constant.

Animals↗
PubMed2001

Proliferation and differentiation of murine haemopoietic progenitor cells in stroma-free culture in the presence of metabolites of chlorinated pesticides.

We have studied the influence of metabolites of chlorinated pesticides (lindane, pentachlorophenol, hexachlorobenzene) on proliferation and differentiation in two stroma-free murine bone marrow culture models, a multipotent progenitor cell line (FDCP-mix) and primary lineage-depleted bone marrow cells. Tetrachlorohydroquinone (Cl(4)pHQ), tetrachloro-p-benzoquinone (Cl(4p)BQ), but not their positional isomers, tetrachlorocatechol (Cl(4)oHQ) and tetrachloro-o-benzoquinone (Cl(4)oBQ), nor 2,4,6-trichlorophenol (2,4,6-Cl(3)P), were much more toxic to FDCP-mix cells cultured under conditions which lead to self-renewal than under conditions which lead to granulocyte-macrophage differentiation. Under the latter conditions, Cl(4)pHQ and Cl(4p)BQ even stimulated growth at intermediate concentration levels. In the primary cell cultures, pronounced differences were observed in the sensitivity between individual developmental pathways and between the different compounds. The percent of cells differentiating into the granulocytic lineage was increased at high concentration levels of each test compound. However, stimulatory effects on the macrophage lineage were observed at intermediate concentration levels of Cl(4)pHQ, Cl(4p)BQ and 2,4,6-Cl(3)P, and differentiation into erythrocytes was stimulated at low concentrations of 2,4,6-Cl(3)P. It is concluded that chlorinated monocyclic pesticides, after biotransformation to quinoid metabolites, may interact directly with haemopoietic progenitor cells with differential effects on self-renewal and differentiation. These mechanisms could lead to myeloplastic disorders.

Animals↗
PubMed2000

Protection by desferrioxamine and other hydroxamic acids against tetrachlorohydroquinone-induced cyto- and genotoxicity in human fibroblasts.

Tetrachlorohydroquinone (TCHQ) has been identified as a major toxic metabolite of the widely used wood preservative pentachlorophenol and has also been implicated in its genotoxicity. We have recently demonstrated that protection by the trihydroxamate iron chelator desferrioxamine (DFO) on TCHQ-induced single-strand breaks in isolated DNA was not the result of its chelation of iron but rather of its efficient scavenging of the reactive tetrachlorosemiquinone (TCSQ) radical. In this study, we extended our research from isolated DNA to human fibroblasts. We found that DFO provided marked protection against both the cyto- and genotoxicity induced by TCHQ in human fibroblasts when it was incubated simultaneously with TCHQ. Pretreatment of the cells with DFO followed by washing also provided marked protection, although less efficiently compared with the simultaneous treatment. Similar patterns of protection were also observed for three other hydroxamic acids (HAs): aceto-, benzo-, and salicylhydroxamic acid. Dimethyl sulfoxide, an efficient hydroxyl radical scavenger, provided only partial protection even at high concentrations. In vitro studies showed that the HAs tested effectively scavenged the reactive TCSQ radical and enhanced the formation of the less reactive and less toxic 2,5-dichloro-3, 6-dihydroxy-1,4-benzoquinone (chloranilic acid). The results of this study demonstrated that the protection provided by DFO and other HAs against TCHQ-induced cyto- and genotoxicity in human fibroblasts is mainly through scavenging of the observed reactive TCSQ radical and not through prevention of the Fenton reaction by the binding of iron in a redox-inactive form.

Cell Line↗
PubMed2001

The non-specific inhibition of enzymes by environmental pollutants: a study of a model system towards the development of electrochemical biosensor arrays.

Previous research has shown that lactate dehydrogenase (LDH) was competitively inhibited by pentachlorophenol (PCP) and a modified assay produced a detection limit of 1 microM (270 microg l(-1)). This work used spectrophotometric rate-determination but in order to move towards biosensor development the selected detection method was electrochemical. The linkage of LDH to lactate oxidase (LOD) provided the electroactive species, hydrogen peroxide. This could be monitored using a screen-printed carbon electrode (SPCE) incorporating the mediator, cobalt phthalocyanine, at a potential of +300 mV (vs. Ag/AgCl). A linked LDH/LOD system was optimised with respect to inhibition by PCP. It was found that the SPCE support material, PVC, acted to reduce inhibition, possibly by combining with PCP. A cellulose acetate membrane removed this effect. Inhibition of the system was greatest at enzyme activities of 5 U ml(-1) LDH and 0.8 U ml(-1) LOD in reactions containing 246 microM pyruvate and 7.5 microM NADPH. PCP detection limits were an EC(10) of 800 nM (213 microg l(-1)) and a minimum inhibition detectable (MID) limit of 650 nM (173 microg l(-1)). The inclusion of a third enzyme, glucose dehydrogenase (GDH), provided cofactor recycling to enable low concentrations of NADPH to be incorporated within the assay. NADPH was reduced from 7.5 to 2 microM. PCP detection limits were obtained for an assay containing 5 U ml(-1) LDH, 0.8 U ml(-1) LOD and 0.1 U ml(-1) GDH with 246 microM pyruvate, 400 mM glucose and 2 microM NADPH. The EC(10) limit was 150 nM (39.9 microg l(-1)) and the MID was 100 nM (26.6 microg l(-1)). The design of the inhibition assays discussed has significance as a model for other enzymes and moves forward the possibility of an electrochemical biosensor array for pollution monitoring.

Biosensing Techniques↗
PubMed2001

Detection of DNA damage in haemocytes of zebra mussel using comet assay.

The aim of the study was to use the comet assay on haemocytes of freshwater mussel, Dreissena polymorpha Pallas, for detection of possible DNA damage after exposure to pentachlorophenol (PCP) and to evaluate the potential application of the comet assay on mussel haemocytes for genotoxicity monitoring of freshwater environment. Zebra mussels were exposed for seven days to different concentrations (10, 80, 100, 150 microg/l) of PCP and in the river Sava downstream from Zagreb municipal wastewater outlet. Significant increase in DNA damage was observed after exposure to PCP at doses of 80 microg/l and higher and after in situ exposure in the river Sava as well. This study confirmed that the comet assay applied on zebra mussel haemocytes may be a useful tool in determining the potential genotoxicity of water pollutants.

Animals↗