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Myeloperoxidase-catalyzed formation of PCDD/F from chlorophenols.

Chlorophenols (CP) are transformed in vitro to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) by a biochemical-catalyzed oxidation. This is shown for 2,4,5-tri-, 2,3,4,6-tetra-and pentachlorophenol with myeloperoxidase recovered from human leucocytes in the presence of hydrogen peroxide. The yield, the reaction, and the PCDD/F-pattern found depend on the CP. The formation rates are in the micromol-per-mol range for all substrates. The experiments confirm the suspicion that a biochemical formation of PCDD/F from precursors such as CPs can take place in the human body and that this metabolic pathway may lead to a higher inner exposure with PCDD/F than is now assumed.

Benzofurans↗

Simulation of long-term environmental dynamics of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans using the dynamic multimedia environmental fate model and its implication to the time trend analysis of dioxins.

Long term environmental fate of polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) were studied using the fugacity-based dynamic multimedia environmental fate model. New formulation about soil wind erosion into air was included into the model. Effect of process assumption, sensitivity analysis for parameters, and tentative validation against the measured sediment core analysis was performed. Mass fluxes between compartments were estimated by the dynamic modeling. From the model estimates, major mass fluxes coming from emission source were shown to go to the soil and water through wet/dry deposition, then go to degradation mainly in the soil and sediment. Major mass fluxes of TCDD and OCDD come from the impurities in CNP (Chlornitrofen) and PCP (Pentachlorophenol) directly into the soil. Consideration about multimedia environmental dynamics using the modeled mass fluxes was shown in the discussion.

Benzofurans↗

Steps towards a European dioxin emission inventory.

The results of a project aiming at collection and evaluation of information regarding the industrial and non-industrial emission sources for dioxins and furans (PCDD/PCDF) in 17 European Countries (EU 15, CH, N) are presented. An overview about national documents covering dioxin emission inventories for the period 1990-1995 is given. Some data on emissions associated with residual materials, waste and waste water are presented additionally. Based on the air emission data contained in these documents the most important emission sources were determined which are assumed to cover about 90% of the overall emissions. For the selected sources and for all 17 countries new estimates of the atmospheric PCDD/PCDF emissions were derived from average emission factors and statistical activity rates for the year 1994. As a result, on the European scale the largest annual PCDD/PCDF emission is assessed to be released from municipal waste incineration, quite closely followed by emissions from iron ore sintering. Considerable releases of dioxins and furans--based, however, on highly uncertain data--are further assessed for domestic burning, accidental fires and (former) use of contaminated wood preservatives (pentachlorophenol). A lower but still significant emission is further assigned to the sector of non-ferrous metal production; particular processes used in this branch proved to generate very high PCDD/PCDF flue gas concentrations.

Air Pollutants↗

Formation of hydroxylated and dimeric intermediates during oxidation of chlorinated phenols in aqueous solution.

The oxidation of selected chlorophenols (2-chlorophenol, 4-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, 2,3,4,6-tetrachlorophenol and pentachlorophenol) was studied in aqueous solutions using UV/H2O2 and O3 methods. The formation of oxidation intermediates was measured to elucidate their importance in the treatment of chlorophenols. Results indicated that chlorophenols can be treated efficiently by the methods studied, but the dechlorination of the compounds was insufficient. Analysis of intermediates in the acetylated extraction fractions showed that hydroxylation of chlorophenols and formation of dimeric products were involved in the oxidation of chlorophenols in both treatment processes. The majority of the intermediates detected were transient, and thus were not detectable after an extended treatment time. The presence of a complicated mixture of intermediates suggests the need for toxicity testing to confirm the detoxifying effect of the chemical oxidation of chlorophenols.

Chlorophenols↗

Secondary exposure to dioxins through exposure to PCP and its derivatives.

This study discusses the potential exposure of occupational workers and general consumers in the European Union (EU) to polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) caused by exposure to pentachlorophenol (PCP) and its derivatives. A synthesis of reported exposure data relating exposure to PCP and its derivatives in an occupational setting has indicated that the PCDD/F intake for exposure in the 1990s averaged over a lifetime is likely to be in the order of 0.02-1 pg I-TEQ kg-1 bw day-1 with actual exposures more likely to occur at the lower end of the range, around the median of 0.16 pg I-TEQ kg-1 bw day-1. Workers who have experienced past exposure to PCP and its derivatives especially in the early to mid 1980s, will have been subjected to higher exposures to PCDD/Fs due to the generally higher concentrations of PCDD/Fs in PCP products at that time. Exposure to PCP and its derivatives via the food chain is judged to be the most significant intake route of PCDD/Fs into consumers. The ingestion of milk and dairy products obtained from cows grazed on pasture dressed with sewage sludge has the potential to raise the average daily intake of PCDD/Fs via the diet by about 40% if all foods consumed derived from sludge amended soil. To the extent quantifiable, exposure to PCP in an occupational setting contributed approximately 16% as a median to the overall background exposure to PCDD/Fs, while for consumers this contribution ranged from 2 to 60% depending on the exposure to sludge amended foods.

Adolescent↗

Examination of an amphibian-based assay using the larvae of Xenopus laevis and Ambystoma mexicanum.

Semistatic acute toxicity tests of amphibian larvae (Xenopus laevis and Ambystoma mexicanum) were conducted at different developmental stages and by different methods to establish a simple amphibian-based assay. Test substance was pentachlorophenol sodium salt (PCP-Na). The endpoint was mortality and the 24-, 48-, 72-, and 96-h LC50 values were calculated by probit analysis. Interspecific differences in larval responses were not clear. Larval sensitivity tended to increase with larval age. Newly hatched larvae were most resistant to PCP-Na. During the tests of well-developed larvae, concentrations of dissolved oxygen and PCP-Na in the test solutions greatly dropped owing to uptake by the larvae. Therefore, middle-developed (2-week-old) larvae were most suitable for the test. Toxicity tests for volatile substances would be also possible using 2-week-old larvae in closed vessels. Test individuals should be kept individually to avoid the effects of poisonous skin secretions released from dead larvae.

Ambystoma mexicanum↗

Activation of tinidazole, an antiprotozoal drug to a mutagen by mammalian liver S9.

Tinidazole was found to display much higher mutagenic activity compared to metronidazole in Salmonella strain TA100 and YG1029. Under anaerobiosis, the specific activity of this nitroimidazole was enhanced, at least, by about 1.75-fold in TA100 and several fold in TA100NR relative to aerobic conditions. The mutagenicity in the latter strain with S9 mix became further increased by 2.5-fold under anaerobiosis, indicating the role of oxygen sensitive bacterial and mammalian S9 nitroreductases in the activation of the drug. The mutagenicity of the drug was slightly lowered in TA100/1,8-DNP6 (O-acetyltransferase deficient), YG1029 (O-acetyltransferase overexpressing) and TA100 in the presence of pentachlorophenol (PCP), an O-acetyltransferase inhibitor. These results rule out the possible involvement of N-acetoxyarylamine pathway in the metabolic activation of these nitroimidazoles.

Animals↗

Direct and indirect effects of sublethal toxicant exposure on population dynamics of freshwater rotifers: a modeling approach.

Ecotoxicological assessments commonly consider the direct effects of anthropogenic toxicants on the survival, growth, and reproduction of organisms. However, toxicants also may have indirect effects on aquatic communities by altering species interactions such as predation and competition. Such indirect effects are often mediated by changes in animal behavior, and contribute to uncertainty in predicting the environmental effects of toxicants. In an attempt to better understand indirect effects, a modeling approach was used to simulate the population dynamics of the predatory rotifer Asplanchna girodi and six prey species of herbivorous rotifers in response to sublethal concentrations (110, 190 or 330 mg/l) of pentachlorophenol (PCP). Simulations were performed assuming three different toxicant effect scenarios: behavioral toxicity alone, reproductive toxicity alone, or both behavioral and reproductive toxicity. Population growth rates (G(r)), maximum population densities (D(max)), and the percent of carrying capacity at D(max) (%K) were calculated for A. girodi and each prey species for each effect scenario and toxicant concentration. Similar qualitative results were obtained regardless of which prey species was exploited by A. girodi. Toxicant effects on predator and prey population densities appeared to be the dominant cause of both direct and indirect toxicant effects, and behavioral toxicity was a redundant endpoint relative to reproductive toxicity. These results suggest that assessing toxicant effects on the population growth of individual species may provide sufficient data to predict toxicant effects on species interactions as well.

Animals↗

Heat shock protein Hsp70 expression and DNA damage in Baikalian sponges exposed to model pollutants and wastewater from Baikalsk Pulp and Paper Plant.

Lake Baikal, a unique habitat for a great number of endemic species, is the largest freshwater reservoir in the world which is still largely unaffected by anthropogenic pollution, except for some shore regions with industrial activity. The expressions of a biomarker of exposure (heat shock protein Hsp70) and a biomarker of effect (DNA single-strand breaks) were measured for the first time in endemic Baikalian sponge species (Baikalospongia intermedia, Lubomirskia fusifera, and Lubomirskia abietina). Tissue cubes of B. intermedia and dissociated cells of L. fusifera and L. abietina reacted to temperature stress (10-16 degrees C above ambient temperature) with a time-dependent increase in expression of Hsp70. In B. intermedia, the effects of model pollutants (lead, copper, and zinc, and the organochlorines tetrachloroguaiacol, TCG, and pentachlorophenol, PCP) and of the wastewater from the final refinement and aeration reservoirs of the Baikalsk Pulp and Paper Plant (BPPP), located at the shore of the southern basin of Lake Baikal, on the expression of Hsp70 and the extent of DNA damage were investigated. It was found that lead and zinc but not copper cause a strong induction of Hsp70 in this sponge, while the frequency of DNA single-strand breaks increased after exposure to all these heavy metals tested. Induction of DNA single-strand breaks was also observed after exposure to TCG and PCP, but these compounds did not (consistently) enhance Hsp70 expression. Wastewater taken from the final water aeration pond of BPPP caused a concentration-dependent increase in Hsp70 expression in B. intermedia. However, there was no difference in the basal levels of Hsp70 between sponges collected in the shallow water at an unpolluted site near Baikalsk City and at a polluted site where the wastewaters of BPPP are discharged into the lake. There was also no clear difference in the wastewater concentration-dependent induction of Hsp70 expression between sponges collected at these sites, indicating no adaptation to continuous stress exposure.

Animals↗

Induction of the multixenobiotic defense mechanism (MXR), P-glycoprotein, in the mussel Mytilus californianus as a general cellular response to environmental stresses.

A multixenobiotic resistance mechanism (MXR) related to the P-glycoprotein multidrug transporter protein (p-gp) has been identified and characterized in several marine invertebrates. p-gp activity and protein titer is induced by exposure to toxins, supporting the suggestion that the role for this transporter is protection from xenobiotics by reducing accumulation of toxins in cells. In this study, we report on the specificity of the induction of the transporter by various chemical and physical stressors. p-gp substrates (including the pesticides pentachlorophenol and chlorthal) as well as non-substrates (including DDE and sodium arsenite) induced p-gp activity and protein titer in the gill tissues of the mussel Mytilus californianus. Similarly, mussels exposed to heat shock of 20 degrees C or 25 degrees C exhibited increased p-gp titer and activity compared to mussels held at ambient (12 degrees C) temperature seawater. Some of the same treatments that induced an increase in p-gp caused a concomitant increase in hsp70, but hsp induction was not always associated with induction of the p-gp protein. These findings suggest that p-gp induction in mussels may be part of a general cellular stress response. This response, however, does not appear to be always coupled with the hsp70 response in mussels.

Journal Article↗

Suppression of superoxide production by chlorothalonil in striped bass (Morone saxatilus) macrophages: the role of cellular sulfhydryls and oxidative stress.

Chlorothalonil (TCIN) is the most commonly applied fungicide in the USA, with substantial use in the Chesapeake Bay area. Little is known about the sublethal toxicity of TCIN to fish, but since it is structurally similar to the immunotoxicant pentachlorophenol, the potential for immunomodulation exists. Previous studies have indicated that in vitro exposure of macrophages to TCIN modulates immunostimulated reactive oxygen species (H(2)O(2)/hypochlorous acid) and NADPH production in striped bass (Morone saxatilus). The goals of this study were to determine if TCIN inhibits superoxide (O(2)(-)) production by macrophage NADPH oxidase, to examine the role of cellular sulfhydryl groups in TCIN-induced macrophage dysfunction, and to identify the extent to which lipid peroxidation contributes to the observed toxic effects. The results of lucigenin-augmented chemiluminescence assays indicated that TCIN suppressed both baseline and stimulated O(2)(-) production in a dose-dependent manner. Similar results were obtained using both the particulate stimulant zymosan and the lipid-soluble stimulant phorbol 12-myristate-13-acetate. Inhibition of glutathione synthesis by pre-treatment with buthionine sulfoximine (BSO) enhanced the suppression of O(2)(-) production. The protection of sulfhydryl groups by culturing macrophages with dithiothreitol (DTT) reduced TCIN-induced macrophage dysfunction. TCIN did not initiate lipid peroxidation in macrophages, as measured by the thiobarbituric acid reactive substances (TBARS) assay, nor did pre-treatment with BSO potentiate lipid peroxidation. Because the observed TCIN-induced suppression of O(2)(-) was modulated by altering cellular sulfhydryl status with BSO and DTT, it is possible that toxicity results from the inhibition of NADPH oxidase activity by TCIN binding to its functional sulfhydryl groups.

Journal Article↗

Effect of pollutants on the ergosterol content as indicator of fungal biomass.

Ergosterol content was determined in 20 white-rot fungi isolates and the values ranged from 2380 to 13060 microg g(-1) fungal biomass. Significant changes of ergosterol content according the physiological stage for Bjerkandera adusta 4312 and Coriolopsis gallica 8260 were found, showing the highest values during the stationary phase. However, in the case of Phanerochaete chrysosporium 3642, no changes were detected during growth. The effect of pollutants, such as heavy metals and fungicides, on the ergosterol content of C. gallica was determined. Heavy metals (Cu 80 ppm, Zn 50 ppm or Cd 10 ppm) and fungicides (thiram 3 ppm or pentachlorophenol 1.5 ppm) at concentrations that reduce the metabolic activity between 18% and 53% (pollutant-stressed cultures) did not affect the ergosterol content. Only the fungicide zineb (25 ppm) reduced significantly the ergosterol content in biomass basis. In soil experiments with Cu (80 ppm) or thiram (10 ppm) after 15 and 30 days of incubation, the ergosterol content in soil was linearly correlated to the fungal biomass C in both polluted and control soil cultures. The ergosterol content was independent of the presence or the absence of pollutants. Thus, these results indicate that ergosterol can be a useful indicator for fungal biomass in polluted soils, and can be applied for monitoring bioremediation processes.

Biodegradation, Environmental↗

Full life-cycle toxicity assessment using rotifer resting egg production: implications for ecological risk assessment.

The majority of standardized toxicity tests incorporate only a fraction of the test organism's life-cycle. However, in natural ecosystems, organisms may be exposed at various times during their life-cycle or throughout their life-cycle. Thus, ecotoxicological data from standardized toxicity tests is of limited ecological relevance. Existing standardized toxicity tests using the freshwater rotifer Brachionus calyciflorus utilize 24-h survival or 48-h asexual reproduction as endpoints, despite evidence that sexual reproduction is more sensitive. A 96-h B. calyciflorus resting egg toxicity test was developed and used to estimate the toxicity of pentachlorophenol (PCP) and copper. Results were compared to a variety of acute and sublethal endpoints for both toxicants. The B. calyciflorus 96-h resting egg production NOEC for PCP of 10 micrograms/l was 20 times lower than the 48-h asexual reproduction no observed effect concentration (NOEC) and 120 times lower than the 24-h acute lethal concentration 50%. The 96-h resting egg production NOEC for copper of 2.8 micrograms/l was 7 times lower than the 48-h asexual reproduction NOEC and nine times lower than the 24-h acute LC50. Resting egg production was a more sensitive indicator of toxicity than several other sublethal endpoints as well. These results indicate that partial life-cycle toxicity tests are not sufficiently sensitive to detect ecologically relevant adverse effects.

Animals↗

Dose-dependent induction of heat shock protein 70 synthesis in Raphidocelis subcapitata following exposure to different classes of environmental pollutants.

In this study an enzyme-linked immunosorbent assay (ELISA) was developed to detect the stress protein Hsp70 in the green alga Raphidocelis subcapitata. Using this ELISA, the response to a variety of pollutants, including ZnCl2, SeO2 (heavy metals), lindane (organochlorine pesticide), pentachlorophenol (PCP, chlorinated hydrocarbon insecticide and fungicide), carbaryl (carbamate pesticide) and sodium dodecyl sulphate (SDS; surfactant) was tested. Our results show that Hsp 70 is produced in a dose-dependent way in response to most chemicals investigated (except PCP) and at concentrations below the range of classical cytotoxicity testing (i.e. growth inhibition, lethality). Still, the potential to induce Hsp70 varied among the pollutants tested, the heavy metals ZnCl2 and SeO2 being the strongest inducers of Hsp70. Combined with the existing literature, these results indicate that Hsp70 in R. subcapitata is a sensitive biomarker for a wide range of environmental pollutants.

Journal Article↗

Reproductive success and chlorinated hydrocarbon contamination in tree swallows (Tachycineta bicolor) nesting along rivers receiving pulp and paper mill effluent discharges.

The insectivorous tree swallow was chosen as an indicator species to investigate the uptake of pulp mill-related chlorinated hydrocarbons from emergent aquatic insects. Nest box populations were monitored for reproductive success at locations upstream and downstream of pulp mills on two river systems in British Columbia, Canada. Also, 16-day-old nestlings were collected and analysed for polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), pesticides, chlorophenols and chloroguaiacols. Most reproductive parameters were not different between populations at upstream and downstream locations, and were equivalent to or greater than those recorded for other tree swallow populations. Chlorinated hydrocarbon contamination of nestlings was low at all sites. The highest tissue concentrations were detected downstream of pulp mills on the Fraser River, where PCDD and PCDF patterns along with the presence of pentachlorophenol (PCP) suggested that the primary source of contaminants was past use of PCP for timber preservation. Although the absolute tissue concentration of contaminants was less, the toxic concentration (as estimated with I-TEQs) was greatest in nestlings downstream of a pulp mill on the Thompson River. The proportionately larger contributions from 2,3,7,8-TCDF and PCB-77 elevated these TEQs in comparison to other populations. Nest success was the one reproductive measure that showed substantial reductions in downstream populations on both rivers; however, there was little indication that nest failures were the direct result of contamination. Failures were largely due to parental abandonment, and, while poor parental attentiveness and nest abandonment have been associated with chlorinated hydrocarbon tissue concentrations in other studies, our 1-year assessment was insufficient to establish any link with pulp mill effluent exposure. Nestling growth models showed some subtle differences in growth patterns between nest box populations on the two rivers, but an association with pulp mill effects on aquatic insect prey availability was not established.

Journal Article↗

Degradation of organochlorine pesticides by meat starter in liquid media and fermented sausage.

The effect of meat starter on the degradation of DDT and lindane was investigated. The insignificant role of Lactobacillus plantarum in degrading p,p'-DDT and lindane presented in tryptone soya broth (TSB) and mineral salt medium (MSM) with or without 120 ppm nitrite was observed.The degradation of DDT and lindane by Micrococcus varians in TSB and MSM with or without nitrite were studied. The results indicated that DDT or lindane were degraded during the incubation period. The reduction in DDT at the end of the incubation period (15 days) was about 24.1 and 32.5% in TSB and MSM without nitrite, respectively. Corresponding values in the same media with nitrite were 37.5 and 46.4%. Regarding the reduction in lindane, it was recorded as 27.9 and 40.0% in TSB and MSM without nitrite, respectively and 38.4 and 48.4% in the same media with nitrite. The results indicated that culture media M. varians metabolized DDT mainly to DDD and lindane mainly to 2,4-, 2,5-, 2,6- and 3,4-dichlorophenol; 2,3,4- and 2,3,5-trichlorophenol; hexachlorobenzene; and pentachlorophenol. The effect of pesticides on the growth rate of meat starter was also investigated. The addition of DDT or lindane resulted in a slight decrease in counts of the strains during the initial incubation in TSB or MSM. Then the microorganisms recovered and began to grow logarithmically, but not as well as in a normal situation. The effect of fermentation stage by meat starter on DDT and lindane in fermented sausage was recorded. The results indicated that during the 72 h of fermentation, the reduction was 10 and 18% of DDT and lindane, respectively. These results confirmed that the fermentation process in meat products reduced pesticide residues and these reductions were due to the activity of meat starter.

Animal Feed↗

Impact of protein binding on the availability and cytotoxic potency of organochlorine pesticides and chlorophenols in vitro.

In vitro toxicity data are generally based on nominal concentrations and thus depend on both activity and availability of a compound. The aims of the present study were to examine the influence of protein binding on the cytotoxicity of selected organochlorine pesticides and chlorophenols in Balb/c 3T3 cell cultures and to determine parameters of protein binding which can be used to estimate protein bound fractions and to model distribution in vitro. EC(50)-values derived from concentration-effect relationships determined in the presence of various concentrations of bovine serum albumin (BSA) were linearly correlated to BSA concentration. Increasing the BSA concentration from about 1.2 to 40 mg/ml increased the EC(50)-values by factors between 3.4 and 34.4. Molar ratios of substance bound to albumin ranged from 0.11 to 2.42. Calculated fractions bound to albumin in the normal growth medium were 0.075-0.17 (p,p'-DDT, p,p'-DDE, dieldrin, lindane), 0.09-0.1 (4-mono- and 2,4-dichlorophenol), 0.68 (2,4,5-trichlorphenol) and almost 1.0 (pentachlorophenol). At 40 mg/ml BSA any compound was largely bound to albumin (fractions bound > or = 0.74). Distribution modelling revealed that the availability of the highly hydrophobic organochlorines additionally was significantly reduced by partitioning into lipids. The results clearly demonstrate that nominal and relative toxic potencies of organochlorine pesticides and chlorophenols determined in vitro are substantially influenced by effects of protein binding on availability.

3T3 Cells↗

Biological Monitoring of Pesticide Exposure: a review. Introduction.

Pesticides are used worldwide in agriculture, industry, public health and for domestic applications: as a consequence, a great part of the population may be exposed to these compounds. In spite of this extensive use, knowledge on the health risks associated with prolonged exposure is rather poor, and major uncertainties still exist. Epidemiological observations in man have so far produced little conclusive information, mainly because of weaknesses in exposure assessment. Therefore, information on the type and levels of exposure is fundamental in order to better understand and characterize risk to human health. Exposure assessment can be carried out via measurement of environmental concentrations, as well as via determination of the chemical or its metabolites in body tissues (biological monitoring). Besides indices of internal dose, biological monitoring also includes measurements of early effects attributable to interaction between the chemical agent and the human body. Biological monitoring has the advantage, over environmental monitoring, of determining the dose actually absorbed via any possible route: differences in absorption can be taken into account. whether they are due to biological variability or to use of protective equipment. When, in some cases, a combination of occupational and non-occupational exposure occurs, this also can be taken into consideration by biological monitoring. Few reference documents have been published on biological monitoring of pesticides. For this reason, the Office of Occupational Health of the World Health Organization gave ICPS a mandate to prepare a monograph specifically addressed to reviewing methods for biological monitoring of pesticide exposure. This review is based on more than 300 studies published over the period 1980-1999. For the most representative chemical classes, the available biological exposure indices are reported. Both indices of internal dose and. when available, of early effects are discussed. The reported tests were used to monitor exposure of pesticide applicators in agriculture and public health, manufacturing and formulating workers. subjects poisoned after accidental exposure or attempted suicide, volunteers involved in pharmacokinetic studies, as well as sub-groups of the general population exposed to environmentally persistent pesticides. Single chapters deal with organophosphorus insecticides, carbamate pesticides, dithiocarbamates, phenoxyacids, quaternary ammonium compounds. coumarin rodenticides, synthetic pyrethroids, organochlorine pesticides, chlorotriazines, and pentachlorophenol.

Animals↗