Polychlorinated biphenyls (pcbs) in fish-eating sea birds—III. Molecular features and metabolic interpretations of PCB isomers and congeners in adipose tissues

1. Enrichment factors have been calculated for several persistent PCB congeners in the adipose tissue for five species of fish-eating sea birds (female razorbills, puffins, guillemots, shags and cormorants) obtained from the same sites during 1978–1984 (see preceding papers).2. The enrichment factor of an individual PCB is expressed as its concentration in the tissue compared with its abundance in commerical mixtures of PCBs or compared with the concentration in the tissue of the abundant congener 2,2',4,4',5,5'-hexachlorobiphenyl (congener 153, IUPAC system of numbering).3. There were no significant differences between the five species in the enrichment factor of individual persistent PCBs compared with congener 153, indicating similar levels of diminished metabolism of this group of congeners.4. Of the 47 individual PCBs identified, ten congeners had enrichment factors of > 1 in all of the species and these accounted for up to 70% of the concentration of total PCBs present. Some of these persistent congeners had approximately coplanar configurations (i.e. non-ortho -substituted congeners). Five congeners, which accounted for about 35% of the total concentration of PCBs in the tissues, shared the molecular feature of chlorine substituents at adjacent meta-para carbon atoms.5. A number of congeners were identified with enrichment factors of <1 compared with their abundance in Aroclor 1260, and very striking differences were observed between the five species in the ratio of non-persistent congeners to the persistent congener 153. These non-persistent congeners share the molecular feature of at least one pair of adjacent unsubstituted meta-para carbon atoms in the rings. This agrees with our molecular “rule” (see preceding papers) that congeners with this structural feature are subjected to metabolism by the cytochrome P-450 component of hepatic microsomal monooxygenases.6. Evidence is presented that this molecular rule applies to the persistence or non-persistence of classes of PCBs in other biological systems and that the complete absence of H atoms at adjacent carbon atoms is an essential structural requirement for the accumulation of PCBs in tissues.7. The persistence or non-persistence of individual PCBs is compared with their ability to induce specific isoforms of the cytochrome P-450 components of hepatic microsomal monooxygenases, and the toxic effects of individual PCBs that accumulate is discussed in terms of the potential environmental hazard that they represent.     

Biodegradation of polychlorinated biphenyls by plant cells

The PCB biodegradative ability of plant cells cultivated in vitro in media containing a mixture of PCB congeners, Delor 103, is demonstrated. For experiments we used submerged cultures of Armoracia rusticana, Solanum aviculare, Atropa bella-donna, transformed hairy root or embryogenic cultures of Solanum nigrum. Transformation of PCB was followed by gas chromatography after cultivations of the above-mentioned cultures with Delor 103 (10 mg 100 ml⁻¹). The overall PCB metabolizing capability and also degradation of individual congeners greatly differed from strain to strain. The highest capability to metabolize PCB was assayed with differentiated cultures of Solanum nigrum. Beside the capability of PCB degradation, total peroxidase activity in the medium and the cell extract was also followed. Differentiated or hairy root cultures exhibiting higher degradation abilities of PCB also showed increase of peroxidase activities.     

Aerobic degradation of polychlorinated biphenyls by Alcaligenes sp. JB1: metabolites and enzymes

In contrast to the degradation of penta-and hexachlorobiphenyls in chemostat cultures, the metabolism of PCBs by Alcaligenes sp. JB1 was shown to be restricted to PCBs with up to four chlorine substituents in resting-cell assays. Among these, the PCB congeners containing ortho chlorine substituents on both phenyl rings were found to be least degraded. Monochloro-benzoates and dichlorobenzoates were detected as metabolites. Resting cell assays with chlorobenzoates showed that JB1 could metabolize all three monochlorobenzoates and dichlorobenzoates containing only meta and para chlorine substituents, but not dichlorobenzoates possessing an ortho chlorine substituent. In enzyme activity assays, meta cleaving 2,3-dihydroxybiphenyl 1,2-dioxygenase and catechol 2,3-dioxygenase activities were constitutive, whereas benzoate dioxygenase and ortho cleaving catechol 1,2-dioxygenase activities were induced by their substrates. No activity was found for pyrocatechase II, the enzyme that is specific for chlorocatechols. The data suggest that complete mineralization of PCBs with three or more chlorine substituents by Alcaligenes sp. JB1 is unlikely.Abbreviations PCB polychlorinated biphenyls - CBA chlorobenzoate - D di - Tr tri - Te tetra - Pe penta- - H hexa     

A Novel Transformation of Polychlorinated Biphenyls by Rhodococcus sp. Strain RHA1

We have characterized a biphenyl degrader, Rhodococcus sp. strain RHA1. Biphenyl-grown cells of strain RHA1 efficiently transformed 45 components in the 62 major peaks of a polychlorinated biphenyl (PCB) mixture of Kanechlors 200, 300, 400, and 500 within 3 days, which includes mono- to octachlorobiphenyls. Among the intermediate metabolites of PCB transformation, di- and trichlorobenzoic acids were identified. The gradual decrease of these chlorobenzoic acids during incubation indicated that these chlorobenzoic acids would also be degraded by this strain. The effect of the position of chlorine substitution was determined by using PCB mixtures that have chlorine substitutions mainly at either the ortho or the meta position. This strain transformed both types of congeners, and strong PCB transformation activity of RHA1 was indicated. RHA1 accumulated 4-chlorobenzoic acid temporally during the transformation of 4-chlorobiphenyl. The release of most chloride in the course of 2,2(prm1)-dichlorobiphenyl degradation was observed. These results suggested that RHA1 would break down at least some PCB congeners into smaller molecules to a considerable extent.     

Metabolism of polychlorinated biphenyls by Solanum nigrum hairy root clone SNC-9O and analysis of transformation products

The study investigates aspects of PCB metabolism by a hairy root culture of Solanum nigrum L. (clone SNC-9O) in vitro. Standard conditions were established for efficient, up to 72% PCB conversion (22 individual PCB congeners examined in commercial mixture Delor 103, 5 g fresh biomass in 100 ml media shaken with 5 mg PCB for 14 days). The conversion products formed from three monochlorobiphenyls were monohydroxychlorobiphenyls and dihydroxychlorobiphenyls, while six dichlorobiphenyls yielded different monohydroxydichlorobiphenyls. Efficiency of the transformation of individual PCB congeners was evaluated together with phytotoxic effect on the clone SNC-9O. Major metabolites of monochlorobiphenyls analysed after extraction from biomass were hydroxylated at the position 4, and 4′, respectively. This revised version was published online in June 2006 with corrections to the Cover Date.     

Degradation of Delor 103, a technical mixture of polychlorinated biphenyls,by selected bacteria

Ability to utilize a technical mixture of polychlorinated biphenyls (PCB), Delor 103, as the sole carbon source, has been tested in 14 bacterial strains. For the five best growing strains (Alcaligenes latus, Alcalgenes eutrophus, Comamonas testosteroni, Micrococcus varians and Pseudomonas putida), the dependence of the degradation of individual PCB congeners on the number of chlorine substituents is discussed.     

The Dehalococcoides population in sediment-free mixed cultures metabolically dechlorinates the commercial polychlorinated biphenyl mixture aroclor 1260

Microbial reductive dechlorination of commercial polychlorinated biphenyl (PCB) mixtures (e.g., Aroclors) in aquatic sediments is crucial to achieve detoxification. Despite extensive efforts over nearly two decades, the microorganisms responsible for Aroclor dechlorination remained elusive. Here we demonstrate that anaerobic bacteria of the Dehalococcoides group derived from sediment of the Housatonic River (Lenox, MA) simultaneously dechlorinate 64 PCB congeners carrying four to nine chlorines in Aroclor 1260 in the sediment-free JN cultures. Quantitative real-time PCR showed that the Dehalococcoides cell titer in JN cultures amended with acetate and hydrogen increased from 7.07 x 10(6) +/- 0.42 x 10(6) to 1.67 x 10(8) +/- 0.04 x 10(8) cells/ml, concomitant with a 64.2% decrease of the PCBs with six or more chlorines in Aroclor 1260. No Dehalococcoides growth occurred in parallel cultures without PCBs. Aroclor 1260 dechlorination supported the growth of 9.25 x 10(8) +/- 0.04 x 10(8) Dehalococcoides cells per mumol of chlorine removed. 16S rRNA gene-targeted PCR analysis of known dechlorinators (i.e., Desulfitobacterium, Dehalobacter, Desulfuromonas, Sulfurospirillum, Anaeromyxobacter, Geobacter, and o-17/DF-1-type Chloroflexi organisms) ruled out any involvement of these bacterial groups in the dechlorination. Our results suggest that the Dehalococcoides population present in the JN cultures also catalyzes in situ dechlorination of Aroclor 1260 in the Housatonic River. The identification of Dehalococcoides organisms as catalysts of extensive Aroclor 1260 dechlorination and our ability to propagate the JN cultures under defined conditions offer opportunities to study the organisms' ecophysiology, elucidate nutritional requirements, identify reductive dehalogenase genes involved in PCB dechlorination, and design molecular tools required for bioremediation applications.     

Phylogenetically Distinct Bacteria Involve Extensive Dechlorination of Aroclor 1260 in Sediment-Free Cultures

Microbial reductive dechlorination of the persistent polychlorinated biphenyls (PCBs) is attracting much attention in cleanup of the contaminated environment. Nevertheless, most PCB dechlorinating cultures require presence of sediment or sediment substitutes to maintain their dechlorination activities which hinders subsequent bacterial enrichment and isolation processes. The information on enriching sediment-free PCB dechlorinating cultures is still limited. In this study, 18 microcosms established with soils and sediments were screened for their dechlorination activities on a PCB mixture – Aroclor 1260. After one year of incubation, 10 out of 18 microcosms showed significant PCB dechlorination with distinct dechlorination patterns (e.g., Process H, N and T classified based on profiles of PCB congeners loss and new congeners formation). Through serial transfers in defined medium, six sediment-free PCB dechlorinating cultures (i.e., CW-4, CG-1, CG-3, CG-4, CG-5 and SG-1) were obtained without amending any sediment or sediment-substitutes. PCB dechlorination Process H was the most frequently observed dechlorination pattern, which was found in four sediment-free cultures (CW-4, CG-3, CG-4 and SG-1). Sediment-free culture CG-5 showed the most extensive PCB dechlorination among the six cultures, which was mediated by Process N, resulting in the accumulation of penta- (e.g., 236-24-CB) and tetra-chlorobiphenyls (tetra-CBs) (e.g., 24-24-CB, 24-25-CB, 24-26-CB and 25-26-CB) via dechlorinating 30.44% hepta-CBs and 59.12% hexa-CBs after three months of incubation. For culture CG-1, dechlorinators mainly attacked double flanked meta-chlorines and partially ortho-chlorines, which might represent a novel dechlorination pattern. Phylogenetic analysis showed distinct affiliation of PCB dechlorinators in the microcosms, including Dehalogenimonas and Dehalococcoides species. This study broadens our knowledge in microbial reductive dechlorination of PCBs, and provides essential information for culturing and stimulating PCB dechlorinators for in situ bioremediation applications.     

Microbial reductive dechlorination of aroclor 1260 in Baltimore harbor sediment microcosms is catalyzed by three phylotypes within the phylum Chloroflexi

The specific dechlorination pathways for Aroclor 1260 were determined in Baltimore Harbor sediment microcosms developed with the 11 most predominant congeners from this commercial mixture and their resulting dechlorination intermediates. Most of the polychlorinated biphenyl (PCB) congeners were dechlorinated in the meta position, and the major products were tetrachlorobiphenyls with unflanked chlorines. Using PCR primers specific for the 16S rRNA genes of known PCB-dehalogenating bacteria, we detected three phylotypes within the microbial community that had the capability to dechlorinate PCB congeners present in Aroclor 1260 and identified their selective activities. Phylotype DEH10, which has a high level of sequence identity to Dehalococcoides spp., removed the double-flanked chlorine in 234-substituted congeners and exhibited a preference for para-flanked meta-chlorines when no double-flanked chlorines were available. Phylotype SF1 had similarity to the o-17/DF-1 group of PCB-dechlorinating bacteria. Phylotype SF1 dechlorinated all of the 2345-substituted congeners, mostly in the double-flanked meta position and 2356-, 236-, and 235-substituted congeners in the ortho-flanked meta position, with a few exceptions. A phylotype with 100% sequence identity to PCB-dechlorinating bacterium o-17 was responsible for an ortho and a double-flanked meta dechlorination reaction. Most of the dechlorination pathways supported the growth of all three phylotypes based on competitive PCR enumeration assays, which indicates that PCB-impacted environments have the potential to sustain populations of these PCB-dechlorinating microorganisms. The results demonstrate that the variation in dechlorination patterns of congener mixtures typically observed at different PCB impacted sites can potentially be mediated by the synergistic activities of relatively few dechlorinating species.     

Establishment of a polychlorinated biphenyl-dechlorinating microbial consortium, specific for doubly flanked chlorines, in a defined, sediment-free medium

Estuarine sediment from Charleston Harbor, South Carolina, was used as inoculum for the development of an anaerobic enrichment culture that specifically dechlorinates doubly flanked chlorines (i.e., chlorines bound to carbon that are flanked on both sides by other chlorine-carbon bonds) of polychlorinated biphenyls (PCBs). Dechlorination was restricted to the para chlorine in cultures enriched with 10 mM fumarate, 50 ppm (173 microM) 2,3,4, 5-tetrachlorobiphenyl, and no sediment. Initially the rate of dechlorination decreased upon the removal of sediment from the medium. However, the dechlorinating activity was sustainable, and following sequential transfer in a defined, sediment-free estuarine medium, the activity increased to levels near that observed with sediment. The culture was nonmethanogenic, and molybdate, ampicillin, chloramphenicol, neomycin, and streptomycin inhibited dechlorination activity; bromoethanesulfonate and vancomycin did not. Addition of 17 PCB congeners indicated that the culture specifically removes double flanked chlorines, preferably in the para position, and does not attack ortho chlorines. This is the first microbial consortium shown to para or meta dechlorinate a PCB congener in a defined sediment-free medium. It is the second PCB-dechlorinating enrichment culture to be sustained in the absence of sediment, but its dechlorinating capabilities are entirely different from those of the other sediment-free PCB-dechlorinating culture, an ortho-dechlorinating consortium, and do not match any previously published Aroclor-dechlorinating patterns.     

Metabolism of [14C]4-chlorobiphenyl by hepatic microsomes isolated from razorbills, pigeons and rats.

1. Analysis of individual PCB-isomers and congeners in extracts of adipose tissue from N = 12 razorbills suggested that 4-chlorobiphenyl was subjected to metabolism. 2. In vitro metabolism studies using [14C]-4-chlorobiphenyl as substrate showed that razorbills metabolise this substrate to [14C]4-chloro-4'-hydroxybiphenyl at an average rate of 20 pmol/mg microsomal protein/min. For comparison, the metabolism of [14C]-4-chlorobiphenyl by pigeons and rats was also studied, and average rates in the formation of [14C]-4-chloro-4'-hydroxybiphenyl of 12 pmol/mg microsomal protein min and 342 pmol/mg microsomal protein min were estimated. 3. A comparison of the hepatic drug metabolising enzyme system of razorbills and pigeons showed similar concentrations of cytochrome P-450, cytochrome b5 and comparable catalytic activities of cytochrome P-450-dependent monooxygenase, when assessed for HHDN epoxidase, PROD, EROD and the Phase II enzymes glutathiones-S-transferase, but were significantly lower, when compared with rats. The results obtained suggest fundamental differences in the catalytic activities of cytochrome P-450-dependent monooxygenase between avian and mammalian species. 4. The present study, however, provides evidence that fish-eating seabirds have the ability to metabolically dispose of certain PCB isomers and congeners, which are amongst the most ubiquitously distributed pollutants in the ecosystem.     

Microbial Reductive Dechlorination of Aroclor 1260 in Baltimore Harbor Sediment Microcosms Is Catalyzed by Three Phylotypes within the Phylum Chloroflexi

The specific dechlorination pathways for Aroclor 1260 were determined in Baltimore Harbor sediment microcosms developed with the 11 most predominant congeners from this commercial mixture and their resulting dechlorination intermediates. Most of the polychlorinated biphenyl (PCB) congeners were dechlorinated in the meta position, and the major products were tetrachlorobiphenyls with unflanked chlorines. Using PCR primers specific for the 16S rRNA genes of known PCB-dehalogenating bacteria, we detected three phylotypes within the microbial community that had the capability to dechlorinate PCB congeners present in Aroclor 1260 and identified their selective activities. Phylotype DEH10, which has a high level of sequence identity to Dehalococcoides spp., removed the double-flanked chlorine in 234-substituted congeners and exhibited a preference for para-flanked meta-chlorines when no double-flanked chlorines were available. Phylotype SF1 had similarity to the o-17/DF-1 group of PCB-dechlorinating bacteria. Phylotype SF1 dechlorinated all of the 2345-substituted congeners, mostly in the double-flanked meta position and 2356-, 236-, and 235-substituted congeners in the ortho-flanked meta position, with a few exceptions. A phylotype with 100% sequence identity to PCB-dechlorinating bacterium o-17 was responsible for an ortho and a double-flanked meta dechlorination reaction. Most of the dechlorination pathways supported the growth of all three phylotypes based on competitive PCR enumeration assays, which indicates that PCB-impacted environments have the potential to sustain populations of these PCB-dechlorinating microorganisms. The results demonstrate that the variation in dechlorination patterns of congener mixtures typically observed at different PCB impacted sites can potentially be mediated by the synergistic activities of relatively few dechlorinating species.     

Biological treatment of transformer oil using commercial mixtures of microorganisms

The biodegradation of six PCB congeners (IUPAC nos. 28, 52, 101, 138, 153, and 168) present in transformer oil using different commercial mixtures of microorganisms (Sybron 1000, Biozyn 301, Biozyn 300, DBC 5, ChZR, NS 20-10, NS 20-20, and NS 20) under anoxic, oxic, or anoxic/oxic treatments was investigated at laboratory scale. Overall, PCB congener biodegradation was observed in all treatments in the ranges of 0–99%, 2–97% and 40–94% under anoxic, oxic, or anoxic/oxic treatments, respectively. The highest biodegradation of total PCB congeners occurred using Sybron under oxic and anoxic conditions (76.0% and 91.3% reduction, respectively; initial PCB concentration, 1417.1 mg l^?1). Also, the highest biodegradation extent of the PCB congeners occurred when the commercially available mixture of microorganisms, Sybron, was used (84.7% reduction) under combined anoxic/oxic conditions. Demonstration that biodegradation of most of the individual PCB congeners was achieved by the commercial mixtures of microorganisms in this study suggests that PCB-impacted environments can sustain populations of these PCB-metabolizing organisms. This is particularly relevant for the development of biostimulation or bioaugmentation strategies for the bioremediation of PCB-contaminated wastes.     

Bacterial degradation of a mixture obtained through the chemical modification of polychlorinated biphenyls by polyethylene glycols

Polychlorinated biphenyls (PCBs), also known by the trade name Sovol, are toxic industrial wastes. They have been subjected to chemical treatment by polyethylene glycols (PEGs) and potassium hydroxide. As a result of the interaction of the Sovol with various molecular mass PEGs (MM_PEG-4 ～ 200, MM_PEG-22 ～ 1000), water-soluble mixtures M1 and M2 containing mono(polyethylene glycol)oxy-derivatives (PCB-PEG-4 and PCB-PEG-22), polychlorobiphenylols, and unreacted PCB congeners (PCB 44, PCB 47, PCB 49, PCB 52, and PCB 66) were obtained. It was shown for the first time that mixtures M1 and M2 are susceptible to bacterial degradation without their fractionation. According to the gas-liquid chromatography with flame-ionization and mass-spectrometric detection, the Rhodococcus wratislaviensis KT112-7 strain degraded all of the chemical compounds occurring in the mixtures. In a 5-day experiment, it was found that the KT112-7 strain decomposes mono(polyethylene glycol)oxy-derivatives completely (by 100%) and polychlorobiphenylols and PCB congeners by 90–95% in the M1 and M2 mixtures. The culture medium did not contain transformation products, whereas free chlorine ions were accumulated (72–94% of the maximum possible amount). Thus, the use of the chemical modification and consecutive bacterial degradation provided an effective destruction of technical PCB mixtures with a high content of highly chlorinated congeners.     

In vitro metabolism of 4-chlorobiphenyl by control and induced rat liver microsomes

The in vitro metabolism, mechanism of metabolism, and macromolecular binding of a monochlorobiphenyl component of commercial polychlorinated biphenyls (PCB) have been investigated. 4-Chlorobiphenyl was metabolized by rat liver microsomes in the presence of NADPH to yield a major metabolite, 4'-chloro-4-biphenylol, and a number of minor metabolites. The metabolism of deuterium-labeled 4-chlorobiphenyl proceeded with the NIH shift of the isotope and no observed isotope effect thus indicating the intermediacy of an arene oxide. Noninduced rat liver microsomes mediated the covalent binding between the 4-chlorobiphenyl and 4'-chloro-4-biphenylol substrates and endogenous microsomal protein. Prior in vivo administration of a commericial PCB preparation, Aroclor 1248 (Monsanto Chemical Co., containing 48 percent by weight of chlorine), resulted in an induced microsomal preparation which significantly increased the substrate-protein binding. The effect of various inhibitors on protein binding was investigated. Aroclor 1248 induced microsomes mediated binding of 4-chlorobiphenyl to endogenous and exogenous nucleic acids, indicating a possible mechanism for the previously reported mutagenic action of this chlorobiphenyl. The spectral properties of Aroclor 1248 induced cytochrome P-450 were investigated and compared with the pentobarbital-induced cytochrome fraction.     

Establishment of a Polychlorinated Biphenyl-Dechlorinating Microbial Consortium, Specific for Doubly Flanked Chlorines, in a Defined, Sediment-Free Medium

Estuarine sediment from Charleston Harbor, South Carolina, was used as inoculum for the development of an anaerobic enrichment culture that specifically dechlorinates doubly flanked chlorines (i.e., chlorines bound to carbon that are flanked on both sides by other chlorine-carbon bonds) of polychlorinated biphenyls (PCBs). Dechlorination was restricted to the para chlorine in cultures enriched with 10 mM fumarate, 50 ppm (173 μM) 2,3,4,5-tetrachlorobiphenyl, and no sediment. Initially the rate of dechlorination decreased upon the removal of sediment from the medium. However, the dechlorinating activity was sustainable, and following sequential transfer in a defined, sediment-free estuarine medium, the activity increased to levels near that observed with sediment. The culture was nonmethanogenic, and molybdate, ampicillin, chloramphenicol, neomycin, and streptomycin inhibited dechlorination activity; bromoethanesulfonate and vancomycin did not. Addition of 17 PCB congeners indicated that the culture specifically removes double flanked chlorines, preferably in the para position, and does not attack ortho chlorines. This is the first microbial consortium shown to para or meta dechlorinate a PCB congener in a defined sediment-free medium. It is the second PCB-dechlorinating enrichment culture to be sustained in the absence of sediment, but its dechlorinating capabilities are entirely different from those of the other sediment-free PCB-dechlorinating culture, an ortho-dechlorinating consortium, and do not match any previously published Aroclor-dechlorinating patterns.     

Demethylation and denitrosation of nitrosamines by cytochrome P-450 isozymes

Metabolism of nitrosamines was studied in a reconstituted monooxygenase system composed of cytochrome P-450 isozymes purified from liver microsomes of ethanol- and phenobarbital-treated rats. The ethanol-induced isozyme (P-450et) was efficient in catalyzing the demethylation of N-nitrosodimethylamine (NDMA), with a Km of 2.4 mM and Vmax of 7.2 nmol min-1 nmol P-450(-1), but less active with N-nitrosomethylbenzylamine and N-nitrosomethylaniline. The phenobarbital-induced form (P-450b) was ineffective in NDMA metabolism but was active in catalyzing the demethylation of N-nitrosomethylaniline, with an estimated Km of 0.08 mM and a Vmax of 7.2 nmol min-1 nmol-1. P-450et also catalyzed the denitrosation of NDMA with a Km of 13.6 mM and a Vmax of 1.36 nmol min-1 nmol-1. With control liver microsomes, multiple Km values were observed for the demethylation and denitrosation of NDMA. Involvement of superoxide radicals in the metabolism of NDMA was suggested by the action of superoxide dismutase, which inhibited the denitrosation by 43 to 73% and the demethylation by 13 to 22% in different monooxygenase systems. The P-450et-dependent NDMA demethylation was strongly inhibited by 2-phenylethylamine and 3-amino-1,2,4-triazole; these compounds were previously believed not to be inhibitors of P-450-dependent reactions but were found to inhibit microsomal NDMA demethylase. The present results establish the role of P-450 in nitrosamine metabolism and help to clarify some of the previous confusion in this area of research.     

Polychlorinated biphenyls (PCBs) in fish-eating sea birds—II. Molecular features of PCB isomers and congeners in adipose tissue of male and female puffins (Fratercula arctica), guillemots (Uria aalga), shags (Phalacrocorax aristotelis) and cormorants (Phalacrocorax carbo) of British and Irish coastal waters

1. The concentration of individual PCBs was measured in adipose tissue of male and female puffins, shags, guillemots and cormorants obtained from the Isle of May and the Saltees islands.2. The concentrations of total PCBs showed positive correlations with that ofp,p'-DDE in the tissues.3. Enrichment factors were calculated by comparing the concentration of an individual PCB in the tissue with its abundance in commercial mixtures of PCBs.4. Of the 47 individual PCBs identified five prominent PCBs had enrichment factors considerably > 1 and accounted for approximately 35% of the total concentration of PCBs present. They shared the common molecular feature of chlorine atoms at adjacent meta-para positions in at least one of the biphenyl rings.5. Many PCBs had enrichment factors of <1, which suggested that they had been subjected to metabolism and presumably excretion. They shared the common feature of the absence of chlorine atoms at the meta-para positions of the biphenyl rings.6. These results support strongly the structural “rules” suggested in the preceding paper (Borlakoglu et al., 1990a) for the tendency of individual PCBs to accumulate or to be subjected to metabolism.     

Degradation of Di- Through Hepta-Chlorobiphenyls in Clophen Oil Using Microorganisms Isolated from Long Term PCBs Contaminated Soil

Present work describes microbial degradation of selected polychlorinated biphenyls (PCBs) congeners in Clophen oil which is used as transformer oil and contains high concentration of PCBs. Indigenous PCBs degrading bacteria were isolated from Clophen oil contaminated soil using enrichment culture technique. A 15 days study was carried out to assess the biodegradation potential of two bacterial cultures and their consortium for Clophen oil with a final PCBs concentration of 100 mg kg⁻¹. The degradation capability of the individual bacterium and the consortium towards the varying range of PCBs congeners (di- through hepta-chlorobiphenyls) was determined using GCMS. Also, dehydrogenase enzyme was estimated to assess the microbial activity. Maximum degradation was observed in treatment containing consortium that resulted in up to 97 % degradation of PCB-44 which is a tetra chlorinated biphenyl whereas, hexa chlorinated biphenyl congener (PCB-153) was degraded up to 90 % by the consortium. This indicates that the degradation capability of microbial consortium was significantly higher than that of individual cultures. Furthermore, the results suggest that for degradation of lower as well as higher chlorinated PCB congeners; a microbial consortium is required rather than individual cultures.     

PCB-153 Shows Different Dynamics of Mobilisation from Differentiated Rat Adipocytes during Lipolysis in Comparison with PCB-28 and PCB-118

Background

Polychlorinated biphenyls (PCBs) are persistent organic pollutants. Due to their lipophilic character, they are preferentially stored within the adipose tissue. During the mobilisation of lipids, PCBs might be released from adipocytes into the bloodstream. However, the mechanisms associated with the release of PCBs have been poorly studied. Several in vivo studies followed their dynamics of release but the complexity of the in vivo situation, which is characterised by a large range of pollutants, does not allow understanding precisely the behaviour of individual congeners. The present in vitro experiment studied the impact of (i) the number and position of chlorine atoms of PCBs on their release from adipocytes and (ii) the presence of other PCB congeners on the mobilisation rate of such molecules.

Methodology/Principal Findings

Differentiated rat adipocytes were used to compare the behaviour of PCB-28, -118 and -153. Cells were contaminated with the three congeners, alone or in cocktail, and a lipolysis was then induced with isoproterenol during 12 hours. Our data indicate that the three congeners were efficiently released from adipocytes and accumulated in the medium during the lipolysis. Interestingly, for a same level of cell lipids, PCB-153, a hexa-CB with two chlorine atoms in ortho-position, was mobilised slower than PCB-28, a tri-CB, and PCB-118, a penta-CB, which are both characterised by one chlorine atom in ortho-position. It suggests an impact of the chemical properties of pollutants on their mobilisation during periods of negative energy balance. Moreover, the mobilisation of PCB congeners, taken individually, did not seem to be influenced by the presence of other congeners within adipocytes.

Conclusion/Significance

These results not only highlight the obvious mobilisation of PCBs from adipocytes during lipolysis, in parallel to lipids, but also demonstrate that the structure of congeners defines their rate of release from adipocytes.