Metabolism of chlorinated biphenyls: use of 3,3'- and 3,5-dichlorobiphenyl as sole sources of carbon by natural species of Ralstonia and Pseudomonas

Ralstonia sp. SA-3, Ralstonia sp. SA-4 and Pseudomonas sp. SA-6 are natural strains with a novel capacity to utilize meta-substituted dichlorobiphenyls (diCBs) hitherto not known to serve as a sole source of carbon and energy for polychlorobiphenyl-degraders. In growth experiments, axenic cultures of isolates grew logarithmically on 3,3'-diCB with generation times that ranged insignificantly (t-test, P>0.05) from 30.4 to 33.8 h. Both 3-chlorobenzoate (3-CBA) and chloride produced as metabolites were recovered in non-stoichiometric quantities. The release of chloride by the cultures lagged substantially, indicating that the initial dioxygenase attack preceded cleavage of carbon-chloride bonds and that chloride must have been released from the chlorinated hydroxypentadienoate. In the case of 3,5-diCB, SA-3 and SA-6 metabolised this substrate primarily to 3,5-CBA. The lack of chloride in the culture media coupled with stoichiometric recovery of 3,5-CBA suggests that growth by these strains occurred predominantly at the expense of the unsubstituted phenyl ring. The unique metabolic properties of these three aerobic isolates point to their potential usefulness as seeds for bioremediation of PCBs polluted environments without the need for repeated inoculation or supplementation by a primary growth substrate such as biphenyl.     

Microbial reductive dechlorination of polychlorinated biphenyls in heat treated and bromoethanesulfonate treated anaerobic sediment slurries

Anaerobic reductive dechlorination of polychlorinated biphenyls (PCBs) in heat treated, bromoethanesulfonate (BES) treated, and untreated slurries of PCB-contaminated upper Hudson River sediment was investigated to better understand the microorganisms that mediate this process. Extensive meta-and para-dechlorination of Aroclor 1242 and 2,3,4-trichlorobiphenyl (2,3,4-CB) occurred in both the untreated and heat treated slurries. Heat treatment of the slurries eliminated methanogenesis, enhanced 2,3,4-CB dechlorination, and caused extensive meta- and para-dechlorination of Aroclor 1242 earlier than in untreated slurries. BES treatment (1 mM) of the slurries caused a 90% reduction in methanogenesis and inhibited metadechlorination of PCB congeners containing 2,3- and 2,5-dichlorophenyl groups. The results suggest that there are at least two distinct microbial PCB reductive dechlorination activities in PCB-contaminated upper Hudson River sediment, a meta-dechlorination activity and a para-dechlorination activity on Aroclor 1242. Both of these microbial activities are apparently not methanogenic and are resistant to or activated by heat treatment. In addition, the meta- but not the para-dechlorinating activity is inhibited by BES treatment.     

Iron-mediated reactions of polychlorinated biphenyls in electrochemical peroxidation process (ECP)

A study was conducted to explore some of the basic processes of polychlorinated biphenyl (PCB) destruction by a new technology termed electrochemical peroxidation process (ECP). ECP represents an enhancement of the classic Fenton reaction (H2O2 + Fe2+) in which iron is electrochemically generated by steel electrodes. Focus was on the extent of adsorption of a mixture of Aroclor 1248 on steel electrodes in comparison to iron filings. Commercially available zero-valent iron filings rapidly adsorbed PCBs from an aqueous solution of Aroclor 1248. Within 4 h, all the PCBs were adsorbed at 1%, 5%, and 10% Fe0 (w/v) concentrations. Little difference in adsorption was found between acidic (2.3) and unamended solutions (pH 5.5), even though significant differences in iron oxidation state and Fe2+ concentrations were measured in solution. PCB adsorption also occurs on steel electrodes regardless of the pH or electric current applied (AC or DC), suggesting the combination of oxidizing (free radical-mediated reactions) and reducing (dechlorination reactions) iron-mediated degradation pathways may be possible. Extraction of the iron powder after 48 h of contact time yielded the progressive recovery of biphenyl with increasing Fe mass(from 0.4% to 3.5%) and changes of the PCB congener-specific pattern as a consequence of dechlorination. A variety of daughter congeners similar to those accumulated during anaerobic microbial dechlorination of Aroclor 1248 in contaminated sediments indicate preferential removal of meta- and para-chlorines.     

Study of the biodegradation process of polychlorinated biphenyls in liquid medium and soil by a new isolated aerobic bacterium (Janibacter sp.)

We have isolated and characterised a novel aerobic bacterial strain, designated MS3-02, belonging to the genus Janibacter sp. The capability of this new strain to degrade polychlorinated biphenyls (PCBs) in a commercial mixture (Aroclor 1242) in liquid medium and in soil (sterile and non sterile soil), under laboratory scale, has been evaluated. MS3-02 was isolated from the soil around of an incinerator, located in the east of Madrid (Spain). Gas-chromatographic analysis showed that MS3-02 was able to reduce most peaks observed in the chromatogram between 70% and 100% after seven days of incubation in a culture mineral medium containing yeast extract, but without the addition of biphenyl. The presence of biphenyl in the culture medium decreased the rate of PCB degradation by this bacterium. Comparing the performance of the MS3-02 in liquid culture medium and in soil, degradation was less efficient in sterile soil and still less efficient in non sterile soil. Under the best conditions (sterile soil and 20 weeks of incubation) MS3-02 was able to reduce, between 50% and 100%, nine of the main gas-chromatographic peaks in Aroclor 1242.     

Aerobic degradation of di- and trichlorobenzenes by two bacteria isolated from polluted tropical soils

Two polychlorinated biphenyl (PCBs)-degrading bacteria were isolated by traditional enrichment technique from electrical transformer fluid (Askarel)-contaminated soils in Lagos, Nigeria. They were classified and identified as Enterobacter sp. SA-2 and Pseudomonas sp. SA-6 on the basis of 16S rRNA gene analysis, in addition to standard cultural and biochemical techniques. The strains were able to grow extensively on dichloro- and trichlorobenzenes. Although they failed to grow on tetrachlorobenzenes, monochloro- and dichlorobenzoic acids, they were able to utilize all monochlorobiphenyls, and some dichlorobiphenyls as sole sources of carbon and energy. The effect of incubation with axenic cultures on the degradation of 0.9 mM 1,4-dichlorobenzene, 0.44 mM 1,2,3- and 0.43 mM 1,3,5-trichlorobenzene in mineral salts medium was studied. Approximately, 80-90% of these xenobiotics were degraded in 200 h, concomitant with cell increase of up to three orders of magnitude, while generation times ranged significantly (P<0.05) from 17-32 h. Catechol 1,2-dioxygenase and catechol 2,3-dioxygenase activities were detected in crude cell-free extracts of cultures pre-grown with benzoate, with the latter enzyme exhibiting a slightly higher activity (0.15-0.17 micromolmin(-1) mg of protein(-1)) with catechol, suggesting that the meta-cleavage pathway is the most readily available catabolic route in the SA strains. The wider substrate specificity of these tropical isolates may help in assessing natural detoxification processes and in designing bioremediation and bioaugmentation methods.     

Catalyzed dehalogenation of Delor 103 by sodium hydridoaluminate

The complete dechlorination of PCB liquid Delor 103 (42.6% Cl) to biphenyl has been effected in toluene with sodium dihydridobis(2-methoxyethoxo)aluminate (SDMA) in the presence of catalytic amounts of Ni(II) and Co(II) 2,4-pentanedionates at elevated temperatures. Changes in PCB congeners distribution during dechlorination were monitored by GC(ECD) and GC/MS, and the scheme of their transformation was proposed on the basis of site selectivity observed. A kinetic analysis of the reaction is presented.     

Dechlorination pathways of ortho-substituted PCBs by UV irradiation in n-hexane and their correlation to the charge distribution on carbon atom

The photodegradations of 22 individual polychlorinated biphenyl (PCB) congeners (including 21 non-coplanar ortho substituted and one non-ortho substituted) by irradiation with ultraviolet lamp in n-hexane solution were studied. Photoproducts were identified by matching their retention times and mass spectra with those of authentic standards. PCB congener with less than two chlorides was photodegraded within half an hour, if more than three chlorine on ring, the photodechlorination time for PCB needs one and half hours or more, sometimes even longer than 15 h. The half-life of PCB degradation by photodechlorination was much shorter than that by anaerobic biological dechlorination. Charge distribution on carbon atom combined with the monitoring products of individual PCB congeners were used to deduce the photodegradation pathways. The higher the charge distribution for carbon to which chlorine is attached, the easier for photodechlorination to occur. A lot of chlorine atoms attached PCB, the dechlorination was found to occur prior to the carbon with higher charge distribution at the benzene ring with more chlorine atoms attached.     

Comparison of an enzyme-linked immunosorbent assay (ELISA) to gas chromatography (GC)--measurement of polychlorinated biphenyls (PCBs) in selected US fish extracts

The analysis of PCBs in fish tissues by immunoassay methods was evaluated using fish collected from a US monitoring program, the National Contaminant Biomonitoring Program of the US Department of Interior, Fish and Wildlife Service. Selected composite whole fish samples, which represented widely varying concentrations and sources of PCBs, were extracted and subjected to congener PCB analysis by gas chromatography (GC) and total PCB analysis using an ELISA (ePCBs) calibrated against technical Aroclor 1248. PCB congener patterns in these fishes were different from the patterns found in commercial Aroclors or their combinations as demonstrated by principal component analysis of normalized GC congener data. The sum of the PCB congeners measured by GC (total-PCBs) ranged from 37 to 4600 ng/g (wet weight). Concentrations of PCBs as determined by the ELISA method were positively correlated with total-PCBs and the ePCBs/total-PCBs ratios for individual samples ranged from 1 to 6. Ratios of ePCBs/total-PCBs for dilutions of Aroclors 1242, 1254, and 1260 and for matrix spikes range from 0.6 for 1242 to 2.5 for 1254 and 1260. These results suggest that higher chlorinated PCB congeners have higher affinity for the anti-PCB antibodies. Partial least squares with latent variable analysis of GC and ELISA data of selected Aroclors and fish samples also support the conclusion that ELISA derived PCB concentrations are dependent on the degree of chlorination.     

Dechlorination of PCBs by electrochemical reduction with aromatic radical anion as mediator

Mediated electrochemical reduction was applied to the dechlorination of polychlorinated biphenyls (PCBs) in tetra-n-butylammonium perchlorate/dimethylformamide solution. Rapid and complete dechlorination was possible with biphenyl or naphthalene as the mediator, whereas the reaction was much slower with anthracene or 9,10-diphenylanthracene. The reaction rate was so high with naphthalene or biphenyl radical anion that differences in reactivity could not be observed between congeners. Side reactions, other than biphenyl formation, could occur depending upon the mediators and the substrates, but contributed less than 10% of the total products in the case of naphthalene-mediated dechlorination. Almost all chloride ion formed in the dechlorination remained in the cathode solution. The order of the reaction was determined to be 0.5 for the substrate and 1 for the mediator (naphthalene) in the dechlorination of 2-chlorobiphenyl; identical to results for the mediated dechlorination of 1-chloronaphthalene. The reaction rate in practical PCB dechlorination could be estimated with the use of the initial concentration of the mediator and chlorine content of the solution provided that the problem of the deactivation of the electrode surface could be solved.     

Polychlorinated biphenyls in surface sediments of Yueqing Bay, Xiangshan Bay, and Sanmen Bay in East China Sea

The spatial distribution and source of polychlorinated biphenyls (PCBs) in 30 surface sediments of Yueqing Bay, Xiangshan Bay, and Sanmen Bay in Eastern China were analysed. Total concentrations of PCBs ranged from 9.33 to 19.60 ng g⁻¹ dry weight for all the sampling stations. The observed PCB levels were lower than those in areas of high urbanisation or contamination in the bay. Low-chlorinated PCBs, dominated by tri-PCB, were identified as the prevalent contaminate of surface sediments, and the top three PCB congeners were lighter chlorinated congeners (PCB 8, PCB 18 and PCB 28). These results were in agreement with the fact that tri-PCB compounds are the dominant contaminants in China. The result of the principal component analysis revealed that all samples were similar in composition to Aroclor 1242, suggesting they might originate from electrical capacitors and transformers. The levels of PCBs were significantly correlated with the total organic carbon in the sediments.     

Development of a new polyclonal antibody for the determination of polychlorinated biphenyls in indoor air by ic-ELISA

A new polyclonal antibody (pAb) was prepared and used for the determination of polychlorinated biphenyls (PCBs) in air samples to promote the application of immunoassay technology in the determination of PCBs. Three PCB congeners immunogen mixture was used to stimulate immune responses in rabbits. The specific pAb to PCBs was obtained and used to develop an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). A standard curve for Aroclor 1248 was prepared using concentrations ranging from 0.1 to 100 μg L^?1. The average IC₅₀ value was 16.21 μg L^?1 and the limit of detection at 10 % inhibition (IC₉₀) was 0.069 μg L^?1. The entire procedure was then evaluated using spiked air samples. The recoveries of Aroclor 1248 at various spiking levels in the air samples ranged from 84 to 113 %, with relative standard deviations of 3 to 6 %. Under optimum conditions, the cross-reactivity profiles of the assays were obtained using three selected congeners, four Aroclor products, and other structurally related compounds of PCBs. The assays were found to be highly specific for PCB congeners and Aroclors 1248 and 1242. The air samples were then analyzed using gas chromatography coupled with high-resolution mass spectrometry to confirm the ic-ELISA results. The attained results demonstrated that the proposed method was an effective and inexpensive technique for the PCBs determination in air samples.     

Isotope dilution analysis of polychlorinated biphenyls (PCBs) in transformer oil and global commercial PCB formulations by high resolution gas chromatography-high resolution mass spectrometry

Special polychlorinated biphenyls (PCBs) standards (native and isotope labeled) were analyzed by isotope dilution method using HRGC-HRMS. Multiple analysis of special PCBs standards by three different laboratories produced the relative response factors (RRFs) and relative standard deviations (RSDs %) was in the average of 0.979 and 3.86, respectively. Additionally, inter-laboratory analysis of various forms of transformer oil revealed the PCBs concentrations were in the following order; PCBs fortified transformer oil (940-1300 ng/g)>PCB polluted transformer oil (490-680 ng/g)>chemically degraded-transformer oil (480-490 ng/g) and PCBs free oil (ND-17 ng/g). Chemical degradation resulted in an order of magnitude decrease in the PCB concentrations. Specifically, higher chlorinated PCBs degraded into lower chlorinated PCBs. Also, composition of PCBs have been determined in PCB formulations from Japan (Kanechlor), Germany (Clophen), USA (Aroclor), Russia (Sovol) and Poland (Chlorofen). Major PCBs (24-PCB congeners) contributed 54-67%, 55-68%, 16-69%, 71% and 72% in Kanechlor, Clophen, Aroclor, Sovol and Chlorofen, respectively to total PCBs. The homologue pattern of Kanechlor, Aroclor and Clophen in technical fromulation was similar (e.g., Kanechlor-300 resembled to those of Clophen A-30 and Aroclor-1242). Furthermore, congener-specific distributions of major PCBs/dioxin-like PCBs and toxic equivalency quantities (TEQ) were calculated. Based on our tentative assumption calculations, cumulative production of five different technical PCB formulations, WHO-TEQ emission was estimated to be approximately 16.05 tons.     

Correlation of gas chromatographic properties of chlorobenzenes and polychlorinated biphenyls with the occurrence of reductive dechlorination by untamed microorganisms

To understand the dechlorination ability of chlorobenzenes (CBs) and polychlorinated biphenyls (PCBs) by untamed microorganisms under anaerobic condition and to correlate gas chromatographic properties with the occurrence of reductive dechlorination, introduction of CBs and PCBs in the culture medium inoculated with microorganisms from sludge and sediment, respectively, were performed. Three kinds of culture media preparing from sludge, river water and a synthetic medium were used in the experiments. HCB was degraded to 1,3,5-trichlorobenzene (1,3,5-TCB) and 1,3-dichlorobenzene (1,3-DCB) in both sludge medium and synthetic medium with inoculated microorganisms. Three PCB congeners including 2,3,4-, 3,4,5- and 2,3,4,5-CBp (chlorinated biphenyl) were not found to be dechlorinated in the river water medium with inoculation culture but to be dechlorinated in the synthetic medium. MNDO methodology was used to compute theoretical dechlorination reaction heats and GC-ECD techniques were used to estimate chromatographic data of CB and PCB congeners. Both CB and PCB congeners showed that dechlorination by untamed microorganisms under anaerobic mixed cultures were more likely to occur when larger amounts of energy were released and greater deltaln RRT value between the parent congener and the daughter product was observed. Deltaln RRT provided a more precise information on the singularity of PCBs ortho-dechlorination in an aspect of thermodynamic favorable rule.     

Microbial dechlorination of three PCB congeners in river sediment.

We investigated the potential for the anaerobic degradation of three PCB congeners (2,3,5,6-CB, 2,3,4,5-CB, and 2,3,4,5,6-CB) in sediments collected from five monitoring sites along the Keelung River in northern Taiwan. Optimal conditions for congener dechlorination were 30 degrees C and pH 7.0. Intermediate 2,3,4,5-CB products were identified as 2,3,5-CB, 2,4,5-CB, and 2,5-CB. Intermediate 2,3,4,5,6-CB products were identified as 2,3,5,6-CB, 2,3,6-CB, and 2,5-CB. For 2,3,5,6-CB, intermediate products were identified as 2,3,6-CB and 2,5-CB. Dechlorination rates for PCB congeners were observed as (fastest to slowest): 2, 3, 4-CB > 2, 3, 4, 5-CB > 2, 3, 4, 5, 6-CB > 2, 3, 5, 6-CB > 2, 2', 3, 3', 4, 4'-CB > 2, 2', 4, 4' 6, 6'-CB > 2, 2', 3, 4, 4', 5, 5'-CB > 2, 2', 3, 3', 4, 4', 5, 5'-CB. Rates decreased for mixtures of the eight congeners. Dechlorination rates for the three primary congeners under different reducing conditions occurred in the order of (fastest to slowest): methanogenic condition > sulfate-reducing condition > nitrate-reducing condition. Under methanogenic and sulfate-reducing conditions, dechlorination rates were enhanced by the addition of lactate, pyruvate, or acetate, but delayed by the addition of manganese oxide, or ferric chloride. Under nitrate-reducing condition, dechlorination rates were delayed by the addition of lactate, pyruvate, acetate, manganese oxide or ferric chloride. Treatment with such microbial inhibitors as bromoethanesulfonic acid (BESA) or molybdate revealed that methanogen and sulfate-reducing bacteria were involved in the dechlorination of these three PCB congeners.     

Dechlorination of polychlorinated biphenyl congeners by anaerobic microorganisms from river sediment.

The microbial dechlorination of seven kinds of polychlorinated biphenyls (PCBs) by anaerobic microorganisms from river sediment was investigated. Dechlorination rates were found to be affected by the chlorine level of PCB congeners; dechlorination rates decreased as chlorine levels increased. Dechlorination rates were fastest under methanogenic conditions and slowest under nitrate-reducing conditions. The addition of individual electron donors (acetate, pyruvate, and lactate) enhanced the dechlorination of PCB congeners under methanogenic and sulfate-reducing conditions but delayed the dechlorination of PCB congeners under nitrate-reducing conditions. PCB congener dechlorination also was delayed by the addition of various polycyclic aromatic hydrocarbons (PAHs) under three reducing conditions and by surfactants, such as brij30, triton SN70, and triton N101. The results suggest that methanogen, sulfate-reducing bacteria, and nitrate-reducing bacteria all are involved in the dechlorination of PCB congeners.     

Evaluation of cumulative PCB exposure estimated by a job exposure matrix versus PCB serum concentrations

Although polychlorinated biphenyls (PCBs) have been banned in many countries for more than three decades, exposures to PCBs continue to be of concern due to their long half-lives and carcinogenic effects. In National Institute for Occupational Safety and Health studies, we are using semiquantitative plant-specific job exposure matrices (JEMs) to estimate historical PCB exposures for workers (n?=?24,865) exposed to PCBs from 1938 to 1978 at three capacitor manufacturing plants. A subcohort of these workers (n?=?410) employed in two of these plants had serum PCB concentrations measured at up to four times between 1976 and 1989. Our objectives were to evaluate the strength of association between an individual worker’s measured serum PCB levels and the same worker’s cumulative exposure estimated through 1977 with the (1) JEM and (2) duration of employment, and to calculate the explained variance the JEM provides for serum PCB levels using (3) simple linear regression. Consistent strong and statistically significant associations were observed between the cumulative exposures estimated with the JEM and serum PCB concentrations for all years. The strength of association between duration of employment and serum PCBs was good for highly chlorinated (Aroclor 1254/HPCB) but not less chlorinated (Aroclor 1242/LPCB) PCBs. In the simple regression models, cumulative occupational exposure estimated using the JEMs explained 14–24 % of the variance of the Aroclor 1242/LPCB and 22–39 % for Aroclor 1254/HPCB serum concentrations. We regard the cumulative exposure estimated with the JEM as a better estimate of PCB body burdens than serum concentrations quantified as Aroclor 1242/LPCB and Aroclor 1254/HPCB.     

PCBQ: Computerized quantification of total PCB and congeners in environmental samples

Computerized methodologies for the quantification of total PCBs, PCB in Aroclor mixtures and individual PCB congeners in environmental samples are presented. The method for total PCBs is based on a multiple-linear regression analysis using data from capillary gas chromatography of Arocolor standards. PCB congeners were identified and their weight percentages determined in Aroclor mixtures by GC/MS. PCB congeners and total PCBs were accurately quantified in predetermined test data and environmental samples.     

Atmospheric PCB congeners across Chicago

We have measured PCBs in 184 air samples collected at 37 sites in the city of Chicago using an innovative system of high-volume air samplers mounted on two health clinic vans. Here we describe results of sampling conducted from November 2006 to November 2007. The samples were analyzed for all 209 PCB congeners using a gas chromatograph with tandem mass spectrometry (GC-MS/MS). The ΣPCBs (sum of 169 peaks) in Chicago ranged from 75 pg m(-3) to 5500 pg m(-3) and primarily varied as a function of temperature. The congener patterns are surprisingly similar throughout the city even though the temperature-corrected concentrations vary by more than an order of magnitude. The average profile resembles a mixture of Aroclor 1242 and Aroclor 1254, and includes many congeners that have been identified as being aryl hydrocarbon receptor (AhR) agonists (dioxin-like) and/or neurotoxins. The toxic equivalence (TEQ) and neurotoxic equivalence (NEQ) in air were calculated and investigated for their spatial distribution throughout the urban-industrial complex of Chicago. The NEQ concentrations are linearly correlated with ΣPCBs while the TEQ concentrations are not predictable. The findings of this study suggest that airborne PCBs in Chicago are widely present and elevated in residential communities; there are multiple sources rather than one or a few locations of very high emissions; the emission includes congeners associated with dioxin-like and neurotoxic effects and congeners associated with unidentified sources.     

Biodegradation of coplanar polychlorinated biphenyls by anaerobic microorganisms from estuarine sediments.

In this study, we investigated the biodegradability of biphenyl and 5 congeners (one non-planar and four coplanar) of polychlorinated biphenyl (PCB). Biphenyl, the non-planar congener 2,3',4',5-tetrachlorobiphenyl (25-34 CB), and the four coplanar congeners 3,3',4,4'-tetrachlorobiphenyl (34-34 CB), 3,4,4',5-tetrachlorobiphenyl (345-4 CB), 3,3',4,4',5-pentachlorobiphenyl (345-34 CB), and 3,3',4,4',5,5'-hexachlorobiphenyl (345-345 CB) were amended at a concentration of 10 mg/L into anoxic sediment slurries collected from the estuaries of the Tansui River and the Erjen River. During 2 years' incubation under sulfidogenic conditions, biphenyl was persistent, while all other chlorinated congeners, except for 345-345 CB, were dechlorinated with or without a lag period in sediment slurries collected from both rivers. Dechlorination of coplanar and non-planar congeners began with para chlorine removal. All para chlorines from the mono-, di-, and trichlorobiphenyl groups could be removed by sediment slurries from both rivers. Microbial communities in sediment from the Erjen River additionally fostered meta-dechlorination activity, but only after removal of all the para chlorines. Addition of Tween 20 (0.05%, v/v) into sediment slurries from the Tansui River did not enhance dechlorination rates or extents, but the addition of toluene- or 3-chlorobenzoate-adapted sediments enhanced dechlorination of 34-34 CB and 345-4 CB.