Influence of quaternary ammonium compounds on the microbial reductive dechlorination of pentachloroaniline

The inhibitory effect of two widely used quaternary ammonium compounds (QACs) – alkyl benzyl dimethyl (AB) and hexadecyl trimethyl (HD) ammonium chloride – on fermentation, methanogenesis and pentachloroaniline (PCA) dechlorination was assessed using a mixed, methanogenic, PCA-dechlorinating culture amended with AB or HD at a concentration range from 5 to 70 μM. PCA dechlorination was inhibited at 5 μM AB and was completely inhibited at 25 or 5 μM by AB or HD, respectively. However, the PCA dechlorination pathway was the same in both the QACs-free and QACs-amended culture series. Fermentation (acidogenesis) and methanogenesis were inhibited by both AB and HD at and above 25 μM but to a lesser degree than PCA dechlorination. Overall, HD resulted in a more severe inhibition of the mixed culture than AB. Adsorption of both QACs to the mixed culture biomass followed the Freundlich isotherm model. The adsorption affinity of HD for the mixed culture biomass was significantly higher than that of AB, which may be related to the observed higher inhibitory effects of HD compared to AB. Both AB and HD were not degraded in the mixed, dechlorinating culture used in this study.     

Anaerobic degradation and methanogenic inhibitory effects of oleic and stearic acids

Oleic acid, an 18 carbon acid with one double bond (C18:1) was degraded anaerobically to palmitic (C16:0) and myristic (C14:0) acid by-products at 21 C by a culture unacclimated to long-chain fatty acids. These by-products were degraded to acetate and ultimately to methane. In comparison, no long-chain fatty acid by-products were observed in unacclimated anaerobic cultures receiving stearic (C18:0) acid although slow removal of stearic acid occurred. Oleic acid concentrations above 30mg l(-1) inhibited acetate degradation but stearic acid up to 100 mg l(-1) did not inhibit aceticlastic methanogenesis. Hydrogenotrophic methanogenesis was slightly inhibited by oleic and stearic acids. A thermodynamic basis for comparing anaerobic C18 acid degradation and predicting by-products is presented.     

Biological and chemical interactions with U(VI) during anaerobic enrichment in the presence of iron oxide coated quartz

Microcosm experiments were performed to understand chemical and biological interactions with hexavalent uranium (U(VI)) in the presence of iron oxide bearing minerals and trichloroethylene (TCE) as a co-contaminant. Interactions of U(VI) and hydrous iron oxide moieties on the mineral oxide surfaces were studied during enrichments for dissimilatory iron reducing (DIRB) and sulfate reducing bacteria (SRB). Microbes enriched from groundwater taken from the Test Area North (TAN) site at the Idaho National Laboratory (INL) were able to reduce the U(VI) in the adsorption medium as well as the iron on quartz surfaces. Early in the experiment disappearance of U(VI) from solution was a function of chemical interactions since no microbial activity was evident. Abiotic removal of U(VI) was enhanced in the presence of carbonate. As the experiment proceeded, further removal of U(VI) from solution was associated with the fermentation of lactate to propionate and acetate. During later phases of the experiment when lactate was depleted from the growth medium in the microcosm containing the DIRB enrichments, U(VI) concentrations in the solution phase increased until additional lactate was added. When additional lactate was added and fermentation proceeded, U(VI) concentrations in the liquid phase again returned to near zero. Similar results were shown for the SRB enrichment but lower uranium concentrations were seen in the liquid phase, while in the enrichment with carbonate a similar increase in uranium concentration was not seen. Chemical and biological interactions appear to be important on the mobilization/immobilization of U(VI) in an iron oxide system when TCE is present as a co-contaminant. Interestingly, TCE present in the microcosm experiments was not dechlorinated which was probably an effect of redox conditions that were unsuitable for reductive dechlorination by the microbial culture tested.     

Comparison of anaerobic dechlorinating enrichment cultures maintained on tetrachloroethene,trichloroethene, cis-dichloroethene and vinyl chloride

An anaerobic mixed microbial culture was enriched from soil and groundwater taken from a site contaminated with trichloroethene (TCE). This enrichment culture was divided into four subcultures amended separately with either perchloroethene (PCE), TCE, cis-dichloroethene (cDCE) or vinyl chloride (VC). In each of the four subcultures, the chlorinated ethenes were rapidly, consistently, and completely converted to ethene at rates of 30-50 micromol/l of culture per day, or an average 160 micro-electron equivalents/l of culture per day. These cultures were capable of sustained and rapid dechlorination of VC, and could not dechlorinate 1,2-dichloroethane, differentiating them from Dehalococcoides ethenogenes, the only known isolate capable of complete dechlorination of PCE to ethene. Chloroform (CF) and 1,1,1-trichloroethane, frequent groundwater co-contaminants with TCE and PCE, inhibited chlorinated ethene dechlorination. Most strongly inhibited was the final conversion of VC to ethene, with complete inhibition occurring at an aqueous CF concentration of 2.5 microM. Differences in rates and community composition developed between the different subcultures, including the loss of the VC enrichment culture's ability to dechlorinate PCE. Denaturing gradient gel electrophoresis of amplified bacterial 16S rRNA gene fragments identified three different DNA sequences in the enrichment cultures, all phylogenetically related to D. ethenogenes. Based on the PCR-DGGE results and substrate utilization patterns, it is apparent that significant mechanistic differences exist between each step of dechlorination from TCE to ethene, especially for the last important dechlorination step from VC to ethene.     

Chitin and corncobs as electron donor sources for the reductive dechlorination of tetrachloroethene

Chitin, corncobs, and a mixture of chitin and corncobs were tested as potential electron donor sources for stimulating the reductive dechlorination of tetrachloroethene (PCE). Semi-batch, sand-packed columns were used to evaluate the donors with aerobic and anaerobic groundwaters containing varying degrees of alkalinity. In all experiments, acetate and butyrate were the dominant fatty acids produced, although propionate, valerate, formate, and succinate were also detected. From a multivariable regression analysis on the data, the presence of chitin, limestone, and dechlorinating culture inoculum were determined to be the most positive predictors of dechlorination activity. Chitin fermentation products supported the degradation of PCE to trichloroethene (TCE), cis-1,2-dichloroethene (DCE), and vinyl chloride (VC), even in columns containing PCE DNAPL, whereas dechlorination activity was not observed in any of the columns containing corncobs alone. The longevity and efficiency of chitin as an electron donor source demonstrates its potential usefulness for passive, in situ field applications.     

Anaerobic degradation of nonionic and anionic surfactants in enrichment cultures and fixed-bed reactors

Anaerobic biodegradation and inhibitory effects of nonionic and anionic surfactants on methanogenic fermentation were tested in incubation experiments with anoxic sediment samples and sewage sludge. Alkylsulfonates and alkylbenzenesulfonates were not degraded but inhibited methanogenesis from sludge constituents at concentrations ≥10 mgl⁻¹. Sodium dodecylsulfate was at least partly degraded after adaptation at concentrations 100 mgl⁻¹ and the sulfate group was reduced to sulfide. The polyethyleneglycol moiety of alkylphenolethoxylates was fermented to methane at concentrations 500 mgl⁻¹ whereas the alkylphenol residue probably remained unchanged. Alkylethoxylates were completely degraded to methane and CO₂ at concentrations up to 1.0 gl⁻¹. Complete anaerobic degradation of this surfactant type to methane, CO₂, and traces of acetate and propionate was demonstrated in a lab scale anaerobic fixed-bed reactor, either with prereduced mineral salts medium or with air-saturated artificial wastewater. This process lends itself as a suited, inexpensive means for treatment of wastewaters containing enhanced loads of nonionic surfactants, e.g. from the surfactant manufacturing or processing industry.     

Temperature dependence of anaerobic TCE-dechlorination in a highly enriched Dehalococcoides-containing culture

Temperature dependence of reductive trichloroethene (TCE) dechlorination was investigated in an enrichment culture (KB-1), using lactate or propionate as electron donors at a temperature interval from 4 to 60 degrees C. Dechlorination was complete to ethene at temperatures between 10 and 30 degrees C (lactate-amended) and between 15 and 30 degrees C (propionate-amended). Dechlorination stalled at cis-1,2-dichloroethene (cDCE) at 4 degrees C (lactate-amended), at and below 10 degrees C (propionate-amended), and at 40 degrees C with both electron donors. No dechlorination of TCE was observed at 50 and 60 degrees C. Concentrations of Dehalococcoides had increased or remained constant after 15 days of incubation at temperatures involving complete dechlorination to ethene. Temperature dependence of dechlorination rates was compared using zero order degradation kinetics and a Monod growth-rate model for multiple electron acceptor usage with competition. Maximum growth rates (mu) and zero order degradation rates were highest for TCE dechlorination at 30 degrees C with lactate as substrate (mu(TCE) of 7.00+/-0.14 days(-1)). In general, maximum growth rates and dechlorination rates of TCE were up to an order of magnitude higher than rates for utilization of cis-dichloroethene (cDCE, mu(c)(DCE) up to 0.17+/-0.02 days(-1)) and vinyl chloride (VC, mu(VC) up to 0.52+/-0.09 days(-1)). Temperature dependence of maximum growth rates and degradation rates of cDCE and VC were similar and highest at 15-30 degrees C, with growth rates on cDCE being lower than on VC. This study demonstrates that bioaugmentation of chlorinated ethene sites may be more efficient at elevated temperatures.     

Effects of anilines and hydantoins on the methanogenic degradation of selected phenols

Five anilines and two hydantoins commonly found in coal conversion wastewaters were evaluated in batch culture, serum bottle tests to determine whether they inhibited phenol- or p-cresoldegrading methanogenic cultures. Working with individual compounds, phenol degradation and subsequent methanogenesis were not affected by 29 mg/l N-methylaniline, 97 mg/l aniline, 300 mg/1 o-methylaniline, 300 mg/1 m-methylaniline or 300 mg/1 p-methylaniline. Neither p-cresol degradation or subsequent methanogenesis was affected by 500 mg/1 hydantoin or 500 mg/l 5,5-dimethylhydantoin. Extended incubation periods showed that none of the three isomers of ring substituted methyl anilines and neither of the two hydantoins were readily degraded to methane.     

Effects of sulfate on anaerobic chloroethene degradation by an enriched culture under transient and steady-state hydrogen supply

Complete anaerobic dechlorination of chlorinated solvents such as trichloroethene (TCE) is essential for bioremediation of chloroethene-contaminated sites. We studied the influence of sulfate on microbial dechlorination of TCE to ethene both under transient and steady-state conditions, encompassing the range of hydrogen (H2) levels commonly found at contaminated sites. The results show that sulfate at a concentration of 2.5 mM limits microbial dechlorination by a mixed anaerobic culture by reducing the rate under steady-state hydrogen supply (a few nM H2), implying a H2 limited dechlorination. Conversely, sulfate did not affect dechlorination when rapid fermentation of lactate resulted in transient buildup of H2 to levels around two orders of magnitude higher compared to steady-state conditions. This has important implications both for optimizing culture conditions for dehalogenating microorganisms and for the efficiency of cleanup strategies. Our findings may contribute to the understanding and bioremediation of chloroethene contaminated environments containing sulfate.     

Biotransformation of alkanoylcholines under methanogenic conditions

Ester quaternary ammonium compounds (esterquats), which are mainly used as active ingredients in fabric softeners and personal care products, are beginning to replace traditional quaternary ammonium compounds. As a result of hydrophobicity and increasing use, esterquats reach anaerobic treatment systems. However, little is known about the fate of esterquats under anaerobic conditions. In the present study, the potential inhibitory effect and biotransformation of two alkanoylcholines - acetylcholine chloride (ACh-Cl) and lauroylcholine chloride (LCh-Cl) - which are simple esterquats, under methanogenic conditions were investigated. ACh-Cl up to 300 mg/L was not inhibitory to a mixed methanogenic culture. In contrast, methanogenesis was inhibited by LCh-Cl above 50 mg/L, primarily caused by the accumulation of lauric acid which resulted from the abiotic hydrolysis of LCh. Below inhibitory concentrations, both ACh and LCh were transformed to methane by the mixed methanogenic culture. Mass spectrometric analysis confirmed that both alkanoylcholines were first abiotically hydrolyzed to choline and the corresponding alkanoic acid, which were then biotically transformed to methane, carbon dioxide, and ammonia. Thus, alkanoylcholine-containing waste streams can be bioprocessed to form methane, but hydrolysis products such as long-chain alkanoic acids may adversely impact the anaerobic bioconversion of alkanoylcholines.     

Methanogenic digestion using mixed substrate of acetic, propionic and butyric acids

Experiments on methanogenic digestion using high concentrations of mixed substrate were conducted. The major intermediate products of anaerobic digestion such as acetic, propionic and butyric acids were mixed in a ratio of 2:1:1 (COD basis), respectively, and used as a substrate for feeding into continuous-flow chemostat reactors maintained at 35°C. These reactors were operated stably at higher feed substrate concentrations and shorter hydraulic retention times (HRT) than those of using a single component of volatile fatty acids as a substrate. At an HRT of 4.43 days, the methanogenesis occurred normally up to a feed substrate concentration of 70,000 mg COD I⁻¹. At a feed substrate concentration of 20,000 mg COD I⁻¹, the methanogenesis occurred normally up to an HRT of 2.91 days and the minimum SRT for microbial populations was calculated to be 2.42 days. An increase in feed substrate concentration adversely affected the propionate degradation strikingly, while a decrease in HRT significantly adversely affected the acetate and propionate degradation. The methane production was 0.301 g⁻¹ COD utilized, and it was independent of the feed substrate concentration and HRT. Bacilli were predominant in all reactors, but sarcinae appeared in the reactors with high feed substrate concentrations and short HRTs. Phenomena in digester failure due to methanogen washout were also observed.     

Enrichment of anaerobic polychlorinated biphenyl dechlorinators from sediment with iron as a hydrogen source

Little is known about anaerobic polychlorinated biphenyl (PCB) dechlorination, although it is believed that some microorganisms are capable of respiring PCBs, gaining energy for growth from PCB dechlorination. If this is the case, the amendment of appropriate electron donors to contaminated sediment should stimulate dechlorination. The effect of elemental iron (Fe0) addition, an easily amended electron donor, on the microbial dechlorination of the PCB congeners 3,4,5-trichlorobiphenyl (3,4,5-CB) and 2,2',3,4,4',5,5'-heptachlorobiphenyl (2,2',3,4,4',5,5'-CB) was investigated in microcosms containing estuarine sediment from Baltimore Harbor. Results showed that the addition of 0.1 g Fe0/g sediment reduced the lag time for removal of doubly flanked para chlorines by approximately 100 days. Because Fe0 is a source of cathodic hydrogen (H2), the effect of direct H2 addition to sediment microcosms was also tested. The addition of 0.001 atm H2 in the headspace generated the same dechlorination activity and reduction in lag time as the addition of 0.1g Fe0/g. Higher concentrations of Fe0 or H2 increased the lag prior to dechlorination. Additional results showed that an alkaline pH (> or = 7.5), high [Fe2+] (3.3 g/L), or HS- (0.1 mg/L total sulfide) inhibited dechlorination. Elevated concentrations of Fe2+, OH-, and HS- are products of Fe0 oxidation or increased microbial activity (methanogenesis, homoacetogenesis, and sulfate reduction), both of which would result from the amendment of large quantities of Fe0 or H2 to sediment. This research shows that not only can PCB dechlorination be stimulated through the addition of electron donor, but implies that the dechlorinators are enriched by the continuous addition of low concentrations of H2, similar to other known dechlorinators, such as the dehalorespirer Dehalococcoides ethenogenes. These results suggest that the direct addition of controlled amounts of Fe0 to sediments may be an effective remediation tool to reduce the lag period prior to dechlorination at PCB-impacted sites. They also suggest that PCB dechlorinators may be enriched using techniques similar to those used with known dehalorespirers.     

Anaerobic toxicity and biodegradability of pulp mill waste constituents

Batch bioassays have been conducted to characterize the response of methanogenic bacteria to several constituents of sulfite evaporator condensate. The results can be grouped into three ranges with increasingly severe consequences to anaerobic reactors: a low concentration, no effect range; a medium concentration range where methanogenesis is temporarily interrupted or slowed down, but may return to normal; and a high concentration range where methanogenesis is permanently inhibited. In some cases the toxicant was metabolized when present in the lower concentration ranges. There was also evidence that mechanisms other than fermentation to methane were significant in accounting for removal of the toxicants from solution. Organisms acclimated to low concentrations of a toxicant are better able to withstand a shock load of that toxicant than are unacclimated organisms.     

Effects of nitrobenzene and zinc on acetate utilizing methanogens

Determination of anaerobic degradation rates and toxic effects of nitrobenzene (NB) on acetate utilizing methanogens was the first objective of this research. Serum bottles were used for anaerobic toxicity assays with an acetate enrichment culture of methanogens. Ten mg/l of nitrobenzene did not inhibit total gas production in the acetate enrichment methanogenic culture. Twenty and thirty mg/l of nitrobenzene caused reversible inhibition of methanogenesis. Batch kinetic experiments showed that 20 mg/l of nitrobenzene was degraded with a first-order rate constant, k, of 0.37 d⁻¹. Acetate was not degraded during the first 7 days when the measured nitrobenzene concentration was higher than about 1 mg/l. The second objective was to determine the effect of zinc on nitrobenzene degradation in methanogenic systems. Ten mg/l of spiked zinc caused a reduction of gas production in the systems with 10 mg/l of nitrobenzene; 20 mg/l of zinc led to failures of systems with 10 and 20 mg/l of nitrobenzene. With 10 and 20 mg/l of added zinc, the k value for nitrobenzene degradation decreased to 0.18 d⁻¹ and 0.14 d⁻¹, respectively. With 20 mg/l of Zn, acetate was not degraded at all even after nitrobenzene concentration reached 0.1 mg/l, indicating toxicity of Zn to methanogenesis. Abiotic control tests with autoclaved culture showed that adsorption alone could remove 60–70% of spiked nitrobenzene in 36 days. However, the samples extracted from solids in the methanogenic test systems showed that nitrobenzene was below the detection limit of 0.1 mg/l, indicating biodegradation of nitrobenzene in these systems. Traces of benzene were seen as an intermediate in the liquid samples. Headspace analysis showed that nitrobenzene and benzene were below detection limits.     

Influence of phase separation on the anaerobic digestion of glucose—I maximum COD-turnover rate during continuous operation

A mineral medium containing 1% of glucose as the main carbon source was subjected to one-phase and to two-phase anaerobic digestion processes under comparable conditions. The one-phase system consisted of an anaerobic up-flow reactor containing both acidogenic as well as methanogenic populations allowing a complete conversion of the carbon source into gaseous end products and biomass.The two-phase system consisted of an acid reactor and a methane reactor connected in series allowing sequential acidogenesis and methanogenesis of the glucose. Performance of the one-phase system and of the methane reactor of the two-phase system is presented by carbon mass balances. By gradually increasing the feed supply to both systems, maximum turnover of COD was determined. Maximum specific sludge loadings of the methanogenic phase of the two-phase system was over 3 times higher than that of the one-phase system. In a second experiment both systems were subjected to overloading, resulting in the accumulation of volatile fatty acids (VFA). In the one-phase system propionate and acetate were formed in considerable amounts. Although acetate disappeared rapidly after cessation of the feed supply, no turnover of propionate was observed within one week. On overloading the methane reactor of the two-phase system accumulation of several fatty acids within the reactor was observed. Rapid conversion of all fatty acids took place immediately after interruption of feed supply.The eco-physiological significance of phase separation is discussed briefly.     

Hydrogen and methane production from household solid waste in the two-stage fermentation process

A two-stage process combined hydrogen and methane production from household solid waste was demonstrated working successfully. The yield of 43 mL H(2)/g volatile solid (VS) added was generated in the first hydrogen production stage and the methane production in the second stage was 500 mL CH(4)/g VS added. This figure was 21% higher than the methane yield from the one-stage process, which was run as control. Sparging of the hydrogen reactor with methane gas resulted in doubling of the hydrogen production. pH was observed as a key factor affecting fermentation pathway in hydrogen production stage. The optimum pH range for hydrogen production in this system was in the range from 5 to 5.5. The short hydraulic retention time (2 days) applied in the first stage was enough to separate acidogenesis from methanogenesis. No additional control for preventing methanogenesis in the first stage was necessary. Furthermore, this study also provided direct evidence in the dynamic fermentation process that, hydrogen production increase was reflected by acetate to butyrate ratio increase in liquid phase.     

Methane microprofiles in a sewage biofilm determined with a microscale biosensor

Microprofiles of the methane concentration in a 3.5-mm-thick sewage outlet biofilm were measured at high spatial and temporal resolution using a microscale biosensor for methane. In the freshly collected biofilm, methane was building up to a concentration of 175 mumol l-1 at 3 mm depth with a total methanogenesis of 0.14 mumol m-2 s-1, as compared to an aerobic respiration (including methane oxidation) of 0.80 mumol m-2 s-1. A model biofilm was established by homogenisation of an in situ biofilm and 12 days of incubation with surplus sodium acetate. The homogenised biofilm was able to maintain 50% of the methanogenic activity in the absence of external electron donor. Oxygen had only a minor effect on the methane production, but aerobic respiration consumed a substantial part of the produced methane and was thus an important control on methane export from the biofilm. A concentration of 2 mmol l-1 nitrate was shown to inhibit methanogenesis only in the upper layer of the biofilm, whereas a further addition of 2 mmol l-1 sulphate inhibited methanogenesis in the entire biofilm. The study demonstrated the power of the methane microsensor in the study of microhabitats with concurrent production and consumption of methane.     

Polyphasic characterization of a PCP-to-phenol dechlorinating microbial community enriched from paddy soil

Dechlorination of PCP has been observed previously under anaerobic condition in paddy soil. However, there is poor information about the dechlorination pathway of PCP and the microbial community associated with the PCP dechlorination in paddy soil. In this study, an anaerobic microbial community dechlorinating PCP was enriched by serial transfers from a paddy soil using a medium containing PCP, lactate and the steam-sterilized paddy soil. The enriched microbial community dechlorinated PCP completely to phenol under the anaerobic condition by a dechlorinating pathway as follows; PCP-->2,3,4,5-tetrachlorophenol-->3,4,5-trichlorophenol-->3,5-dichlorophenol-->3-chlorophenol-->phenol. Intermediate products such as 3-chlorophenol were not accumulated, which were immediately dechlorinated to phenol. The enriched microbial community was characterized physiologically by testing the effects of electron donors and electron acceptors on the dechlorinating activity. The dechlorinating activity was promoted with lactate, pyruvate, and hydrogen as electron donors but not with acetate. Electron acceptors, nitrate and sulphate, inhibited the dechlorinating activity competitively but not iron (III). The microbial group associated with the anaerobic dechlorination was characterized by the effect of specific inhibitors on the PCP dechlorination. Effects of specific metabolic inhibitors and antibiotics indicated the involvement of Gram-positive spore-forming bacteria with the PCP dechlorinating activity, which was represented as bacteria of phylum Firmicutes. The structure of the microbial community was characterized by fluorescence in situ hybridization, quinone profiling, and PCR-DGGE (denaturing gel gradient electrophoresis). The combined results indicated the predominance of Clostridium species of phylum Firmicutes in the microbial community. Desulfitobacterium spp. known as anaerobic Gram-positive spore-forming bacteria dechlorinating PCP were not detected by PCR using a specific primer set. These indicated a probable presence of novel anaerobic Gram-positive spore-forming bacteria dechlorinating PCP in the microbial community.     

Phosphine generation by mixed- and monoseptic-cultures of anaerobic bacteria

A microbial basis for bioreductive generation of phosphine is proposed, which could account at least in part for the presence of this toxic gas in natural anaerobic environments and in sewage and landfill gases. Phosphine generation under anaerobic growth conditions was dependent upon both the culture inoculum source (animal faeces) and enrichment culture conditions. Phosphine was detected in headspace gases from mixed cultures under conditions promoting fermentative growth of mixed acid and butyric acid bacteria, either in the presence or absence of methane generation. Monoseptic cultures of certain mixed acid fermentors (Escherichia coli, Salmonella gallinarum, and Salmonella arizonae) and solvent fermentors (Clostridium sporogenes, Clostridium acetobutyricum and Clostridium cochliarium) also generated phosphine. Such fermentative bacteria participate in the multi-stage process of methanogenesis in nature. Generation of phosphine by these bacteria, rather than by methanoarchaea themselves, could explain the apparent correlation between methanogenesis and the formation of phosphine in nature.     

Biomonitoring of benzene and 1,3-butadiene exposure and early biological effects in traffic policemen

The objective of this study was to determine benzene and 1,3-butadiene exposure through ambient air and personal air monitoring, as well as through biomarkers of exposure, and to evaluate the potential health risk of exposure through the use of biomarkers of early biological effects in central Bangkok traffic policemen. Ambient air concentrations of benzene and 1,3-butadiene at the roadsides were significantly higher than in police offices used as control sites (p < 0.001). Traffic policemen had a significantly higher exposure to benzene (median 38.62 μg/m³) and 1,3-butadiene (median 3.08 μg/m³) than office policemen (median 6.17 μg/m³ for benzene and 0.37 μg/m³ for 1,3-butadiene) (p < 0.001). Biomarkers of benzene exposure, blood benzene, and urinary metabolite, trans, trans-muconic acid were significantly higher in traffic policemen than office policemen (p < 0.001). No significant difference between traffic and office policemen was found in urinary benzene metabolite, S-phenyl mercapturic acid, or in urinary 1,3-butadiene metabolite, monohydroxy-butenyl mercapturic acid. Biomarkers of early biological effects, 8-hydroxy-2′-deoxyguanosine in leukocytes (8-OHdG), DNA-strand breaks, and DNA-repair capacity, measured as an increase in gamma ray-induced chromosome aberrations were significantly higher in traffic policemen than controls (p < 0.001 for 8-OHdG, p < 0.01 for tail length, p < 0.001 for olive tail moment, p < 0.05 for dicentrics and p < 0.01 for deletions). Multiple regression model including individual exposure, biomarkers of exposure, ages and years of work as independent variables showed that only the levels of individual 1,3-butadiene exposure were significantly associated with 8-OHdG and olive tail moment at p < 0.0001 indicating more influence of 1,3-butadiene on DNA damage. These results indicated that traffic policemen, who are exposed to benzene and 1,3-butadiene at the roadside in central Bangkok, are potentially at a higher risk for development of diseases such as cancer than office policemen.