The use of Magallanic peat as non-conventional sorbent for EDTA removal from wastewater

Kraft mills are responsible for large volumes discharges of highly polluted effluents. Application of new bleaching processes (i.e. total chlorine-free (TCF) process) is already a feasible option to reduce environmental impacts. The current trend in the increase in the production of TCF pulp will proportionally increase the consumption of chelating agents. The most commonly used chelants, ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DPTA) are supposed to be relatively persistent substances, poorly degradable in biological treatment facilities and are subsequently considered as environmentally critical compounds. Adsorption could be used as a treatment technique to remove recalcitrant compounds from wastewaters. However, in most cases, sorbent and regeneration costs can make the whole process not economically feasible. The goal of this study was to evaluate the use of Magallanic peat as non-conventional sorbent for EDTA removal from wastewater. Adsorption studies were carried out considering a 2(3) factorial design. pH, temperature and sorbent/sorbate (S/S) relationship effects were evaluated in EDTA adsorption onto Magallanic peat. In addition, adsorption isotherm constants were determined according to the Langmuir and Freundlich models. The results showed that the optimal conditions for EDTA adsorption onto Magallanic peat were 20 degrees C, acid pH (4.0) and a low sorbent/sorbate ratio (0.1/100). At these conditions Magallanic peat showed an adsorption capacity for EDTA (Cs(sat)) of 128.2mg/g, comparable and even better than activated carbon (Cs(sat) 56.5mg/g). EDTA adsorption data at 60 degrees C obtained are not shown due to Magallanic peat degradation phenomena.     

Molecular weight distribution of Pinus radiata kraft mill wastewater treated by anaerobic digestion

Kraft mill is responsible for massive discharge of highly polluted effluents. The main characteristics of this effluent are high toxicity and low biodegradability due to tannin, lignin and chlorophenol compounds. The composition may vary dramatically depending, for instance, on the utilised feedstock and process. The purpose of this work was to investigate the molecular weight distribution of Pinus radiata kraft pulping wastewater treated by anaerobic digestion by using two types of anaerobic reactors: fixed bed and sludge blanket. Anaerobic sludge blanket (UASB) and anaerobic filter (AF) were operated. In both reactors, the total alkalinity ranged between 1.0 and 1.5 g CaCO3/l, while the organic load rate (OLR) was increasing during operation from 1.2 to 3.3 gCOD/l d. COD and total phenolic compounds (UV215) removal ranged between 30-50% and 13-20%, respectively, while the BOD5 removal ranged 60-90%. However only a partial biodegradation (10-43%) of tannin and lignin was observed. Results from ultrafiltration analyses indicated that the fraction with a molecular weight (MW) < 1000, COD and colour decreased after anaerobic treatment, but the total phenolic compounds increased. In the 1000 < MW < 10,000 fraction, there was no change in COD, UV215 and colour. In the > 10,000 MW fraction, colour and COD fraction increased by 14% and 5%, respectively, after anaerobic treatment. It can be concluded from this study, that treatment with UASB or AF reactors is not enough, under the conditions tested, for a large COD removal from Pinus radiata wastewater.     

Treatment of recycled wastewaters from fishmeal factory by an anaerobic filter

Fishmeal industries processes produce effluents with high load organic matter. These effluents, after recycling and physical-chemical pretreatment, have a high organic content (5-6 g COD/l), proteins (3-5 g/l), salinity close to sea water, sodium chloride (30 g/l) and sulphate (1-3.3 g/l). An anaerobic filter was used for the treatment of this wastewater, with marine sediment as anaerobic inoculum. Anaerobic filter removed up to 70% of the influent COD concentrations at organic loading rates (OLR) of 9.5 and 14.3 (g/l d) and sulphate up to 80% at OLR of 7.1 and 14.3 (g/l d) whereas the pH ranged between 7.0 and 7.5. These results show that anaerobic filter systems are applicable to recycled wastewaters from fishmeal.     

Biodegradation of chlorinated phenolic compounds

Chlorophenolic compounds are generated from a number of industrial manufacturing processes including pulp and paper manufacture. These compounds are found to be toxic and recalcitrant and hence their discharge into the environment must be regulated. Slow and partial degradation of chlorophenols under aerobic and anaerobic natural environment has been observed. Aerobic biodegradation of chlorophenols proceeds through the formation of catechols while under anaerobic conditions, reductive dehalogenation is the preferred metabolic pathway. Number and position of chlorine substituents on the phenolic ring has influence on the rate and extent of biodegradation of chlorophenols. In engineered systems, acclimatization of biomass to chlorophenols markedly enhances the biodegradation ability by reducing the initial lag phase and by countering inhibition. Partial removal of chlorophenols between 40-60% is usually observed in aerobic and anaerobic processes. Removal can be enhanced by a combination of aerobic and anaerobic operations.     

Novel application of oxygen-transferring membranes to improve anaerobic wastewater treatment

Anaerobic biological wastewater treatment has numerous advantages over conventional aerobic processes; anaerobic biotechnologies, however, still have a reputation for low-quality effluents and operational instabilities. In this study, anaerobic bioreactors were augmented with an oxygen-transferring membrane to improve treatment performance. Two anaerobic bioreactors were fed a synthetic high-strength wastewater (chemical oxygen demand, or COD, of 11,000 mg l(-1)) and concurrently operated until biomass concentrations and effluent quality stabilized. Membrane aeration was then initiated in one of these bioreactors, leading to substantially improved COD removal efficiency (> 95%) compared to the unaerated control bioreactor (approximately 65%). The membrane-augmented anaerobic bioreactor required substantially less base addition to maintain circumneutral pH and exhibited 75% lower volatile fatty acid concentrations compared to the unaerated control bioreactor. The membrane-aerated bioreactor, however, failed to improve nitrogenous removal efficiency and produced 80% less biogas than the control bioreactor. A third membrane-augmented anaerobic bioreactor was operated to investigate the impact of start-up procedure on nitrogenous pollutant removal. In this bioreactor, excellent COD (>90%) and nitrogenous (>95%) pollutant removal efficiencies were observed at an intermediate COD concentration (5,500 mg l(-1)). Once the organic content of the influent wastewater was increased to full strength (COD = 11,000 mg l(-1)), however, nitrogenous pollutant removal stopped. This research demonstrates that partial aeration of anaerobic bioreactors using oxygen-transferring membranes is a novel approach to improve treatment performance. Additional research, however, is needed to optimize membrane surface area versus the organic loading rate to achieve the desired effluent quality.     

Batch and continuous culture-based selection strategies for acetic acid tolerance in xylose-fermenting Saccharomyces cerevisiae

Acetic acid tolerance of Saccharomyces cerevisiae is crucial for the production of bioethanol and other bulk chemicals from lignocellulosic plant-biomass hydrolysates, especially at a low pH. This study explores two evolutionary engineering strategies for the improvement of acetic acid tolerance of the xylose-fermenting S. cerevisiae RWB218, whose anaerobic growth on xylose at pH 4 is inhibited at acetic acid concentrations >1 g L(-1) : (1) sequential anaerobic, batch cultivation (pH 4) at increasing acetic acid concentrations and (2) prolonged anaerobic continuous cultivation without pH control, in which acidification by ammonium assimilation generates selective pressure for acetic acid tolerance. After c. 400 generations, the sequential-batch and continuous selection cultures grew on xylose at pH≤4 with 6 and 5 g L(-1) acetic acid, respectively. In the continuous cultures, the specific xylose-consumption rate had increased by 75% to 1.7 g xylose g(-1) biomass h(-1) . After storage of samples from both selection experiments at -80 °C and cultivation without acetic acid, they failed to grow on xylose at pH 4 in the presence of 5 g L(-1) acetic acid. Characterization in chemostat cultures with linear acetic acid gradients demonstrated an acetate-inducible acetic acid tolerance in samples from the continuous selection protocol.     

Toxic effects of thiol-reactive compounds on anaerobic biomass

The objective of this study was to characterize the toxic effects of three well known thiol-reactive electrophilic compounds, N-ethylmaleimide (NEM), pentachlorophenol (PCP) and 1-chloro-2,4-dinitrobenzene (CDNB) on anaerobic biotransformation process. The work was part of a larger investigation on potassium efflux as a possible response mechanism of anaerobic microorganisms to the presence of thiol-reactive organic compounds and the interference of such compounds on the reductive dehalogenation process. Using anaerobic toxicity assay (ATA) and granular anaerobic biomass from a full-scale upflow anaerobic sludge blanket (UASB) reactor, inhibitory concentrations of these compounds that reduced the microbial activity of granular biomass to 50% of a control (IC50) were determined to be 592, 0.97, and 450 mg/l for NEM, PCP, and CDNB, respectively. Toxicity of NEM was also tested on anaerobic biomass from a municipal wastewater treatment plant digester and slightly lower IC50 of 532 mg/l was obtained. The results presented here indicate that anaerobic biomass can acclimate to the three thiol-reactive compounds studied and recover from inhibition as long as the toxicant concentration is below a threshold level. That threshold concentration was found to be 500 mg/l for NEM on biomass from the municipal digester, 1 mg/l for PCP, and 500 mg/l for CDNB, both on granular biomass. Granular anaerobic biomass showed recovery even at NEM concentrations of 1000 mg/l.     

A kinetic analysis and experimental validation of an integrated system of anaerobic filter and biological aerated filter

An anaerobic/aerobic filter (AF/BAF) system was developed treating dairy wastewater. The influent was blended with recirculated effluent to allow for pre-denitrification in the AF followed by nitrification in the BAF. The recirculation ratio ranged 100-300%. The average chemical oxygen demand (COD) removal efficiency was 79.8-86.8% in the AF and the average total nitrogen removal efficiency was 50.5-80.8% in the AF/BAF system. Steady-state mass balances on the AF were used to analyze removal kinetics in the AF. The kinetic model values for effluent COD in the AF were overestimated as compared with experimental data. The integrated suspended and attached biomass growth rates in the AF were estimated. The specific growth rate of the integrated biomass at each recirculation ratios was 0.6213, 0.6647, and 1.20831/day, respectively. The increase in specific growth rate corresponded to increases in biomass sloughing as the recirculation ratio increased.     

Anaerobic degradation of adsorbable organic halides (AOX) from pulp and paper industry wastewater

Adsorbable organic halides (AOX) are generated in the pulp and paper industry during the bleaching process. These compounds are formed as a result of reaction between residual lignin from wood fibres and chlorine/chlorine compounds used for bleaching. Many of these compounds are recalcitrant and have long half-life periods. Some of them show a tendency to bioaccumulate while some are proven carcinogens and mutagens. Hence, it is necessary to remove or degrade these compounds from wastewater. Physical, chemical and electrochemical methods reported to remove AOX compounds are not economically viable. Different types of aerobic, anaerobic and combined biological treatment processes have been developed for treatment of pulp and paper industry wastewater. Maximum dechlorination is found to occur under anaerobic conditions. However, as these processes are designed specifically for reducing COD and BOD of wastewater, they do not ensure complete removal of AOX. This paper reviews the anaerobic biological treatments developed for pulp and paper industry wastewater and also reviews the specific micro-organisms reported to degrade AOX compounds under anaerobic conditions, their nutritional and biochemical requirements. It is imperative to consider these specific micro-organisms while designing an anaerobic treatment for efficient removal of AOX.     

Effect of sulfur compounds on biological reduction of nitric oxide in aqueous Fe(II)EDTA2- solutions.

Biological reduction of nitric oxide (NO) in aqueous solutions of EDTA chelated Fe(II) is one of the main steps in the BioDeNOx process, a novel bioprocess for the removal of nitrogen oxides (NOx) from polluted gas streams. Since NOx contaminated gases usually also contain sulfurous pollutants, the possible interferences of these sulfur compounds with the BioDeNOx process need to be identified. Therefore, the effect of the sulfur compounds Na2SO4, Na2SO3, and H2S on the biological NO reduction in aqueous solutions of Fe(II)EDTA2- (25 mM, pH 7.2, 55 degrees C) was studied in batch experiments. Sulfate and sulfite were found to not affect the reduction rate of Fe(II)EDTA2- complexed NO under the conditions tested. Sulfide, either dosed externally or formed during the batch incubation out of endogenous sulfur sources or the supplied sulfate or sulfite, influences the production and consumption of the intermediate nitrous oxide (N2O) during Fe(II)EDTA2- bound NO reduction. At low concentrations (0.2 g VSS/l) of denitrifying sludge, 0.2 mM free sulfide completely inhibited the nitrosyl-complex reduction. At higher biomass concentrations (1.3-2.3 g VSS/l), sulfide (from 15 microM to 0.8 mM) induced an incomplete NO denitrification with N2O accumulation. The reduction rates of NO to N2O were enhanced by anaerobic sludge, presumably because it kept FeEDTA in the reduced state.     

Phytoextraction of copper from contaminated soil by Elsholtzia splendens as affected by EDTA, citric acid, and compost

Phytoextraction of copper (Cu) from contaminated soils greatly depends on the metal bioavailability in the soils and metal uptake ability of the plant. In this study, the effects of chelators [ethylenediamine tetraacetic acid (EDTA), citric acid (CA)] and compost amendments on Cu phytoextraction potential by a tolerant and accumulating plant species (E. splendens) were examined in two types of contaminated soils, ie., the mined soil from Cu-mined area (MS) and a paddy soil polluted by Cu refining (PS). The results showed that EDTA application at 2.5-5.0 mmol kg(-1) increased phytoextraction of Cu by four- and eight-fold from both MS and PS, respectively, which is mainly attributed to increased H2O extractable Cu in the soil. The Cu amount extracted by the shoots of E. splendens reached 800-1000 microg Cu plant(-1) from the MS and 400-700 microg Cu plant(-1) from the PS at EDTA application rates of 2.5-5.0 mmol kg(-1). The application of CA at 5.0 mmol kg(-1) had minimal effects on Cu extractability in both soils and slightly decreased Cu extraction efficiency by E. splendens. Plant biomass production was enhanced by CA at 0.25 mmol L(-1) in nutrient solution, but inhibited by CA at 5.0 mmol kg(-1) in both MS and PS. Increasing the compost rate significantly decreased H2O extractable Cu in the MS, but raised H2O-extractable Cu in the PS, which resulted mainly front the reduced exchangeable Cu in the MS and the increased exchangeable and organic fractions of Cu in the PS by compost. At high compost rate (5%), the shoots of E. splendens extracted 3.6-fold higher Cu from the PS than from the MS. These results indicate that, among the soil amendments, efficiency of Cu phytoextraction is enhanced mostly by 2.5-5.0 mmol kg(-1) EDTA, followed by 5% (w:w) compost, whereas < 5.0 mmol kg(-1) CA has minimal effects on Cu phytoextraction by E. splendens in the PS. As for the MS, only 2.5-5.0 mmol kg(-1) EDTA can elevate the efficiency of Cu, while 5% compost amendment and < 5.0 mmol kg(-1) CA application have no marked effects on Cu phytoextraction by E. splendens.     

Flocculants effect in biomass retention in a UASB reactor treating dairy manure

The performance and biomass retention of an upflow anaerobic sludge bed (UASB) reactor treating liquid fraction of dairy manure has been investigated at several organic loading rates. Two identical UASB reactors were employed. The biomass of one UASB reactor (FBR) had previously been treated with a cationic polyacrylamide, the other reactor was operated as a control reactor (CR). At 3 and 2 days of HRT both reactors functioned similarly, but at 1.5 days HRT some differences were observed between both effluents. Mean COD(T) removal percentages were 83.4% and 76.5%; COD(VFA) values in effluents were 977 and 2682 mg l(-1) for the FBR and the CR respectively. The VSS initial value in both reactors was 25.66 g VSS, whereas after the experiment the quantities were 31.83 g VSS in the FBR and 23.18 g VSS in the CR reactors. Polymer addition resulted in a higher degree of biomass retention and better performance in the FBR reactor.     

Mesophilic anaerobic treatment of sludge from saline fish farm effluents with biogas production

The mesophilic anaerobic treatment of sludge from saline fish farm effluents (total solids (TS): 8.2-10.2 wt%, chemical oxygen demand (COD): 60-74 g/l, sodium (Na): 10-10.5 g/l) was carried out in continuously stirred tank reactors (CSTRs) at 35 degrees C. COD stabilization between 36% and 55% and methane yields between 0.114 and 0.184 l/g COD added were achieved. However, the process was strongly inhibited, presumably by sodium, and unstable, with propionic acid being the main compound of the volatile fatty acids (VFA). When diluting the sludge 1:1 with tap water (Na: 5.3 g/l), the inhibition could be overcome and a stable process with low VFA concentrations was achieved. The results of the study are used to make recommendations for the configuration of full-scale treatment plants for the collected sludge from one salmon farming licence and to estimate the energy production from these plants.     

Growth and production of biomass of Rhodovulum sulfidophilum in sardine processing wastewater

AIMS: Rhodovulum sulfidophilum was grown in sardine processing wastewater to assess growth characteristics for the production of bacterial biomass with simultaneous reduction of chemical oxygen demand. METHODS AND RESULTS: Growth characteristics were compared in diluted and undiluted, settled and non-settled wastewater growing in anaerobic light and aerobic dark conditions; and also at different agitation speeds. The highest biomass (8.75 g l(-1)) and a reduction in chemical oxygen demand of 71% were obtained in unsettled, undiluted wastewater after 120 h culture with 15% inoculum. In settled wastewater, highest biomass (7.64 g l(-1)) and a COD reduction of 77% was also obtained after 120 h. Total biomass was higher (4.34 g l(-1)) after 120 h culture in anaerobic light compared to (3.23 g l(-1)) in aerobic dark growth. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Better performance, mean of total biomass (6.97 g l(-1) after 96 h), total carotenoids (4.24 mg g(-1) dry cell from 24 h) and soluble protein (431 microg ml(-1) after 96 h) were obtained from aerobic dark culture at 300 rev min(-1). The COD reduction, however, was lower (69%) after 96 h culture. Thus, the benefits in the production of bacterial biomass in non-sterilized sardine processing wastewater with the reduction of chemical oxygen demand could be achieved.     

Post-treatments of anaerobic effluents

Post-treatments are necessary if anaerobic effluents need to be discharged into surface waters, because anaerobic digestion alone is not able to produce effluents that can meet the discharge standards applied in most industrialized countries, particularly for suspended solids, particulate COD, nitrogen, phosphorus and sulphides. This paper has the aim to present some results obtained in the recent years in our laboratory, where different comprehensive processes that include anaerobic digestion have been studied. Discussion will regard: 1) the ANANOX (ANaerobic-ANoxic-OXic) process for the treatment of municipal wastewater; 2) a process studied for the biological removal of C, N and P from piggery wastewater that has a hybrid anaerobic/anoxic reactor as the first treatment step; 3) the use of a Sequencing Batch Reactor for the post-treatment of digested cheese whey mixed with cheese factory cleaning waters.Abbreviations ABR Anaerobic Baffled Reactor - Bv organic volumetric loading rate (gCOD·L^-1·d^-1) - BV bed volumes - F/M food to microorganism ratio or sludge loading rate (gCOD·g.VSS^-1·d^-1) - HRT hydraulic retention time (t) - JHB University of Johannesburg nutrient removal process - p.e. person equivalent - Qrd recycle for denitrification flow rate - Qrs sludge recycle flow rate - SBR Sequencing Batch Reactor - TKN Total Kjeldahl Nitrogen - VCF Volumetric Concentration Factor (vol. permeate/vol. retentate)     

Physiology of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures

The physiology of Saccharomyces cerevisiae CBS 8066 was studied in anaerobic glucose-limited chemostat cultures in a mineral medium supplemented with ergosterol and Tween 80. The organism had a mu max of 0.31 h-1 and a Ks for glucose of 0.55 mM. At a dilution rate of 0.10 h-1, a maximal yield of 0.10 g biomass (g glucose)-1 was observed. The yield steadily declined with increasing dilution rates, so a maintenance coefficient for anaerobic growth could not be estimated At a dilution rate of 0.10 h-1, the yield of the S. cerevisiae strain H1022 was considerably higher than for CBS 8066, despite a similar cell composition. The major difference between the two yeast strains was that S. cerevisiae H1022 did not produce acetate, suggesting that the observed difference in cell yield may be ascribed to an uncoupling effect of acetic acid. The absence of acetate formation in H1022 correlated with a relatively high level of acetyl-CoA synthetase. The uncoupling effect of weak acids on anaerobic growth was confirmed in experiments in which a weak acid (acetate or propionate) was added to the medium feed. This resulted in a reduction in yield and an increase in specific ethanol production. Both yeasts required approximately 35 mg oleic acid (g biomass)-1 for optimal growth. Lower or higher concentrations of this fatty acid, supplied as Tween 80, resulted in uncoupling of dissimilatory and assimilatory processes.     

Impact of chelator-induced phytoextraction of cadmium on yield and ionic uptake of maize

Enhanced phytoextraction uses soil chelators to increase the bioavailability of heavy metals. This study tested the effectiveness of ethylenediaminetetraacetic acid (EDTA) and citric acid in enhancing cadmium (Cd) phytoextraction and their effects on the growth, yield, and ionic uptake of maize (Zea mays). Maize seeds of two cultivars were sown in pots treated with 15 (Cd₁₅) or 30 mg Cd kg^?1 soil (Cd₃₀). EDTA and citric acid at 0.5 g kg^?1 each were applied 2 weeks after germination. Results demonstrated that the growth, yield per plant, and total grain weight were reduced by exposure to Cd. EDTA increased the uptake of Cd in shoots, roots, and grains of both maize varieties. Citric acid did not enhance the uptake of Cd, rather it ameliorated the toxicity of Cd, as shown by increased shoot and root length and biomass. Cadmium toxicity reduced the number of grains, rather than the grain size. The maize cultivar Sahiwal-2002 extracted 1.6% and 3.6% of Cd from soil in both Cd+ EDTA treatments. Hence, our study implies that maize can be used to successfully phytoremediate Cd from soil using EDTA, without reducing plant biomass or yield.     

Phenol degradation and colour removal in submerged culture of Pleurotus sajor-caju with paper mill effluents

The fungus Pleurotus sajor-caju secretes phenol-oxidases that enable the use of recalcitrant compounds as substrates. The residues of paper manufacture contain high lignin levels, which gives the effluents a characteristic brownish colour. To test the potential of P. sajor-caju cultures on reducing these parameters, we used 90% of raw effluents from medium consistency oxygen delignification and bleaching stages plus 10% of mineral solution and different levels of glucose (5-15 g L^-1) as substrate. We observed a greater fungal biomass in cultures using effluent than in controls. Cultures containing 10 to 15 g L^-1 of glucose resulted in about 42% colour reduction. The polyphenol content was also reduced by 58.9% by the 13th day of culture. In addition, we observed the secretion of laccases (211.44 U mL^-1 and 45.98 U mL^-1 using ABTS and syringaldazine, respectively) and peroxidases (6.11 U mL^-1-ABTS) both peaking at the 7th day of culture and with similar kinetics of production in different glucose concentrations.     

Whole-cell hybridization of Methanosarcina cells with two new oligonucleotide probes.

Two new oligonucleotide probes targeting the 16S rRNA of the methanogenic genus Methanosarcina were developed. The probes have the following sequences (Escherichia coli numbering): probe SARCI551, 5'-GAC CCAATAATCACGATCAC-3', and probe SARCI645, 5'-TCCCGGTTCCAAGTCTGGC-3'. In situ hybridization with the fluorescently labelled probes required several modifications of standard procedures. Cells of Methanosarcina mazeii S-6 were found to lyse during the hybridization step if fixed in 3% formaldehyde and stored in 50% ethanol. Lysis was, however, not observed with cells fixed and stored in 1.6% formaldehyde-0.85% NaCl. Extensive autofluorescence of the cells was found upon hybridization in the presence of 5 mM EDTA, but successful hybridization could be obtained without addition of this compound. The mounting agent Citifluor AF1, often used in conjugation with the fluorochrome fluorescein, was found to wash the labelled probes out of the cells. Stable labelling could be obtained with rhodamine-labelled probes when the specimen was mounted in immersion oil, and high hybridization intensities of the Methanosarcina cells were found even in the presence of biomass from an anaerobic reactor. The inherent high autofluorescence of the biomass could be lowered by use of a highly specific narrow-band filter. The probes were found to be specific for Methanosarcina and useful for detection of this genus in samples from anaerobic reactors.     

Bacterial metabolism of chlorinated dehydroabietic acids occurring in pulp and paper mill effluents

Chlorinated dehydroabietic acids are formed during the chlorine bleaching of wood pulp and are very toxic to fish. Thus, destruction of these compounds is an important function of biological treatment systems for pulp and paper mill effluents. In this study, 12 strains of diverse, aerobic resin acid-degrading bacteria were screened for the ability to grow on chlorinated dehydroabietic acids as sole organic substrates. All seven strains of the class Proteobacteria able to use dehydroabietic acid were also able to use a mixture of 12- and 14-chlorodehydroabietic acid (Cl-DhA). None of the strains used 12,14-dichlorodehydroabietic acid. Sphingomonas sp. strain DhA-33 grew best on Cl-DhA and simultaneously removed both Cl-DhA isomers. Ralstonia sp. strain BKME-6 was typical of most of the strains tested, growing more slowly on Cl-DhA and leaving higher residual concentrations of Cl-DhA than DhA-33 did. Strains DhA-33 and BKME-6 mineralized (converted to CO(inf2) plus biomass) 32 and 43%, respectively, of carbon in Cl-DhA consumed. Strain DhA-33 produced a metabolite from Cl-DhA, tentatively identified as 3-oxo-14-chlorodehydroabietin, and both strains produced dissolved organic carbon which may include unidentified metabolites. Cl-DhA removal was inducible in both DhA-33 and BKME-6, and induced DhA-33 cells also removed 12,14-dichlorodehydroabietic acid. Based on activities of strains DhA-33 and BKME-6, chlorinated DhAs, and potentially toxic metabolite(s) of these compounds, are relatively persistent in biological treatment systems and in the environment.