Biological Activity in a Heavily Organohalogen-Contaminated River Sediment (8 pp)

Background, Aims and Scope Sediments of the Spittelwasser creek are highly polluted with organic compounds and heavy metals due to the discharge of untreated waste waters from the industrial region of Bitterfeld-Wolfen, Germany over the course of more than one century. However, relatively few data have been published about the chloroorganic contamination of the sediment. This paper reports on the content of different (chloro)organic compounds with special emphasis on polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), and chlorobenzenes. Existing concepts for the remediation of Spittelwasser sediment include the investigation of natural attenuation processes, which largely depend on the presence of an intact microbial food web. In order to gain more insight in terms of biological activity, we analyzed the capacity of sediment microflora to degrade organic matter by measuring the activities of extracellular hydrolytic enzymes involved in the biogeochemical cycling of carbon, nitrogen, phosphorus and sulfur. Furthermore, the detection of physiologically active bacteria in the sediment, particularly of those known for their capability to reductively dehalogenate organochlorine compounds, illustrates the potential for intrinsic bioremediation processes. Methods PCDD/F and chlorobenzenes were analyzed by gas chromatography(GC)/mass spectrometry and GC/flame ionization detection, respectively. The activities of hydrolytic enzymes were determined from freshly sampled sediment layers using 4-methylumbelliferyl (MUF) or 7-amino-4-methylcoumarin-conjugated model compounds and kinetic fluorescence measurements. Physiologically active bacteria from different sediment layers were microscopically visualized by fluorescence in situ hybridization (FISH). Specific bacteria were identified by 16S rRNA gene amplification and sequencing. Results and Discussion The PCDD/F congener profile was dominated by dibenzofurans. In addition, the presence of specific tetra and pentachlorinated dibenzofurans supported the assumption that extensive magnesium production was one possible source for the high contamination. A range of other chloroorganic compounds, including several isomers of chlorobenzenes, hexachlorocyclohexane and 1,1,1-trichloro-2,2-bis (p-chloro-phenyl)ethane (DDT), was present in the sediment. Activities of extracellular hydrolytic enzymes showed a strong decrease in those sediment layers that were characterized by high contents of absorbable organic halogen (AOX), indicating disturbed organic matter decay. Interestingly, an abnormal increase of cellulolytic enzyme activities below the organochlorine-rich layers was observed, possibly caused by residual cellulose from discharges of sulfite pulping wastes. FISH revealed physiologically active bacteria in most sediment layers from the surface down to the depth of about 60 cm, including members of Desulfitobacterium (D.) and Sulfurospirillum. The presence of D. dehalogenans was confirmed by its partial 16S rRNA gene sequence. Conclusions Results of chemical sediment analyses demonstrated high loads of organochlorine compounds, particularly of PCDD/F. Several years after stopping the waste water discharge to Spittelwasser creek, this sediment remains a main source for pollution of the downstream river system by way of the ongoing mobilization of sediment during high floods. As indicated by our enzyme activity measurements, the decomposition potential for organic matter is low in organochlorine-rich sediment layers. In contrast, the comparably higher enzyme activities in less organochlorine-polluted sediment layers as well as the presence of physiologically active bacteria suggest a considerable potential for natural attenuation. Recommendations and Perspectives From our data we strongly recommend to explore the degradative capacity of sediment microorganisms and the limits for in situ activity towards specific sediment pollutants in more detail. This will give a sound basis for the integration of bioremediation approaches into general concepts to reduce the risk that permanently radiates from this highly contaminated sediment. Submission Editor: Dr. Henner Hollert (Henner.Hollert@urz.uniheidelberg.de)     

Evaluation of ecological disturbance and intrinsic bioremediation potential of pulp mill-contaminated lake sediment using key enzymes as probes

A rapid protocol was developed to measure 10 different enzymic activities from a large number of 1-cm-sliced freshly collected lake sediments. Layers heavily polluted by organic halogens (4900 mg Cl kg(-1)) revealed severe depression of phosphatase, sulfatase, leucine-aminopeptidase, chitinase, acetate esterase and butyrate esterase activities as compared to layers above and below the most polluted zone. alpha-Glucosidase, beta-glucosidase, beta-xylosidase and palmitate esterase were less affected. Methane oxidation potential was dramatically depressed in the polluted strata whereas tetrachloromethane dehalogenating activity was observed in the polluted sediment only. The sediment layers formed after the chlorine discharges into the lake had diminished to 1/10, and showed restoration of the activities close to those observed in non-recipient sediment, in spite of the persisting presence of >1000 mg of organic chlorine (kg dry wt)(-1). We conclude that certain enzymic activities involved in breakdown or oxidation of organic matter in the sediments are useful probes for assessing the degree of ecological damage and its potential for restoration in recipient lakes of industrial discharges.     

Soil contamination at abandoned sawmill areas

Contamination of soil by technical chlorophenol formulation at wood preserving sites was studied. The examined soils contained 50 to 1000 mg of polychlorinated phenols (CPs), 1 to 50 mg of polychlorinated phenoxyphenols (“pre-dioxins”, PCPPs) and 0.1 to 5 mg of polychlorinated dibenzofurans (PCDFs) per kg of dry soil. CPs were found to be mobile, leaching deep into the soil, while PCPPs and PCDFs accumulated in the top soil. All three classes of contaminants were stable in soil.     

Chlorinated phenols as contaminants of soil and water in the vicinity of two Finnish sawmills

Contamination of soil and water in the vicinity of two sawmills using preservative against blue staining fungi that contained chlorophenols was studied. The soil around the treatment basins was found to contain up to 70 mg chlorophenols per kg and that in the storage area for treated lumber up to 6 mg per kg. Contamination extended to a depth of at least 2 meters near the treatment basins. Surface water inside the sawmill area was found to contain the same chlorophenols as those used in wood preservation, plus some additional isomers. The ground water and lake water around the sawmill areas was found to be contaminated by wood preservative.     

Pyrene degradation in forest humus microcosms with or without pine and its mycorrhizal fungus

The mineralization potential of forest humus and the self-cleaning potential of a boreal coniferous forest environment for polycyclic aromatic hydrocarbon (PAH) compounds was studied using a model ecosystem of acid forest humus (pH = 3.6) and pyrene as the model compound. The matrix was natural humus or humus mixed with oil-polluted soil in the presence and absence of Scots pine (Pinus sylvestris L.) and its mycorrhizal fungus (Paxillus involutus). The rates of pyrene mineralization in the microcosms with humus implants (without pine) were initially insignificant but increased from Day 64 onward to 47 microg kg(-1) d(-1) and further to 144 microg kg(-1) d(-1) after Day 105. In the pine-planted humus microcosms the rate of mineralization also increased, reaching 28 microg kg(-1) d(-1) after Day 105. The 14CO2 emission was already considerable in nonplanted microcosms containing oily soil at Day 21 and the pyrene mineralization continued throughout the study. The pyrene was converted to CO2 at rates of 0.07 and 0.6 microg kg(-1) d(-1) in the oily-soil implanted microcosms with and without pine, respectively. When the probable assimilation of 14CO2 by the pine and ground vegetation was taken into account the most efficient microcosm mineralized 20% of the 91.2 mg kg(-1) pyrene in 180 d. The presence of pine and its mycorrhizal fungus had no statistically significant effect on mineralization yields. The rates of pyrene mineralization observed in this study for forest humus exceeded the total annual deposition rate of PAHs in southern Finland. This indicates that accumulation in forest soil is not to be expected.     

Chemical and microbiological hazards associated with recycling of anaerobic digested residue intended for agricultural use

In the present study, three full-scale biogas plants (BGP) were investigated for the concentration of heavy metals, organic pollutants, pesticides and the pathogenic bacteria Bacillus cereus and Escherichia coli in the anaerobically digested residues (ADR). The BGPs mainly utilize source-separated organic wastes and industrial food waste as energy sources and separate the ADR into an ADR-liquid and an ADR-solid fraction by centrifugation at the BGP. According to the Norwegian standard for organic fertilizers, the ADR were classified as quality 1 mainly because of high zinc (132-422 mg kg(-1) DM) and copper (23-93 mg kg(-1) DM) concentrations, but also because of high cadmium (0.21-0.60 mg kg(-1) DM) concentrations in the liquid-ADR. In the screening of organic pollutants, only DEHP (9.7-62.1 mg kg(-1)) and ∑ PAH 16 (0.2-1.98 mg kg(-1) DM) were detected in high concentrations according to international regulations. Of the 250 pesticides analyzed, 11 were detected, but only imazalil (<0.30-5.77 mg kg(-1) DM) and thiabendazol (<0.14-0.73 mg kg(-1) DM) were frequently detected in the ADR-fiber. Concentrations of imazalil and thiabendazol were highest during the winter months, due to a high consumption of citrus fruits in Norway in this period. Ten percent of the ADR-liquid samples contained cereulide-producing B. cereus, whereas no verotoxigenic E. coli was detected. The authors conclude that the risk of chemical and bacterial contamination of the food chain or the environment from agricultural use of ADR seems low.     

Sediment accumulation of organic halogens in pristine forest lakes

Sediment accumulation of organic halogen was studied in two forest lakes, one pristine and one which received 30 m³ of biologically purified bleaching wastewater from a kraft pulp mill in 1979 equivalent to ca. 2 kg of adsorbable organic halogen (AOX). Lake sediments were dated with²¹⁰Pb,¹³⁴Cs and¹³⁷Cs and the annual deposition rates of organic halogens and organic matter were calculated. Organic bound halogen contents of the sediment aged 150 years was 180 μg Cl g^?1 d.w. in both lakes. The concentration of organic bound halogen at the topmost 6 cm of the sediments (less than 20-years-old) ranged from 45 to 80 μg Cl g^?1 d.w. This suggests that solvent extractable halogen had enriched in the older sediment layers. The deposition of extractable organic halogen (EOX) in the lakes in 1950’s was 4 to 5 mg Cl m^?2 a^?1. Since then, the depositon of EOX doubled in both lakes. The deposition of organic matter increased concomitantly from 50 g m^?2 a^?1 to 110 g m^?2 a^?1 in Lake Mustalampi and from 35 g m^?2 a^?1 to 62 g m^?2 a^?1 in Lake Pyylampi suggesting that the increase in the deposition of organic halogen followed the increase in the deposition of organic matter. Of the 2 kg of organic halogen discharged into the lake, 5% or less was detected in the sediment in tetrahydrofuran extractable form 15 years later.     

Nitrification in polluted soil fertilized with fast- and slow-releasing nitrogen: a case study at a refinery landfarming site

The nitrifying activity and the effect of fertilization with urea and methylene urea were studied in a landfarming site. The site has been operative over 20 years and maintained by heavy nitrogen fertilization. The landfarming soil contained 4-6% (w/w) oil. The nitrate accumulation was 20-50mg NO3-N day(-1)kg(-1) observed after methylene urea fertilization of 889 g Nm(-2). Nitrification ex situ (in laboratory conditions) was 8.8 mg NO3-N day(-1) kg(-1) in the presence of 380 mg kg(-1) NH4+-N. The half-saturation concentration of nitrification was more than 200 mg NH4+-N kg(-1). The results show that nitrification was active in soil with high oil concentration. Urea fertilization of 893 g Nm(-2) caused an increase of soil NH4+-N concentration up to 5500 mg kg(-1) and pH>8.5. This led to inhibition of nitrification, which persisted after NH4+ concentration decreased below 200mg NH4+ kg(-1).     

Bleached kraft pulp mill discharged organic matter in recipient lake sediment

Environmental properties of organic matter contained halogen and sulfur were studied in sediments of bleached kraft pulp mill effluent (BKME) recipient lakes and 2 m³ outdoor enclosures (mesocosms). The BKME contributed to 1% (v/v) of the total water flow in the lake downstream of the pulp mill where the sediments contained 1.7 to 4 mg of tetrahydrofuran extractable organic halogen (EOX-Cl) and 0.6 to 0.8 mg of tetrahydrofuran extractable organic sulfur (EOS-S) g⁻¹ of organic matter. Upstream sediment contained 0.03 mg of EOXCl and 0.7 mg of EOS-S g⁻¹ of organic matter. EOX was a better indicator for the influence of BKME in the recipient sediment than EOS. The polarity of BKME contained EOX corresponded to log K^ow of < 1, and that of the downstream sediment contained EOX to > 4.5. HP-SEC analysis of the molecular weight distribution (MWD) of the EOX showed a peak between 300 to 600 g mol⁻¹ for the BKME and between 1000 to 2000 g mol⁻¹ for the downstream sediment. The MWD of the BKME contained EOS peaked at 300 to 1000 g mol⁻¹, and that of the downstream sediment contained EOS at 1000 to 5000 g mol⁻¹. These results indicate that BKME contained organic halogen and sulfur undergo major structural transformations when incorporated into sediment. The biota-to-sediment accumulation factor (BSAF) of EOX from sediments formed downstream of the mill and in the mesocosms to the lipids ofLumbriculus variegatus was 0.4 to 0.7. This is of a similar order of magnitude to the BSAF reported for 2,3,7,8-tetrachlorodibenzop-dioxin and 2,3,7,8-tetrachlorodibenzofuran.