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Influence of diet on the bioaccumulation of PCBs.

The levels of several chlorinated biphenyls (PCBs) congeners were analysed in muscle of flounder (Platichthys flesus), which were kept in semi-field condition for 1 month. The influence of resuspended sediments and two diets (worms and shrimps) on PCB accumulation in these fish was tested. Highest concentrations were among PCB 138 and 153 in both abiotic (sediment and suspended particulate matter (SPM)) and biotic compartments (shrimps, worms and flounder). Whereas all PCB congeners vary in concentration between polluted and non-polluted sediment, SPM and mussels, only high chlorinated congeners (101, 105, 153, 180 and 187) are significantly different between the worms and shrimps. Differences in bioaccumulation of PCBs by fish induced by differences in food composition were not found. For the low chlorinated compounds, however, pollution exposure might account for the higher concentration found in exposed fish.

Animals↗

From sediment bioassay to fish biomarker--connecting the dots using simple trophic relationships.

Two common problems in applying and interpreting invertebrate bioassays and fish biomarkers in sediment toxicology are the wide gap between significant effects concentrations determined by these two approaches, and a general lack of ecological context. We have devised an exposure system that is able to reconcile much of the disparity between invertebrate bioassay and fish biomarker results by incorporating realistic ecological processes based on deposit feeding and predator-prey interactions. This system relates the disturbance of interest (sediment contamination) to biologically meaningful effects in a resource of interest (marine flatfish) via a realistic contaminant vector (a deposit-feeding polychaete worm). In this pilot study, polychaetes (Armandia brevis) were exposed for 28 days to clean sediments supplemented with benzo(a)pyrene (BaP), para-para dichlorodiphenyldichloroethylene (pp'DDE), Aroclor 1254, or field sediments collected from two sites in Puget Sound, Washington, contaminated predominantly with polcyclic aromatic hydrocarbons (PAHs) or chlorinated compounds. Exposed worms were then fed live to juvenile English sole (Pleuronectes vetulus) for 10 or 12 days. At the end of the exposure period, fish were measured for length and weight, sacrificed, and preserved for either routine histopathology and immunohistochemical analysis of cytochrome P450 1A induction, or 32P post-labeling determination of hepatic PAH-DNA adducts. Growth of predatory flatfish was lower than reference in all but one of eight groups fed contaminant-exposed polychaetes; however, statistically significant reductions in growth were only observed in three of these eight groups, at least in part due to low statistical power. Juvenile sole from all contaminant-exposed groups showed increased expression of CYP1A, and fish exposed to BaP-exposed worms showed clear evidence of hepatic PAH-DNA adducts. This method allows the concurrent evaluation of sediment contamination at multiple biological and ecological levels. These results indicate that sediments determined to be nontoxic by common invertebrate bioassays may have the potential to cause adverse effects at higher trophic levels.

Animals↗

Natural attenuation of trichloroethylene in fractured shale bedrock.

This paper describes one of the first well-documented field examples of natural attenuation of trichloroethylene (TCE) in groundwater in a fractured shale bedrock. The study was carried out adjacent to a former waste burial site in Waste Area Grouping 5 (WAG5) on the Oak Ridge Reservation, Oak Ridge, TN. A contaminant plume containing TCE and its daughter products were detected downgradient from the buried waste pits, with most of the contamination occurring in the upper 6 m of the bedrock. The monitoring well array consists of a 35-m-long transect of multilevel sampling wells, situated along a line between the waste pits and a seep which discharges into a small stream. Concentrations of volatile organic carbons (VOCs) were highest in the waste trenches and decreased with distance downgradient towards the seep. Sampling wells indicated the presence of overlapping plumes of TCE, cis-dichloroethylene (cDCE), vinyl chloride (VC), ethylene, ethane, and methane, with the daughter products extending further downgradient than the parent (TCE). This type of distribution suggests anaerobic biodegradation. Measurements of redox potential at the site indicated that iron-reduction, sulfate reduction, and potentially methanogensis were occurring and are conducive to dechlorination of TCE. Bacteria enrichment of groundwater samples revealed the presence of methanotrophs, methanogens, iron-reducing bacteria and sulfate-reducing bacteria, all of which have previously been implicated in anaerobic biodegradation of TCE. 16S rDNA sequence from DNA extracted from two wells were similar to sequences of organisms previously implicated in the anaerobic biodegradation of chlorinated solvents. The combined data strongly suggest that anaerobic biodegradation of the highly chlorinated compounds is occurring. Aerobic biodegradation may also be occurring in oxygenated zones, including near a seep where groundwater exits the site, or in the upper bedrock during seasonal fluctuations in water table elevation and oxygen levels.

Biodegradation, Environmental↗

Evaluation of cadmium and other metal losses from various municipal wastes during incineration disposal.

The Japanese intake much more Cd through food than people in other countries, this may be partly caused by the activity of municipal waste incineration disposal. The present study was carried out to examine the behavior of Cd and to identify the other lost metals during municipal waste incineration. Thirty-eight waste samples, including food scraps (FS), animal wastes (AW), horticulture wastes (HW) and sewage sludges (SS) were collected from various places in Japan. Treatment temperatures of 500, 850, and 1000 degrees C were conducted to these samples in laboratory, and thirty-nine kinds of metals were examined. The results showed that average Cd losses were 69% (2.4 mg kg-1) and 74% (2.6 mg kg-1) at 850 and 1000 degrees C, respectively. Furthermore, the losses of 20 other metals were also found, i.e. Sn lost at 500 degrees C, K, Mg, Na, Bi, Cr, Ge, Li, Pb, Sn, Tl, Zn lost at 850 degrees C, and nine more metals, Al, Be, Cs, Nb, Sb, Sr, Th, Y, Zr lost at 1000 degrees C. The lost metals were considered to be transferred into the combustion flue gas, mainly in the forms of metallic chloride compounds, e.g. CdCl2, SnCl4, SnCl2, ZnCl2, PbCl2, etc. On the other hand, no significant losses for Ca, Fe, Ag, Ba, Co, Cu, Ga, Hf, Mn, Mo, Ni, Rb, Sc, Ta, Ti, U, V, W were found at 0.05 probability level. Total metal losses could be calculated at 9.9 and 20 g kg-1, in which major metal (Al, K, Mg, Na) losses were 9.5, 19 g kg-1 and minor and trace metal losses were 0.4, 0.6 g kg-1 at 850 and 1000 degrees C, respectively. Among the four types of waste materials, Cd losses were 0.8-3.7 mg kg-1, occupied about 60-80% at 850 degrees C, and 0.9-3.9 mg kg-1, occupied about 65-85% at 1000 degrees C. The total major metal losses from the four types of waste materials could be calculated at 6.5-15 g kg-1 at 850 degrees C, and 11-31 g kg-1 at 1000 degrees C, minor and trace metal losses were 0.03-1.3 g kg-1 at 850 degrees C, and 0.3-1.5 g kg-1 at 1000 degrees C, respectively.

Agriculture↗

Contaminant risks from biosolids land application: contemporary organic contaminant levels in digested sewage sludge from five treatment plants in Greater Vancouver, British Columbia.

This study examines the potential for environmental risks due to organic contaminants at sewage sludge application sites, and documents metals and various potential organic contaminants (volatile organics, chlorinated pesticides, PCBs, dioxins/furans, extractable petroleum hydrocarbons, PAHs, phenols, and others) in current production biosolids from five wastewater treatment plants (WWTPs) within the Greater Vancouver Regional District (GVRD). There has been greater focus in Europe, North America and elsewhere on metals accumulation in biosolids-amended soil than on organic substances, with the exception of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans. Another objective, therefore, was to evaluate the extent to which management of biosolids re-use based on metal/metalloid levels coincidentally minimizes environmental risks from organic contaminants. Historical-use contaminants such as chlorophenols, PCBs, and chlorinated pesticides were not detected at environmentally relevant concentrations in any of the 36 fresh biosolids samples, and appear to have virtually eliminated from sanitary collection system inputs. The few organic contaminants found in freshly produced biosolids samples that exhibited high concentrations relative to British Columbia and Canadian soil quality benchmarks included p-cresol, phenol, phenanthrene, pyrene, naphthalene, and heavy extractable petroleum hydrocarbons (HEPHs--nCl9-C34 effective carbon chain length). It was concluded that, with the exception of these petroleum hydrocarbon constituents or their microbial metabolites, the mixing of biosolids with uncontaminated soils during land application and based on the known metal concentrations in biosolids from the Greater Vancouver WWTPs investigated provides adequate protection against the environmental risks associated with organic substances such as dioxins and furans, phthalate esters, or volatile organics. Unlike many other organic contaminants, the concentrations of petroleum hydrocarbon derived substances in biosolids has not decreased within the last decade or more in the WWTPs studied, and--unlike persistent chlorinated compounds--the associated PAHs and other hydrocarbon constituents merit careful consideration, especially in the context of repeated land-application of biosolid.

Canada↗

Loss of heterozygosity on chromosome 6 in dichloroacetic acid and trichloroacetic acid-induced liver tumors in female B6C3F1 mice.

Dichloroacetic acid (DCA) and trichloroacetic acid (TCA) are major metabolites of tetrachloroethylene (PCE) and trichloroethylene (TCE) and are found in chlorinated drinking water. All four chlorinated compounds are liver carcinogens in B6C3F1 mice. It has previously been reported that approximately 20% of hepatic tumors induced by PCE exhibited loss of heterozygosity (LOH) on chromosome 6, suggesting the presence of a tumor suppressor gene. In the current investigation, we determined whether TCA or DCA also induced LOH on chromosome 6. Liver tumors were initiated in 15 day old female B6C3F1 mice with N-methyl-N-nitrosourea (MNU) and promoted with 20 mmol/l DCA or TCA in their drinking water. Twenty-four and thirty-seven liver tumors promoted by DCA and TCA, respectively, were examined for LOH using 4 polymorphic loci on chromosome 6. Ten of 37 (27%) tumors (7 of 27 carcinomas and 3 of 10 adenomas) promoted by TCA exhibited LOH at least for two loci on chromosome 6. All 10 tumors that exhibited LOH, lost the C57BL/6J allele at both the D6mit9 loci, while two also lost at least one of the C3H/HeJ alleles. No LOH on chromosome 6 was observed in the 24 liver tumors promoted by DCA. The LOH on chromosome 6 in TCA but not in DCA-promoted tumors supports it as an active metabolite of PCE and demonstrates different pathogenesis at least for some of the DCA and TCA-promoted liver cancer.

Animals↗

Intrinsic bioremediation of trichloroethylene and chlorobenzene: field and laboratory studies.

Activities at a former fire training area at Robins Air Force Base in Georgia, USA resulted in contamination of groundwater with a mixture of trichloroethylene (TCE) and chlorobenzene (CB). Results from the field investigation suggest that intrinsic bioremediation process is occurring, which caused the decrease in TCE and CB concentrations, and increase in TCE degradation byproducts [e.g., dichloroethylene isomers (DCEs), vinyl chloride (VC)] concentrations. Contaminated groundwater samples collected from this site were used to conduct microbial enumeration tests, and used as the inocula for microcosm establishment. Results from the microbial enumeration study indicate that methanogenesis was the dominant biodegradation pattern within the source and mid-plume areas, and the aerobic biodegradation process dominated the downgradient area. Laboratory microcosm experiments were conducted to evaluate the feasibility of using CB as the primary substrate to enhance the intrinsic biodegradation of TCE. Microcosm results suggest that CB can serve as the primary substrate (electron donor), and enhance TCE biodegradation to less-chlorinated compounds under both aerobic cometabolism and reductive dechlorination conditions.

Chlorobenzenes↗

The relative embryotoxicity of 1,3-dichloro-2-propanol on primary chick embryonic cells.

1,3-Dichloro-2-propanol (1,3-DCP) is a chlorinated compound used in the fabrication of industrial products such as hard resins, celluloid or paints. It has also been detected in instant soups and soy sauce. 1,3-DCP has been associated with major necrosis of the liver in humans [Chem.-Bio. Interact. 80 (1991) 73]. In humans and laboratory animals, 1,3-DCP is metabolised to dichloroacetone (1,3-DCA) by cytochromes P450 2E1 and 1A2 [J. University Occup. Environ. Health 14 (1992) 13]. 1,3-DCA is a hepatotoxin. We suggest that 1,3-DCA could be embryotoxic at doses that do not cause adverse maternal hepatic damage. To investigate the embryotoxic effects of 1,3-DCA, we have adapted a micromass culture method from Atterwill and colleagues [1992. A tiered system for in vitro neurotoxicity testing. In: Zbinden, G. (Ed.), The Brain in Bits and Pieces. Verlag M.T.C., Vollikon, pp. 89-91], using chick midbrain cells and from Wiger et al. [Pharmacol. Toxicol. 62 (1988) 32] using chick mesenchymal cells. The basis of the micromass system is that embryotoxins in vitro are likely to affect development and differentiation of disaggregated neuronal and limb bud micromass cultures. The endpoints chosen for the midbrain assay are resazurin reduction (viability), total protein content (cell number), morphological quantification of neuronal cultures (neuronal projection number) and of limb bud cultures (cartilage nodule number). Preliminary results using chick whole embryo cultures indicated that 1,3-DCA had an inhibitory effect on whole chick embryo development. We also found that embryonic derived cells were sensitive to 1,3-DCA but not 1,3-DCP at concentrations above 1 microM, suggesting a potential teratogenic effect of 1,3-DCA. The exposure to 1,3-DCP is not limited to industrial settings, and hence a better knowledge of its effects and tissue specific actions on embryonic-derived cells would be beneficial.

Animals↗

Retrospective study of 273 deaths due to poisoning at Sir Salimullah Medical College from 1988 to 1997.

In Bangladesh all poisoning death cases are recorded as unnatural death and medico-legal autopsy is routine. This paper investigates the pattern, trend and incidence of unnatural poisoning deaths attended in Sir Salimullah Medical College Mortuary, Dhaka, Bangladesh from January 1, 1988 to December 31, 1997. A total of 2534 medico-legal autopsy cases were carried out during this period and 273 deaths by poisoning constituted 10.8%. The autopsies done were predominantly male (54.6%). The 13-24-year-old age group was the highest victim group (48.4%). The urban incidence was 60.8%. The main reasons behind taking poison were diseases, familial dispute, alleged insanity and marital discord. Organo-chlorine compounds were the main killer (51.6%) followed by organo-phosphorous compounds (37.7%). This study shows that poison death victims are gradually increasing, in terms of growth rate male death is higher than female, and maximum number of deaths occur below 24 years of age. The trend of poisoning with organo-chloride compounds is being replaced by organo-phosphorous compound use.

Adolescent↗

Sonochemical treatment of natural ground water at different high frequencies: preliminary results.

Naturally occurring ground water polluted by highly volatile chlorinated compounds, with 1,2-dichloroethane as the main contaminant, was sonicated with high frequency ultrasound (361, 620 and 1086 kHz) in order to investigate if results from studies performed with single model compounds in water can be applied to a natural ground water. It was shown that the treated highly volatile halogenated hydrocarbons are almost completely destroyed by ultrasound in natural ground water within 60 min. It also transpired that the naturally occurring inorganic components and gases in the ground water play an important role.

Fresh Water↗

Vanadium haloperoxidases.

The nature of the oxidized halogen intermediate in vanadium bromoperoxidase has recently been shown to depend on the nature of the organic substrate. For example, in the presence of indoles, vanadium bromoperoxidase does not release a freely diffusible oxidized halogen intermediate (such as HOBr+/-BR2+/-Br3-). Regioselective investigations are, therefore, now feasible.

Binding Sites↗

Determination and mass spectrometric investigation of a new mixed halogenated persistent component in fish and seal.

An unknown component that caused an intense signal in sample extracts of fish tissue was enriched and investigated using a variety of mass spectrometric techniques coupled to gas chromatographic separation. With the help of electron capture negative ion mass spectrometry (ECNI-MS) and electron impact mass spectrometry (EI-MS) it was established that the component carries 2Br and 3Cl atoms and forms a molecular ion at m/z 396. A concentrated solution of this mixed halogenated compound (MHC-1) was investigated by gas chromatography interfaced to electron impact high-resolution mass spectrometry (GC/EI-HRMS). Using full scan and SIM techniques, the molecular formula of MHC-1 was established to be C10H13Br2Cl3. This points toward MHC-1 having a monoterpene backbone. No chemical with this molecular formula has been synthesized, but two components with this composition have been earlier isolated from marine algae.

Alkenes↗

Formation of PCDDs, PCDFs, and coplanar PCBs from polyvinyl chloride during combustion in an incinerator.

Exhaust gases from the combustion of poly(vinyl chloride) (PVC), polyethylene (PE), polystyrene (PS), poly(ethylene terephthalate) (PET), and theirvarious mixtureswere analyzed for PCDDs, PCDFs, and coplanar PCBs by gas chromatography/mass spectrometry (GC/MS) in order to investigate the role of PVC in these chlorinated compounds. Total amounts of dioxins (PCDDs + PCDFs) found in the samples were 11.7 ng/g PE alone, 1.17 ng/g from PS alone, 25.3 ng/g from PET alone, 448 ng/g from PE with PVC, 140 ng/g from PS with PVC, 126 ng/g from PET with PVC, 824 ng/g from PVC alone under low-CO level, and 8,920 ng/g from PVC alone under high-CO level. CO level in high-CO level condition was 880 ppm which was 20 times greater than that in low-CO level condition. Formation of coplanar PCBs ranged from 0.095 ng/g (PE alone) to 77 ng/g (PVC alone under high-CO level). There is a clear correlation between dioxin formation and chloride content. PCDFs composed 80% (PET + PVC)--98% (PET alone) of the total dioxins formed in the exhaust gases. The results indicate that PVC contributes significantly to the formation of PCDDs, PCDFs, and coplanar PCBs from mixtures of plastics upon combustion.

Benzofurans↗

Nucleophilic aromatic substitution reactions of chloroazines with bisulfide (HS-) and polysulfides (Sn2-).

Reactions of bisulfide and polysulfides with chloroazines (important constituents of agrochemicals and textile dyes) were examined in aqueous solution at 25 degrees C. For atrazine, rates are first-order in polysulfide concentration, and polysulfide dianions are the principal reactive nucleophiles; no measurable reaction occurs with HS-. Second-order rate constants for reactions of an array of chloroazines with polysulfides are several orders of magnitude greater than for reactions with HS-. Transformation products indicate the substitution of halogen(s) by sulfur. Ring aza nitrogens substantially enhance reactivity through a combination of inductive and mesomeric effects, and electron-withdrawing or electron-donating substituents markedly enhance or diminish reactivity, respectively. The overall second-order nature of the reaction, the products observed, and reactivity trends are all consistent with a nucleophilic aromatic substitution (S(N)Ar) mechanism. Rate constants for reactions with HS- and Sn2- (n = 2-5) correlate only weakly with lowest unoccupied molecular orbital energies, suggesting that the electrophilicity of a chloroazine is not the sole determinant of its reactivity. When second-order rate constants are extrapolated to HS- and Sn2- concentrations reported in salt marsh pore waters, half-lives of minutes to years are obtained. Polysulfides in particular could play an important role in effecting abiotic transformations of chloroazines in hypoxic marine waters.

Azo Compounds↗

Hydrogen abstraction and decomposition of bromopicrin and other trihalogenated disinfection byproducts by GC/MS.

Tribromonitromethane (bromopicrin), dibromochloronitromethane, bromodichloronitromethane, and trichloronitromethane (chloropicrin) have been identified as drinking water disinfection byproducts (DBPs). They are thermally unstable and decompose under commonly used injection port temperatures (200-250 degrees C) during gas chromatography (GC) or GC/mass spectrometry (GC/MS) analysis. The major decomposition products are haloforms (such as bromoform), which result from the abstraction of a hydrogen atom from the solvent bythermally generated trihalomethyl radicals. A number of other products formed by radical reactions with the solvent and other radicals were also detected. The trihalonitromethanes also decompose in the hot GC/MS transfer line, and the mass spectra obtained are mixed spectra of the undecomposed parent compound and decomposition products. This can complicate the identification of these compounds by GC/MS. Trihalomethyl compounds that do not have a nitro group, such as tribromoacetonitrile, carbon tetrabromide, methyl tribromoacetate, and tribromoacetaldehyde, do not decompose or only slightly decompose in the GC injection port and GC/MS transfer line. The brominated trihalomethyl compounds studied also showed H/Br exchange by some of their fragment ions. This H/Br exchange also makes the identification of these compounds in drinking water more difficult. The extent of H/Br exchange was found to depend on the mass spectrometer ion source temperature, and it is proposed that the internal surface of the ion source is involved in this process.

Bromine Compounds↗

Polychlorinated dibenzo-p-dioxin and dibenzofuran concentration profiles in sediments and flood-plain soils of the Tittabawassee River, Michigan.

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in sediments and flood-plain soils collected along the Tittabawassee River in Michigan ranged from 102 to 53,600 pg/g, dry wt. Mean PCDD/PCDF concentrations in downstream sediment and soil were from 10- to 20-fold greater than those found at locations upstream of Midland, Michigan. Concentrations of PCDD/PCDF in sediments and flood-plain soils from the Tittabawassee watershed were comparable to those found in industrialized areas such as the Housatonic and lower Passaic Rivers in the U.S. Concentrations of PCDDs/PCDFs in soil and sediment were not correlated with total organic carbon (TOC) in sediments or soils. OCDD and 2,3,7,8-TeCDF were the predominant congeners in sediment/soil collected from locations downstream of Midland, Michigan. Principal component analysis of the PCDD/PCDF congener profile suggested the presence of sources originating from a mixture of chlorophenol and other chlorinated compound production. Mass balance analysis of TCDD equivalents (TCDD-EQs) derived from H4IIE-luc bioassay of sediment extracts and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) estimated from instrumental analysis suggested that PCDDs/PCDFs were the major dioxin-like compounds present in sediments. A significant correlation existed between bioassay-derived TCDD-EQs and instrumentally measured TEQs (r2 = 0.94).

Benzofurans↗

TCE diffusion to the atmosphere in phytoremediation applications.

The phytoremediation of trichloroethylene (TCE) and other chlorinated compounds has been studied over the past decade, and full-scale systems are in place. The results regarding TCE fates and removal pathways are inconclusive and conflicting, particularly the results regarding volatilization to the atmosphere. Research presented here demonstrates that TCE is taken up by trees and volatilized to the atmosphere. TCE diffusion along the transpiration pathway is shown to be the primary process for TCE volatilization, although volatilization can occur from both stems and leaves. Two concurrent processes influence the eventual fate: transport with transpiration stream through xylem tissues and diffusion from transpiration stream to atmosphere. TCE diffusion flux invariably decreased with height for trees planted in soil or grown hydroponically. In both laboratory experiments and field sampling, TCE concentrations in the transpiration stream (e.g., xylem tissues) decreased with elevation. In field samples, TCE concentrations also decreased in the radial direction, providing fundamental evidence for diffusion. The TCE concentrations in tissues responded linearly to the exposure concentrations at the roots, while TCE diffusion from tree stems was influenced by concentration and transpiration rates.

Biodegradation, Environmental↗

Hydrogen thresholds as indicators of dehalorespiration in constructed treatment wetlands.

Anaerobic degradation of cis-1,2-dichloroethene (cis-1,2-DCE) and 1,2-dichloroethane (1,2-DCA) was studied in microcosms derived from a laboratory-scale upflow treatment wetland system used to biodegrade chlorinated compounds present in groundwater from a Superfund site. Dechlorination kinetics of cis-1,2-DCE (0.94-1.57 d(-1)) and 1,2-DCA (0.15-0.71 d(-1)) were rapid, and degradation proceeded to completion with ethene or ethane as terminal dechlorination products. Hydrogen concentrations, measured simultaneously during dechlorination, were significantly different for the two compounds, approximately 2.5 nM for cis-1,2-DCE and 38 nM for 1,2-DCA. Methanogenesis proceeded during the degradation of 1,2-DCA when H2 concentrations were high but not during the dechlorination of cis-1,2-DCE when H2 concentrations were below published thresholds for methanogenesis. A 16S rRNA gene-based approach indicates that microorganisms closely related to Dehalococcoides ethenogenes were present and that they were distributed throughout the bottom, middle, and top of the upflow treatment wetland system. These results coupled with consideration of hydrogen thresholds, degradation kinetics, daughter products, and measurements of methanogenesis strongly suggest that halorespirers were responsible for dechlorination of cis-1,2-DCE and that 1,2-DCA dechlorination was co-metabolic, likely mediated by acetogens or methanogens. Rapid dechlorination potential was distributed throughout the wetland bed, both within and below the rhizosphere, indicating that reductive dechlorination pathways can be active in anaerobic environments located in close spatial proximity to aerobic environments and plants in treatment wetland systems.

Bacteria, Anaerobic↗