Phyto/rhizoremediation studies using long-term PCB-contaminated soil

Purpose  

Polychlorinated biphenyls (PCBs) represent a large group of recalcitrant environmental pollutants, differing in the number of chlorine atoms bound to biphenyl ring. Due to their excellent technological properties, PCBs were used as heat-transfer media, for filling transformers and condensers, as paint additives, etc. With increasing knowledge of their toxicity, transfer to food chains and accumulation in living organisms, their production ended in most countries in the 1970s and in 1984 in the former Czechoslovakia. But even a quarter of century after the PCB production ceased, from contaminated areas, the volatile PCBs evaporate and contaminate much larger areas even at very distant parts of the world. For this reason, PCBs still represent a global problem. The main method of PCB removal from contaminated environment is at present the expensive incineration at high temperatures. With the aim of finding effective alternative approaches, we are studying biological methods for PCB removal from the environment. In this paper, we summarise 10 years of studies using long-term PCB-contaminated soil from a dumpsite in South Bohemia, targeted for the use of plants (phytoremediation) and their cooperation with microorganisms in the root zone (rhizoremediation).     

The effect of ryegrass (Lolium perenne) on decrease of PAH content in long term contaminated soil

The biodegradation of polycyclic aromatic hydrocarbons in microecosystems containing long-term contaminated soil was investigated. Soil was contaminated by different chemicals, including PAHs since World War II. Aging of the soil was expected to act as a principal factor limiting biodegradation. Half of the microecosystems contained ryegrass (Lolium perenne) and long-term selected natural soil microflora originally present in contaminated soil. The others contained contaminated soil with natural microflora only. Half of the microecosystems in each parallel experiment was fertilised with N-P-K fertiliser. Cultivation was carried out at 12 and 18 months in a greenhouse with a natural photoperiod and the ability to degrade 15 chosen PAH was investigated. For analysis, the soil from each pot was divided into three horizontal layers for mutual comparison among layers and each layer was further divided into four equal samples. Soil extracts were analysed using HPLC. After a one-year-cultivation period the content of the monitored PAHs declined to 50%. Mostly, there were no significant differences between the microecosystems. Best degraded were fluoranthene and pyrene, which were the major contaminants present in original soil. Also, other compounds were successfully degraded, even benzo[a]pyrene and benzo[ghi]perylene. Dibenz[a,h]anthracene and indeno[1,2,3-cd]pyrene were the only PAHs, examined that showed no significant degradation. Although some differences between the soil layers were detected, no conclusive trends could be found. However, significantly lower concentrations of PAHs were determined mostly in the bottom layer of the analysed profiles. In vegetated microecosystems the decline of PAHs concentrations was more remarkable after 18 months cultivation.     

Plant metabolites of polychlorinated biphenyls in hairy root culture of black nightshade Solanum nigrum SNC-9O

The present study is intended to determine metabolites of 12 dichlorinated, seven trichlorinated, five tetrachlorinated and one pentachlorinated PCB congener transformed by black nightshade (Solanum nigrum) hairy root culture SNC-9O. Free hydroxylated PCB metabolites were identified based on the mass spectra characteristics after gas chromatography separation. The number of metabolites decreases with an increasing number of chlorine atoms per molecule of PCB. Dichlorinated PCBs lead always to at least two metabolites. In the case of PCB 9 some metabolites could be identified by comparing their RF values due to available standards. The 2',5'-dichloro-2-biphenylol, 2',5'-dichloro-3-biphenylol and 2',5'-dichloro-4-biphenylol, present as the main metabolite, were found in biomass of SNC-9O hairy root culture. Two monochlorinated biphenylols were found in biomass of SNC-9O degrading PCB 9 congener. It was the only case when metabolites with decreased number of chlorine atoms compared to parent PCB were found. Trichlorinated PCBs mostly lead to a lower number of metabolites but tetrachlorinated and pentachlorinated PCBs mostly did not give any metabolites. In the media, only traces of metabolites were found in sporadic cases, so exudation of unbound biphenylols from the cells is not expected.     

Decolorization of RBBR by plant cells and correlation with the transformation of PCBs

An extracellular H2O2-requiring Remazol Brilliant Blue R (RBBR) decolorizing enzyme activity was detected after cultivation of cells of various plant species both in liquid medium and when growing on agar plates containing RBBR. Level of the enzyme activity was compared with the ability to metabolize polychlorinated biphenyls (PCBs). The ability to decolorize RBBR was tested in the presence and absence of PCBs. The cultures with high PCB-transforming activity proved to exhibit RBBR oxidase much more resistant towards the influence of PCBs. In addition low activities of lignin peroxidase (LiP) and manganese dependent peroxidase (MnP) were detected in medium and in plant cells. No correlation of MnP and LiP activities with PCB degradation could be found. The RBBR decolorization could be used as a rough screening method for plant cultures able to metabolize PCBs.     

Metabolites of 2,2'-dichlorobiphenyl and 2,6-dichlorobiphenyl in hairy root culture of black nightshade Solanum nigrum SNC-9O

The hairy root culture of black nightshade (Solanum nigrum) SNC-9O was exposed to 2,2′-dichlorobiphenyl (PCB 4) and 2,6-dichlorobiphenyl (PCB 10) to follow the metabolites produced. The analytical standards of 4-hydroxy-2,2′-dichlorobiphenyl, 5′-hydroxy-2,2′-dichlorobiphenyl, 4-hydroxy-2,6-dichlorobiphenyl, 2-hydroxy-2′,6′-dichlorobiphenyl, 3-hydroxy-2′,6′-dichlorobiphenyl and 4-hydroxy-2′,6′-dichlorobiphenyl have been synthesized. Hydroxy-metabolites of both PCB 4 and PCB 10 were present in the biomass. These appeared mainly as conjugates rather than as free hydroxy-PCBs, both maintained in plant cells. The concentrations of non-conjugated hydroxy-PCBs ranged between 0.9 and 35.2 μg kg⁻¹ of biomass fresh weight and the concentration of the conjugated ones ranged between 2.0 and 113.0 μg kg⁻¹ depending on the position of hydroxyl. The para- position of biphenyl (4 or 4′) seems to be preferred for hydroxylation. Methoxy-PCBs and hydroxy-methoxy-PCBs have also been identified in plant cells. Hydroxyl in the meta-position (3, 3′, 5 or 5′) appears to be preferred for methylation in hydroxy-PCBs. Hydroxy-methoxy-PCBs have occurred in the conjugated form as well.     

Removal of 4-chlorobenzoic acid from spiked hydroponic solution by willow trees (<Emphasis Type="Italic">Salix viminalis</Emphasis>)

Background  

Chlorobenzoic acids (CBA) are intermediate products of the aerobic microbial degradation of PCB and several pesticides. This study explores the feasibility of using basket willows, Salix viminalis, to remove 4-CBA from polluted sites, which also might stimulate PCB degradation.     

Dry Wetlands: Nutrient Dynamics in Ephemeral Constructed Stormwater Wetlands

Environmental Management - Constructed stormwater wetlands (CSWs) are used to address contaminants in urban stormwater such as nitrogen (N) and phosphorus (P), but their performance is variable....