Genotoxicity assessment of soils from wastewater irrigation areas and bioremediation sites using the Vicia faba root tip micronucleus assay

Genotoxicity potential of soils taken from wastewater irrigation areas and bioremediation sites was assessed using the Vicia faba root tip micronucleus assay. Twenty five soils were tested, of which 8 were uncontaminated soils and taken as the control to examine the influence of soil properties; 6 soils were obtained from paddy rice fields with a history of long-term wastewater irrigation; 6 soils were obtained from bioremediation sites to examine effects of bioremediation; and 5 PAH-contaminated soils were used to examine methodological effects between direct soil exposure and exposure to aqueous soil extracts on micronuclei (MN) frequency () in the V. faba root tips. Results indicate that soil properties had no significant influences on MN frequencies (p > 0.05) when soil pH varied between 3.4 to 7.6 and organic carbon between 0.4% and 18.6%. The MN frequency measured in these control soils ranged from 1.6‰ to 5.8‰. MN frequencies in soils from wastewater irrigation areas showed 2- to 48-fold increase as compared with the control. Soils from bioremediation sites showed a mixed picture: MN frequencies in some soils decreased after bioremediation, possibly due to detoxification; whereas in other cases remediated soils induced higher MN frequencies, suggesting that genotoxic substances might be produced during bioremediation. Exposure to aqueous soil extracts gave a higher MN frequency than direct exposure in 3 soils. However, the opposite was observed in the other two soils, suggesting that both exposure routes should be tested in case of negative results from one route. Data obtained from this study indicate that the MN assay is a sensitive assay suitable for evaluating genotoxicity of soils.

Field-scale bioremediation of soil contaminated with crude oil

Field-scale remediation of oil-contaminated soils from the Liaohe Oil Fields in China was examined using composting biopiles in windrow technology. Micronutrient-enriched chicken excrement and rice husk were applied as nutrition and a bulking agent. The lipase activities of indigenous micro-organisms were analyzed, and three indigenous fungi with high lipase activities was identified. An inoculum consisting of the three indigenous fungi and one introduced (exotic) fungus was applied to four different types of oil-contaminated soils. The results showed that the inoculum of indigenous fungi increased both the total colony-forming units (TCFU) and increased the rate of degradation of total petroleum hydrocarbons (TPH) in all contaminated soils but at different rates. In sharp contrast to other studies, the introduction of exotic micro-organisms did not improve the remediation, and suggests that inoculation of oil-contaminated sites with nonindigenous species is likely to fail. On the other hand, indigenous genera of microbes were found to be very effective in increasing the rate of degradation of TPH. The degradation of TPH was mainly controlled by the compositions of aromatic hydrocarbons and asphaltene and resin. Between 38 to 57% degradation of crude oils (with densities ranging from 25,800 to 77,200 mg/kg dry weight) in contaminated soils was achieved after 53 days of operation. The degradation patterns followed typical first-order reactions. We demonstrate that the construction and operation of field-scale composting biopiles in windrows with passive aeration is a cost-effective bioremediation technology.

Acid drainage, limnology and bioremediation of western Victorian coal mine lakes

Rehabilitation of Alcoa's Anglesea open cut brown coal mine to a healthy lake has many environmental challenges. The study of regional acid drainage, limnology of Wenslydale Coal Mine Lake and passive bioremediation of acid mine water has shown that a healthy lake can be created.

Degradation mechanisms of benzo[a]pyrene and its accumulated metabolites by biodegradation combined with chemical oxidation

A high degradation extent of benzo[a]pyrene (BaP) should not be considered as the sole desirable criterion for the bioremediation of BaP-contaminated soils because some of its accumulated metabolites still have severe health risks to human. Two main metabolites of BaP, benzo[a]pyrene-1,6-quinone (BP1,6-quinone) and 3-hydroxybenzo[a]pyrene (3-OHBP) were identified by high performance liquid chromatography (HPLC) with standards. This study was the first time that degradation of both BaP and the two metabolites was carried out by chemical oxidation and biodegradation. Three main phases during the whole degradation process were proposed.

Hydrogen peroxide–zinc (H₂O₂–Zn), the fungus – Aspergillus niger and the bacteria – Zoogloea sp. played an important role in the different phases. The degradation parameters of the system were also optimized, and the results showed that the effect of degradation was the best when fungus–bacteria combined with H₂O₂–Zn, the concentration range of BaP in the cultures was 30–120 mg/l, the initial pH of the cultures was 6.0. However, as co-metabolites, phenanthrene significant inhibited the degradation of BaP. This combined degradation system compared with the conventional method of degradation by domestic fungus only, enhanced the degradation extent of BaP by more than 20% on the 12 d. The highest accumulation of BP1,6-quinone and 3-OHBP were reduced by nearly 10% in the degradation experiments, which further proved that the combined degradation system was more effective as far as joint toxicity of BaP and its metabolites are concerned.

Biodegradation of benzo[a]pyrene in soil by Mucor sp. SF06 and Bacillus sp. SB02 co-immobilized on vermiculite

Two indigenous microorganisms, Bacillus sp. SB02 and Mucor sp. SF06, capable of degrading polycyclic aromatic hydrocarbons (PAHs) were co-immobilized on vermiculite by physical adsorption and used to degrade benzo[a] pyrene (BaP). The characteristics of BaP degradation by both free and co-immobilized microorganism were then investigated and compared. The removal rate using the immobilized bacterial-fungal mixed consortium was higher than that of the freely mobile mixed consortium. 95.3% of BaP was degraded using the co-immobilized system within 42 d, which was remarkably higher than the removal rate of that by the free strains. The optimal amount of inoculated co-immobilized system for BaP degradation was 2%. The immobilized bacterial-fungal mixed consortium also showed better water stability than the free strains. Kinetics of BaP biodegradation by co-immobilized SF06 and SB02 were also studied. The results demonstrated that BaP degradation could be well described by a zero-order reaction rate equation when the initial BaP concentration was in the range of 10—200 mg/kg. The scanning electronic microscope (SEM) analysis showed that the co-immobilized microstructure was suitable for the growth of SF06 and SB02. The mass transmission process of co-immobilized system in soil is discussed. The results demonstrate the potential for employing the bacterial-fungal mixed consortium, co-immobilized on vermiculite, for in situ bioremediation of BaP.

Influence of chemical oxidant on degradation of benzo[a]pyrene metabolites by the bacterium-Zoogloea sp.

It is neither comprehensive nor appropriate that the bioremediation of a benzo[a]pyrene (BaP)-contaminated environment be assessed only by its high degradation extent because its metabolites' chemical structures are similar to the parent compound and maybe equally toxic. Therefore, further degradation of BaP metabolites is significant. Three methods, combining the Zoogloea sp. with potassium permanganate, combining the Zoogloea sp. with H₂O₂, Zoogloea sp. alone, were investigated to degrade cis-BP4,5-dihydrodiol and cis-BP7,8-dihydrodiol, which are the metabolites of BaP formed by bacterium-Zoogloea sp. Optimum parameters of degradation in the best method are that: of the three methods, coupling the Zoogloea sp. and KMnO₄ is the best; compared with cis-BP7,8-dihydrodiol, cis-BP4,5-dihydrodiol is the more liable to be accumulated in pure cultures; the degradation effect of the two metabolites is optimal when the initial concentration of KMnO₄ in the cultures is 0.05%; initial concentration of cis-BP4,5-dihydrodiol and cis-BP7,8-dihydrodiol is 4 mg L⁻¹, 8 mg L⁻¹, respectively; cometabolic substance is salicylic acid or sodium succinate. The degradation extent of cis-BP4,5-dihydrodiol and cis-BP7,8-dihydrodiol using combining the Zoogloea sp. and KMnO₄ reach 76.1% and 85.9% after 12 days of cultivation, respectively, which were more than twice compared with conventional method.

Surfactant-mediated biodegradation of polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) are toxic environmental pollutants that are known or suspected carcinogens or mutagens. Bioremediation has been used as a general way to eliminate them from the contaminated sites or aquifers, but their biodegradation is rather limited due to their low bioavailability because of their sparingly soluble nature. Surfactant-mediated biodegradation is a promising alternative. The presence of surfactants can increase the solubility of PAHs and hence potentially increase their bioavailability. However, inconclusive results have been reported on the effects of surfactant on the biodegradation of PAHs. In this work, surfactant-mediated biodegradation of PAHs is reviewed.

Effect of a commercial alcohol ethoxylate surfactant (C11–15E7) on biodegradation of phenanthrene in a saline water medium by Neptunomonas naphthovorans

Biodegradation of poorly soluble polycyclic aromatic hydrocarbons (PAHs) has been a challenge in bioremediation. In recent years, surfactant-enhanced bioremediation of PAH contaminants has attracted great attention in research. In this study, biodegradation of phenanthrene as a model PAHs solubilized in saline micellar solutions of a biodegradable commercial alcohol ethoxylate nonionic surfactant was investigated. The critical micelle concentration (CMC) of the surfactant and its solubilization capacity for phenanthrene were examined in an artificial saline water medium, and a type of marine bacteria, Neptunomonas naphthovorans, was studied for the biodegradation of phenanthrene solubilized in the surfactant micellar solutions of the saline medium. It is found that the solubility of phenanthrene in the surfactant micellar solutions increased linearly with the surfactant concentrations, but, at a fixed phenanthrene concentration, the biodegradability of phenanthrene in the micellar solutions decreased with the increase of the surfactant concentrations. This was attributed to the reduced bioavailability of phenanthrene, due to its increased solubilization extent in the micellar phase and possibly lowered mass transfer rate from the micellar phase into the aqueous phase or into the bacterial cells. In addition, an inhibitory effect of the surfactant on the bacterial growth at high surfactant concentrations may also play a role. It is concluded that the surfactant largely enhanced the solubilization of phenanthrene in the saline water medium, but excess existence of the surfactant in the medium should be minimized or avoided for the biodegradation of phenanthrene by Neptunomonas naphthovorans.

A Ramsar wetland in crisis the Coorong, Lower Lakes and Murray Mouth, Australia

The state of global freshwater ecosystems is increasingly parlous with water resource development degrading high-conservation wetlands. Rehabilitation is challenging because necessary increases in environmental flows have concomitant social impacts, complicated because many rivers flow between jurisdictions or countries. Australia's MurrayDarling Basin is a large river basin with such problems encapsulated in the crisis of its Ramsar-listed terminal wetland, the Coorong, Lower Lakes and Murray Mouth. Prolonged drought and upstream diversion of water dropped water levels in the Lakes below sea level (20092010), exposing hazardous acid sulfate soils. Salinities increased dramatically (e.g. South Lagoon of Coorong>200gL-1, cf. modelled natural 80gL-1), reducing populations of waterbirds, fish, macroinvertebrates and littoral plants. Calcareous masses of estuarine tubeworms (Ficopomatus enigmaticus) killed freshwater turtles (Chelidae) and other fauna. Management primarily focussed on treating symptoms (e.g. acidification), rather than reduced flows, at considerable expense (≥AU$2 billion). We modelled a scenario that increased annual flows during low-flow periods from current levels up to one-third of what the natural flow would have been, potentially delivering substantial environmental benefits and avoiding future crises. Realisation of this outcome depends on increasing environmental flows and implementing sophisticated river management during dry periods, both highly contentious options.

Physiological metal uptake by Nostoc punctiforme

Trace metals are required for many cellular processes. The acquisition of trace elements from the environment includes a rapid adsorption of metals to the cell surface, followed by a slower internalization. We investigated the uptake of the trace elements Co²⁺, Cu²⁺, Mn²⁺, Ni²⁺, and Zn²⁺ and the non-essential divalent cation Cd²⁺ in the cyanobacterium Nostoc punctiforme. For each metal, a dose response study based on cell viability showed that the highest non-toxic concentrations were: 0.5 μM Cd²⁺, 2 μM Co²⁺, 0.5 μM Cu²⁺, 500 μM Mn²⁺, 1 μM Ni²⁺, and 18 μM Zn²⁺. Cells exposed to these non-toxic concentrations with combinations of Zn²⁺ and Cd²⁺, Zn²⁺ and Co²⁺, Zn²⁺ and Cu²⁺ or Zn²⁺ and Ni²⁺, had reduced growth in comparison to controls. Cells exposed to metal combinations with the addition of 500 μM Mn²⁺ showed similar growth compared to the untreated controls. Metal levels were measured after one and 72 h for whole cells and absorbed (EDTA-resistant) fractions and used to calculate differential uptake rates for each metal. The differences in binding and internalisation between different metals indicate different uptake processes exist for each metal. For each metal, competitive uptake experiments using ⁶⁵Zn showed that after 72 h of exposure Zn²⁺ uptake was reduced by most metals particularly 0.5 μM Cd²⁺, while 2 μM Co²⁺ increased Zn²⁺ uptake. This study demonstrates that N. punctiforme discriminates between different metals and favourably substitutes their uptake to avoid the toxic effects of particular metals.

The effect of concentrations and properties of phenanthrene, pyrene, and benzo(a)pyrene on desorption in contaminated soil aged for 1 year

Purpose : The choice and timing of microorganisms added to soils for bioremediation is affected by the dominant bioavailable contaminants in the soil. However, changes to the concentration of bioavailable PAHs in soil are not clear, especially when several PAHs coexist. This study investigated the effects of PAH concentration and chemical properties on desorption in meadow brown soil after a 1-year aging period, which could reflect changes of PAH bioavailability during bioremediation. Materials and methods : Based on the percentage of different molecular weights in a field investigation, high-level contaminated soil (HCS) and low-level contaminated soil (LCS) were prepared by adding phenanthrene (PHE), pyrene (PYR) and benzo(a)pyrene (BaP) to uncontaminated meadow brown soil. The concentrations of HCS and LCS were 250 mg kg−1 (PHE, PYR, and BaP: 100, 100, and 50 mg kg−1) and 50 mg kg−1 (PHE, PYR, and BaP: 20, 20, and 10 mg kg−1) respectively. The soils were aged for 1 year, after which desorption was induced by means of a XAD-2 adsorption technique over a 96-h period. Results and discussion : The range of the rapidly desorbing fraction (F rap) for PHE, PYR, and BaP in HCS and LCS was from 1.9 to 27.8 %. In HCS, desorption of PYR was most difficult, and the rate constant of very slow desorption (K vs) of PYR was 8 orders of magnitude lower than that of BaP, which had similar very slow desorbing fractions (49.8 and 50.5 %, respectively). However, in LCS, desorption of PYR was the easiest; the Kvs of PYR was 8–10 orders of magnitude higher than those of PHE and BaP. In HCS, the time scale for release of 50 % of the PAHs was ranked as BaP > PYR > PHE, while in LCS this was BaP > PHE > PYR. Conclusions : The combined effect of PAH concentrations and properties should be taken into account during desorption. The desorption of PAH did not always decrease with increasing molecular weight, and the desorption of four-ring PAHs might be special. These results are useful for screening biodegrading microbes and determining when they should be added to soils based on the dominant contaminants present during different periods, thus improving the efficiency of soil bioremediation.

Oil-spill cleanup: the influence of acetylated curaua fibers on the oil-removal capability of magnetic composites

A magnetic resin based on cardanol, furfural, and curaua fibers was prepared and characterized. The material could be used in oil-spill cleanup processes, because of its aromatic/aliphatic balance. The resin was prepared through bulk polycondensation of cardanol and furfural in the presence of curaua fibers and maghemite nanoparticles. Hydrophobicity of the curaua fibers was improved by acetylation, increasing the oil-absorbing capability of the composites. The obtained magnetic composites were studied by Fourier-transform infrared spectroscopy, X-ray diffraction, and thermogravimetric analysis. Degree of cure, magnetic force, and oil-removal capability tests were also performed. The results show that the composites possess an elevated cure degree in addition to a considerable magnetic force. The materials exhibit a good oil removal capability in the presence of a magnetic field, which is improved by the use of acetylated curaua. In the best case, the composite filled with maghemite and curaua can remove 12 parts of oil from water.

Inquisition of Microcystis aeruginosa and Synechocystis nanowires: characterization and modelling

Identification of extracellular conductive pilus-like structures (PLS) i.e. microbial nanowires has spurred great interest among scientists due to their potential applications in the fields of biogeochemistry, bioelectronics, bioremediation etc. Using conductive atomic force microscopy, we identified microbial nanowires in Microcystis aeruginosa PCC 7806 which is an aerobic, photosynthetic microorganism. We also confirmed the earlier finding that Synechocystis sp. PCC 6803 produces microbial nanowires. In contrast to the use of highly instrumented continuous flow reactors for Synechocystis reported earlier, we identified simple and optimum culture conditions which allow increased production of nanowires in both test cyanobacteria. Production of these nanowires in Synechocystis and Microcystis were found to be sensitive to the availability of carbon source and light intensity. These structures seem to be proteinaceous in nature and their diameter was found to be 4.5-7 and 8.5-11 nm in Synechocystis and M. aeruginosa, respectively. Characterization of Synechocystis nanowires by transmission electron microscopy and biochemical techniques confirmed that they are type IV pili (TFP) while nanowires in M. aeruginosa were found to be similar to an unnamed protein (GenBank : CAO90693.1). Modelling studies of the Synechocystis TFP subunit i.e. PilA1 indicated that strategically placed aromatic amino acids may be involved in electron transfer through these nanowires. This study identifies PLS from Microcystis which can act as nanowires and supports the earlier hypothesis that microbial nanowires are widespread in nature and play diverse roles.

Metal and metalloid containing natural products and a brief overview of their applications in biology, biotechnology and biomedicine

Bioinorganic natural product chemistry is a relatively unexplored but rapidly developing field with enormous potential for applications in biology, biotechnology (especially in regards to nanomaterial development, synthesis and environmental cleanup) and biomedicine. In this review the occurrence of metals and metalloids in natural products and their synthetic derivatives are reviewed. A broad overview of the area is provided followed by a discussion on the more common metals and metalloids found in natural sources, and an overview of the requirements for future research. Special attention is given to metal hyperaccumulating plants and their use in chemical synthesis and bioremediation, as well as the potential uses of metals and metalloids as therapeutic agents. The potential future applications and development in the field are also discussed.

Enzymes for food waste remediation and valorisation

Industrial food processing operations are among the largest contributors of environmental waste. The use of enzymes in food waste remediation and valorisation is the central theme of this chapter. Specifically, the chapter first discusses the sources and values of food waste in food supply chains. Different strategies for bioremediation of wastewater using enzymes are then reviewed. The last part of the chapter focuses on waste valorisation—how to turn food waste into valuable products using enzymes—including a discussion of emerging trends in this field.