Bacterial tolerance strategies against lead toxicity and their relevance in bioremediation application

Among heavy metals, lead (Pb) is a non-essential metal having a higher toxicity and without any crucial known biological functions. Being widespread, non-biodegradable and persistent in every sphere of soil, air and water, Pb is responsible for severe health and environmental issues, which need appropriate remediation measures. However, microbes inhabiting Pb-contaminated area are found to have evolved distinctive mechanisms to successfully thrive in the Pb-contaminated environment without exhibiting any negative effects on their growth and metabolism. The defensive strategies used by bacteria to ameliorate the toxic effects of lead comprise biosorption, efflux, production of metal chelators like siderophores and metallothioneins and synthesis of exopolysaccharides, extracellular sequestration and intracellular bioaccumulation. Lead remediation technologies by employing microbes may appear as potential advantageous alternatives to the conventional physical and chemical means due to specificity, suitability for applying in situ condition and feasibility to upgrade by genetic engineering. Developing strategies by designing transgenic bacterial strain having specific metal binding properties and metal chelating proteins or higher metal adsorption ability and using bacterial activity such as incorporating plant growth-promoting rhizobacteria for improved Pb resistance, exopolysaccharide and siderophores and metallothionein-mediated immobilization may prove highly effective for formulating bioremediation vis-a-vis phytoremediation strategies.

     

The effect of zeolite on the toxicity of lead to fungi

In order to determine whether clinoptilolite, a naturally occurring zeolite, had any ameliorative effect on lead (Pb) toxicity to fungi, a series of growth experiments were performed. Three fungi, Aspergillus niger, Botrytis cinerea, and Fusarium culmorum, were grown on appropriately amended solid agar media, and their linear extension rates determined. B. cinerea was 25% inhibited, as compared to a control, at 100 mg dm(-3) Pb, and completely inhibited at 1000 mg dm(-3) Pb. F. culmorum was completely inhibited, and A. niger 97% inhibited at 1000 mg dm(-3) Pb. The addition of 3% clinoptilolite partially removed this inhibition in the case of A. niger and B. cinerea and almost completely removed it for F. culmorum. At a constant 500 mg dm(-3) Pb, increasing concentrations of clinoptilolite increased the linear extension rate of F. culmorum and B. cinerea, close to the rates achieved by the untreated controls. A. niger was not inhibited markedly at this Pb concentration. The evidence suggests that the Pb is adsorbed by the clinoptilolite which reduces the availability, and hence toxicity, of the metal to the fungi.     

Predicting molybdenum toxicity to higher plants: Estimation of toxicity threshold values

Four plant species (oilseed rape, Brassica napus L.; red clover, Trifolium pratense L.; ryegrass, Lolium perenne L.; and tomato, Lycopersicon esculentum L.) were tested on ten soils varying widely in soil properties to assess molybdenum (Mo) toxicity. A larger range (66-fold-609-fold) of added Mo concentrations resulting in 50% inhibition of yield (ED₅₀) was found among soils than among plant species (2-fold-38-fold), which illustrated that the soils differed widely in the expression of Mo toxicity. Toxicity thresholds based on soil solution Mo narrowed the variation among soils compared to thresholds based on added Mo concentrations. We conclude that plant bioavailability of Mo in soil depends on Mo solubility, but this alone did not decrease the variability in observed toxicity enough to be used in risk assessment and that other soil properties influencing Mo toxicity to plants need to be considered.     

Biotrickling filtration of nitric oxide

A biotrickling filter with blast-furnace slag packings (sizes = 20-40 mm and specific surface area = 120 m2/m3) was utilized to treat NO in an air stream. The operational stability, as well as the effects of gas empty-bed retention time (EBRT) and nutrient addition on the removal ability of NO, were tested. Approximately six weeks were required for the development of a biofilm for NO degradation, and a two-week organic carbon deficiency resulted in the detachment of biofilms from the packing surfaces. A steady removal rate of 80% was attained at specified influent NO concentrations of 892 to 1237 ppm and an EBRT of 118 sec. The effluent NO concentration diminished exponentially with enlarging EBRT, with influent NO concentrations of 203-898 ppm, and EBRTs of 25 to 118 sec. Nutrient addition is essential for efficient removal of the influent NO. Mass ratios of C: P: N = 7: 1: 30 and NaHCO3: NO-N = 6.3 could be used for practical applications.     

Protective role of zeolite on short- and long-term lead toxicity in the teleost fish Heteropneustes fossilis.

The high ion-exchange capacity of zeolite (sodium aluminium silicate) enhances the removal of lead from water, thus decreasing its availability to fish. Zeolites are very important in the field of environmental preservation due to the low cost and ecological compatibility. Zeolites can adsorb metallic ions by cation exchange reactions. Continuous exposure of the teleost fish Heteropneustes fossilis to sublethal concentrations of lead nitrate in water solution for short (35 days) and long (120 days) periods decreased both the soluble protein, RNA and glycogen contents in the liver and the body weight, but increased the cholesterol content. The presence of zeolite in the exposure solution decreased all of the adverse effects. In fish exposed to zeolite as feed additive, all the parameters improved in comparison to control fish, indicating that zeolites can be used safely in biological systems.     

Combined phytoremediation of metal-working fluids with maize plants inoculated with different microorganisms and toxicity assessment of the phytoremediated waste

The aim of this study was to validate the effectiveness of a phytoremediation procedure for metal-working fluids (MWFs) with maize plants growing in hydroponic culture in which the roots grow on esparto fibre and further improve bioremediation potential of the system with root beneficial bacteria, seeking a synergistic effect of the plant–microorganism combination. Chemical oxygen demand (COD), pH, total and type of hydrocarbons measured after phytoremediation indicated that the process with maize plants was successful, as demonstrated by the significant decrease in the parameters measured. This effect was mainly due to the plant although inoculated microorganisms had a relevant effect on the type of remaining hydrocarbons. The success of the phytoremediation process was further confirmed by two toxicity tests, one of them based on chlorophyll fluorescence measurements on maize plants and another one based on cyanobacteria, using a bioluminescent toxicity bioassay; both tests demonstrated that the phytoremediated waste was significantly less toxic than the initial non-phytoremediated MWFs.     

The influence of time on lead toxicity and bioaccumulation determined by the OECD earthworm toxicity test

Internationally agreed standard protocols for assessing chemical toxicity of contaminants in soil to worms assume that the test soil does not need to equilibrate with the chemical to be tested prior to the addition of the test organisms and that the chemical will exert any toxic effect upon the test organism within 28 days. Three experiments were carried out to investigate these assumptions. The first experiment was a standard toxicity test where lead nitrate was added to a soil in solution to give a range of concentrations. The mortality of the worms and the concentration of lead in the survivors were determined. The LC50s for 14 and 28 days were 5311 and 5395 microgPb g(-1)soil respectively. The second experiment was a timed lead accumulation study with worms cultivated in soil containing either 3000 or 5000 microgPb g(-1)soil. The concentration of lead in the worms was determined at various sampling times. Uptake at both concentrations was linear with time. Worms in the 5000 microg g(-1) soil accumulated lead at a faster rate (3.16 microg Pb g(-1)tissue day(-1)) than those in the 3000 microg g(-1) soil (2.21 microg Pb g(-1)tissue day(-1)). The third experiment was a timed experiment with worms cultivated in soil containing 7000 microgPb g(-1)soil. Soil and lead nitrate solution were mixed and stored at 20 degrees C. Worms were added at various times over a 35-day period. The time to death increased from 23 h, when worms were added directly after the lead was added to the soil, to 67 h when worms were added after the soil had equilibrated with the lead for 35 days. In artificially Pb-amended soils the worms accumulate Pb over the duration of their exposure to the Pb. Thus time limited toxicity tests may be terminated before worm body load has reached a toxic level. This could result in under-estimates of the toxicity of Pb to worms. As the equilibration time of artificially amended Pb-bearing soils increases the bioavailability of Pb decreases. Thus addition of worms shortly after addition of Pb to soils may result in the over-estimate of Pb toxicity to worms. The current OECD acute worm toxicity test fails to take these two phenomena into account thereby reducing the environmental relevance of the contaminant toxicities it is used to calculate.     

Melatonin and nitric oxide enhance cadmium tolerance and phytoremediation efficiency in Catharanthus roseus (L.) G. Don

Environmental Science and Pollution Research - In this study, a pot experiment was performed to evaluate the effects of foliar spray with sodium nitroprusside (200 μM SNP) and melatonin (100...     

The use of maize and poplar in chelant-enhanced phytoextraction of lead from contaminated agricultural soils

Chelant-enhanced phytoextraction of heavy metals is an emerging technological approach for a non-destructive remediation of contaminated soils. The main objectives of this study were (i) to assess the extraction efficiency of two different synthetic chelating agents (ethylenediaminetetraacetic acid (EDTA) and ethylenediaminedisuccinic acid (EDDS)) for desorbing Pb from two contaminated agricultural soils originating from a mining and smelting district and (ii) to assess the phytoextraction efficiency of maize (Zea mays) and poplar (Populus sp.) after EDTA application. EDTA was more efficient than EDDS in desorbing and complexing Pb from both soils, removing as much as 60% of Pb. Maize exhibited better results than poplar when extracting Pb from the more acidic (pH approximately 4) and more contaminated (up to 1360 mg Pb kg(-1)) agricultural soil originating from the smelting area. On the other hand, poplars proved to be more efficient when grown on the near-neutral (pH approximately 6) and less contaminated (up to 200 mg Pb kg(-1)) agricultural soil originating from the mining area. Furthermore, the addition of EDTA led to a significant increase of Pb content especially in poplar leaves, proving a strong translocation rate within the poplar plants.     

Predicting molybdenum toxicity to higher plants: Influence of soil properties

The effect of soil properties on the toxicity of molybdenum (Mo) to four plant species was investigated. Soil organic carbon or ammonium-oxalate extractable Fe oxides were found to be the best predictors of the 50% effective dose (ED₅₀) of Mo in different soils, explaining > 65% of the variance in ED₅₀ for four species except for ryegrass (26-38%). Molybdenum concentrations in soil solution and consequently plant uptake were increased when soil pH was artificially raised because sorption of Mo to amorphous oxides is greatly reduced at high pH. The addition of sulphate significantly decreased Mo uptake by oilseed rape. For risk assessment, we suggest that Mo toxicity values for plants should be normalised using soil amorphous iron oxide concentrations.     

Consumption of biogenic nitric oxide in hydrated soil

An experimental study was conducted in order to determine the relationship of nitric oxide (NO) consumption to water-filled pore space in soil. A test system that included the capability to blend gases, test soil samples, and analyze off-gases was used to conduct the study. The experimental set consisted of three replicates at five different levels of soil water content and three different levels of soil nitrogen in a sandy loam soil: unamended soil, soil fertilized at 56.2 kg N per ha (50 lb N acre(-1)), and soil fertilized at 112.3 kg N per ha (100 lb N acre(-1)). The average NO consumption rates were 7.1x10(-13) g-NO cm(-3) soil, 3.5x10(-11) g-NO cm(-3) soil, and 1.5x10(-10) g-NO cm(-3) soil, respectively.     

Comparison of sewage sludge toxicity to plants and invertebrates in three different soils

Understanding the effect of soil type on the overall toxicity of sewage sludge is one of the most important issues concerning environmental risks associated with the sewage sludge land application. The aim of the study was to determine the influence of different soils (sandy, loamy and OECD soil) on sewage sludges toxicity in relation to plants (Lepidium sativum, Sorghum saccharatum, Sinapis alba) and an invertebrate species (Heterocypris incongruens). The most evident negative influence of sewage sludges on root growth was observed in the case of OECD soil. The EC(50) values determined on the basis of the root growth inhibition of all tested plants were in the range 0.1-6.4%, 0.03-9.4% and 6.6-22.1% (% of sewage sludgekg(-1) soil) for OECD, sandy and loamy soil, respectively. Soil type also affects the sewage sludge toxicity in relation to H. incongruens. The LC(50) (mortality) values ranged from 0.26% to 11.5% depending on the sludge tested. For EC(50) (growth inhibition) values ranged from 10.7% to 36.2%.     

Uptake of polycyclic aromatic hydrocarbons by maize plants

Roots and above-ground parts (tops) of maize plants, comprising cuticles, leaves and stems, have been exposed separately to polycyclic aromatic hydrocarbons (PAHs) by means of air-tight bicameral exposure devices. Maize roots and tops of plants directly accumulate PAHs from aqueous solutions and from air in proportion to exposure levels. Root and leaf concentration factors (log RCF and log LCF) are log-linear functions of log-based octanol-water partition coefficient (log Kow) and log-based octanol-air partition coefficient (log Koa). The PAHs' concentrations among cuticles, leaves and stems display good correlations with each other. PAH concentrations in each part of the plant tested correlated positively with atmospheric PAHs' concentrations. Comparisons between PAHs' concentrations of root epidermis and root tissue showed similar correlations. Bulk concentrations of contaminants in various plant tissues differed greatly, but these differences disappeared after normalization to lipid contents suggesting lipid-based partitioning of PAHs among maize tissues.     

Lead tolerance in plants: strategies for phytoremediation

Lead (Pb) is naturally occurring element whose distribution in the environment occurs because of its extensive use in paints, petrol, explosives, sludge, and industrial wastes. In plants, Pb uptake and translocation occurs, causing toxic effects resulting in decrease of biomass production. Commonly plants may prevent the toxic effect of heavy metals by induction of various celular mechanisms such as adsorption to the cell wall, compartmentation in vacuoles, enhancement of the active efflux, or induction of higher levels of metal chelates like a protein complex (metallothioneins and phytochelatins), organic (citrates), and inorganic (sulphides) complexes. Phyotochelains (PC) are synthesized from glutathione (GSH) and such synthesis is due to transpeptidation of γ-glutamyl cysteinyl dipeptides from GSH by the action of a constitutively present enzyme, PC synthase. Phytochelatin binds to Pb ions leading to sequestration of Pb ions in plants and thus serves as an important component of the detoxification mechanism in plants. At cellular level, Pb induces accumulation of reactive oxygen species (ROS), as a result of imbalanced ROS production and ROS scavenging processes by imposing oxidative stress. ROS include superoxide radical (O₂ ^.?), hydrogen peroxide (H₂O₂) and hydroxyl radical (^·OH), which are necessary for the correct functioning of plants; however, in excess they caused damage to biomolecules, such as membrane lipids, proteins, and nucleic acids among others. To limit the detrimental impact of Pb, efficient strategies like phytoremediation are required. In this review, it will discuss recent advancement and potential application of plants for lead removal from the environment.     

Physiological responses,tolerance, and remediation strategies in plants exposed to metalloids

Environmental Science and Pollution Research - Metalloids are a subset of particular concern to risk assessors and toxicologists because of their well-documented potential hazards to plant system....     

The role of humic acid in the toxicity of arsenite to the diatom Navicula sp.

Dissolved organic matter (DOM) affects arsenite [As(III)] toxicity by altering its sorption equilibrium at the cell wall interface. A better understanding of such mechanism is of great importance to assess As(III) ecotoxicity in aquatic systems. Batch experiments were conducted to study the effects of DOM on the regulation of As(III) sorption and toxicity in the diatom Navicula sp. The influence of humic acid (HA) on As(III) toxicity was assessed by measuring algal growth, chlorophyll a, and reactive oxygen species (ROS), whereas As(III) mobility across the cell wall was estimated by determining the concentration of intracellular, cell-wall-bound, and free As(III) ions in cell media. Results showed that the effects of HA on arsenite toxicity varied depending on various combinations of As(III)-HA concentrations. EC₅₀ had an approximate threefold increase from 8.32 (HA-free control) to 22.39 μM (at 20 mg L^?1 HA) when Navicula sp. was exposed to 1.0–100.0 μM of As(III), compared to an overall low complexation ratio of HA-As(III) in a range of 0.91–6.00 %. The cell wall-bound and intracellular arsenic content decreased by 19.8 and 20.3 %, respectively, despite the lower arsenite complexation (2.10?±?0.16 % of the total As). Meanwhile, intracellular ROS was decreased by 12.6 % in response to 10.0 μM As(III) and 10 mg L^?1 HA vs. the HA-free control. The significant contrast indicated that complexation alone could not explain the HA-induced reduction in arsenite toxicity and other factors including HA–cell surface interactions may come into play. Isotherms describing adsorption of HA to the Navicula sp. cells combined with morphological data by scanning electron microscopy revealed a protective HA floccule coating on the cell walls. Additional Fourier transform infrared spectroscopic data suggested the involvement of carboxylic groups during the adsorption of both HA and As(III) on the Navicula sp. cell surface. Collective data from this study suggest that cell wall-bound HA can moderate As(III) toxicity through the formation of a protective floccule coating occupying As(III) sorption sites and decreased effective functional groups capable of binding As(III). Our findings imply that As(III) toxicity can be alleviated due to the increased hindrance to cellular internalization of As(III) in the presence of naturally abundant DOM in water.     

The changes of nitric oxide production during the growth of Microcystis aerugrinosa

This study characterized the changes of nitric oxide (NO) production during the growth of Microcystis aerugrinosa, a cyanobacterium which usually cause cyanobacterial blooms. Results showed a drastic NO release accompanying with cell density and Chl-a content sharp rises when M. aerugrinosa grew from fifth day to sixth day. Moreover, high N:P ratio accelerated the cyanobacterial growth and NO burst. Sodium nitroprusside, an exogenous NO donor, promoted M. aerugrinosa growth with the optimal concentration of 0.1 mg/L. Experiments by supplementing with sodium nitrite and l-arginine demonstrated NO production in M. aerugrinosa cells was mainly through nitrate reductase (NR) pathway while minorly through NO synthase pathway. All these data suggested M. aerugrinosa produced increasing NO during its growth mainly by NR pathway, during which NO positively regulated the growth of M. aerugrinosa.     

Emissions of nitric oxide from 79 plant species in response to simulated nitrogen deposition

To assess the potential contribution of nitric oxide (NO) emission from the plants grown under the increasing nitrogen (N) deposition to atmospheric NO budget, the effects of simulated N deposition on NO emission and various leaf traits (e.g., specific leaf area, leaf N concentration, net photosynthetic rate, etc.) were investigated in 79 plant species classified by 13 plant functional groups. Simulated N deposition induced the significant increase of NO emission from most functional groups, especially from conifer, gymnosperm and C(3) herb. Moreover, the change rate of NO emission was significantly correlated with the change rate of various leaf traits. We conclude that the plants grown under atmospheric N deposition, especially in conifer, gymnosperm and C(3) herb, should be taken into account as an important biological source of NO and potentially contribute to atmospheric NO budget.     

Uptake and accumulation of lead by plants from the Bo Ngam lead mine area in Thailand

A field survey of terrestrial plants growing on Bo Ngam lead mine area, Thailand, was conducted to identify species accumulating exceptionally high concentrations of lead. Plant and soil samples were collected from five areas. Lead concentrations in surface soil ranged from 325 to 142,400 mg/kg. The highest lead concentration in soil was found at the ore dressing plant area and lowest at a natural pond area. In different areas, the concentrations of lead in plants were different when comparing various study sites. A total of 48 plant species belonging to 14 families were collected from five sampling sites. Twenty-six plant species had lead concentrations more than 1000 mg/kg in their shoots. Three species (Microstegium ciliatum, Polygala umbonata, Spermacoce mauritiana) showed extremely high lead concentrations in their shoots (12,200-28,370 mg/kg) and roots (14,580-128,830 mg/kg).