Uptake of lead by bacteria isolated from industrial effluents and their potential use in bioremediation of wastewater

Due to rising populations and human activities, heavy metals (HM) toxicity has become a serious problem for all life forms. The present study deals with isolating and identifying lead-resistant bacteria from contaminated wastewater of tanneries effluents. Two isolated strains were identified as Bacillus cereus (ID1), and Bacillus sp. (ID3), and both strains resisted a 25 mM concentration of Lead nitrate (Pb (NO₃)₂). After four days of treatment, Bacillus cereus (ID1) showed 80% lead uptake, and Bacillus sp. (ID3) showed 88%. Lead uptake was confirmed by Energy dispersive X-Ray (EDX) analysis. Fourier transform infrared spectroscopy (FTIR) showed that structural alterations had occurred in functional groups of the treated samples compared to the controls. Our research indicates that these Bacillus strains may be useful in bioremediating heavy metals from polluted environments. Further investigation into the processes involved in the uptake and homeostasis of heavy metals by these strains is required, as is the identification of the genes and enzymes responsible for Pb-bioremediation.     

Dual augmentation for aerobic bioremediation of MTBE and TCE pollution in heavy metal-contaminated soil

In this work we isolated from soil and characterized several bacterial strains capable of either resisting high concentrations of heavy metals (Cd²⁺ or Hg²⁺ or Pb²⁺) or degrading the common soil and groundwater pollutants MTBE (methyl-tert-butyl ether) or TCE (trichloroethylene). We then used soil microcosms exposed to MTBE (50 mg/l) or TCE (50 mg/l) in the presence of one heavy metal (Cd 10 ppm or Hg 5 ppm or Pb 50 or 100 ppm) and two bacterial isolates at a time, a degrader plus a metal-resistant strain. Some of these two-membered consortia showed degradation efficiencies well higher (49–182% higher) than those expected under the conditions employed, demonstrating the occurrence of a synergetic relationship between the strains used. Our results show the efficacy of the dual augmentation strategy for MTBE and TCE bioremediation in the presence of heavy metals.     

Environmental control of metabolic fluxes in thermotolerant methylotrophic Bacillus strains

Recently we have isolated a number of thermotolerant, spore-forming methylotrophic bacilli in pure culture. With a methanol-limited chemostat culture of strain Ts1, incremental increases in the incubation temperature from 45°C to 62.5°C revealed an optimum with respect to growth yield of 52.5°C, and a maximum of 62.5°C. Similar investigations revealed a pH optimum of 7.5 and a broad growth rate optimum with respect to growth yield. The organism displayed a low maintenance energy requirement and high growth yield (attained simultaneosly with high growth rates) during growth on methanol. Under all conditions of methanol limitation, substrate was oxidized solely to biomass and CO₂ and carbon recoveries greater than 90% were manifest. Our data suggested that this resulted from an ability of the organism to precisely adjust its catabolic and anabolic pathways to suit prevailing growth conditions. These results are discussed in relation to previously reported data on thermophiles in both batch and chemostat culture.     

土壤污染的生物修复技术研究进展

土壤污染是当今面临的一个重要环境问题。常规的土壤污染治理技术,如物理及化学治理技术,由于其技术要求高或经济成本昂贵,对土壤结构的扰动破坏严重,因而对其大规模的推广应用存在较大问题。而生物修复技术已被证明是一项非常有应用前景的新技术,成为土壤污染治理研究领域的一个热点,本文综述了近年来有机物及重金属土壤污染的生物修复机理和研究进展,并对其治理技术的最新研究动态、存在问题及发展趋势做了初步的讨论。     

Genetics of carbon metabolism in methylotrophic bacteria

Abstract The application of genetic techniques to the methylotrophic bacteria has greatly enhanced studies of these important organisms. Two methylotrophic systems have been studied in some detail, the serine cycle for formaldehyde assimilation and the methanol oxidation system. In both cases, genes have been cloned and mapped in Methylobacterium species (facultative serine cycle methanol-utilizers). In addition, methanol oxidation genes have been studied in an autotrophic methanol-utilizer ( Paracoccus denitrificans ) and three methanotrophs ( Methylosporovibrio methanica, Methylomonas albus and Methylomonas sp. A4). Although much remains to be learned in these systems, it is becoming clear that the order of C₁ genes has been conserved to some extent in methylotrophic bacteria, and that many C₁ genes are loosely clustered on the chromosome. Operons appear to be rare, but some examples have been observed. The extension of genetic approaches to both the obligate and facultative methylotrophs holds much promise for the future in understanding and manipulating the activities of these bacteria.     

Metabolic fluxes during the growth of thermotolerant methylotrophic Bacillus strains in methanol-sufficient chemostat cultures

Growth of the thermotolerant methylotrophic Bacillus strain TS1 in methanol-limited chemostat culture showed that the substrate was oxidized solely to biomass and CO₂. When a pulse of methanol was added to the growth vessel anabolism could be shown to be dissociated from catabolism for a transient period of time. Present data shows that when the organism was grown with a limitation other than carbon, some of the substrate was channelled into metabolite over-production. When the organism was grown under N-limitation 2-oxoglutarate accumulated in the culture medium in small amounts whilst acetate accumulated under all carbon excess conditions. Although the average carbon recovery was 92%, analysis of the culture filtrates for other metabolites failed to show significant amounts of any individual product above those detected in carbon-limited growth comditions. The results are discussed in relation to published data.     

The use of macrophytes in bioremediation

The development of reedbed technology for bioremediation is reviewed. The future development, potential and implementation of these systems are considered in detail together with the cost effectiveness and ease of maintenance.     

Fungal degradation of chlorpyrifos by Verticillium sp. DSP in pure cultures and its use in bioremediation of contaminated soil and pakchoi

Pesticides residues in soils and on vegetables are a public safety concern. Pretreatment with microorganisms degrading pesticides has the potential to alleviate the conditions. For this purpose, the degradation characteristics of chlorpyrifos by an isolated fungal strain Verticillium sp. DSP in pure cultures, soil, and on pakchoi (Brassica chinensis L.) were investigated. Degradation rate of chlorpyrifos in the mineral salts medium was proportional to the concentrations of chlorpyrifos ranging from 1 to 100 mg l⁻¹. The rate of degradation for chlorpyrifos (1 mg l⁻¹) in the mineral salts medium was 1.12 and 1.04 times faster at pH 7.0 than those at pHs 5.0 and 9.0, and the degradation at 35 °C was 1.15 and 1.12 times faster, respectively, than those at 15 and 20 °C. The addition of the fungal strain DSP into the contaminated soils was found to significantly increase the degradation of chlorpyrifos. Degradation rates of chlorpyrifos in inoculated soils were 3.61, 1.50 and 1.10 times faster in comparison with the sterilized soil, previously chlorpyrifos-untreated soil, and previously chlorpyrifos-treated soil under laboratory conditions. In contrast to the controls, the half-lives of chlorpyrifos were significantly shortened by 10.9% and 17.6% on treated pakchoi, 12.0% and 37.1% in inoculated soils, respectively, in the greenhouse and open field. The results indicate that the fungal strain DSP can be used successfully for the removal or detoxification of chlorpyrifos residues in/on contaminated soil and vegetable.     

How synthetic biology can help bioremediation

The World Health Organization reported that “an estimated 12.6 million people died as a result of living or working in an unhealthy environment in 2012, nearly 1 in 4 of total global deaths”. Air, water and soil pollution were the significant risk factors, and there is an urgent need for effective remediation strategies. But tackling this problem is not easy; there are many different types of pollutants, often widely dispersed, difficult to locate and identify, and in many cases cost-effective clean-up techniques are lacking. Biology offers enormous potential as a tool to develop microbial and plant-based solutions to remediate and restore our environment. Advances in synthetic biology are unlocking this potential enabling the design of tailor-made organisms for bioremediation.In this article, we showcase examples of xenobiotic clean-up to illustrate current achievements and discuss the limitations to advancing this promising technology to make real-world improvements in the remediation of global pollution.     

Electron transport chains in organohalide-respiring bacteria and bioremediation implications

In situ remediation employing organohalide-respiring bacteria represents a promising solution for cleanup of persistent organohalide pollutants. The organohalide-respiring bacteria conserve energy by utilizing H₂ or organic compounds as electron donors and organohalides as electron acceptors. Reductive dehalogenase (RDase), a terminal reductase of the electron transport chain in organohalide-respiring bacteria, is the key enzyme that catalyzes halogen removal. Accumulating experimental evidence thus far suggests that there are distinct models for respiratory electron transfer in organohalide-respirers of different lineages, e.g., Dehalococcoides, Dehalobacter, Desulfitobacterium and Sulfurospirillum. In this review, to connect the knowledge in organohalide-respiratory electron transport chains to bioremediation applications, we first comprehensively review molecular components and their organization, together with energetics of the organohalide-respiratory electron transport chains, as well as recent elucidation of intramolecular electron shuttling and halogen elimination mechanisms of RDases. We then highlight the implications of organohalide-respiratory electron transport chains in stimulated bioremediation. In addition, major challenges and further developments toward understanding the organohalide-respiratory electron transport chains and their bioremediation applications are identified and discussed.     

Comparison of bioremediation strategies for soil impacted with petrochemical oily sludge

Different bioremediation techniques (natural attenuation, biostimulation and bioaugmentation) in contaminated soils with two oily sludge concentrations (1.5% and 6.0%) in open and closed microcosms systems were assessed during 90 days. The results showed that the highest biodegradation rates were obtained in contaminated soils with 6% in closed microcosms. Addition of microbial consortium and nutrients in different concentrations demonstrated higher biodegradation rate of total petroleum hydrocarbons (TPH) than those of the natural attenuation treatment. Soils treated in closed microcosms showed highest removal rate (84.1 ± 0.9%) when contaminated at 6% and bacterial consortium and nutrients in low amounts were added. In open microcosms, the soil contaminated at 6% using biostimulation with the highest amounts of nutrients (C:N:P of 100:10:1) presented the highest degradation rate (78.7 ± 1.3%). These results demonstrate that the application of microbial consortium and nutrients favored biodegradation of TPH present in oily sludge, indicating their potential applications for treatment of the soils impacted with this important hazardous waste.     

Effect of petrochemical sludge concentrations on microbial communities during soil bioremediation

Qualitative and quantitative changes of microbial communities in soil microcosms during bioremediation were determined throughout one year. The soil was contaminated with 0%, 2.5%, 5%, 10% (wt/wt) of petrochemical sludge containing polynuclear aromatic hydrocarbons. We analyzed the hydrocarbon concentration in the microcosms, the number of cultivable bacteria using CFU and most probable number assays, the community structure using denaturing gradient gel electrophoresis, and the metabolic activity of soil using dehydrogenase activity and substrate-induced respiration assays. After one year of treatment, the chemical analysis suggested that the hydrocarbon elimination process was over. The biological analysis, however, showed that the contaminated microcosms suffered under long-term disturbance. The number of heterotrophic bacteria that increased after sludge addition (up to 10(8)-10(9) cells ml(-1)) has not returned to the level of the control soil (2-6 x 10(7) cells ml(-1)). The community structure in the contaminated soils differed considerably from that in the control. The substrate-induced respiration of the contaminated soils was significantly lower (approximately 10-fold) and the dehydrogenase activity was significantly higher (20-40-fold) compared to the control. Changes in the community structure of soils depended on the amount of added sludge. The species, which were predominant in the sludge community, could not be detected in the contaminated soils.     

Soil chromium bioremediation: Synergic activity of actinobacteria and plants

The aim of this work was to evaluate a strategy to reduce the bioavailable chromium fraction in soil, using a Cr(VI) resistant microorganism, Streptomyces sp. MC1, under non sterile conditions, with maize plants as bioindicator and/or bioremediator.Soil samples were contaminated with 100, 200 and 400 mg kg⁻¹ of Cr(VI) or Cr(III). Bioavailable chromium (35%) was only detected in samples with Cr(VI). Soil samples with Cr(VI) 200 mg kg⁻¹ were inoculated with Streptomyces sp. MC1, and bioavailable chromium decreased up to 73%.Zea mays seedlings were planted in soil samples contaminated with chromium. Plantlets accumulated chromium mainly as Cr(III), and biomass decreased up to 88%. Streptomyces sp. MC1 was inoculated in soil samples contaminated with 200 mg kg⁻¹ of Cr(VI) and Z.mays seedlings were planted.Streptomyces sp. MC1 caused Z.mays biomass increase (57%), chromium accumulation and bioavailable chromium decreased up to 46% and 96%, respectively.This work constitutes the first contribution of cooperative action between actinobacteria and Z.mays in the bioremediation of Cr(VI) contaminated soil. The large removal capacity of bioavailable chromium by Streptomyces sp. MC1 and Z.mays infers that they could be successfully applied together in bioremediation of soils contaminated with Cr(VI).     

Environmental regulation of alcohol metabolism in thermotolerant methylotrophic Bacillus strains

The thermotolerant methylotroph Bacillus sp. C1 possesses a novel NAD-dependent methanol dehydrogenase (MDH), with distinct structural and mechanistic properties. During growth on methanol and ethanol, MDH was responsible for the oxidation of both these substrates. MDH activity in cells grown on methanol or glucose was inversely related to the growth rate. Highest activity levels were observed in cells grown on the C₁-substrates methanol and formaldehyde. The affinity of MDH for alcohol substrates and NAD, as well as V _max, are strongly increased in the presence of a M _r 50,000 activator protein plus Mg²⁺-ions [Arfman et al. (1991) J Biol Chem 266: 3955–3960]. Under all growth conditions tested the cells contained an approximately 18-fold molar excess of (decameric) MDH over (dimeric) activator protein. Expression of hexulose-6-phosphate synthase (HPS), the key enzyme of the RuMP cycle, was probably induced by the substrate formaldehyde. Cells with high MDH and low HPS activity levels immediately accumulated (toxic) formaldehyde when exposed to a transient increase in methanol concentration. Similarly, cells with high MDH and low CoA-linked NAD-dependent acetaldehyde dehydrogenase activity levels produced acetaldehyde when subjected to a rise in ethanol concentration. Problems frequently observed in establishing cultures of methylotrophic bacilli on methanol- or ethanol-containing media are (in part) assigned to these phenomena.Abbreviations MDH NAD-dependent methanol dehydrogenase - ADH NAD-dependent alcohol dehydrogenase - A1DH CoA-linked NAD-dependent aldehyde dehydrogenase - HPS hexulose-6-phosphate synthase - G6Pdh glucose-6-phosphate dehydrogenase     

Mineralization of hexachlorocyclohexane in soil during solid-phase bioremediation

Soil containing hexachlorocyclohexane (HCH) was spiked with ¹⁴C--HCH and then subjected to bioremediation in bench-scale microcosms to determine the rate and extent of mineralization of the ¹⁴C-labeled HCH to ¹⁴CO₂. The soil was treated using two different DARAMEND amendments, D6386 and D6390. The amendments were previously found to enhance natural HCH bioremediation as determined by measuring the disappearance of parent compounds under either strictly oxic conditions (D6386), or cycled anoxic/oxic conditions (D6390). Within 80 days of the initiation of treatment, mineralization was observed in all of the strictly oxic microcosms. However, mineralization was negligible in the cycled anoxic/oxic microcosms throughout the 275-day study, even after cycling was ceased at 84 days and although significant removal (up to 51%) of indigenous -HCH (146 mg/kg) was detected by GC with electron capture detector. Of the amended, strictly oxic treatments, only one, in which 47% of the spiked ¹⁴C-HCH was recovered as ¹⁴CO₂, enhanced mineralization compared with an unamended treatment (in which 34% recovery was measured). Other oxic treatments involving higher amendment application rates or auxiliary carbon sources were inhibitory to mineralization. Thus, although HCH degradation occurs during the application of either oxic or cycled anoxic/oxic DARAMEND treatments, mineralization of -HCH may be inhibited depending on the amendment and treatment protocol.     

Novel static composting method for bioremediation of olive mill waste

This paper presents results obtained on the evaluation of static composting process aimed at bioremediation of the hazardous solid olive mill waste (OMW). The static composting process carried out in gas-permeable polyethylene bags followed the fluctuating temperature and oxygen profiles similar to those seen in aerated composting systems. Static composting resulted in apparent increases and decreases in values for total nitrogen and C:N ratios respectively during the process. The amount of nitrogen (>3%) in the composting end product was in agreement with the Italian legislation (Decreto Legislativo 29 aprile 2010, n. 75) specification for nitrogen fertilizer. A gradual decrease in polyphenols during the storage of compost resulted in a non-phytotoxic composted organic matter high in humic substances. Different respirometric tests also stated high biological stability of the end compost product.     

Uptake of heavy metals by <Emphasis Type="Italic">Stylonychia mytilus</Emphasis> and its possible use in decontamination of industrial wastewater

The heavy metal resistant ciliate, Stylonychia mytilus, isolated from industrial wastewater has been shown to be potential bioremediator of contaminated wastewater. The ciliate showed tolerance against Zn²⁺ (30 μg/mL), Hg²⁺ (16 μg/mL) and Ni²⁺ (16 μg/mL). The metal ions slowed down the growth of the ciliate as compared with the culture grown without metal stress. The reduction in cell population was 46% for Cd²⁺, 38% for Hg²⁺, 23% for Zn²⁺, 39% for Cu²⁺ and 51% for Ni²⁺ after 8 days of metal stress. S. mytilus reduced 91% of Cd²⁺, 90% of Hg²⁺ and 98% of Zn²⁺ from the medium after 96 h of incubation in a culture medium containing 10 μg/mL of the respective metal ions. Besides this, the ciliate could also remove 88% of Cu²⁺ and 73% Ni²⁺ from the medium containing 5 μg/mL of each metal after 96 h. The ability of Stylonychia to take up variety of heavy metals from the medium could be exploited for metal detoxification and environmental clean-up operations.