Bioaugmentation and biostimulation strategies to improve the effectiveness of bioremediation processes

Bioremediation, involving bioaugmentation and/or biostimulation, being an economical and eco-friendly approach, has emerged as the most advantageous soil and water clean-up technique for contaminated sites containing heavy metals and/or organic pollutants. Addition of pre-grown microbial cultures to enhance the degradation of unwanted compounds (bioaugmentation) and/or injection of nutrients and other supplementary components to the native microbial population to induce propagation at a hastened rate (biostimulation), are the most common approaches for in situ bioremediation of accidental spills and chronically contaminated sites worldwide. However, many factors like strain selection, microbial ecology, type of contaminant, environmental constraints, as well as procedures of culture introduction, may lead to their failure. These drawbacks, along with fragmented literature, have opened a gap between laboratory trials and on-field application. The present review discusses the effectiveness as well as the limitations of bioaugmentation and biostimulation processes. A summary of experimental studies both in confined systems under controlled conditions and of real case studies in the field is presented. A comparative account between the two techniques and also the current scenario worldwide for in situ biotreatment using bioaugmentation and biostimulation, are addressed.     

Bioaugmentation for enhancing biological wastewater treatment

The literature on bioaugmentation products has been reviewed. Their manufacture and method of use is explained. The various applications are listed and the independent investigations, as opposed to manufacturers accounts, at laboratory and full scale are reviewed. The economics and kinetic modelling are also discussed. In laboratory investigations bioaugmentation often failed, whereas at full scale it was often successful, probably due to the imposition of steady state at laboratory scale. Most products require a period of acclimatisation before working; this and other possible reasons for failure are discussed.     

Anaerobic fluidised bed for the purification of effluents from chemical and mechanical pulping

Anaerobic treatment has seldom been used for wastewaters from the pulp and paper industry and other branches of the chemical industry. Escape of volatile pollutants to the atmosphere, which always occurs during aerobic treatment, is avoided, and much less sludge is being produced than in an aerobic process. The greatest obstacle for using anaerobic treatment in the pulp and paper industry is the large wastewater volume, which necessitates short hydraulic detention times, because the treatment is to occur in an enclosed space. We used solid carrier particles to prevent wash-out of biomass from the reactor at high hydraulic loading, and an up-flow system in order to be able to use very small carrier particles, maximizing the surface area for biomass attachment. In this paper we describe and discuss the results obtained with this type of anaerobic reactor (fluidised bed) at bench and semitechnical scale for wastewaters from pressurized ground wood pulping and paper manufacture, sulphite pulp evaporator condensate and bleach waste. Earlier work with Kraft pulp bleaching effluent and thermomechanical pulping wastewater and evaporator condensates using anaerobic reactors is also discussed. The results obtained thus far show that there are several wastewater streams from the pulping industry, where 60 to 90% of the dissolved organic pollutants (measured as COD_Cr or TOC) was biodegraded within 4 to 24 h. The high strength waste streams (COD_Cr 2000 mg O₂ 1⁻¹) allowed organic space load of 4 to 10 kg COD_Cr m⁻³ reactor volume d⁻¹. With low strength wastes the hydraulic loading was the limiting factor.     

Robustness of optimal mixed strategies

 Mixed strategies, or variable phenotypes, can evolve in fluctuating environments when at the time that a strategy is chosen the consequences of that decision are relatively uncertain. In a previous paper, we have shown several examples of explicit forms of optimal mixed strategies when an environmental distribution and payoff function are given. In many of these examples, the mixed strategy has a continuous distribution. In a recent study, however, Sasaki and Ellner proved that, if the distribution of the environmental parameter is modified in certain ways, the exact ESS distribution becomes discrete rather than continuous. This forces us to take a closer look at the robustness of optimal mixed strategies. In the current paper we prove that such strategies are indeed robust against small perturbations of the environmental distribution and/or the payoff function, in the sense that the optimal strategy distribution for the perturbed system, converges weakly to the optimal strategy distribution for the unperturbed system as the magnitude of the perturbation goes to zero. Furthermore, we show that the fitness difference between the two strategies converges to zero. Thus, although optimal strategies in ‘ideal’ and perturbed systems can be qualitatively different, the difference between the distributions (in a measure theoretic sense) is small. Received: 27 October 1996 / Revised version: 5 March 1997     

Different strategies for the chemical synthesis of lignans

This paper summarises the results of three projects. The first is concerned with developing general routes for the synthesis of lignans. In particular, two routes involving tandem conjugate addition reactions and Diels Alder reactions respectively that have been used to synthesise podophyllotoxin derivatives are described. The second project is concerned with the asymmetric synthesis of lignans and involves the application of these reactions, with the introduction of a menthyloxy group as a chiral auxiliary, to achieve the asymmetric synthesis of podophyllotoxin derivatives. The third project is concerned with the attempted biomimetic syntheses of podophyllotoxin derivatives using oxidative coupling reactions. Attention is focussed primarily on the use of hypervalent iodine reagents, which yield stegane and isostegane derivatives rather than podophyllotoxin derivatives. Other examples of biaryl coupling leading to stegane and isostegane derivatives are included, and other examples of lignan synthesis involving hypervalent iodine reagents are also described. Abbreviations: DDQ – 2,3-dichloro-5,6-dicyano-1,4-benzoquinone; DMAD – dimethyl acetylenedicarboxylate (dimethyl butynedioate); PIDA – phenyliodonium diacetate, iodobenzene diacetate; PIFA – phenyliodonium bis(trifluoroacetate), [bis(trifluoroacetoxy) iodobenzene]; Ra-Ni – Raney nickel; TBAF – tetrabutylammonium fluoride; TBDMS –t-butyldimethylsilyl; TFA – trifluoroacetic acid; TFAA – trifluoroacetic anhydride; TFE – 2,2,2-trifluoroethanol; TTFA – thallium(III) trifluoroacetate.     

Bioaugmentation of activated sludge towards 3-chloroaniline removal with a mixed bacterial population carrying a degradative plasmid

A bioaugmentation approach combining several strategies was applied to achieve degradation of 3-chloroaniline (3CA) in semicontinuous activated sludge reactors. In a first step, a 3CA-degrading Comamonas testosteroni strain carrying the degradative plasmid pNB2 was added to a biofilm reactor, and complete 3CA degradation together with spread of the plasmid within the indigenous biofilm population was achieved. A second set of reactors was then bioaugmented with either a suspension of biofilm cells removed from the carrier material or with biofilm-containing carrier material. 3CA degradation was established rapidly in all bioaugmented reactors, followed by a slow adaptation of the non-bioaugmented control reactors. In response to variations in 3CA concentration, all reactors exhibited temporary performance breakdowns. Whereas duplicates of the control reactors deviated in their behaviour, the bioaugmented reactors appeared more reproducible in their performance and population dynamics. Finally, the carrier-bioaugmented reactors showed an improved performance in the presence of high 3CA influent concentrations over the suspension-bioaugmented reactors. In contrast, degradation in one control reactor failed completely, but was rapidly established in the remaining control reactor.     

Mutation-selection equilibrium in games with mixed strategies

We develop a new method for studying stochastic evolutionary game dynamics of mixed strategies. We consider the general situation: there are n pure strategies whose interactions are described by an n×n payoff matrix. Players can use mixed strategies, which are given by the vector (p₁,…,p_n). Each entry specifies the probability to use the corresponding pure strategy. The sum over all entries is one. Therefore, a mixed strategy is a point in the simplex S_n. We study evolutionary dynamics in a well-mixed population of finite size. Individuals reproduce proportional to payoff. We consider the case of weak selection, which means the payoff from the game is only a small contribution to overall fitness. Reproduction can be subject to mutation; a mutant adopts a randomly chosen mixed strategy. We calculate the average abundance of every mixed strategy in the stationary distribution of the mutation-selection process. We find the crucial conditions that specify if a strategy is favored or opposed by selection. One condition holds for low mutation rate, another for high mutation rate. The result for any mutation rate is a linear combination of those two. As a specific example we study the Hawk-Dove game. We prove general statements about the relationship between games with pure and with mixed strategies.     

Growth and mineral accumulation in Eucalyptus camaldulensis seedlings irrigated with mixed industrial effluents

Effects of mixed industrial effluents on growth, dry matter accumulation and mineral nutrient in Eucalyptus camaldulensis seedlings were studied. The objective was to evaluate the adaptability of E. camaldulensis to effluent, tolerance to excess/deficiency of mineral elements and ultimately to determine suitable combinations of industrial/municipal effluent for their use in biomass production in dry areas. Different irrigation treatments were: T(1): good water; T(2): municipal effluent; T(3): textile effluent; T(4): steel effluent; T(5): textile effluent+municipal effluent in 1:1 ratio; T(6): steel effluent+municipal effluent in 1:2 ratio; T(7): steel+textile+municipal effluent in 1:2:2 ratio; and T(8): steel+textile effluent in 1:2 ratio. High concentrations of metal ions and low concentrations of Ca, Mg, K, Na, N and P in soil and seedlings of T(4) resulted in mortality of the seedlings within a few days. Addition of the textile/municipal effluent increased the survival time of the seedlings for two to three months in T(6), T(7) and T(8) treatments. Among the remaining treatments, the seedlings of T(2) attained 131 cm height, 1.97 cm collar diameter, 19 total branches and produced 158 g seedling(-1) of dry biomass at the age of 10 months. The seedling of T(3) produced the least growth and biomass. Growth equivalent to that of the seedlings of T(1) treatment was achieved when municipal effluent was mixed with textile effluent (T(5)). There was a decrease in soil pH, EC, SOC, NH(4)-N, NO(3)-N, PO(4)-P and basic cations and increase in the concentration of Cu, Fe, Mn and Zn with T(4) treatment. The reverse trend was observed in T(3) where a high concentration of Na might have reduced Mg and micronutrient concentration in seedlings potentially affecting root and leaf growth. Mixing of effluents may be useful in tree irrigation to increase biomass productivity, which is evidenced by improved growth in T(5) and survival in T(6), T(7) and T(8) treatments. Further, reduction of toxic concentration of metal ions in effluents may be helpful for a long-term field application.     

Destruction of cyanide in gold mill effluents: biological versus chemical treatments

In gold mining, cyanide has been the preferred lixiviant worldwide since 1887. Although cyanide can be destroyed and recovered by several processes, it is still widely discussed and examined due to its potential toxicity and environmental impact. Biological treatment of cyanide is a well-established process and has been commercially used at gold mining operations in North America. Biological treatment processes facilitate growth of microorganisms that are essential for the treatment. The present review describes the advances in the use of biological treatment for the destruction of cyanide in gold mill effluents.     

Current chemical tagging strategies for proteome analysis by mass spectrometry

Proteomics, the analysis of the protein complement of a cell or an organism, has grown rapidly as a subdiscipline of the life sciences. Mass spectrometry (MS) is one of the central detection techniques in proteome analysis, yet it has to rely on prior sample preparation steps that reduce the enormous complexity of the protein mixtures obtained from biological systems. For that reason, a number of so-called tagging (or labeling) strategies have been developed that target specific amino acid residues or post-translational modifications, enabling the enrichment of subfractions via affinity clean-up, resulting in the identification of an ever increasing number of proteins. In addition, the attachment of stable-isotope-labeled tags now allows the relative quantitation of protein levels of two samples, e.g. those representing different cell states, which is of great significance for drug discovery and molecular biology. Finally, tagging schemes also serve to facilitate interpretation of MS/MS spectra, therefore assisting in de novo elucidation of protein sequences and automated database searching. This review summarizes the different application fields for tagging strategies for today's MS-based proteome analysis. Advantages and drawbacks of the numerous strategies that have appeared in the literature in the last years are highlighted, and an outlook on emerging tagging techniques is given.     

Development of Fungal Inocula for Bioaugmentation of Contaminated Soils

This report describes novel fungal inocula for bioaugmentation of soils contaminated with hazardous organic compounds. The inocula are in the form of pelleted solid substrates coated with a sodium alginate suspension of fungal spores or mycelial fragments and incubated until overgrown with the mycelium of selected lignin-degrading fungi. The organisms evaluated were Phanerochaete chrysosporium (BKM F-1767, ATCC 42725), P. sordida (HHB-8922-Sp), Irpex lacteus (Mad-517, ATCC 11245), Bjerkandera adusta (FP-135160-Sp, ATCC 62023), and Trametes versicolor (MD-277). The pelleted fungal inocula resisted competition and proliferation from indigenous soil microbes, were lower in moisture content than current fungal inocula, and had sufficient mechanical strength to allow handling and introduction into the soil without a change in the mechanical consistency of the pellets. Inoculated at a rate of 3% in artificially contaminated nonsterile soil, I. lacteus, B. adusta, and T. versicolor removed 86, 82, and 90%, respectively, of the pentachlorophenol in 4 weeks. A mathematical model was developed to explain moisture distribution in a hydrogel-coated pelleted substrate.     

Evolution: vertebrate reproductive strategies get mixed up

The mangrove killifish is the only vertebrate known to have a mixed-mating strategy, where hermaphrodites reproduce by either self-fertilisation or cross-breeding. New molecular evidence from this species reveals that occasional cross-breeding between common hermaphroditic individuals and rare pure males results in an injection of genetic variation into otherwise highly homozygous 'clonal' lineages.     

Endophyte-assisted phytoremediation: mechanisms and current application strategies for soil mixed pollutants

Abstract

Phytoremediation uses plants and associated microbes to remove pollutants from the environment and is considered a promising bioremediation method. Compared with well-described single contaminant treatments, the number of studies reporting phytoremediation of soil mixed pollutants has increased recently. Endophytes, including bacteria and fungi, exhibit beneficial traits for the promotion of plant growth, stress alleviation, and biodegradation. Moreover, endophytes either directly or indirectly assist host plants to survive high concentrations of organic and inorganic pollutants in the soil. Endophytic microorganisms can also regulate the plant metabolism in different ways, exhibiting a variety of physiological characteristics. This review summarizes the taxa and physiological properties of endophytic microorganisms that may participate in the detoxification of contaminant mixtures. Furthermore, potential biomolecules that may enhance endophyte mediated phytoremediation are discussed. The practical applications of pollutant-degrading endophytes and current strategies for applying this valuable bio-resource to soil phytoremediation are summarized.     

Bioaugmentation for bioremediation: the challenge of strain selection

Despite its long-term use in bioremediation, bioaugmentation of contaminated sites with microbial cells continues to be a source of controversy within environmental microbiology. This largely results from its notoriously unreliable performance record. In this article, we argue that the unpredictable nature of the approach comes from the initial strain selection step. Up until now, this has been dictated by the search for catabolically competent microorganisms, with little or no consideration given to other essential features that are required to be functionally active and persistent in target habitats. We describe how technical advances in molecular biology and analytical chemistry, now enable assessments of the functional diversity and spatial distribution of microbial communities to be made in situ. These advances now enable microbial populations, targeted for exploitation, to be differentiated to the cell level, an advance that is bound to improve microbial selection and exploitation. We argue that this information-based approach is already proving to be more effective than the traditional 'black-box' approach of strain selection. The future perspectives and opportunities for improving selection of effective microbial strains for bioaugmentation are also discussed.     

Evaluation of different chemical strategies for conjugation of oligonucleotides to peptides

We developed novel assays for high-throughput detection of one or many kinases or proteases. The assays use hundreds of different peptide substrates, each covalently linked to an oligonucleotide tag. After incubation with sample, the pool of substrates is hybridized to a microarray containing oligonucleotides complementary to the tag sequences. We screened several specific chemistries for the conjugation based on the following criteria: easy derivatization of oligonucleotides and peptides; high efficiency of the conjugation reaction; good stability of the conjugates; and satisfactory conjugate performance in our assays. We have validated selected method during the successful generation of thousands oligonucleotide-peptide conjugates.     

两种农业种植模式对重金属土壤的修复潜力

植物修复农田土壤重金属污染需要经历一个长期的过程,而大部分用来修复的植物都不具备利用价值,不能给当地农民带来经济收入。因此,一些农作物由于其较大的生物量和一定的经济价值,在植物修复土壤重金属污染的应用中受到广泛关注。是在重金属(Cu、Zn、Pb、Cd、As、Hg)复合污染的郴州矿区废弃农田种植油菜、玉米和油葵,研究油菜-玉米和油-油葵两种种植模式对土壤重金属污染的修复潜力。实验结果表明:三种作物在复合污染土壤中对重金属都表现出一定的耐性及吸收积累能力。向日葵的根和叶中重金属Cd、Cu的含量都很高,其中Cd在向日葵的各个部位的富集系数(BCF)及Cu在向日葵的根和叶的富集系数(BCF)都大于1。两种轮作模式对作物的产量没有明显的影响,收获得到的干物质量都很高,每年每公顷分别为油菜16.6t、玉米25.29t、油葵22.5t。两种种植模式都可以对土壤中的重金属进行有效的提取,油菜-油葵种植模式下提取重金属Cu、Pb、Cd、As的量较高,分别为:Cu 2408g hm~(-2)a~(-1),Pb 2027g hm~(-2)a~(-1),Cd 658.5g hm~(-2)a~(-1),As 250g hm~(-2)a~(-1),油菜-玉米模式下Zn和Hg的提取量较高,分别为Zn 4987g hm~(-2)a~(-1)和Hg 7.92g hm~(-2)a~(-1);对于多种重金属复合污染的土壤来说,油菜-油葵的种植模式要优于油菜-玉米的种植模式。总的来说,利用3种作物两两轮作的种植模式,在不影响作物产量的前提下大大的提高了作物对重金属的提取总量。3种作物在收获以后又可以用做工业原料,这就使得当地农民充分利用矿区废弃农田修复污染的同时又能从中获得一定经济效益。