Changes induced by heavy metals in the plant-associated microbiome of Miscanthus x giganteus.

Miscanthus x giganteus is a high biomass producing plant with tolerance to heavy metals. This makes Miscanthus interesting to be used for phytoremediation of heavy metal contaminated areas coupled with energy production. Since plant performance in metal polluted areas is impaired, their growth and phytoremediation effect can be improved with bacterial assistance. To identify positive and negative responders of M. x giganteus associated microbiome influenced by Cd, Pb and Zn stress compared to non-contaminated controls, we designed a greenhouse experiment. Structure of the bacterial community in three rhizocompartments, namely rhizosphere, rhizoplane and root endosphere was analysed using an isolation independent molecular approach based on 16S rRNA gene barcoding. Furthermore, quantitative PCR (qPCR) was used for bacterial biomass estimation. Our results indicated that biomass and total bacterial diversity in rhizosphere, rhizoplane and root endosphere did not significantly change despite of substantial root uptake of heavy metals. Overall, we detected 6621 OTUs, from which 171 were affected by metal addition. Whereas Streptomyces and Amycolatopsis taxa were negatively affected by the heavy metal treatment in endosphere, taxa assigned to Luteolibacter in rhizosphere and rhizoplane (log2 fold change 1.9-4.1) and Micromonospora in endosphere (log2 fold change 10.2) were found to be significantly enriched and highly abundant (0.1-3.7% relative abundance) under heavy metal stress. Those taxa might be of key importance for M. x giganteus performance under heavy metal pollution and might be interesting candidates for the development of new bioinocula in the future to promote plant growth and phytoremediation in heavy metal contaminated soils.

Complete Genome Sequences of Two Plant-Associated Pseudomonas putida Isolates with Increased Heavy-Metal Tolerance.

We report here the complete genome sequences of two Pseudomonas putida isolates recovered from surface-sterilized roots of Sida hermaphrodita The two isolates were characterized by an increased tolerance to zinc, cadmium, and lead. Furthermore, the strains showed typical plant growth-promoting properties, such as the production of indole acetic acid, cellulolytic enzymes, and siderophores.

Antimicrobial, antiadhesive and antibiofilm potential of lipopeptides synthesised by Bacillus subtilis, on uropathogenic bacteria.

The aim of this study was to investigate the antimicrobial effect of lipopeptide biosurfactants from surfactin, iturin and fengycin families, synthesised by the Bacillus subtilis I'1a strain, on uropathogenic bacteria, including the effects on planktonic growth, processes of biofilm formation and dislodging. Antimicrobial activity was tested against 32 uropathogenic strains belonging to 12 different species of Gram-negative and Gram-positive bacteria. The sensitivity of 25 tested bacterial strains to the B. subtilis I'1a filtrate was confirmed by an agar diffusion assay. None of the strains seemed to be sensitive to pure surfactin at concentrations ranging from 0.1 mg × ml(-1) to 0.4 mg ml(-1). After the treatment of uropathogens with B. subtilis lipopeptides, the metabolic activity of planktonic cells was inhibited by 88.05±3.96% in the case of 21 studied uropathogens, the process of biofilm formation was reduced by 88.15±4.77% in the case of 24 uropathogens and mature biofilms of 18 strains were dislodged by about 81.20±4.72%. Ten strains of uropathogenic bacteria were selected to study the antimicrobial activity of surfactin (concentrations 0.1, 0.2 and 0.4 mg × ml(-1)). Surfactin had no influence on the metabolic activity of planktonic forms of uropathogens, however, biofilms of 5 tested strains were reduced by 64.77±9.05% in the presence of this biosurfactant at the concentration 0.1 mg × ml(-1). The negative effect of the compound on the biofilm formation process was observed at all concentrations used. The above-described results were fully confirmed by CLSM. It could suggest that synergistic application of biosurfactants could be efficient in uropathogen eradication.

Biosurfactant mediated biosynthesis of selected metallic nanoparticles.

Developing a reliable experimental protocol for the synthesis of nanomaterials is one of the challenging topics in current nanotechnology particularly in the context of the recent drive to promote green technologies in their synthesis. The increasing need to develop clean, nontoxic and environmentally safe production processes for nanoparticles to reduce environmental impact, minimize waste and increase energy efficiency has become essential in this field. Consequently, recent studies on the use of microorganisms in the synthesis of selected nanoparticles are gaining increased interest as they represent an exciting area of research with considerable development potential. Microorganisms are known to be capable of synthesizing inorganic molecules that are deposited either intra- or extracellularly. This review presents a brief overview of current research on the use of biosurfactants in the biosynthesis of selected metallic nanoparticles and their potential importance.

Assessment of the chemical, microbiological and toxicological aspects of post-processing water from underground coal gasification.

The purpose of this paper is to provide a comprehensive characterisation (including chemical, microbiological and toxicological parameters) of water after the underground coal gasification (UCG) process. This is the first report in which these parameters were analysed together to assess the environmental risk of the water generated during the simulation of the underground coal gasification (UCG) process performed by the Central Mining Institute (Poland). Chemical analysis of the water indicated many hazardous chemical compounds, including benzene, toluene, ethylbenzene, xylene, phenols and polycyclic aromatic hydrocarbons (PAHs). Additionally, large quantities of inorganic compounds from the coal and ashes produced during the volatilisation process were noted. Due to the presence of refractory and inhibitory compounds in the post-processing water samples, the microbiological and toxicological analyses revealed the high toxicity of the UCG post-processing water. Among the tested microorganisms, mesophilic, thermophilic, psychrophilic, spore-forming, anaerobic and S-oxidizing bacteria were identified. However, the number of detected microorganisms was very low. The psychrophilic bacteria dominated among tested bacteria. There were no fungi or Actinomycetes in any of the water samples. Preliminary study revealed that hydrocarbon-oxidizing bacteria were metabolically active in the water samples. The samples were very toxic to the biotests, with the TU50 reaching 262. None of biotests was the most sensitive to all samples. Cytotoxicity and genotoxicity testing of the water samples in Vicia uncovered strong cytotoxic and clastogenic effects. Furthermore, TUNEL indicated that all of the water samples caused sporadic DNA fragmentation in the nuclei of the roots.

Environmental applications of biosurfactants: recent advances.

Increasing public awareness of environmental pollution influences the search and development of technologies that help in clean up of organic and inorganic contaminants such as hydrocarbons and metals. An alternative and eco-friendly method of remediation technology of environments contaminated with these pollutants is the use of biosurfactants and biosurfactant-producing microorganisms. The diversity of biosurfactants makes them an attractive group of compounds for potential use in a wide variety of industrial and biotechnological applications. The purpose of this review is to provide a comprehensive overview of advances in the applications of biosurfactants and biosurfactant-producing microorganisms in hydrocarbon and metal remediation technologies.

Estimation of the environmental risk posed by landfills using chemical, microbiological and ecotoxicological testing of leachates.

The leachates from 22 municipal solid waste (MSW) landfill sites in Southern Poland were characterized by evaluation of chemical, microbiological and ecotoxicological parameters. Chemical analyses were mainly focused on the identification of the priority hazardous substances according to Directive on Priority Substances, 2008/105/EC (a daughter directive of the WFD) in leachates. As showed, only five substances (Cd, Hg, hexachlorobutadiene, pentachlorobenzene and PAHs) were detected in the leachates. The compounds tested were absent or present at very low concentrations. Among them, only PAHs were found in all samples in the range from 0.057 to 77.2 µg L⁻¹. The leachates were contaminated with bacteria, including aerobic, psychrophilic and mesophilic bacteria, coliform and fecal coliforms, and spore-forming-bacteria, including Clostridium perfringens, and with filamentous fungi. From the analysis of specific microorganism groups (indicators of environmental pollution by pathogenic or opportunistic pathogenic organisms) it can be concluded that the landfill leachates showed sanitary and epidemiological hazard. In the ecotoxicological study, a battery of tests comprised of 5 bioassays, i.e. Microtox(®), Spirotox, Rotoxkit F™, Thamnotoxkit F™ and Daphtoxkit F™ magna was applied. The leachate samples were classified as toxic in 13.6%, highly toxic in 54.6% and very highly toxic in 31.8%. The Spirotox test was the most sensitive bioassay used. The percentage of class weight score was very high - above 60%; these samples could definitely be considered seriously hazardous and acutely toxic to the fauna and microflora. No correlations were found between the toxicity values and chemical parameters. The toxicity of leachate samples cannot be explained by low levels of the priority pollutants. It seems that other kinds of xenobiotics present in the samples at subacute levels gave the high aggregate toxic effect. The chemical, ecotoxicological and microbiological parameters of the landfill leachates should be analyzed together to assess the environmental risk posed by landfill emissions.

Characterization of bacterial communities at heavy-metal-contaminated sites.

The microbial community in soil samples from two long-term contaminated sites was characterized by using culture-dependent and culture-independent methods. The two sites investigated contained high amounts of heavy metals and were located in the upper Silesia Industrial Region in southern Poland. The evaluation of the aerobic soil microbial population clearly demonstrated the presence of considerable numbers of viable, culturable bacteria at both sites. A high fraction of the bacterial population was able to grow in the presence of high amounts of metals, i.e. up to 10 mM Zn²(+), 3 mM Pb²(+) or 1 mM Cu²(+). Site 1 contained significantly (P < 0.05) lower bacterial numbers growing in the presence of 10 mM Zn²(+) than site 2, while the opposite was observed for bacteria tolerating 1 mM Cu²(+). This coincided with the contents of these two metals at the two sites. Ecophysiological (EP) indices for copiotrophs (r-strategists) and oligotrophs (K-strategists) pointed to high bacterial diversity at both sites. Fluorescence in situ hybridization (FISH) analysis indicated that Actinobacteria and Proteobacteria represent the physiologically active fraction of bacteria at the two sites. Shannon diversity (H') indices for FISH-detected bacterial phylogenetic groups were not significantly different at the two sites.

Ecotoxicological and microbiological characterization of soils from heavy-metal- and hydrocarbon-contaminated sites.

The aims of this study were to characterize soils from industrial sites by combining physicochemical, microbiological, and ecotoxicological parameters and to assess the suitability of these assays for evaluation of contaminated sites and ecological risk assessment. The soil samples were taken from long-term contaminated sites containing high amounts of heavy metals (sites 1 and 2) or petroleum hydrocarbons (site 3) located in the upper Silesia Industrial Region in southern Poland. Due to soil heterogeneity, large differences between all investigated parameters were measured. Microbiological properties revealed the presence of high numbers of viable hetrotrophic microorganisms. Soil enzyme activities were considerably reduced or could not be detected in contaminated soils. Activities involved in N turnover (N mineralization and nitrification) were significantly (P < 0.05) higher in samples from the metal-contaminated sites than in samples from the hydrocarbon-contaminated site, whereas the opposite was observed for phosphatase activity. The Microtox test system appeared to be the most appropriate to detect toxicity and significant differences in toxicity between the three sites. The Ostracodtoxkit test was the most appropriate test system to detect toxicity in the hydrocarbon-contaminated soil samples. Correlation analysis between principal components (obtained from factor analysis) determined for physicochemical, microbiological, and ecotoxicological soil properties demonstrated the impact of total and water-extractable contents of heavy metals on toxicity.

Reduction of petroleum hydrocarbons and toxicity in refinery wastewater by bioremediation.

The aim of the study was to investigate petroleum waste remediation and toxicity reduction by five bacterial strains: Ralstonia picketti SRS (BP-20), Alcaligenes piechaudii SRS (CZOR L-1B), Bacillus subtilis (I'-1a), Bacillus sp. (T-1), and Bacillus sp. (T'-1), previously isolated from petroleum-contaminated soils. Petroleum hydrocarbons were significantly degraded (91%) by the mixed bacterial cultures in 30 days (reaching up to 29% in the first 72 h). Similarly, the toxicity of the biodegraded petroleum waste decreased 3-fold after 30 days. This work shows the influence of bacteria on hydrocarbon degradation and associated toxicity, and its dependence on the specific microorganisms present. The ability of these mixed cultures to degrade hydrocarbons and reduce toxicity makes them candidates for environmental restoration applications at other hydrocarbon-contaminated environments.

Surface active properties of bacterial strains isolated from petroleum hydrocarbon-bioremediated soil.

Two bacterial strains identified as Ralstonia picketti (BP 20) and Alcaligenes piechaudii (CZOR L-1B) were isolated from petroleum hydrocarbon-contaminated soil following bioremediation treatment. The surface active properties, e.g. surface tension, emulsification and foamability of their culture filtrates were evaluated. Bacterial cell-surface hydrophobicity (BAH) as measured by analyzing cell affinity towards aliphatic and aromatic compounds was also determinated. The bacteria grew in liquid cultures containing 1% (v/v) of crude oil as carbon and energy source at 30 degrees C under aerobic conditions. The surface tensions were reduced to 61 mN/m and 55 mN/m by Ralstonia picketti and Alcaligenes piechaudii, respectively. The emulsification index (EI24) was almost 100% for all tested compounds except diesel oil. The stability of the emulsions was deteminated at 4 degrees C, 45 degrees C and 65 degrees C. The emulsions were stable at 4 degrees C. Ralstonia picketti was better foam inducer (FV = 50 ml) compared to Alcaligenes piechaudii (FV = 10 ml). The BAH measurements revealed higher adhesion of Alcaligenes piechaudii cells towards different hydrocarbons compared to Ralstonia picketti cells. The strains were found to have a surface hydrophobicity in the following order: aliphatic hydrocarbons, BTEX, and PAHs. The ability to adhere to bulk hydrocarbon is mostly a characteristic of hydrocarbon-degrading bacteria. The strains were found to be better emulsifiers than surface tension reducers. They produce water-soluble extracellular bioemulsifiers. Both bacterial isolates have good properties to use them, mainly in the petroleum industry, e.g. in enhanced oil recovery and in bioremediation processes-primarily due to their emulsification property, i.e. emulsion forming and stabilizing capacity.