In vitro cytokine stimulation assay for glycolipid biosurfactant from Rhodococcus ruber: role of monocyte adhesion

Glycolipid biosurfactant (GLB) from Rhodococcus ruber IEGM 231 was found to stimulate tumor necrosis factor-α (TNF-α), interleukin (IL) -1β and IL-6 production when applied as an ultrasonic emulsion to the adherent human peripheral blood monocyte culture. However, a lack of cytokine-stimulating activity was registered with the GLB applied as a hydrophobic film coating in 24-well culture plates, indicating that it may have been due to its inhibitory effect on monocyte adhesion. The mode of GLB application may therefore play an important role in in vitro assay of immunostimulatory activity of this compound as well as other bacterial glycolipids. Additionally, GLB from R. ruber displayed no cytotoxicity against human lymphocytes and therefore could be proposed as a potential immunomodulating and antitumor agent

Oil spill problems and sustainable response strategies through new technologies

Crude oil and petroleum products are widespread water and soil pollutants resulting from marine and terrestrial spillages. International statistics of oil spill sizes for all incidents indicate that the majority of oil spills are small (less than 7 tonnes). The major accidents that happen in the oil industry contribute only a small fraction of the total oil which enters the environment. However, the nature of accidental releases is that they highly pollute small areas and have the potential to devastate the biota locally. There are several routes by which oil can get back to humans from accidental spills, e.g. through accumulation in fish and shellfish, through consumption of contaminated groundwater. Although advances have been made in the prevention of accidents, this does not apply in all countries, and by the random nature of oil spill events, total prevention is not feasible. Therefore, considerable world-wide effort has gone into strategies for minimising accidental spills and the design of new remedial technologies. This paper summarizes new knowledge as well as research and technology gaps essential for developing appropriate decision-making tools in actual spill scenarios. Since oil exploration is being driven into deeper waters and more remote, fragile environments, the risk of future accidents becomes much higher. The innovative safety and accident prevention approaches summarized in this paper are currently important for a range of stakeholders, including the oil industry, the scientific community and the public. Ultimately an integrated approach to prevention and remediation that accelerates an early warning protocol in the event of a spill would get the most appropriate technology selected and implemented as early as possible-the first few hours after a spill are crucial to the outcome of the remedial effort. A particular focus is made on bioremediation as environmentally harmless, cost-effective and relatively inexpensive technology. Greater penetration into the remedial technologies market depends on the harmonization of environment legislation and the application of modern laboratory techniques, e.g. ecogenomics, to improve the predictability of bioremediation

Potential risks of antibiotic resistant bacteria and genes in bioremediation of petroleum hydrocarbon contaminated soils

Bioremediation represents a sustainable approach to remediating petroleum hydrocarbon contaminated soils. One aspect of sustainability includes the sourcing of nutrients used to stimulate hydrocarbon-degrading microbial populations. Organic nutrients such as animal manure and sewage sludge may be perceived as more sustainable than conventional inorganic fertilizers. However, organic nutrients often contain antibiotic residues and resistant bacteria (along with resistance genes and mobile genetic elements). This is further exacerbated since antibiotic resistant bacteria may become more abundant in contaminated soils due to co-selection pressures from pollutants such as metals and hydrocarbons. We review the issues surrounding bioremediation of petroleum-hydrocarbon contaminated soils, as an example, and consider the potential human-health risks from antibiotic resistant bacteria. While awareness is coming to light, the relationship between contaminated land and antibiotic resistance remains largely under-explored. The risk of horizontal gene transfer between soil microorganisms, commensal bacteria and/or human pathogens needs to be further elucidated, and the environmental triggers for gene transfer need to be better understood. Findings of antibiotic resistance from animal manures are emerging, but even fewer bioremediation studies using sewage sludge have made any reference to antibiotic resistance. Resistance mechanisms, including those to antibiotics, have been considered by some authors to be a positive trait associated with resilience in strains intended for bioremediation. Nevertheless, recognition of the potential risks associated with antibiotic resistant bacteria and genes in contaminated soils appears to be increasing and requires further investigation. Careful selection of bacterial candidates for bioremediation possessing minimal antibiotic resistance as well as pre-treatment of organic wastes to reduce selective pressures (e.g., antibiotic residues) are suggested to prevent environmental contamination with antibiotic-resistant bacteria and genes

Effects of Nickel Nanoparticles on Rhodococcus Cell Surface Morphology and Nanomechanical Properties

Nickel nanoparticles (NPs) are used for soil remediation and wastewater treatment due to their high adsorption capacity against complex organic pollutants. However, despite the growing use of nickel NPs, their toxicological towards environmental bacteria have not been sufficiently studied. Actinobacteria of the genus Rhodococcus are valuable bioremediation agents degrading a range of harmful and recalcitrant chemicals. Both positive and negative effects of metal ions and NPs on the biodegradation of organic pollutants by Rhodococcus were revealed, however, the mechanisms of such interactions, in addition to direct toxic effects, remain unclear. In the present work, the influence of nickel NPs on the viability, surface topology and nanomechanical properties of Rhodococcus cells have been studied. Bacterial adaptations to high (up to 1.0 g/L) concentrations of nickel NPs during prolonged (24 and 48 h) exposure were detected using combined confocal laser scanning and atomic force microscopy. Incubation with nickel NPs resulted in a 1.25–1.5-fold increase in the relative surface area and roughness, changes in cellular charge and adhesion characteristics, as well as a 2–8-fold decrease in the Young’s modulus of Rhodococcus ruber IEGM 231 cells. Presumably, the treatment of rhodococcal cells with sublethal concentrations (0.01–0.1 g/L) of nickel NPs facilitates the colonization of surfaces, which is important in the production of immobilized biocatalysts based on whole bacterial cells adsorbed on solid carriers. Based on the data obtained, cell surface functionalizing with NPs is possible to enhance adhesive and catalytic properties of bacteria suitable for environmental applications

Responses to Ecopollutants and Pathogenization Risks of Saprotrophic Rhodococcus Species

Under conditions of increasing environmental pollution, true saprophytes are capable of changing their survival strategies and demonstrating certain pathogenicity factors. Actinobacteria of the genus Rhodococcus, typical soil and aquatic biotope inhabitants, are characterized by high ecological plasticity and a wide range of oxidized organic substrates, including hydrocarbons and their derivatives. Their cell adaptations, such as the ability of adhering and colonizing surfaces, a complex life cycle, formation of resting cells and capsule-like structures, diauxotrophy, and a rigid cell wall, developed against the negative effects of anthropogenic pollutants are discussed and the risks of possible pathogenization of free-living saprotrophic Rhodococcus species are proposed. Due to universal adaptation features, Rhodococcus species are among the candidates, if further anthropogenic pressure increases, to move into the group of potentially pathogenic organisms with “unprofessional” parasitism, and to join an expanding list of infectious agents as facultative or occasional parasites

Draft genome sequence of propane- and butane-oxidizing actinobacterium Rhodococcus ruberRhodococcus\ ruber IEGM 231

International audienceWe report a draft genome sequence of $Rhodococcus\ ruber$ IEGM 231, isolated from a water spring near an oil-extracting enterprise (Perm region, Russian Federation). This sequence provides important insights into the genetic mechanisms of propane and n -butane metabolism, organic sulfide and beta-sitosterol biotransformation, glycolipid biosurfactant production, and heavy metal resistance in actinobacteria

Adhesion of Rhodococcus bacteria to solid hydrocarbons and enhanced biodegradation of these compounds

Adhesive activities of hydrocarbon-oxidizing Rhodococcus bacteria towards solid hydrocarbons, effects of adhesion on biodegradation of these compounds by rhodococcal cells and adhesion mechanisms of Rhodococcus spp. were studied in this work. It was shown that efficiency of Rhodococcus cells' adhesion to solid n-alkanes and polycyclic aromatic hydrocarbons (PAHs) varied from 0.0 to 10.6·106 CFU/cm2. R. erythropolis IEGM 212 and R. opacus IEGM 262 demonstrated the highest (≥ 4.3·106 CFU/cm2) adhesion. The percentage biodegradation of solid hydrocarbons (n-hexacosane and anthracene as model substrates) by Rhodococcus cells was 5 to 60% at a hydrocarbon concentration of 0.2% (w/w) after 9 days and strongly depended on cell adhesive activities towards these compounds (r ≥ 0.71, p < 0.05). No strict correlation between the adhesive activities of rhodococcal cells and physicochemical properties of bacteria and hydrocarbons was detected. Roughness of the cell surface was a definitive factor of Rhodococcus cell adhesion to solid hydrocarbons. Specific appendages with high adhesion force (≥ 0.6 nN) and elastic modulus (≥ 6 MPa) were found on the surface of Rhodococcus cells with high surface roughness. We hypothesized that these appendages participated in the adhesion process

Biosynthesis and Characterization of Gold Nanoparticles Produced Using Rhodococcus Actinobacteria at Elevated Chloroauric Acid Concentrations

The growing industrial and medical use of gold nanoparticles (AuNPs) requires environmentally friendly methods for their production using microbial biosynthesis. The ability of actinobacteria of the genus Rhodococcus to synthesize AuNPs in the presence of chloroauric acid (HAuCl4) was studied. The effect of elevated (0.8&ndash;3.2 mM) concentrations of HAuCl4 on bacterial viability, morphology, and intracellular accumulation of AuNPs by different Rhodococcus species was shown. An increase in surface roughness, a shift of the zeta potential to the positive region, and the formation of cell aggregates of R. erythropolis IEGM 766 and R. ruber IEGM 1135 during nanoparticle synthesis were revealed as bacterial adaptations to toxic effects of HAuCl4. The possibility to biosynthesize AuNPs at a five times higher concentration of chloroauric acid compared to chemical synthesis, for example, using the citrate method, suggests greater efficiency of the biological process using Rhodococcus species. The main parameters of biosynthesized AuNPs (size, shape, surface roughness, and surface charge) were characterized using atomic force microscopy, dynamic and electrophoretic light scattering, and also scanning electron microscopy in combination with energy-dispersive spectrometry. Synthesized by R. erythropolis spherical AuNPs have smaller (30&ndash;120 nm) dimensions and are positively (12 mV) charged, unlike AuNPs isolated from R. ruber cells (40&ndash;200 nm and &minus;22 mV, respectively). Such differences in AuNPs size and surface charge are due to different biomolecules, which originated from Rhodococcus cells and served as capping agents for nanoparticles. Biosynthesized AuNPs showed antimicrobial activity against Gram-positive (Micrococcus luteus) and Gram-negative (Escherichia coli) bacteria. Due to the positive charge and high dispersion, the synthesized by R. erythropolis AuNPs are promising for biomedicine, whereas the AuNPs formed by R. ruber IEGM 1135 are prone to aggregation and can be used for biotechnological enrichment of gold-bearing ores

Bioaccumulation of molybdate ions by alkanotrophic Rhodococcus leads to significant alterations in cellular ultrastructure and physiology

Alkanotrophic Rhodococcus strains from the Regional Specialised Collection of Alkanotrophic Microorganisms (acronym IEGM, www.iegmcol.ru) were screened for accumulation and sorption of MoO42- ions. Morphological and ultrastructural changes observed in bacterial cells during their cultivation in the molybdenum-containing medium are described. The species peculiarities, growth substrate preferences, and other physiological features allowing for the efficient removal of molybdate ions from the culture medium are discussed. Bioinformatics analysis of genes and proteins responsible for resistance to and accumulation of molybdenum was carried out using the sequenced R. ruber IEGM 231 and other published Rhodococcus genomes. n-Hexadecane growing strains with high (up to 85 %) accumulative activity and resistance to elevated (up to 20.0 mM) molybdenum concentrations were selected, which can be used for bioremediation of environments co-contaminated with heavy metals and hydrocarbons. Transmission electron microscopy and energy dispersive X-ray spectroscopy (TEM-EDX) revealed the ability of Rhodococcus not only to accumulate, but also to chemically convert soluble toxic molybdenum into insoluble compounds detected in the form of electron-dense nanoparticles