Ecological criteria for assessing the content of petroleum hydrocarbons in the main soils of coniferous-deciduous forests and forest steppe.

The effect of pollution of Albicluvisols/Retisols, Calcaric Leptosols, Luvic Phaeozems, Greyzamic Phaeozems and Folic Fluvisols with oil (Solovatovsky oil field, Perm region) added in amounts of 1, 2, 3 and 5 g oil/kg of soil on the organisms was studied in a model laboratory experiment. Oil addition showed phytotoxic effects on root length in Triticum aestivum L., Lepidium sativum L., Picea obovata Ledeb. and Pinus sylvestris L. in all soils. However, oil contamination of Calcaric Leptosols and Greyzamic Phaeozems led to growth stimulation in Picea obovata seedlings. A remarkable shift in the diversity and number of colony-forming units of heterotrophic and oil-oxidizing bacteria was detected in all soil types. The maximum decrease in biodiversity (45%) was noted for heterotrophic bacteria in Luvic Phaeozems. Aqueous extracts from all oil-contaminated soils had a toxic effect on Chlorella vulgaris Beijer, causing an increase in biomass by more than 30%, but did not show acute toxicity on Daphnia magna Straus. Oil addition in the range of 1-3 g oil/kg soil posed no environmental risk to human health. However, oil addition at 5 g oil/kg of soil led to an increase in the level of carcinogenic risk to children to the threshold values of acceptable risk and ranged from 0.95 × 10-4 for Greyzamic Phaeozems and Folic Fluvisols to 1.098 × 10-4 for Luvic Phaeozems. Our results suggest that the reaction of test organisms to oil pollution depends on the soil type, and their complex application makes it possible to identify the most sensitive factor and assess the dangerous level of pollution.

Effects of Monoacyltrehalose Fraction of Rhodococcus Biosurfactant on the Innate and Adaptive Immunity Parameters In Vivo.

The biosurfactant monoacyltrehalose fraction isolated from Rhodococcus ruber IEGM 231 actinobacterium suppresses antibody production, bactericidal potential, and production of IL-1ß by mouse peritoneal cells after intraperitoneal and intramuscular injection and stimulates the production of IL-10 after intraperitoneal injection. The data of in vitro experiments attest to an important role of bacterial glycolipids in the regulation of the functions of splenocytes and peritoneal macrophages.

Effects of Glycolipid Rhodococcus Biosurfactant on Innate and Adaptive Immunity Parameters In Vivo.

The glycolipid biosurfactant complex from actinobacterium Rhodococcus ruber IEGM 231 inhibits the innate and adaptive immunity parameters after intraperitoneal and intramuscular injection. Marked suppression of antibody production, bactericidal potential, and production of proinflammatory cytokines by peritoneal macrophages, detected in vivo, do not agree with the previously detected immunostimulatory activity of biosurfactants towards the immunocompetent cell cultures; this fact indicates an important role of the cell environment in the formation of immune response under the effect of bacterial glycolipids.

Effect of glycolipid Rhodococcus biosurfactant on secretory activity of neutrophils in vitro.

Glycolipid biosurfactant synthesized by nonpathogenic strain Rhodococcus ruber IEGM231 modulated the production of ROS and IL-8 by peripheral blood neutrophils in spontaneous and stimulated cultures. Secretion of IL-1ß Ð¸ TNF-α by neutrophils in the presence of biosurfactant changed insignificantly.

[Adsorptive immobilization of rhodococcal cells in hydrophobized derivatives of wide-pore polyacrylamide cryogel].

Adsorption of Rhodococcus ruber cells on columns with polyacrylamide cryogel (CryoPAAG) partially hydrophobized by different quantities (0.2, 1, and 5 mol %) of chemically grafted n-dodecane residues has been studied. The adsorption capacity (1.1 x 10(9) cells/g) of gel carrier for rhodococcal cells and the optimal content (1 mol %) of hydrophobizing groups were determined. The respirometric method showed the high catalytic activity and functional stability of immobilized bacterial cells. Respiratory activity of immobilized rhodococci in the presence of a model mixture of oil hydrocarbons exceeded the respective parameter for free cells by 12-17%. Viability of rhodococcal cells adsorptionally fixed in hydrophobized cryoPAAG was maintained at a level of 93-95% after a half-year period of storage. The results may be used for development of immobilized biocatalyst for directed transformation of hydrocarbon compounds and biological purification of oil-polluted water.

Modulation of cytokine secretion and oxidative metabolism of innate immune effectors by Rhodococcus biosurfactant.

The glycolipid biosurfactant complex from Rhodococcus ruber IEGM 231 had a stimulatory effect on the production of IL-12, IL-18, and reactive oxygen species by cells of the innate immunity. This effect depended on the composition of cell cultures and presence of LPS. It was primarily observed in non-stimulated cultures. The glycolipid biosurfactant complex had little effect on IL-10 secretion by monocytes and mononuclear cells.

In vitro immunomodulating activity of biosurfactant glycolipid complex from Rhodococcus ruber.

The biosurfactant glycolipid complex synthesized by Rhodococcus ruber actinobacteria is not toxic and exhibits no appreciable effect on proliferative activity of peripheral blood leukocytes. In the monocyte fraction, the biosurfactant activates the production of IL-1beta and TNF-alpha cytokines without modifying the production of IL-6. In the mononuclear fraction, the glycolipid biosurfactant exhibited no effects on the production of IL-1beta, TNF-alpha, and IL-6. These results indicate good prospects for further studies of immunomodulating and antitumor activities of biosurfactant drug.

Alkanotrophic Rhodococcus ruber as a biosurfactant producer.

In this report we examined the structure and properties of surface-active lipids of Rhodococcus ruber. Most historical interest has been in the glycolipids of Rhodococcus erythropolis, which have been extensively characterised. R. erythropolis has been of interest due to its great metabolic diversity. Only recently has the metabolic potential of R. ruber begun to be explored. One major difference in the two species is that most R. ruber strains are able to oxidise the gaseous alkanes propane and butane. In preparation for investigation of the effects of gas metabolism on biosurfactant production, we set out to characterise the biosurfactants produced during growth on liquid n-alkanes and to compare these with R. erythropolis glycolipids.

Recovery of Rhodococcus biosurfactants using methyl tertiary-butyl ether extraction.

In the present study, we proposed methyl tertiary-butyl ether (MTBE) as a solvent for extraction of biosurfactants from Rhodococcus bacterial cultures. After comparison with other well known solvent systems used for biosurfactant extraction, it was found that MTBE was able to extract crude surfactant material with high product recovery (10 g/l), efficiency (critical micelle concentration (CMC), 130-170 mg/l) and good functional surfactant characteristics (surface and interfacial tensions, 29 and 0.9 mN/m, respectively). The isolated surfactant complex contained 10% polar lipids, mostly glycolipids possessing maximal surface activity. Ultrasonic treatment of the extraction mixture increased the proportion of polar lipids in crude extract, resulting in increasing surfactant efficiency. Due to certain characteristics of MTBE, such as relatively low toxicity, biodegradability, ease of downstream recovery, low flammability and explosion safety, the use of this solvent as an extraction agent in industrial scale biosurfactant production is feasible.

Identification and environmental detection of Rhodococcus species by 16S rDNA-targeted PCR.

Bacteria of the genus Rhodococcus can degrade a wide range of organic pollutants and catalyse many useful biotransformations. There is a need for improved tests to identify Rhodococcus species. PCR-based methods for species identification offer advantages in terms of speed and accuracy over traditional methods and can allow direct detection of microbes in environmental samples., PCR tests, using primers targeted at species-specific sequences in the 16S rRNA gene, were successfully developed for R. globerulus, R. erythropolis, R. opacus and R. ruber. These tests gave positive results with all or most strains of target species but did not generally cross-react with other species. Cases of apparent cross-reaction were shown to be due to prior misclassification of strains of R. opacus as R. erythropolis and of strains of R. ruber as R. rhodochrous. A simple and rapid method for the extraction and purification of DNA from soil was developed and successfully applied to the PCR detection of indigenous R. erythropolis in an environmental sample. Cell lysis in the samples was achieved by lysozyme and sarkosyl treatment, aided by freeze-thaw cycles. Removal of humic compounds inhibitory to PCR was accomplished by CTAB treatment with solvent extraction and, if necessary, passage of extracts through Sepharose CL-6B in a spun-column format. Extracts prepared using a tris-EDTA buffer were much clearer than those prepared using a sodium phosphate buffer, indicating lower levels of humic compounds. A detection limit of 104 cfu g-1 of soil was achieved and the use of a secondary PCR allowed detection of 1 cfu g-1.