Vladimir P. Beškoski

Ex situ bioremediation of a soil contaminated by mazut (heavy residual fuel oil)--a field experiment.

Mazut (heavy residual fuel oil)-polluted soil was exposed to bioremediation in an ex situ field-scale (600 m(3)) study. Re-inoculation was performed periodically with biomasses of microbial consortia isolated from the mazut-contaminated soil. Biostimulation was conducted by adding nutritional elements (N, P and K). The biopile (depth 0.4m) was comprised of mechanically mixed polluted soil with softwood sawdust and crude river sand. Aeration was improved by systematic mixing. The biopile was protected from direct external influences by a polyethylene cover. Part (10 m(3)) of the material prepared for bioremediation was set aside uninoculated, and maintained as an untreated control pile (CP). Biostimulation and re-inoculation with zymogenous microorganisms increased the number of hydrocarbon degraders after 50 d by more than 20 times in the treated soil. During the 5 months, the total petroleum hydrocarbon (TPH) content of the contaminated soil was reduced to 6% of the initial value, from 5.2 to 0.3 g kg(-1) dry matter, while TPH reduced to only 90% of the initial value in the CP. After 150 d there were 96%, 97% and 83% reductions for the aliphatic, aromatic, and nitrogen-sulphur-oxygen and asphaltene fractions, respectively. The isoprenoids, pristane and phytane, were more than 55% biodegraded, which indicated that they are not suitable biomarkers for following bioremediation. According to the available data, this is the first field-scale study of the bioremediation of mazut and mazut sediment-polluted soil, and the efficiency achieved was far above that described in the literature to date for heavy fuel oil.

Biodegradation of petroleum sludge and petroleum polluted soil by a bacterial consortium: a laboratory study

This article presents a study of the efficiency and degradation pattern of samples of petroleum sludge and polluted sandy soil from an oil refinery. A bacterial consortium, consisting of strains from the genera Pseudomonas, Achromobacter, Bacillus and Micromonospora, was isolated from a petroleum sludge sample and characterized. The addition of nitrogen and phosphorus nutrients and a chemical surfactant to both the samples and bioaugmentation to the soil sample were applied under laboratory conditions. The extent of biodegradation was monitored by the gravimetric method and analysis of the residual oil by gas chromatography. Over a 12-week experiment, the achieved degree of TPH (total petroleum hydrocarbon) degradation amounted to 82–88% in the petroleum sludge and 86–91% in the polluted soil. Gas chromatography–mass spectrometry was utilized to determine the biodegradability and degradation rates of n-alkanes, isoprenoids, steranes, diasteranes and terpanes. Complete degradation of the n-alkanes and isoprenoids fractions occurred in both the samples. In addition, the intensities of the peaks corresponding to tricyclic terpenes and homohopanes were decreased, while significant changes were also observed in the distribution of diasteranes and steranes.

Perfluorinated compounds in sediment samples from the wastewater canal of Pančevo (Serbia) industrial area

Perfluoroalkyl sulfonates (PFSAs) and perfluoroalkyl carboxylates (PFCAs) were analyzed in sediment samples from the wastewater canal draining the industrial complex of Pančevo, Serbia (oil refinery, petrochemical plant, and fertilizer factory). The canal is directly connected to Europes second largest river, the Danube, which drains its water into the Black Sea. Perfluorooctane sulfonate (PFOS) up to 5.7ngg(-1) dry weight (dw) and total Perfluorinated compounds (PFCs) up to 6.3ngg(-1) dw were detected. Compared to other reports, high levels of PFOS were found, even though PFCs are not used in the industrial production associated with this canal. The PFOS concentration in water was recalculated using the adsorption coefficient, KOC from literature. Using the average output of wastewater from the canal, a mass load of 1.38kg PFOS per year discharged in the Danube River has been calculated, which undoubtedly points to the contribution to global persistent organic pollution of surface waters originating from this industrial place.

Bioremediation of soil polluted with crude oil and its derivatives: Microorganisms, degradation pathways, technologies

The contamination of soil and water with petroleum and its products occurs due to accidental spills during exploitation, transport, processing, storing and use. In order to control the environmental risks caused by petroleum products a variety of techniques based on physical, chemical and biological methods have been used. Biological methods are considered to have a comparative advantage as cost effective and environmentally friendly technologies. Bioremediation, defined as the use of biological systems to destroy and reduce the concentrations of hazardous waste from contaminated sites, is an evolving technology for the removal and degradation of petroleum hydrocarbons as well as industrial solvents, phenols and pesticides. Microorganisms are the main bioremediation agents due to their diverse metabolic capacities. In order to enhance the rate of pollutant degradation the technology optimizes the conditions for the growth of microorganisms present in soil by aeration, nutrient addition and, if necessary, by adding separately prepared microorganisms cultures. The other factors that influence the efficiency of process are temperature, humidity, presence of surfactants, soil pH, mineral composition, content of organic substance of soil as well as type and concentration of contaminant. This paper presents a review of our ex situ bioremediation procedures successfully implemented on the industrial level. This technology was used for treatment of soils contaminated by crude oil and its derivatives originated from refinery as well as soils polluted with oil fuel and transformer oil.

Synthesis and characterization of a new type of levan-graft-polystyrene copolymer.

Novel macromolecular graft copolymers were synthesized by reaction of the hydroxyl groups of the microbial polysaccharide levan, produced using Bacillus licheniformis, with polystyrene (Lev-g-PS). Synthesis was performed by the free radical reaction using potassium persulfate (PPS) as initiator. The prepared copolymer was characterized by FTIR, SEM, TG/DTA, XRD and (13)C NMR. The influence of the different conditions (reaction temperature, air or nitrogen atmosphere, reaction time, type of amines and ascorbic acid (AA) concentration) on the grafting reaction was investigated. Results showed that maximum percentage of grafting (58.1%) was achieved at a reaction temperature 70°C, in a nitrogen atmosphere and using dimethylethanolamine (DMEA) as the amine activator. On the basis of the obtained results, the likely reaction mechanism was proposed. Synthesized copolymers have better thermal stability in comparison with their initial components. Copolymers such as Lev-g-PS could potentially have many applications, such as compatibilizers and material for membranes.

Cadmium specific proteomic responses of a highly resistant Pseudomonas aeruginosa san ai

Pseudomonas aeruginosa san ai is a promising candidate for bioremediation of cadmium pollution, as it resists a high concentration of up to 7.2 mM of cadmium. Leaving biomass of P. aeruginosa san ai exposed to cadmium has a large biosorption potential, implying its capacity to extract heavy metal from contaminated medium. In the present study, we investigated tolerance and accumulation of cadmium on protein level by shotgun proteomics approach based on liquid chromatography and tandem mass spectrometry coupled with bioinformatics to identify proteins. Size exclusion chromatography was used for protein prefractionation to preserve native forms of metalloproteins and protein complexes. Using this approach a total of 60 proteins were observed as up-regulated in cadmium-amended culture. Almost a third of the total numbers of up-regulated were metalloproteins. Particularly interesting are denitrification proteins which are over expressed but not active, suggesting their protective role in conditions of heavy metal exposure. P. aeruginosa san ai developed a complex mechanism to adapt to cadmium, based on: extracellular biosorption, bioaccumulation, the formation of biofilm, controlled siderophore production, enhanced respiration and modified protein profile. An increased abundance of proteins involved in: cell energy metabolism, including denitrification proteins; amino acid metabolism; cell motility and posttranslational modifications, primarily based on thiol-disulfide exchange, were observed. Enhanced oxygen consumption of biomass in cadmium-amended culture versus control was found. Our results signify that P. aeruginosa san ai is naturally well equipped to overcome and survive high doses of cadmium and, as such, has a great potential for application in bioremediation of cadmium polluted sites.

Microbial Polysaccharides: Between Oil Wells, Food and Drugs

The field of microbial polysaccharides is an immensely developing field over the last two decades as a result of its wide-ranging applications in different areas of science and technology. Microbial exopolysaccharides (MPSs) such as xanthan, dextran, gellan, pullulan and levan have been commercially used in their natural or modified state for many years. A large number of these natural polymer applications is a consequence of their excellent physical and chemical properties, based on their capacity to alter the basic properties of water (e.g. thickening or gelling). In addition, these polymers have secondary functions, such as emulsification, suspension, stabilization, encapsulation, flocculation, film forming, binding and coating. MPSs, and particularly exopolysaccharides, have many other novel properties to offer, and the discovery of immune modulation and bifidogenic effect of some of them should provide other applications. This work focuses on the more recent developments in the extent of application of microbial polysaccharides in the various fields which makes these polymers promising and versatile materials in the future, and also in our investigations of these natural products.

Gas Chromatography in Environmental Sciences and Evaluation of Bioremediation

Both soil and water become contaminated by oil and oil derivatives due to accidental spills in their exploitation, transportation, processing, storing and utilization. In 2010, 3.91 billion tons of crude oil was produced (BP Statistical Review, 2011) and estimations are that annually 0.1% of produced petroleum is released into the environment (Ward et al., 2003) as a result of anthropogenic activities.

Degradation of methyl-phenanthrene isomers during bioremediation of soil contaminated by residual fuel oil

Phenanthrene and methyl-phenanthrenes are major aromatic pollutants originating in particular from fuel oil. Phenanthrene is usually degraded faster than methyl-phenanthrenes under geological and environmental conditions. Here, we report a preferential and accelerated biodegradation of methyl-phenanthrenes versus phenanthrene in soil contaminated by fuel oil. The polluted soil was mixed with sawdust and sand to form a homogenized biopile. The biopile was continuously sprayed with microbial consortia isolated from crude oil–contaminated soil and treated by biosurfactants and nutritive substances for biostimulation. During a 6-month bioremediation experiment, a steady increase in the relative abundance of phenanthrene compared to methyl-phenathrenes was observed by gas chromatography–mass spectrometry. The increase was the highest for trimethyl-phenanthrenes, with a phenanthrene/trimethyl-phenanthrenes ratio increasing from 0.42 to 2.45. By contrast, the control, non-stimulated samples showed a ratio decrease from 0.85 to 0.11. Moreover, the results showed that the level of degradability depends on the number of methyl groups.

Brachybacterium sp. CH-KOV3 isolated from an oil-polluted environment–a new producer of levan

Various microorganisms isolated from polluted environments, such as Pseudomonas sp. and Micrococcus sp. can synthesize exopolysaccharides (EPSs) which are natural, non-toxic and biodegradable polymers. EPSs play a key role in protection of microbial cells under various external influences. For humans, these substances have potential use in many industries. EPSs can be applied as a flavor or a fragrance carrier, an emulsifier, a stabilizer, a prebiotic, an antioxidant or an antitumor agent. In this study, we characterized an environmental microorganism that produces EPS, optimized EPS production by this strain and characterized the EPS produced. Isolate CH-KOV3 was identified as Brachybacterium paraconglomeratum. The sucrose level in the growth medium greatly influenced EPS production, and the highest yield was when the microorganism was incubated in media with 500g/L of sucrose. The optimal temperature and pH were 28°C and 7.0, respectively. The nuclear magnetic resonance (NMR) results and GC-MS analysis confirmed that the residues were d-fructofuranosyl residues with β-configuration, where fructose units are linked by β-2,6-glycosidic bonds, with β-2,1-linked branches. All these data indicate that the investigated EPS is a levan-type polysaccharide. Thus, it was concluded that Brachybacterium sp. CH-KOV3 could constitute a new source for production of the bioactive polysaccharide, levan.