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Persistent organic pollutants (POPs) are lipophilic, constant and bioaccumulative toxic compounds. In general, they are considered resistant to biological, photolytic, and chemical degradation with polychlorinated biphenyls (PCBs) belonging to these chemicals. PCBs were never produced in Serbia, but they were imported and mainly used in electrical equipment, transformers, and capacitors. Our study aimed to analyse sequential multi-stage aerobic/anaerobic microbial biodegradation of PCBs present in the river sediment from the area known for long-term pollution with these chemicals. The study with an autochthonous natural microbial community (NMC model system) and NMC augmented with allochthonous hydrocarbon-degrading (AHD) microorganisms (isolated from location contaminated with petroleum products) (NMC-AHD model system) was performed in order to estimate the potential of these microorganisms for possible use in future bioremediation treatment of these sites. The laboratory biodegradation study lasted 70 days, after which an overall >33 % reduction in the concentration of total PCBs was observed. This study confirmed the strong potential of the NMC for the reduction of the level of PCBs in the river sediment under alternating multi-stage aerobic/anaerobic conditions.

Within a 30-day incubation laboratory study, the polycyclic aromatic hydrocarbon (PAH) degradation profile of two bacteria, Planomicrobium sp. RNP01 and Rhodococcus sp. RNP05 were studied by three microtiter plate assays to reveal the combination of certain biological and biochemical characteristics which are reliable indicators in evaluation of bacterial biodegradation abilities. The three assays, which are hydrocarbon growth assay, 2,6-DCPIP assay and dehydrogenase activity assay revealed that Rhodococcus sp. RNP05 exhibited better potential for PAH degradation than Planomicrobium sp. RNP01. Differences between initial and final optical density and specific growth rate constants were significantly higher (r = 0.995, P < 0.05) in case of Rhodococcus sp. RNP05 on all tested substrates, as compared to Planomicrobium sp. RNP01. This was confirmed by GC-FID analyses. Dehydrogenase activity of Rhodococcus sp. RNP05 was higher (r = 0.9995, P < 0.05) than Planomicrobium sp. RNP01 and correlated positively with the hydrocarbon growth assay (r = 0.999, P < 0.05, for Rhodococcus sp. RNP05, r = 0.986, P < 0.05 for Planomicrobium sp. RNP01). This study has shown that the combination of these assays could be used for determining the bioremediation potential of PAHs in petroleum contaminated soil with the ability of screening a large number of bacterial strains. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III43004]

Cerium oxide (CeO2) nanoparticles (CONPs) are interesting biomaterials with various applications in biomedicine, cosmetics and the pharmaceutical industry, but with limited practical application because of their low stability in aqueous media. The aim of this study was to obtain CONPs with increased stability by coating the particles. Microbial exopolysaccharides (levan, pullulan) and glucose were used to prepare CONPs under different synthesis conditions. Coating was attempted by adding the carbohydrates during (direct coating) or after (subsequent coating) the synthesis of CONPs. The obtained nanoparticles were characterized by X-Ray diffraction analysis, Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The suspension stability of the uncoated and coated CONPs in aqueous media was evaluated by measuring the hydrodynamic size, zeta potential and turbidity. The FT-IR spectra revealed the differences between coated CONPs and showed the success of subsequent coating with carbohydrates. Coating with carbohydrates improved the stability the CONP suspension by decreasing the size of aggregated particles. The suspensions of levan- and glucose-coated CONPs had the best stability. In this study, CONPs were prepared using non-toxic materials, which were completely environmentally friendly. The obtained results open new horizons for CONP synthesis, improving their biological applications. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III45012 and Grant no. III43004]

In the present work, four derivatives of salicylaldehyde (salicylaldehyde-hydrazone, phenylhydrazone, semicarbazone and thiosemicarbazone) were synthesized using both conventional (95% ethanol) and green (crude glycerol from biodiesel production) solvents. The obtained compounds were identified by elemental microanalysis, as well as FTIR, UV/Vis and 1H-NMR spectroscopic methods. Yields of 93–98% of the compounds in crude glycerol were achieved within 10–25 min. The derivatives of salicylaldehyde and crude glycerol were used as a nitrogen and carbon source, respectively, in the medium for antibiotic (hexaene H-85 and azalomycine B) production by Streptomyces hygroscopicus CH-7. The highest concentrations of hexaene H-85 and azalomycine B were achieved in the medium containing salicylaldehyde-thiosemicarbazone (198 and 69 μg/cm3, respectively). Derivatives of salicylaldehyde also impacted the strain morphology. In the media with salicylaldehyde- phenylhydrazone and salicylaldehyde-thiosemicarbazone, S. hygroscopicus CH-7 grew like large dispersive pellets with long twisted filaments that produced the highest yield of the antibiotics. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III 45001]

The effect of isatin derivatives as a nitrogen source on antibiotic (Hexaene H-85 and Azalomycine B) production by Streptomyces hygroscopicus CH-7 was studied. Isatin-3-hydrazone, 5-chloroisatin-3-hydrazone, isatin-3-tosylhydrazone, 5-chloroisatin-3-tosylhydrazone, isatin-3-(4`-hidroxy)benzoilhydrazone and 5-chloroisatin-3-(4`-hidroxy)benzoilhydrazone were synthesized in a crude glycerol, obtained during the biodiesel production from edible sunflower oil. The highest concentration of Hexaene H-85 is achieved with 5-chloroisatin-3-hydrazone (197 mg/cm3) in medium, while isatin-3-hydrazone has a greatest impact on Azalomycine B production (72 μg/cm3). [Projekat Ministarstva nauke Republike Srbije, br. III 45001]

The group of polycyclic aromatic hydrocarbons (PAHs) are very hazardous environmental pollutants because of their mutagenic, carcinogenic and toxic effects on living systems. The aim of this study was to examine and compare the ability and efficiency of selected bacterial isolates obtained from oil-contaminated areas to biodegrade PAHs. The potential of the bacteria to biodegrade various aromatic hydrocarbons was assessed using the 2,6-dichlorophenol-indophenol assay. Further biodegradation of PAHs was monitored by gravimetric and gas-chromatographic analysis. Among the eight bacterial isolates, identified on the basis of 16S rDNA sequences, two isolates, Planomicrobium sp. RNP01 and Rhodococcus sp. RNP05, had the ability to grow on and utilize almost all examined hydrocarbons. Those isolates were further examined for biodegradation of phenanthrene and pyrene, as single substrates, and as a mixture, in vitro for ten days. After three days, both isolates degraded a significant amount phenanthrene, which has a simpler chemical structure than pyrene. Planomicrobium sp.RNP01 commenced biodegradation of pyrene in the PAH mixture only after it had almost completly degraded phenanthrene. The isolated and characterized bacteria, Planomicrobium sp. RNP01 and Rhodococcus sp. RNP05, have shown high bioremediation potential and are likely candidates to be used for degradation of highly toxic PAHs in contaminated areas. [Projekat Ministarstva nauke Republike Srbije, br. III43004]

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.

The paper presents results of the ex situ bioremediation of soil contaminated by mazut (heavy residual fuel oil) in the field scale (600 m3). A treatment-bed (thickness 0.4 m) consisted of mechanically mixed mazut-contaminated soil, softwood sawdust as the additional carbon source and crude river sand, as bulking and porosity increasing material. The inoculation/reinoculation was conducted periodically using a biomass of a consortium of zymogenous microorganisms isolated from the bioremediation substrate. The biostimulation was performed through addition of nutritious substances (N, P and K). The aeration was improved by systematic mixing of the bioremediation system. After 50 days, the number of hydrocarbon degraders increased 100 times. Based on the changes in the group composition, the average biodegradation rate during bioremediation was 24 mg/kg/day for the aliphatic fraction, 6 mg/kg/day for the aromatic fraction, and 3 mg/kg/day for the nitrogen-sulphuroxygen compounds (NSO)-asphaltene fraction. In the saturated hydrocarbon fraction, gas chromatography-mass spectrometry (GC-MS) in the single ion-monitoring mode (SIM) was applied to analyse isoprenoids pristane and phytane and polycyclic molecules of sterane and triterpane type. Biodegradation occurred during the bioremediation process, as well as reduction of relative quantities of isoprenoids, steranes, tri- and tetracyclic terpanes and pentacyclic terpanes of hopane type.

Bioremediation, a process that utilizes the capability of microorganism to degrade toxic waste, is emerging as a promising technology for the treatment of soil and groundwater contamination. The technology is very effective in dealing with petroleum hydrocarbon contamination. The aim of this study was to examine the composition of the microbial consortium during the ex situ experiment of bioremediation of soil heavily contaminated with crude oil and its products from the Oil Refinery Pančevo, Serbia. After a 5.5-month experiment with biostimulation and bioventilation, the concentration of the total petroleum hydrocarbons (TPH) had been reduced from 29.80 to 3.29 g/kg (89 %). In soil, the dominant microorganism population comprised Gram-positive bacteria from actinomycete-Nocardia group. The microorganisms which decompose hydrocarbons were the dominant microbial population at the end of the process, with a share of more than 80 % (range 107 CFU/g). On the basis of the results, it was concluded that a stable microbial community had been formed after initial fluctuations.

Four new air-stable mixed-ligand Co(II) complexes having the general formula [Co2(Y)tpmc]Z3×q(H2O/CH3CN) (HY = N-methylglycine/N,N-dimethylglycine, Z = - 4BF , qH2O=4 or 3; HY=S-norvaline/S-valine Z=-ClO4 , qCH3CN = 0.5; qH2O = 0.5; tpmc = N,N',N'',N'''-tetrakis(2-pyridylmethyl)- -1,4,8,11-tetraazacyclotetradecane) were prepared. The composition, some physical and chemical properties and their tentative geometries were evaluated based on elemental analysis (C, H, N), conductometric and magnetic measurements, spectroscopic data (UV/Vis, IR) and cyclic voltammetry. The data were compared with earlier described analogous complexes containing the macrocyclic ligand and aliphatic aminocarboxylates. It is assumed that all complexes are binuclear with an exo coordination mode of the octaazamacrocyclic pendant ligand in the boat conformation. In addition, two -N-(CH2)2-N- portions of the cyclam ring within the tpmc ligand and Co(II) ions in the high-spin state are most probably bridged via oxygen atoms from the anion of the aminocarboxylate/derivatives, whereas nitrogen atoms rest uncoordinated. In all cases, a combined chelate-bridged coordination is proposed as the most probable. The complexes were electrochemically stable in the potential range -1.0 to 1.0 V. They were also preliminary assayed toward some microorganisms together with the ligands, starting simple salts and solvents as test substances. In some cases, certain antimicrobial activity of the complexes was detected.

Investigations of heavy metals from lignite ash, which is a coal combustion by-product, have for years been very interesting from different aspects of environmental protection. In this study, the possibility of microbiological ash leaching, with a strong oxidation agent, bacterially generated Fe2(SO4)3 was investigated in order to minimize the concentration of toxic ash elements. Leaching experiment by the shake flask testing technique was performed during a period of 20 d at a temperature of 28°C. The results obtained show 5- to 22-fold (for Cd and Ni approx. 5, Zn approx. 6, Mn approx. 8, Cr approx. 13 and Cu approx. 22) higher metal leaching efficiency in the suspension with bacterially generated Fe2(SO4)3 than in the control suspension. .

The association patterns of xylene and of total phosphorus with sediment constituents in an alluvial aquifer consisting of clays and sands which had been contaminated by a spill of technical grade xylene were investigated with the aim of assessing the dependence of in situ xylene degradation on the P-availability. The investigated alluvial sediments of the Makiš Plain (Sava river alluvion, Quaternary age, Serbia and Montenegro) are relatively poor in P. The most reliable associations were proposed by analyzing the quality of the correlations of xylene- and of P-contents with 32 sediment characteristics. The association patterns depend on the mineralogical and granulometrical composition of sediments, the structural characteristics of the organicmatter (N/C-ratio) and the redox-conditions. Differing association patterns in the clays and the sand had been developed, and these characteristics influence the intrinsic xylene degradation. The main proof for the occurrence of xylene degradation is the percentul share of bacteria which oxidize xylene as a single carbon-source.

The effects of methyl oleate and Tween 80, hexaene H-85 production stimulators, on the fatty acid composition of the total lipids of S. hygroscopicus CH-7 were investigated. Besides oleic acid in the biomass, an increase of the branched chain fatty acid contents was observed: from 1.6 to 2.1 fold in the medium with methyl oleate and 2.5-2.9 in the medium with Tween 80 compared to the base medium. These two additives act differently on the growth of the producing strain and the uptake of valine, an essential nutrient. A decreased proteolytic activity and similar cellular fatty acid profiles indicate that the main effect of the added compounds is to facilitate the excretion of antibiotics to the outer medium, thus increasing biosynthesis.