Soil bacterial strains with heavy metal resistance and high potential in degrading diesel oil and <Emphasis Type="Italic">n</Emphasis>-alkanes

Four bacterial strains, capable of degrading diesel oil, n-alkanes or hexadecane, were isolated from soils contaminated with petroleum oil and identified. Strains of Pseudomonas sp., Pseudomonas putida TPHK-1 and Pseudomonas aeruginosa TPHK-4, were more efficient in degrading high concentrations of the hydrocarbons than the other two strains, Stenotrophomonas maltophilia TPHK-2 and Acenitobacter sp. TPHK-3. P. putida TPHK-1 exhibited tolerance to very high concentrations of heavy metals such as cadmium, lead, zinc and copper. The innate ability of P. putida TPHK-1, as evidenced by the amplified genes alkB1 and alkB2 that encode alkane hydroxylases, and cat12o and cat23o coding for catechol dioxygenase, in degrading diesel oil in the presence of heavy metals is far greater than that of the strains reported in the literature. Heavy metal tolerance coupled with rapid degradation of hydrocarbons, even at high concentrations, suggests that P. putida TPHK-1 has a great potential in remediating soils contaminated with mixtures of hydrocarbons and heavy metals.     

Petrological considerations for the demineralization of Rajmahal coals with <Emphasis Type="Italic">Pseudomonas mendocina</Emphasis> B6-1

The present study entails the results of the petrological coniderations for demineralization of Rajmahal Gondwana coals with Psudomonas mendocina B6-1. Inertinite group macerals are the dominant constituents of these coals, followed by vitrinite group, while liptinite occurs in low concentration. The amount of Mineral matter is moderately high. The concentration of major, minor and trace elements is high when compared with Clarke values. After the treatment of these coals with Pseudomonas mendocina strain B6-1, a significant reduction in the elemental content of oxygen, hydrogen and sulphur was observed. A gradual reduction of pyrite phase due to bioleaching was identified and its signatures were reflected in the XRD spectra and FTIR absorption bands. Over 5 % reduction in the ash content and decrease of major, minor and trace element to variable degrees were also noticed. Fe, As, and Ca positively correlate with the ash removal percentage indicating that the samples with high concentration of these elements were prone to demineralization with Pseudomonas mendocina strain B6-1 whereas Ni, Zn, Cr and Cu, negatively correlate with the ash removal percentage and shows that their enrichment impeded the process of demineralization. With increasing concentration of vitrinite the removal of major/minor/trace elements also increased which is attributed to the possible association of these elements with the mineral matter occurring as superficial mounting and superficial blanketing over the vitrinite macerals. This could facilitate the bacterial access to the elements to act upon and remove it to the variable extent. Some minerals occur intergrown with inertinites causing restricted bacterial action owing to the nonexposure of the mineral particles and less surface area available to the bacteria for bioleaching. Sulfur removal strongly relates with increase in inertinite content and decrease in liptinite content. This appears that sulfur associated with liptinites have shown difficult removal condition. The maximum removal of the trace elements like Cd, Cu, Co, Zn and Pb was observed from the ‘banded dull coal’ samples of the Rajmahal basin while maximum removal of major/minor elements like Fe, Ca and Mg was noticed from the ‘banded bright coal’ samples. However, maximum removal of Mn and As was also observed in the samples of ‘banded coal’.     

Production of biosurfactant by Nocardia otitidiscaviarum and its role in biodegradation of crude oil

Nocardia otitidiscaviarum microbial type cultural collection 6471 isolated from oil contaminated Alang India seawater was examined for production of surface active compound. Isolate shows halos and α-heamolysis on cetyl trimethyl ammonium bromide and blood agar respectively indicating the production of biosurfactant. Biosurfactant was extracted by precipitation and was partially purified. Partially purified product was characterized by thin layer chromatography and Infra-red spectroscopy and was identified as glycolipid. Sugar present in glycolipid was rhamnose and hence, biosurfactant was quantified as rhamnose equivalent. Role of cell-surface hydrophobicity and emulsification activity in correlation with biosurfactant production was examined. Correlation between biosurfactant production, growth and crude oil degradation was also examined and showed positive correlation at significant level 0.001 and 0.01 respectively. Thus, this is a first report on a marine strain of Nocardia otitidiscaviarum microbial type culture collection 6471, which can be a potential candidate for restoration of oil contaminated marine environment.     

Assessment of bacterial diversity associated with crude oil-contaminated soil samples from Assam

In this study, we are reporting bacterial diversity in crude oil-contaminated soil of Assam, India. Integration of physiological community profiling, culture-dependent and culture-independent (metagenome) approaches, was employed to obtain a complete picture of the total bacterial diversity. Samples collected from 10 sites contaminated with crude oil ranging from 0.22 to 89.36% were analysed, and altogether 160 culturable bacteria were isolated (117 Gram-positive and 43 Gram-negative bacteria). Molecular identification showed the predominance of genera Lysinibacillus, Alcaligenes, Bacillus, Clostridium, Enterobacter and Pseudomonas. Conversely, denaturing gradient gel electrophoresis (DGGE) profiles of 16S rDNA phylotypes showed the predominance of Sphingomonas, Ralstonia, Sphingobium, Massilia, Acinetobacter and Pseudomonas. Both culture-dependent and culture-independent approaches resulted in 11 genera of which Bacillus and Pseudomonas were the key inhabitants creating most favourable bacterial milieu in the presence of crude oil contaminants. Metabolic fingerprints data depicted in PCA plot demonstrated that sites CTF-D-1 and Core-10 were most diverged. It was further confirmed that variations of bacterial species dominance in different sites were due to origin of hydrocarbon contamination. We here claim that the present findings is a first-hand report on combined physiological community profiling, culture-based and culture-independent approaches in assessing total bacterial diversity in crude oil-contaminated soil of Assam.     

Isolation of <Emphasis Type="Italic">Bacillus cereus</Emphasis> from botanical soil and subsequent biodegradation of waste engine oil

Waste engine oil causes a vital environmental pollution when it spill during change and transportation and products of waste engine oil causes lethal effects to the living systems. Thus, abiotic and biotic approaches are being extensively used for removal of waste engine oil pollution. Therefore in present study, waste engine oil degradation was accomplished by a new bacterial culture, isolated from the soil by an enrichment technique. Morphological, biochemical and gene sequence analysis revealed that isolate was Bacillus cereus. Subsequently, biodegradation potential of B. cereus for waste engine oil was studied. Experimental variables, such as pH, substrate concentration, inoculum size, temperature and time on the biodegradation, were checked in mineral salt medium. The biodegradation efficiency of B. cereus was determined by gravimetry, UV–visible spectrophotometry and gas chromatography. In addition, waste engine oil was also characterized by GC–MS and FTIR for its major constituents, which showed total 38 components in waste engine oil, including hopanes, benzopyrene, long-chain aliphatic hydrocarbons, dibenzothiophenes, biphenyl and their derivatives. Results of successive biodegradation indicated that B. cereus was capable to degrade 1% of waste engine oil with 98.6% degradation potential at pH 7 within 20 days. Hence, B. cereus presents an innovative tool for removing the engine oil from the contaminated area.     

Bacterial consortium expressing surface displayed,intra- and extracellular lipases and pseudopyronine B for the degradation of oil

The lipA gene, encoding a solvent-tolerant extracellular lipase from Proteus sp. SW1, was displayed on the cell surface of Escherichia coli by fusing it to an antigen 43 anchoring motif. The display of LipA on the Escherichia coli cell surface was directly confirmed by immunofluorescence microscopy and flow cytometry. After 6 days of incubation in media containing 1 % used cooking oil, an Escherichia coli strain expressing surface displayed lipase was able to degrade 27 % of the oil. The biosurfactant, pseudopyronine B, was purified from culture supernatants of Pseudomonas sp. SL31. Its critical micelle concentration was determined to be 1400 mg/l, and the surfactant was stable within a temperature range from 0 to 120 °C and a pH range of 3–11. Pseudopyronine B-containing crude media extracts efficiently removed up to 51 % of the cadmium from contaminated water. We demonstrated the oil degradation ability of the mixed culture of four bacterial strains, namely the recombinant Escherichia coli expressing cell surface displayed lipase (pKKJlipA), His-tagged lipase (pETlipA), extracellular lipase-producing Proteus sp. SW1, and pseudopyronine B-producing Pseudomonas sp. SL31 by culturing in LB media containing 1 % oil. The consortium degraded 29 % of oil in one day and reached 84 % after 7 days.     

Development of a two-stage washing and biodegradation system to remediate octachlorinated dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin-contaminated soils

A two-stage system for octachlorinated dibenzo-p-dioxin (OCDD)-contaminated soil remediation was developed. Soil washing using emulsified oil (EO) was applied in the first stage for OCDD extraction followed by the second stage of bioremediation using P. mendocina NSYSU for remaining OCDD biodegradation. The major tasks included (1) determination of optimal soil washing conditions for OCDD extraction by EO, (2) evaluation of feasibility of OCDD biodegradation by P. mendocina NSYSU under aerobic cometabolic conditions using EO as the primary substrate, and (3) assessment of the effectiveness of OCDD removal using the two-stage system. During the soil washing stage, EO with two different oil-to-water ratios (1:50 and 1:200) and pore volumes were tested with initial soil OCDD concentration of 21,000 µg/kg. Results indicate that EO could effectively improve the solubility and desorption of OCDD in soils. Up to 74% of OCDD removal could be obtained after washing with 60 PVs of EO and dilution factor of 50. After the soil washing process, enriched P. mendocina NSYSU solution was added into the reactor to enhance the aerobic biodegradation of remaining OCDD in soils. P. mendocina NSYSU could use adsorbed EO globules as substrates and caused significant OCDD degradation via the aerobic cometabolic mechanism. Approximately 82% of the remaining OCDD could be removed after 50 days of operation, and P. mendocina NSYSU played important roles in OCDD biodegradation. Up to 87% of OCDD was removed through the EO washing and biodegradation process. The two-stage system is a potential technology to remediate dioxin-contaminated soils.     

Isolation and characterization of a novel native Bacillus strain capable of degrading diesel fuel

The ability of native bacteria to utilize diesel fuel as the sole carbon and energy source was investigated in this research. Ten bacterial strains were isolated from the oil refinery field in Tehran, Iran. Two biodegradation experiments were performed in low and high (500 and 10000 ppm, respectively) concentration of diesel fuel for 15 days. Only two isolates were able to efficiently degrade the petroleum hydrocarbons in the first test and degraded 86.67% and, 80.60 % of diesel fuel, respectively. The secondary experiment was performed to investigate the toxicity effect of diesel fuel at high concentration (10000 ppm). Only one strain was capable to degrade 85.20 % of diesel fuel at the same time (15 days). Phenotype and phylogeny analysis of this strain was characterized and identified as diesel-degrading bacteria, based on gram staining, biochemical tests, 16S rRNA gene sequence analysis. These results indicate that this new strain was Bacillus sp. and could be considered as Bacillus Cereus with 98 % 16 S rRNA gene sequence similarity. The results indicate that native strains have great potential for in situ remediation of diesel-contaminated soils in oil refinery sites.     

Induced degradation of crude oil mediated by microbial augmentation and bulking agents

Different bacterial and fungal strains, isolated from petroleum hydrocarbon-contaminated soil, were tested, in isolation as well as in combination, for their ability to degrade total petroleum hydrocarbon (TPH) in soil samples spiked with crude oil (2, 5 or 10 %, w/w) for 30 days. The selected combination of bacterial and fungal isolates, i.e., Pseudomonas stutzeri BP10 and Aspergillus niger PS9, exhibited the highest efficiency of TPH degradation (46.7 %) in soil spiked with 2 % crude oil under control condition. Further, when this combination was applied under natural condition in soil spiked with 2 % (w/w) crude oil along with inorganic fertilizers (NPK) and different bulking agents such as rice husk, sugarcane, vermicompost or coconut coir, the percent degradation of TPH was found to be maximum (82.3 %) due to the presence of inorganic fertilizers and rice husk as bulking agent. Further, results showed that the presence of NPK and bulking agents induced the activity of degradative enzymes, such as catalase (0.718 m mol H₂O₂ g^?1), laccase (0.77 µmol g^?1), dehydrogenase (37.5 µg g^?1 h^?1), catechol 1, 2 dioxygenase (276.11 µ mol g^?1) and catechol 2, 3 dioxygenase (15.15 µ mol g^?1) as compared to control (without bioaugmentation). It was inferred that the selected combination microbes along with biostimulants could accentuate the crude oil degradation as evident from the biostimulant-induced enhanced activity of degradative enzymes.     

Evaluation of petroleum hydrocarbon biodegradation in shallow groundwater by hydrogeochemical indicators and C, S-isotopes

Biodegradation is one of the main natural attenuation processes in groundwater contaminated with petroleum hydrocarbons. In this work, preliminary studies have been carried out by analyzing the concentrations of total petroleum hydrocarbons (TPH), dissolved inorganic carbon (DIC), dominant terminal electron accepters or donors, as well as δ ¹³C_DIC and δ ³⁴S_SO4, to reveal the biodegradation mechanism of petroleum hydrocarbons in a contaminated site. The results show that along groundwater flow in the central line of the plume, the concentrations of electron acceptors, pH, and E _h increased but TPH and DIC decreased. The δ ¹³C_DIC values of the contaminated groundwater were in the range of ?14.02 to ?22.28 ‰_PDB and ?7.71 to 8.36 ‰_PDB, which reflected a significant depletion and enrichment of ¹³C, respectively. The increase of DIC is believed to result from the non-methanogenic and methanogenic biodegradation of petroleum hydrocarbon in groundwater. Meanwhile, from the contaminated source to the downgradient of the plume, the ³⁴S in the contaminated groundwater became more depleted. The Rayleigh model calculation confirmed the occurrence of bacterial sulfate reduction as a biodegradation pathway of the petroleum hydrocarbon in the contaminated aquifers. It was concluded that stable isotope measurements, combined with other biogeochemical measurements, can be a useful tool to prove the occurrence of the biodegradation process and to identify the dominant terminal electron-accepting process in contaminated aquifers.     

Biosynthesis and genetics of polyhydroxyalkanoates by newly isolated <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> IFS and 30N using inexpensive carbon sources

Poly(3-hydroxyalkanoates) (PHAs) are the bioplastics that are stored in many genera of bacteria as carbon and energy storage polyester granules. PHAs have established themselves as strong competitors to conventional plastics. This study reports the isolation of PHA-accumulating bacteria from local environment and their PHA characterization. Two potential strains identified as Pseudomonas aeruginosa strain IFS (Accession no. JQ041638) and P. aeruginosa strain 30N (Accession no. JQ041639) based on 16S rRNA gene sequence identity were cultivated under nitrogen limited conditions to study their PHA biosynthesis capabilities. The strain IFS and strain 30N produced 1.36 and 1.40 gl^?1 dry biomass with percentage PHA contents of 44.85 and 45.74%, respectively, when grown on glucose as carbon source. The PHA was identified as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by fourier transform infrared spectroscopy (FTIR) and gas chromatography coupled with mass spectrometry (GC–MS). The PHA synthase genes of these strains were isolated, sequenced and analyzed using bioinformatic tools that showed they belonging to type 2 PHA synthases and presented their evolutionary relationships with PHA synthases of other Pseudomonas species. The experimental results of this study highlight the importance of these strains for future use of bacterial biopolymer production growing on simple and inexpensive carbon sugars.     

The efficiency of Penicillium commune for bioremoval of industrial oil

Among all environmental contaminations, industrial oil is one of the major pollutants of soil, water, and air. There are different chemical, physical, and biological methods to remove all types of oil pollutions. One of the common biological methods is to utilize the microorganisms like yeast, fungi or bacteria. Previous studies concerning the biodegradation of an aromatic compound in industrial waste water by Aspergillus niger have been reported. In this study, we tried to identify an oil-derived microorganism and evaluate its efficacy on self-removal of industrial oil. Firstly, the strain of isolated fungus from various bulks of used oil was defined via colonial identification and DNA sequencing. Secondly, bioremoval activity of defined fungus (Penicillium commune) was evaluated using gas chromatography–mass spectrometry. The optimum conditions in biological elimination of oil including the incubation time, pH level of culture, and amount of reagents were determined. In the best condition, a removal rate of 95.4 % was obtained.     

Microbial diversity in contaminated soils along the T22 trench of the Chernobyl experimental platform

The diversity of bacterial communities exposed to radioactive contamination in Chernobyl soils was examined by a combination of molecular and culture-based approaches. A set of six radioactive soil samples, exhibiting high levels of ¹³⁷Cs contamination, were collected from the T22 trench. Three samples were also collected in nearby soils with low contamination. Complex bacterial community structures were observed in both highly and weakly contaminated samples, using a molecular approach targeting the 16S rRNA gene. However, the presence of specific populations within samples from highly contaminated soils could not be revealed by statistical analysis of the DGGE profiles. More than 200 culturable isolates, representative of dominant morphotypes, were grouped into 83 Operational Taxonomic Units (OTUs) and affiliated to Firmicutes, Actinobacteria, Alpha-, Beta-, Gamma-Proteobacteria and Bacteroïdetes. No specific pattern linked to contamination was observed for these culturable bacteria. The results show that both highly and weakly contaminated soils host a wide diversity of bacteria, suggesting that long term exposure to radionuclides does not lead to the extinction of bacterial diversity.     

Bacterial degradation of crude oil: Comparison of field and experimental data

It has been reported that the composition of crude oils in the subsurface may be altered by bacterial action to the extent that oil correlations (Winters and Williams, 1969) and the value of the crude (Evans et al., 1971) are severely affected. Experimental documentation of these effects is provided by this study.A crude oil was degraded in a 21-day laboratory experiment by a culture of four aerobic bacteria isolated from an oil-contaminated soil. The progress of the experiment was measured by the changes induced in the chemical composition of the oil fraction boiling above 270°C. These changes were similar to the variations in composition found in the MC5 oils of Saskatchewan, Canada.Normal paraffins through to at least nC₃₄ were severely depleted although the attack was temporarily blocked at nC₂₅ (Jobson et al., 1972). The position of this blockage is a function of the isolate employed. The isoprenoids, pristane and phytane, were metabolised after the disappearance of the n-paraffins. Lower-ring naphthenes and aromatics were attacked at the same time as the lighter normal paraffins and before the heavier ones.The more condensed cyclic hydrocarbons were apparently unaffected. Additional non-hydrocarbon NSO, and particularly asphaltene (both defined under section “Methods”), compounds were formed by the metabolism of the hydrocarbons.The residual oil after attack was heavier by approximately 30° API than the initial crude oil.     

Production of biosurfactant from Iranian oil fields by isolated Bacilli

Crude oil and water samples were collected from selected Iranian oil reservoirs. Experimental works were carried out in laboratory conditions. The samples have been grown on PYG medium and incubated at 30–80 °C. Thirty-six mesophile and thermophile bacterial strains have been isolated. All the isolates were able to grow at aerobic condition. Batch growth kinetic studies were carried out in a 500 ml. shake flask. Out of 36 isolated strains from 24 crude oil and water samples, 35 strains were gram positive rod. Shaped spore forming bacteria and one strain was coccid form. Eight out to 35 bacillus species were capable of producing surfactant. Production of biosurfactant was found to be cell growth associated. The ability of surfactant producing bacteria indicated by reduction of surface tension (ST) and interfacial tension (IFT) of the supernatant. Eight strains obtained the IFT reduction in crude oil, hexadecane, sucrose, glucose, fructose and mannose medium as a sole source of carbon and energy at 40 °C by 15–30 mN/m. Results showed that all the bacteria are producing more surfactant when glucose is the carbon source. Further screening of biosurfactant producer showed that three of the isolated strains resulted the maximum ST and IFT. Effect of temperature on these three isolates investigated at 30–80 °C, above 50 °C surfactant production was dramatically reduced. The isolated strains had the capacity to produce the surfactant at 3–5% NaCl a wide rang of pH (6.5–8.5).     

Bioremediation of crude oil by <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> in the presence of different concentration nanoparticles and produced biosurfactant

This research work focuses on testing the bacterial strain Bacillus licheniformis for the bioremediation capacity of the crude oil. A biosurfactant and two different nanoparticles with different concentrations (0.05, 0.1, 0.2 g/100 ml) were applied separately to enhance the biodegradation process. The optimum biodegradation of crude oil was demonstrated at 60% of microcosms containing biosurfactant and nanoparticles after 7 days. The bacterial strain is highly potential to consume the total paraffins (iso- and n-paraffins) in crude oil samples. Accordingly, the best biodegradation of total paraffins was observed in microcosms containing (0.2 g) of Fe₂O₃, Zn₅(OH)₈Cl₂ (nps) and biosurfactant separately. Additionally, the consumption of specific member rings of polyaromatics depends on the type and the concentration of nanoparticles. Thus, this bacterial strain was considered as a good candidate to be applied in the bioremediation process of petroleum-contaminated sites using biosurfactant and specific concentration of (Fe₂O₃ and Zn₅OH₈Cl₂) nanoparticles.     

Bioremediation of 2,4,6-trinitrotoluene-contaminated groundwater using unique bacterial strains: microcosm and mechanism studies

Groundwater at many military factory, munition storage and maneuver sites is contaminated by explosives chemicals that were released into the subsurface. The 2,4,6-trinitrotoluene (TNT) is among the most common explosive pollutants. In this study, two TNT-degrading strains, isolated from TNT-contaminated soils and wastewater sludge, were applied for TNT biodegradation. Based on the 16S rDNA sequence analyses, these two bacterial strains were identified as Achromobacter sp. and Klebsiella sp. via biochemical and DNA analyses. Microcosm study was conducted to evaluate the feasibility and efficiency of TNT biodegradation under aerobic conditions. Results indicate that TNT degradation by-products were detected in microcosms (inoculated with Achromobacter sp. and Klebsiella sp.) with cane molasses addition. Klebsiella sp. and Achromobacter sp. used TNT as the nitrogen source and caused completely removal of TNT. Two possible TNT biodegradation routes could be derived: (1) part of the TNT was transformed to nitrotoluene then transformed to nitrobenzene followed by the nitro substitute process, and trinitrobenzene, dinitrobenzene, and nitrobenzene were detected; and (2) TNT was transformed via the nitro substitute mechanism, and dinitrotoluene followed by nitrotoluene isomers were detected. The initial TNT degradation involved the reduction or removal of the nitro substitute to an amino derivative or free nitrite. Results show that the second route was the dominant TNT biodegradation pathway. The produced by-products were also degraded without significant accumulation during the degradation process. These findings would be helpful in designing a practical system inoculated with isolated TNT degradation strains for the treatment of TNT-contained groundwater.     

Optimization of calcium-based bioclogging and biocementation of sand

Bioclogging and biocementation can be used to improve the geotechnical properties of sand. These processes can be performed by adsorption of urease-producing bacterial cells on the sand grain surfaces, which is followed by crystallization of calcite produced from the calcium salt and urea solution due to bacterial hydrolysis of urea. In this paper, the effect of intact cell suspension of Bacillus sp. strain VS1, suspension of the washed bacterial cells, and culture liquid without bacterial cells on microbially induced calcite precipitation in sand was studied. The test results showed that adsorption/retention of urease activity on sand treated with washed cells of Bacillus sp. strain VS1 was 5–8 times higher than that treated with culture liquid. The unconfined compressive strength of sand treated with the suspension of washed cells was 1.7 times higher than that treated with culture liquid. This difference could be due to fast inactivation of urease by protease which was present in the culture liquid. The adsorption of bacterial cells on sand pretreated with calcium, aluminum, or ferric salts was 29–37 % higher as compared with that without pretreatment. The permeability of sand varied with the content of precipitated calcium. For bioclogging of sand, the content of precipitated calcium had to be 1.3 % (w/w) or higher. The shear strength of biotreated sand was also dependent on the content of precipitated calcium. To achieve an unconfined compressive strength of 1.5 MPa or higher, the content of precipitated calcium in the treated sand had to be 4.2 % (w/w) or higher. These data can be used as the reference values for geotechnical applications such as bioclogging for reducing the permeability of sand and biocementation for increasing the shear strength of soil.     

Production of rhamnolipids by Pseudomonas aeruginosa growing on carbon sources

Arhamnolipid producing bacterium, Pseudomonas aeruginosa was previously isolated from Iranian oil over years. Isolated strain was identified by morphological, biochemical, physiological and 16 sr RNA (1). Glycolipid production by isolated bacterium using sugar beet molasses as a carbon and energy source was investigated. Biosurfactant production was quantified by surface tension reduction, Critical Micelle Dilution (CMD), Emulsification Capacity (EC), and Thin Layer Chromatogeraphy. biosurfactants during growth on waste Dates as the primary carbon and nitrogen sources, respectively. After 48 h of growth the culture supernatant fluid had a rhamnose concentration of 0.18 g/L and surface tension was reduced to 20 mN/m ( %).( reduced the interfacial tension against crude oil from 21 mN/m to 0,47 mN/m) Result from the study showed that the growth of the bacteria using molasses as carbon sources is growth-associated. The specific production rate of rhamnolipid with 2 %, 4 %, 6 %, 8 % and 10 % of molasses are 0.00065; 4.556; 8.94; 8.85; and 9.09. respectively The yield of rhamnolipid per biomass with 2%,4%,6%,8% and 10% molasses are 0.003;0.009;0.053;0.041 and 0.213 respectively. The production of rhamnolipid (0.0531 g rhamnolipid/g biomass) is higher compare to the culture grown in aerobic condition (0.04 g rhamnolipid/g biomass).The rhamnolipids were able to form stable emulsions with n-alkanes, aromatics, crude oil and olive oil. These studies indicate that renewable, relatively inexpensive and easily available resources can be used for important biotechnological processes.     

Vicia sativa–rhizospheric bacteria interactions to improve phenol remediation

The aim of this study was to evaluate whether the interaction of Vicia sativa with a bacterial strain capable of using phenol as sole carbon and energy sources can moderate adverse effects of this pollutant in plant tissues. A bacterial strain identified as Bacillus sp., isolated from a heavily polluted environment, was inoculated at different stages of growth. In root elongation assay, inoculated seeds showed higher values of relative root elongation and germination index than uninoculated ones in the presence of high phenol concentrations. Thus, common vetch–Bacillus sp. association could be important at the first stages of development allowing this plant to grow in highly polluted environments. Besides, phenol removal was largely accelerated in phenol-spiked soils, after 48 h of treatment with uninoculated/inoculated plants rather than by adsorption or biodegradation of the bacterial strain. Peroxidase and ascorbate peroxidase activities increased significantly in uninoculated plants, while superoxide dismutase activity, chlorophyll, malondialdehyde, and H₂O₂ levels of aerial parts remained unaltered in uninoculated/inoculated plants treated with the pollutant, demonstrating that the efficient response to oxidative damage did not depend on the inoculation.