Biodegradation of diesel fuel by a microbial consortium in the presence of 1-alkoxymethyl-2-methyl-5-hydroxypyridinium chloride homologues

Fast development of ionic liquids as gaining more and more attention valuable chemicals will undoubtedly lead to environmental pollution. New formulations and application of ionic liquids may result in contamination in the presence of hydrophobic compounds, such as petroleum mixtures. We hypothesize that in the presence of diesel fuel low-water-soluble ionic liquids may become more toxic to hydrocarbon-degrading microorganisms. In this study the influence of 1-alkoxymethyl-2-methyl-5-hydroxypyridinium chloride homologues (side-chain length from C₃ to C₁₈) on biodegradation of diesel fuel by a bacterial consortium was investigated. Whereas test performed for the consortium cultivated on disodium succinate showed that toxicity of the investigated ionic liquids decreased with increase in side-chain length, only higher homologues (C₈–C₁₈) caused a decrease in diesel fuel biodegradation. As a result of exposure to toxic compounds also modification in cell surface hydrophobicity was observed (MATH). Disulphine blue active substances method was employed to determine partitioning index of ionic liquids between water and diesel fuel phase, which varied from 1.1 to 51% for C₃ and C₁₈ homologues, respectively. We conclude that in the presence of hydrocarbons acting as a solvent, the increased bioavailability of hydrophobic homologues is responsible for the decrease in biodegradation efficiency of diesel fuel.     

Effects of rhamnolipids on cell surface hydrophobicity of PAH degrading bacteria and the biodegradation of phenanthrene

The effects of rhamnolipids produced by Pseudomonas aeruginosa ATCC9027 on the cell surface hydrophobicity (CSH) and the biodegradation of phenanthrene by two thermophilic bacteria, Bacillus subtilis BUM and P. aeruginosa P-CG3, and mixed inoculation of these two strains were investigated. Rhamnolipids significantly reduced the CSH of the hydrophobic BUM and resulted in a noticeable lag period in the biodegradation. However, they significantly increased the CSH and enhanced the biodegradation for the hydrophilic P-CG3. In the absence of rhamnolipids, a mixed inoculation of BUM and P-CG3 removed 82.2% of phenanthrene within 30 days and the major contributor of the biodegradation was BUM (rapid degrader) while the growth of P-CG3 (slow degrader) was suppressed. Addition of rhamnolipids promoted the surfactant-mediated-uptake of phenanthrene by P-CG3 but inhibited the uptake through direct contact by BUM. This resulted in the domination of P-CG3 during the initial stage of biodegradation and enhanced the biodegradation to 92.7%.     

Enhanced biodegradation of phenanthrene using different inoculum types in a microbial fuel cell

Environmental pollution by petroleum hydrocarbons from contaminated groundwater and soils is a serious threat to human health. Microbial fuel cells (MFCs) could be employed in the treatment of these recalcitrant pollutants with concomitant bioelectricity generation. In this study, the use of MFCs in biodegradation of phenanthrene, a model hydrocarbon, was investigated with respect to its biodegradation rate, biodegradation efficiency, and power production using a range of inocula (Shewanella oneidensis MR1 14063, Pseudomonas aeruginosa NCTC 10662, mixed cultures, and combinations thereof). All the inocula showed high potentials for phenanthrene degradation with a minimum degradation efficiency of 97%. The best overall performing inoculum was anaerobically digested sludge supplemented with P. aeruginosa NCTC 10662, having a degradation rate, maximum power density and chemical oxygen demand removal efficiency of 27.30 μM/d, 1.25 mW/m² and 65.6%, respectively. Adsorption of phenanthrene on the carbon anode was also investigated; it conformed to a Type II adsorption isotherm and could be modelled using a modified Brunauer, Emmett and Teller model with a maximum monolayer capacity of 0.088 mg/cm². This work highlights the possibility of using MFCs to achieve high degradation rates of phenanthrene through co‐metabolism and could potentially be used as a replacement of permeable reactive barriers for remediation of hydrocarbon‐contaminated groundwater.     

Biodegradation of diesel fuel hydrocarbons by mangrove fungi from Red Sea Coast of Saudi Arabia

Mangrove sediments were collected from major mangrove stands on the Red Sea Coast of Saudi Arabia. Forty five isolates belonging to 12 genera were purified and five isolates as well as their consortium were found to be able to grow in association with petroleum oil as sole carbon source under in vitro conditions. The isolated strains were identified based on internal transcribed spacer (ITS) rDNA sequence analysis. The fungal strains with the greatest potentiality to degrade diesel oil, without developing antagonistic activity, were identified as Alternaria alternata, Aspergillus terreus, Cladosporium sphaerospermum, Eupenicillium hirayamae and Paecilomyces variotii. As compared to the controls, these fungi accumulated significantly higher biomass, produced extracellular enzymes and liberated larger volumes of CO₂. These observations with GC–MS data confirm that these isolates displayed rapid diesel oil bioremoval and when used together as a consortium, there was no antagonistic activity.     

Application of biocathode in microbial fuel cells: cell performance and microbial community

Instead of the utilization of artificial redox mediators or other catalysts, a biocathode has been applied in a two-chamber microbial fuel cell in this study, and the cell performance and microbial community were analyzed. After a 2-month startup, the microorganisms of each compartment in microbial fuel cell were well developed, and the output of microbial fuel cell increased and became stable gradually, in terms of electricity generation. At 20 ml/min flow rate of the cathodic influent, the maximum power density reached 19.53 W/m3, while the corresponding current and cell voltage were 15.36 mA and 223 mV at an external resistor of 14.9 Omega, respectively. With the development of microorganisms in both compartments, the internal resistance decreased from initial 40.2 to 14.0 Omega, too. Microbial community analysis demonstrated that five major groups of the clones were categorized among those 26 clone types derived from the cathode microorganisms. Betaproteobacteria was the most abundant division with 50.0% (37 of 74) of the sequenced clones in the cathode compartment, followed by 21.6% (16 of 74) Bacteroidetes, 9.5% (7 of 74) Alphaproteobacteria, 8.1% (6 of 74) Chlorobi, 4.1% (3 of 74) Deltaproteobacteria, 4.1% (3 of 74) Actinobacteria, and 2.6% (2 of 74) Gammaproteobacteria.     

Effect on cell surface hydrophobicity and susceptibility of Helicobacter pylori to medicinal plant extracts

Effects on aqueous extracts of medicinal plants on ten Helicobacter pylori strains were studied by the salt aggregation test to determine the possibility to modulate their cell surface hydrophobicity and by an agar diffusion assay for detection of antimicrobial activity. It was established that aqueous extracts of bearberry and cowberry leaves enhance cell aggregation of all H. pylori strains tested by the salt aggregation test, and the extract of bearberry possessed a remarkable bacteriostatic activity. Pure tannic acid showed a result similar to that of bearberry and cowberry extracts which contained a large amount of tannins. In contrast, extracts of wild camomile and pineapple-weed, which blocked aggregation of H. pylori, contained small amounts of tannins and did not reveal any antimicrobial activity. Tannic acid seems to be the component of bearberry and cowberry aqueous extracts with the highest activity to decrease cell surface hydrophobicity as well as in antibacterial activity against H. pylori.     

Supersusceptibility to hydrophobic antimicrobial agents and cell surface hydrophobicity in Branhamella catarrhalis

To clarify the cause of the supersusceptibility of Branhamella catarrhalis to macrolide antibiotics, which are well-known to be inactive to most Gram-negative bacteria, we determined its cell surface hydrophobicity by the partition experiment between water and hydrocarbons. Its cell surface was found to be markedly more hydrophobic than that of Escherichia coli or Pseudomonas aeruginosa cells. This suggested that the outer membrane of B. catarrhalis plays no role as a diffusion barrier towards hydrophobic agents.     

Adsorption of dirhamnolipid on four microorganisms and the effect on cell surface hydrophobicity

In this study, adsorption of dirhamnolipid biosurfactant on a Gram-negative Pseudomonas aeruginosa, two Gram-positive Bacillus subtilis, and a yeast, Candida lipolytica, was investigated, and the causality between the adsorption and change of cell surface hydrophobicity was discussed. The adsorption was not only specific to the microorganisms but also depended on the physiological status of the cells. Components of the biosurfactant with different rhamnosyl number or aliphatic chain length also exhibited slight difference in adsorption manner. The adsorption indeed caused the cell surface hydrophobicity to change regularly; however, the changes depended on both the concentrations of rhamnolipid solutions applied and the adsorbent physiological conditions. Orientation of rhamnolipid monomers on cell surface and micelle deposition are supposed to be the basic means of adsorption to change cell hydrophobicity at low and high rhamnolipid concentrations, respectively. This study proposed the possibility to modify cell surface hydrophobicity with biosurfactant of low concentrations, which may be of importance in in situ soil remediation. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Influence of fluconazole at subinhibitory concentrations on cell surface hydrophobicity and phagocytosis of Candida albicans

Cell surface hydrophobicity (CSH) status influences virulence of Candida albicans and decreases the susceptibility of yeast cells to phagocytic killing. We tested whether subinhibitory concentrations of fluconazole, which is widely used in the treatment and prophylaxis of candidiasis, affect CSH and the susceptibility of C. albicans to enzymatic digestion by glucanase and to phagocytic killing. Treatment of yeast cells with subinhibitory fluconazole concentrations resulted in greater phagocytosis. This effect was independent of CSH but may be related to increased cell wall porosity resulting from alterations in the cell envelope. The use of subinhibitory concentrations of fluconazole in patients with competent phagocytes may contribute to resistance to candidiasis regardless of yeast CSH status.     

Transmembrane diffusion of hydrophobic antimicrobial agents and cell surface hydrophobicity in Bacteroides fragilis

The transmembrane diffusion of hydrophobic antimicrobial agents, e.g. lincomycin and clindamycin, was examined in Bacteroides fragilis which is sensitive to these agents. The results showed that these agents penetrate efficiently through the outer membrane. Cell surface hydrophobicity measured by the partition assay between water and p-xylene revealed that the cell surface of B. fragilis is more hydrophobic than that of Salmonella typhimurium or Pseudomonas aeruginosa. Furthermore, treatment with low concentrations of surfactant caused cell lysis. These results suggest that the cell surface hydrophobicity in B. fragilis plays an important role in the efficient transmembrane penetration of hydrophobic compounds. This efficiency explains the susceptibility of B. fragilis to hydrophobic antimicrobial agents.     

Simultaneous decolorization of azo dye and bioelectricity generation using a microfiltration membrane air-cathode single-chamber microbial fuel cell

Electricity generation from readily biodegradable organic substrates accompanied by decolorization of azo dye was investigated using a microfiltration membrane air-cathode single-chamber microbial fuel cell (MFC). Batch experiment results showed that accelerated decolorization of active brilliant red X-3B (ABRX3) was achieved in the MFC as compared to traditional anaerobic technology. Biodegradation was the dominant mechanism of the dye removal, and glucose was the optimal co-substrate for ABRX3 decolorization, while acetate was the worst one. Confectionery wastewater (CW) was also shown to be a good co-substrate for ABRX3 decolorization and a cheap fuel source for electricity generation in the MFC. Low resistance was more favorable for dye decolorization than high resistance. Suspended sludge (SS) should be retained in the MFC for accelerated decolorization of ABRX3. Electricity generation was not significantly affected by the ABRX3 at 300 mg/L, while higher concentrations inhibited electricity generation. However, voltage can be recovered to the original level after replacement with anodic medium not containing azo dye.     

Adsorption of monorhamnolipid and dirhamnolipid on two <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> strains and the effect on cell surface hydrophobicity

Previously, adsorption feature of a dirhamnolipid biosurfactant on diverse microbial cells was studied and the effect of the adsorption on cell surface hydrophobicity was compared. In this paper, the adsorption behavior of a monorhamnolipid and a dirhamnolipid on cells of two Pseudomonas aeruginosa strains was investigated in order to further reveal the influence of biosurfactant structure and cell property on the adsorption and the relation between the adsorption and cell surface hydrophobicity. Experimental results showed that the adsorption capacity of all the cells to monorhamnolipid was much stronger than to dirhamnolipid, and the rhamnolipid-sourced P. aeruginosa cells, no matter grown on glucose or hexadecane, released extra dirhamnolipid when aqueous concentration of dirhamnolipid was too high. Length of surfactant alkyl chain as well as the type of carbon source used to cultivate the cell adsorbents had only minor influence on the adsorption. The adsorption was assumed to be driven by polar interaction between the rhamnolipid molecules and the cell surface chemical groups. The directional orientation of the rhamnolipid molecules with hydrophobic moiety extending to the environment may account for the rapid increase of cell surface hydrophobicity at low aqueous concentrations of the surfactant, while the stable or decreased cell hydrophobicity was probably the consequence of multiple surfactant layer formation or hemimicelle accumulation.     

Encystment of zoospores of the fungus,Phytophthora cinnamomi,is induced by specific lectin and monoclonal antibody binding to the cell surface

Summary Only two of a number of macromolecules that bind to the surface of zoospores of the dieback fungus,Phytophthora cinnamomi, induce encystment when added to a suspension of actively swimming zoospores. One, the lectin Concanavalin A (ConA), binds to the entire surface of the zoospores including the surface of both flagella. Within 10 minutes more than 70% of the cells have encysted in the presence of 5 g/ml ConA. This encystment is inhibited by preincubation of the lectin with its hapten sugar, -methyl-D-mannoside. The other effective molecule, a monoclonal antibody designated Zf-1, is one of 35 that have been raised to components on the surface of zoospores and cysts ofP. cinnamomi. The antigen for Zf-1 occurs only on the surface of the two flagella. Purified Zf-1 at 15 g/ml causes encystment of 75% of the zoospores in 13minutes. To show that the induction of encystment by these two probes is not due simply to the presence of protein either in solution or bound to the zoospore a number of other proteins were tested, including other antibodies that bind to the zoospore surface. None of these other molecules caused encystment even at concentrations greater than 200 g/ml. The results are consistent with the surface components that bind ConA and Zf-1 being involved in the critical step of triggering encystment at the surface of a potential host during infection.