Bioremediation of diesel contaminated soils : an investigation into the effects of biosurfactant and organic matter amendments

The aim of this study was to investigate the effects of biosurfactants and organic matter amendments on the bioremediation of diesel contaminated soil. Two strains of Pseudomonas aeruginosa with the ability to produce biosurfactant were isolated from a water and soil sample in Co. Sligo. The first strain, Isolate A, produced a biosurfactant which contained four rhamnose containing compounds, when grown in proteose peptone glucose ammonium salts medium with glucose as the carbon source. Two of the components were identified as rhamnolipid 1 and 2 whilst the other two components were unidentified. The second strain, Isolate GO, when grown in similar conditions produced a biosurfactant which contained only rhamnolipid 2. The type of aeration system used had a significant effect on the abiotic removal of diesel from soil. Forced aeration at a rate of 120L 02/kg soil/ hour resulted in the greatest removal. Over a 112 day incubation period this type o f aeration resulted in the removal o f 48% o f total hexane extractable material. In relation to bioremediation of the diesel contaminated sandy soil, amending the soil with two inorganic nutrients, KH2PO4 and NÜ4N03, significantly enhanced the removal of diesel, especially the «- alkanes, when compared to an unamended control. The biosurfactant from Isolate A and a biosurfactant produced by Pseudomonas aeruginosa NCIMB 8628 (a known biosurfactant producer), when applied at a concentration of three times their critical micelle concentration, had a neutral effect on the biodégradation o f diesel contaminated sandy soil, even in the presence o f inorganic nutrients. It was deduced that the main reason for this neutral effect was because they were both readily biodegraded by the indigenous microorganisms. The most significant removal of diesel occurred when the soils were amended with two organic materials plus the inorganic nutrients. Amendment of the diesel contaminated soil with spent brewery grain (SBG) removed significantly more diesel than amendment with dried molassed sugar beet pulp (DMSBP). After a 108 day incubation period, amendment of the diesel contaminated soil with DMSBP plus inorganic nutrients and SBG plus inorganic nutrients resulted in 72 and 89% removal of diesel range organics (DRO), in comparison to 41% removal of DRO in an inorganic nutrient amended control. The first order kinetic model described the degradation of the different diesel components with high correlation and was used to calculate Vi lives. The V2 life, of the total «-alkanes in the diesel was reduced from 40 days in the control to 8.5 and 5.1 days in the presence of DMSBP and SBG, respectively. The V2 life o f the unresolved complex mixture (UCM) in the diesel contaminated soil was also significantly reduced in the presence o f the two organics. DMSBP and SBG addition reduced UCM V2 life to 86 and 43 days, respectively, compared to 153 days in the control. The component of diesel whose removal was enhanced the greatest through the organic material amendments was the isoprenoid, pristane, a compound which until recently was thought to be nonbiodegradable and was used as an inert biomarker in oil degradation studies. The V2 life of pristane was reduced from 533 days in the nutrient amended control to 49.5 and 19.5 days in DMSBP and SBG amended soils. These results indicate that the addition o f the DMSBP and SBG to diesel contaminated soil stimulated diesel biodégradation, probably by enhancing the indigenous diesel degrading microbial population to degrade diesel hydrocarbons, whilst the addition o f biosurfactants had no enhanced effect on the bioremediation process.

Synchronous reproduction may facilitate introgression in a hybrid mussel (Mytilus) population

Mussel populations on the Irish Atlantic coast comprise an interbreeding mixture of the blue mussel, Mytilus edulis (L.) and the Mediterranean mussel, Mytilus galloprovincialis (Lmk.). The occurrence of hybrid genotypes varies between sites but can be as high 80%. This study compares the reproductive cycle of M. edulis, M. galloprovincialis and their hybrids to determine if the extensive hybridisation observed at Irish Atlantic coast sites is linked to spawning synchrony between the two taxa. Mussels (40-45 mm size class) were collected monthly from a sheltered shore in Galway Bay from January to December 2005. Two major spawning events (March- June and September-October) were observed and gametogenesis took place throughout the year. The spawning cycles of the three taxa were largely overlapping. Small differences were observed in the timing of peak spawning which occurred in March and October in M. galloprovincialis and in May-June and September in M.edulis. Spawning of hybrid individuals was intermediate between the parental genotypes. Fecundity was slightly higher in M. galloprovincialis females compared to the other taxa (up to 30% difference, p<0.05). This apparent advantage is not shared by the sexes and is likely being offset by high numbers of hybrid genotypes releasing gametes during peak spawning of M. galloprovincialis. There was no evidence for increased mortality in hybrid males; sex ratios did not deviate from the 1:1 ratio. The results show that in this region of the hybrid zone the timing of reproduction does not present a barrier to gene flow between M. edulis and M. galloprovincialis. Nonetheless, small differences in the timing of peak spawning may increase the likelihood of conspecific fertilisation at certain times of the year. Hybrids outnumber the parental genotypes, undergo complete gametogenesis and show no evidence of depressed fitness (i.e. hybrids are reproductively competent suggesting a high degree of introgression.