Effects of temperature and fertilization on total vs. active bacterial communities exposed to crude and diesel oil pollution in NW Mediterranean Sea.

The dynamics of total and active microbial communities were studied in seawater microcosms amended with crude or diesel oil at different temperatures (25, 10 and 4 degrees C) in the presence/absence of organic fertilization (Inipol EAP 22). Total and hydrocarbon-degrading microbes were enumerated by fluorescence microscopy and Most Probable Number (MPN) method, respectively. Total (16S rDNA-based) vs. active (16S rRNA) bacterial community structure was monitored by Capillary-Electrophoresis Single Strand Conformation Polymorphism (CE-SSCP) fingerprinting. Hydrocarbons were analyzed after 12 weeks of incubation by gas chromatography-mass spectrometry. Total and hydrocarbon-degrading microbial counts were highly influenced by fertilization while no important differences were observed between temperatures. Higher biodegradation levels were observed in fertilized microcosms. Temperature and fertilization induced changes in structure of total bacterial communities. However, fertilization showed a more important effect on active bacterial structure. The calculation of Simpson's diversity index showed similar trends among temperatures whereas fertilization reduced diversity index of both total and active bacterial communities.

Comparative mesocosm study of biostimulation efficiency in two different oil-amended sub-antarctic soils.

Biological treatment has become increasingly popular as a remediation method for soils and groundwater contaminated with petroleum hydrocarbon, chlorinated solvents, and pesticides. Bioremediation has been considered for application in cold regions such as Arctic and sub-Arctic climates and Antarctica. Studies to date suggest that indigenous microbes suitable for bioremediation exist in soils in these regions. This paper reports on two case studies at the sub-Antarctic Kerguelen Island in which indigenous bacteria were found that were capable of mineralizing petroleum hydrocarbons in soil contaminated with crude oil and diesel fuel. All results demonstrate a serious influence of the soil properties on the biostimulation efficiency. Both temperature elevation and fertilizer addition have a more significant impact on the microbial assemblages in the mineral soil than in the organic one. Analysis of the hydrocarbons remaining at the end of the experiments confirmed the bacterial observations. Optimum temperature seems to be around 10 degrees C in organic soil, whereas it was higher in mineral soil. The benefit of adding nutrients was much stronger in mineral than in the organic soil. Overall, this study suggests that biostimulation treatments were driven by soil properties and that ex situ bioremediation for treatment of cold contaminated soils will allow greater control over soil temperature, a limiting factor in cold climates.

Marinomonas polaris sp. nov., a psychrohalotolerant strain isolated from coastal sea water off the subantarctic Kerguelen islands.

Two aerobic, psychrohalotolerant, motile bacterial isolates, CK13T and CK16, isolated from sea-water samples collected off the subantarctic Kerguelen island, were characterized by using a polyphasic taxonomic approach. On the basis of 16S rRNA gene sequence data, the strains were 99.6% similar and exhibited 93-97% similarity with the seven recognized species of Marinomonas. The most closely related species were Marinomonas pontica and Marinomonas primoryensis, with 97 and 96 % similarity at the 16S rRNA gene sequence level, respectively. DNA-DNA hybridization values between strain CK13T and M. pontica and M. primoryensis were only 58 and 40%, respectively. The major fatty acids present in strain CK13T were iso-C(16:0), C(16:0), C(16:1)omega7c and C(18:1)omega7c. The DNA G+C content of strain CK13T was 41.2 mol%. Phosphatidylethanolamine and phosphatidylglycerol were identified as the predominant phospholipids. All the above characteristics support the affiliation of strain CK13T to the genus Marinomonas. Phylogenetic analysis and phenotypic and genotypic distinctiveness confirmed that strains CK13T and CK16 are members of a novel species of the genus Marinomonas, for which the name Marinomonas polaris sp. nov. is proposed. The type strain is CK13T (=MTCC 6645T=DSM 16579T=JCM 12522T).

Degradation of petroleum hydrocarbons in two sub-antarctic soils: influence of an oleophilic fertilizer.

In order to determine the long-term effects of fertilizer on the degradation rate and the toxicity of hydrocarbons in sub-Antarctic soils contaminated by petroleum hydrocarbons, a field study was initiated in December 2000 on two different soils of the Kerguelen Islands (69 degrees 42'E, 49 degrees 19'S). The number of hydrocarbon-degrading bacteria (HDB) increased greatly after crude-oil and diesel-fuel contamination, and the fertilizer addition had a favorable effect on HDB growth and activity. Hydrocarbon-degrading bacteria counts remained high until the end of the experiment although the total hydrocarbon content in all contaminated soils was reduced to 80 to 90% of their initial value after 330 d. Degradation of n-alkanes was enhanced significantly in the presence of the fertilizer, while the degradation of polycyclic aromatic hydrocarbons (PAHs) was only barely enhanced. Toxicity results showed a noticeable reduction with time, although toxicity remained present and important in both soils at the end of the experiment. In addition, fertilized plots showed a toxic signal greater than unfertilized ones. Overall results clearly show that fertilizer addition improves the rate of degradation of both oil contaminants. However, remaining toxic residues may constitute a drawback of the fertilizer-assisted biodegradation process at low temperatures.