Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous organic pollutants that harbour two or more fused benzene rings (Kennish, 1992). Diverse anthropogenic activities, such as the combustion of fossil fuels and oil spills, are responsible for its accumulation in natural environments (Kennish, 1992). Many high-molecular-mass PAHs (i.e. containing from four to seven benzene rings) are mutagenic, carcinogenic or teratogenic for a wide range of organisms including fish, amphibians, birds and mammals (Eisler, 1987).Micro-organisms that are able to degrade PAHs have been isolated from various environments (Juhasz & Naidu, 2000). However, high-molecular-mass PAHs present high hydrophobicity and have a tendency to adsorb to the organic fraction of soil and sediments (Gauthier et al., 2003). These characteristics are responsible for their low bioavailability to micro-organisms and the resulting persistence in natural environments (Hughes et al., 1997). Twoliquid-phase (TLP) cultures increase bioavailability and biodegradation of hydrophobic substrates (Déziel et al., 1999) and the microbial degradation occurs at the interface and/or in the aqueous phase (Gauthier et al., 2003).Using a TLP culture containing silicon oil as the waterimmiscible phase, we isolated a bacterial strain capable of degrading PAHs, designated strain CEE_131 T , from the Corcubion Ria in north-western Spain, a coastal system that was highly exposed to the Prestige oil spill in 2002.
Strain CEE_131T harbours 16S rRNA gene sequence heterogeneity. Although not exhaustively studied, 16S rRNA gene heterogeneity at the species or strain level is a common phenomenon (Bosshard et al., 2002). Unexpectedly high levels of intraspecific variation of 16S rRNA gene sequences in GenBank have been found previously (Clayton et al., 1995). Other studies have dealt with heterogeneity in 16S rRNA gene sequences (Bricker, 2000;Cilia et al., 1996; Liefting et al., 1996; Martinez-Murcia Abbreviations: CE-SSCP, capillary electrophoresis single-strand conformation polymorphism; PAH, polycyclic aromatic hydrocarbon; TLP, two-liquid-phase. The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strain CEE_131T are FM955223-FM955227.A micrograph of sectioned cells of strain CEE_131 T obtained by using transmission electron microscopy and graphs showing the effects of temperature, pH and salinity on the growth of the strain are available as supplementary material with the online version of this paper.