Pseudomonas diversity in crude-oil-contaminated intertidal sand samples obtained after the Prestige oil spill.
The Galicia seashore, in northwestern Spain, was one of the shorelines affected by the Prestige oil spill in November 2002. The diversity of autochthonous Pseudomonas populations present at two beaches (Carnota municipality) was analyzed using culture-independent and culture-dependent methods. The first analysis involved the screening of an rpoD gene library. The second involved the isolation of 94 Pseudomonas strains that were able to grow on selective media by direct plating or after serial enrichments on several carbon sources: biphenyl, gentisate, hexadecane, methylnaphthalene, naphthalene, phenanthrene, salicylate, xylene, and succinate. Eight denitrifying Pseudomonas strains were also isolated by their ability to grow anaerobically with nitrate. The calculated coverage index for Pseudomonas species was 89% when clones and isolates were considered together, and there were 29 phylospecies detected. The most abundant were members of the species P. stutzeri, P. putida, P. anguilliseptica, and P. oleovorans. Thirty-one isolates could not be identified at the species level and were considered representatives of 16 putative novel Pseudomonas species. One isolate was considered representative of a novel P. stutzeri genomovar. Concordant results were obtained when the diversities of the cloned DNA library and the cultured strains were compared. The clone library obtained by the rpoD PCR method was a useful tool for evaluating Pseudomonas communities and also for microdiversity studies of Pseudomonas populations.
Microbiologia, Departament de Biologia, Institut Mediterrani d'Estudis Avançats (CSIC-UIB), Universitat de les Illes Balears, 07122 Palma de Mallorca, Illes Balears, Spain.
Magdalena
M
Mulet
Zoyla
Z
David
Balbina
B
Nogales
Rafael
R
Bosch
Jorge
J
Lalucat
Elena
E
García-Valdés
D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D003470:Culture Media; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D049308:Dioxygenases / Q000235:genetics; D004190:Disasters; D014644:Genetic Variation; D019015:Geologic Sediments / Q000737:chemistry / Q000382:microbiology*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics; D010578:Petroleum; D010802:Phylogeny; D011549:Pseudomonas / Q000145:classification* / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012808:Sigma Factor / Q000235:genetics; D012822:Silicon Dioxide / Q000737:chemistry*; D013030:Spain; D013045:Species Specificity; D014877:Water Pollution, Chemical
D003470:Culture Media; D009097:Multienzyme Complexes; D010578:Petroleum; D012336:RNA, Ribosomal, 16S; D012808:Sigma Factor; D012822:Silicon Dioxide; D049308:Dioxygenases; C038703:naphthalene dioxygenase; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases