In the present study, acrylic coupons with a thin layer of oil on the surface were incubated in the coastal water of Trindade Island, Brazil, for 60 days. The microorganisms adhered to the coupons were isolated using enrichment medium with hexadecane and naphthalene as the sole carbon and energy source. A total of 15 bacterial isolates were obtained, and the ability of these isolates to use different hydrocarbons as the source of carbon and energy was investigated. None of the isolates produced biosurfactants under our experimental conditions. Subsequently, identification methods such as partial sequencing of the 16S rRNA gene and analysis of fatty acids (MIDI) profile were employed. Among the 15 isolates, representatives of Actinobacteria, Firmicutes, and Alphaproteobacteria were detected. The isolates Rhodococcus rhodochrous TRN7 and Nocardia farcinica TRH1 were able to use all the hydrocarbons added to the culture medium (toluene, octane, xylene, naphthalene, phenanthrene, pyrene, hexadecane, anthracene, eicosane, tetracosane, triacontane, and pentacontane). Polymerase chain reaction amplification of the DNA isolated by employing primers for catechol 2,3-dioxygenase, alkane dehydrogenase and the alpha subunit of hydroxylating dioxygenases polycyclic aromatic hydrocarbon rings genes demonstrated that various isolates capable of utilizing hydrocarbons do not exhibit genes of known routes of catabolism, suggesting the existence of unknown catabolic pathways in these microorganisms. Our findings suggest that the microbiota associated to the coast of tropical oceanic islands has the ability to assist in environmental regeneration in cases of accidents involving oil spills in its shore. Thus, it motivates studies to map bioremediation strategies using the autochthonous microbiota from these environments.
We aimed to isolate biosurfactant-producing bacteria in high salt conditions from uncontaminated soils on the Brazilian oceanic island, Trindade. Blood agar medium was used for the isolation of presumptive biosurfactant-producing bacteria. Confirmation and measurements of biosurfactant production were made using an oil-spreading method. The isolates were identified by fatty acid profiles and partial 16S rRNA gene sequence analysis. A total of 14 isolates obtained from the 12 soil samples were found to produce biosurfactants. Among them, two isolates stood out as being able to produce biosurfactant that is increasingly active in solutions containing up to 175 g L(-1) NaCl. These high salt tolerant biosurfactant producers are affiliated to different species of the genus Bacillus. Soil organic matter showed positive correlation with the number of biosurfactant-producing bacteria isolated from our different sampling sites. The applied approach successfully recovered and identified biosurfactant-producing bacteria from non-contaminated soils. Due to the elevated salt tolerance, as well as their capacity to produce biosurfactants, these isolates are promising for environmental biotechnological applications, especially in the oil production chain.
Controversy surrounding bacterial phylogenies has become one of the most important challenges for microbial ecology. Comparative analyses with nucleotide databases and phylogenetic reconstruction of the amplified 16S rRNA genes from DGGE (Denaturing Gradient Gel Electrophoresis) excised bands have been used by several researchers for the identification of organisms in complex samples. Here, we individually analyzed DGGE-excised 16S rRNA gene bands from 10 certified bacterial strains of different species, and demonstrated that this kind of approach can deliver erroneous outcomes to researchers, besides causing/emphasizing errors in public databases.
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