Bioremediation of hydrocarbon-contaminated soil from Carlini Station, Antarctica: effectiveness of different nutrient sources as biostimulation agents

Logistics and scientific activities carried out in Antarctic stations entail the risk of contamination by fuels. Among remediation strategies, biostimulation significantly improves the efficiency of hydrocarbons (HCs) removal. A 1-year-long field trial was performed in mesocosms filled with soil chronically contaminated with HCs. Three nutrient sources were evaluated as biostimulation agents: inorganic salts (with and without aeration by mixing), a slow-release granular fertilizer (Nitrofoska®) and a commercial bioremediation product (OSEII®). Their performance was assessed considering the number of culturable bacteria, the changes induced in the structure of bacterial communities, the HCs removal efficiencies and the estimation of the abiotic and biodegradative losses of HCs. The soil indigenous microbiota reduced the concentration of hydrocarbons by up to 50% in 50 days and 87% in 365 days depending on the biostimulation agent used. OSEII® (a mixture of surfactants, nutrients, and enzymes) performed better in the medium term, promoting bacterial growth and rapidly inducing changes in the structure of bacterial community, and Nitrofoska® proved to be more efficient for long-term processes, less affecting the size and structure of the microbiota. A mixed strategy combining the fastest action of commercial products acting during summer with slow-release fertilizers acting throughout the year is proposed as a long-term bioremediation treatment for Antarctic areas where the temperature rises above the freezing point and the ground is free of snow shortly during summer. This study highlights the importance of conducting research to develop remediation processes compatible with the Antarctic Treaty, exploiting the metabolic potential of the indigenous microbiota.Fil: Villalba Primitz, Julia Elena Rosina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Vázquez, Susana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; ArgentinaFil: Ruberto, Lucas Adolfo Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Lo Balbo, Alfredo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Mac Cormack, Walter Patricio. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentin

Bioremediation of hydrocarbon-contaminated soils in cold regions: Development of a pre-optimized biostimulation biopile-scale field assay in Antarctica

Bioremediation proved to be an effective approach to deal with soil contamination, especially in isolated, cold environments such as Antarctica. Biostimulation, involving the addition of macronutrients -mainly nitrogen and phosphorous- is considered the simplest and cheapest bioremediation process. Optimizing the levels of these nutrients is a key step prior to the application of a biostimulation strategy. In this work, N and P levels, optimized by Response Surface Methodology (RSM) at lab-scale, were applied to an Antarctic hydrocarbon contaminated soil. The process was performed on-site, using high density polyethylene geomembranes (800 μm) to isolate treated soil from the surroundings and under environmental conditions at Carlini station (Antarctica) during 50 days. Two 0.5 ton biopiles were used as experimental units; a control biopile (CC), and a biostimulated system (BS), amended with N and P. At the end of the assay, hydrocarbon removal was significantly higher in BS system compared to CC (75.79% and 49.54% respectively), showing that the applied strategy was effective enough to perform a field-assay in Antarctica that significantly reduce soil contamination levels; and proving that RSM represents a fundamental tool for the optimization of nutrient levels to apply during bioremediation of fuel contaminated cold soils.Fil: Martinez Alvarez, Lucas Manuel. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; ArgentinaFil: Ruberto, Lucas Adolfo Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Lo Balbo Alfredo. Universidad de Buenos Aires; ArgentinaFil: Mac Cormack, Walter Patricio. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentin

Hydrocarbon removal and bacterial community structure in on-site biostimulated biopile systems designed for bioremediation of diesel-contaminated Antarctic soil

Several studies have shown that biostimulation can promote hydrocarbon bioremediation processes in Antarctic soils. However, the effect of the different nutrient sources on hydrocarbon removal heavily depends on the nutrients used and the soil characteristics. In this work, using a sample of chronically contaminated Antarctic soil that was exposed to a fresh hydrocarbon contamination, we analyzed how a complex organic nutrient source such as fish meal (FM) and a commercial fertilizer (OSEII) can affect hydrocarbon biodegradation and bacterial community composition. Both amended and unamended (control) biopiles were constructed and controlled at Carlini Station and sampled at days 0, 5, 16, 30 and 50 for microbiological, chemical and molecular analyses. FM caused a fast increase in both total heterotrophic and hydrocarbon degrading bacterial counts. These high values were maintained until the end of the assay, when statistically significant total hydrocarbon removal (71 %) was detected when compared with a control system. The FM biopile evidenced the dominance of members of the phylum Proteobacteria and a clear shift in bacterial structure at the final stage of the assay, when an increase of Actinobacteria was observed. The biopile containing the commercial fertilizer evidenced a hydrocarbon removal activity that was not statistically significant when compared with the untreated system and exhibited a bacterial community that differed from those observed in the unamended and FM-amended biopiles. In summary, biostimulation using FM in biopiles significantly enhanced the natural hydrocarbon-degradation activity of the Carlini station soils in biopile systems and caused significant changes in the bacterial community structure. The results will be considered for the future design of soil bioremediation protocols for Carlini Station and could also be taken into account to deal with diesel-contaminated soils from other cold-climate areas.Fil: Dias, Romina Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Ruberto, Lucas Adolfo Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Virología; Argentina. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; ArgentinaFil: Calabró López, Roberto Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Virología; ArgentinaFil: Lo Balbo, Alfredo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: del Panno, Maria Teresa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Mac Cormack, Walter Patricio. Universidad de Buenos Aires; Argentina. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentin

Bacterial Community Dynamics during Bioremediation of Diesel Oil-Contaminated Antarctic Soil

The effect of nutrient and inocula amendment in a bioremediation field trial using a nutrient-poor Antarctic soil chronically contaminated with hydrocarbons was tested. The analysis of the effects that the treatments caused in bacterial numbers and hydrocarbon removal was combined with the elucidation of the changes occurring on the bacterial community, by 16S rDNA-based terminal restriction fragment length polymorphism (T-RFLP) typing, and the detection of some of the genes involved in the catabolism of hydrocarbons. All treatments caused a significant increase in the number of bacteria able to grow on hydrocarbons and a significant decrease in the soil hydrocarbon content, as compared to the control. However, there were no significant differences between treatments. Comparison of the soil T-RFLP profiles indicated that there were changes in the structure and composition of bacterial communities during the bioremediation trial, although the communities in treated plots were highly similar irrespective of the treatment applied, and they had a similar temporal dynamics. These results showed that nutrient addition was the main factor contributing to the outcome of the bioremediation experiment. This was supported by the lack of evidence of the establishment of inoculated consortia in soils, since their characteristic electrophoretic peaks were only detectable in soil profiles at the beginning of the experiment. Genetic potential for naphthalene degradation, evidenced by detection of nahAc gene, was observed in all soil plots including the control. In treated plots, an increase in the detection of catechol degradation genes (nahH and catA) and in a key gene of denitrification (nosZ) was observed as well. These results indicate that treatments favored the degradation of aromatic hydrocarbons and probably stimulated denitrification, at least transiently. This mesocosm study shows that recovery of chronically contaminated Antarctic soils can be successfully accelerated using biostimulation with nutrients, and that this causes a change in the indigenous bacterial communities and in the genetic potential for hydrocarbon degradation.Fil: Vázquez, Susana Claudia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Nogales, Balbina. Universidad de las Islas Baleares; EspañaFil: Ruberto, Lucas Adolfo Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; ArgentinaFil: Hernández, Edgardo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; ArgentinaFil: Christie Oleza, Joseph. Universidad de las Islas Baleares; EspañaFil: Lo Balbo, Alfredo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; ArgentinaFil: Bosch, Rafael. Universidad de las Islas Baleares; EspañaFil: Lalucat, Jorge. Universidad de las Islas Baleares; EspañaFil: Mac Cormack, Walter Patricio. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentin

Bacterial Community Dynamics during Bioremediation of Diesel-Oil Contaminated Antarctic Soil

The effect of nutrient and inocula amendment in a bioremediation field trial using a nutrient-poor Antarctic soil chronically contaminated with hydrocarbons was tested. The analysis of the effects that the treatments caused in bacterial numbers and hydrocarbon removal was combined with the elucidation of the changes occurring on the bacterial community, by 16S rDNA-based terminal restriction fragment length polymorphism (T-RFLP) typing, and the detection of some of the genes involved in the catabolism of hydrocarbons. All treatments caused a significant increase in the number of bacteria able to grow on hydrocarbons and a significant decrease in the soil hydrocarbon content, as compared to the control. However, there were no significant differences between treatments. Comparison of the soil T-RFLP profiles indicated that there were changes in the structure and composition of bacterial communities during the bioremediation trial, although the communities in treated plots were highly similar irrespective of the treatment applied, and they had a similar temporal dynamics. These results showed that nutrient addition was the main factor contributing to the outcome of the bioremediation experiment. This was supported by the lack of evidence of the establishment of inoculated consortia in soils, since their characteristic electrophoretic peaks were only detectable in soil profiles at the beginning of the experiment. Genetic potential for naphthalene degradation, evidenced by detection of nahAc gene, was observed in all soil plots including the control. In treated plots, an increase in the detection of catechol degradation genes (nahH and catA) and in a key gene of denitrification (nosZ) was observed as well. These results indicate that treatments favored the degradation of aromatic hydrocarbons and probably stimulated denitrification, at least transiently. This mesocosm study shows that recovery of chronically contaminated Antarctic soils can be successfully accelerated using biostimulation with nutrients, and that this causes a change in the indigenous bacterial communities and in the genetic potential for hydrocarbon degradation.(© 2008 Springer Science+Business Media, LLC.)This research was supported by grants from the Argentinean Antarctic Institute (IAA no. 42), the National Agency for Scientific and Technical Researches (PICTO 11555), and the University of Buenos Aires (UBACyT U007) as well as by a short-term travel fellowship from the Argentinean Research Council (CONICET). Research at the University of the Balearic Islands (UIB) was supported by grants VEM2003-20565 and CTM2005-01783 from the Spanish Ministry of Education and Science (MEC). B.N. was supported by a contract from the program “Ramón y Cajal” from MEC, and J.C-O by a fellowship of the Balearics Autonomous GovernmentPeer Reviewe