The potential groundwater impacts of biodiesel releases have received limited attention despite the increasing probability of such events. In this work, microcosms were prepared with unacclimated sediment and groundwater from the Ressacada Experimental Site (Florianopolis, Santa Catarina, Brazil) and spiked with 54.8 mg/L of pure soybean or castor oil biodiesel (B100). Oxygen was purged from the microcosms to mimic commonly anoxic and hypoxic conditions at fuel‐impacted sites; low background concentrations of nitrate (1.2 to 2.5 mg/L) and sulfate (2.2 to 3.0 mg/L) were present. Biodegradation was assessed by the removal of fatty acid methyl esters and hydrocarbon components relative to sterile controls. Approximately 80% of soybean biodiesel was biotransformed in 41 d, compared to only 40% of castor oil biodiesel removed in 90 d. The higher persistence of castor biodiesel was attributed to its higher viscosity and lower bioavailability. Additional microcosms were prepared similarly to assess the impact of biodiesel on hydrocarbon degradation. These microcosms were spiked with benzene (2.9 mg/L) and toluene (0.8 mg/L) with or without soybean biodiesel (54.8 mg/L). The biodiesel had an inhibitory effect, increasing the time required to remove toluene from 25 to 34 d. Similarly, 45% of benzene was removed in the presence of biodiesel within 34 d, compared to 90% in the absence of biodiesel. Overall, we postulate that the relatively high viscosity of biodiesel is conducive to limited migration potential and a smaller but longer lasting inhibitory region of influence, compared to that exerted by more soluble, more mobile, and readily degradable biofuels such as ethanol. However, controlled release studies are needed to test this hypothesis and characterize the complex dynamics of such releases.

Field experiments were conducted to assess the potential for anaerobic biostimulation to enhance BTEX biodegradation under fermentative methanogenic conditions in groundwater impacted by a biodiesel blend (B20, consisting of 20 % v/v biodiesel and 80 % v/v diesel). B20 (100 L) was released at each of two plots through an area of 1 m(2) that was excavated down to the water table, 1.6 m below ground surface. One release was biostimulated with ammonium acetate, which was added weekly through injection wells near the source zone over 15 months. The other release was not biostimulated and served as a baseline control simulating natural attenuation. Ammonium acetate addition stimulated the development of strongly anaerobic conditions, as indicated by near-saturation methane concentrations. BTEX removal began within 8 months in the biostimulated source zone, but not in the natural attenuation control, where BTEX concentrations were still increasing (due to source dissolution) 2 years after the release. Phylogenetic analysis using quantitative PCR indicated an increase in concentration and relative abundance of Archaea (Crenarchaeota and Euryarchaeota), Geobacteraceae (Geobacter and Pelobacter spp.) and sulfate-reducing bacteria (Desulfovibrio, Desulfomicrobium, Desulfuromusa, and Desulfuromonas) in the biostimulated plot relative to the control. Apparently, biostimulation fortuitously enhanced the growth of putative anaerobic BTEX degraders and associated commensal microorganisms that consume acetate and H2, and enhance the thermodynamic feasibility of BTEX fermentation. This is the first field study to suggest that anaerobic-methanogenic biostimulation could enhance source zone bioremediation of groundwater aquifers impacted by biodiesel blends.

A influência do biodiesel de soja na biodegradação dos hidrocarbonetos monoaromáticos benzeno e tolueno foi estudada sob condições anaeróbias em dois microcosmos montados com água subterrânea sintética, inóculo metanogênico, benzeno, tolueno e biodiesel. Na presença de biodiesel não foi observada biodegradação do benzeno e do tolueno. Com a biodegradação do biodiesel ocorreu a formação de acetato e metano, uso do sulfato e um aumento de 45 vezes no número de bactérias sulfato-redutoras.Esses resultados mostraram que, na mistura com benzeno e tolueno, o biodiesel foi biodegradado tanto sob condições de sulfato-redução quanto metanogênicas e que sua presença estimulou o crescimento da biomassa. Palavras