RESUMO
The gradual introduction of biodiesel in the Brazilian energy landscape has primarily occurred through its blending with conventional petroleum diesel (e.g., B20 (20% biodiesel) and B5 (5% biodiesel) formulations). Because B20 and lower-level blends generally do not require engine modifications, their use as transportation fuel is increasing in the Brazilian distribution networks. However, the environmental fate of low-level biodiesel blends and pure biodiesel (B100) is poorly understood and the ecotoxicological-safety endpoints of biodiesel-contaminated environments are unknown. Using laboratory microcosms consisting of closed reactor columns filled with clay loam soil contaminated with pure biodiesel (EXPB100) and a low-level blend (EXPB5) (10% w/v), this study presents soil ecotoxicity assessement and dynamics of culturable heterotrophic bacteria. Most-probable-number (MPN) procedures for enumeration of bacteria, dehydrogenase assays and soil ecotoxicological tests using Eisenia fetida have been performed at different column depths over the course of incubation. After 60 days of incubation, the ecotoxicity of EXPB100-derived samples showed a decrease from 63% of mortality to 0% while EXPB5-derived samples exhibited a reduction from 100% to 53% and 90% on the top and at the bottom of the reactor column, respectively. The dehydrogenase activity of samples from EXPB100 and EXPB5 increased significantly compared to pristine soil after 60 days of incubation. Growth of aerobic bacterial biomass was only observed on the top of the reactor column while the anaerobic bacteria exhibited significant growth at different column depths in EXPB100 and EXPB5. These preliminary results suggest the involvement of soil indigenous microbiota in the biodegradation of biodiesel and blends. However, GC-FID analyses for quantification of fatty acid methyl esters (FAMEs) and aliphatic hydrocarbons and targeted sequencing of 16S rRNA tags using illumina platforms will provide important insights into the profiles and underlying mechanisms of (bio)diesel biodegradation in soil environments.
