Bacterial Bioreporters for the Real-Time Monitoring of Oil Bioremediation in Marine Microcosms

Francesca Crisafi; Renata Denaro

doi:7080274

Hydrocarbons are common contaminants in aquatic habitats, in particular in the Mediterranean sea, because of the number of refineries distributed along the coast and the intensive oil transportation by ship. One of the main challenges of bioremediation is to define effective protocols to overcome in situ the limiting factors of microbial degradation activity. Monitoring the effectiveness of bioremediation is very complex and time-consuming. The aim of our study was to validate bacterial bioreporters as innovative tools to evaluate in real time the efficiency of bio-treatment of oil-polluted samples. Three treatments (Aeration; Biostimulation, and Bioaugmentation) were carried out on artificially contaminated seawater with oil at microcosm scale. The treatments were monitored for 7 days by the use of an Escherichia coli-based alkane luciferase bioreporter (5150) and of an E. coli-based strain constitutively expressing luciferase (5463). Hydrocarbons were monitored by GC-FID analyses while in parallel, quantitative functional gene expression was carried out by real-time PCR with the aim to verify specific microbial consortium activity. The effectiveness of the treatments was estimated as follows: bioaugmentation>biostimulation> no-treatment. The results by the bioreporters were coherent with chemical analyses and Real-time PCR. During the early period of the experiment we observed a different response of the reporters to the oil input and treatments, after that phase, we were able to detect bioreporter induction at significantly reduced rates. Our analyses have also provided evidence for an inhibitory effect of high cell densities in the medium, in particular after 24h of the treatments. In conclusion, for monitoring the efficacy of bioremediation it is necessary to have a significant technical and methodological framework, which includes chemical, biological, engineering approaches. Bacterial bioreporters are versatile tools that can help to monitor the trend of the treatment, can substantially improve the systematic protocols, both in terms of time as well as economic aspects.

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