Maria Nikolopoulou

Enhanced bioremediation of crude oil utilizing lipophilic fertilizers combined with biosurfactants and molasses

Many research studies have demonstrated the feasibility and efficacy of fertilization with nitrogen and phosphorus to combat oil spills in marine environments. Rapid dilution of water-soluble nutrients can be overcome by oleophilic formulations that retain optimal nutrient concentrations at the oil-water interface where biodegradation occurs. Previous work has demonstrated that biodegradation processes are enhanced by the addition of lipophilic fertilizers of natural origin (uric acid and lecithin). In this work, we examined the effectiveness of these nutrients in combination with biosurfactants (rhamnolipids) and molasses (source of C and vitamins) to enhance the biodegradation by naturally occurring microorganisms. It was found that the use of biosurfactants resulted in an increased removal of petroleum hydrocarbons (96% removal of C19-C34 n-alkanes within a period of 18 days) as well as in a reduction of the lag phase (almost 80% removal was achieved within the first week of biosurfactant application).

Evaluation of autochthonous bioaugmentation and biostimulation during microcosm-simulated oil spills.

Oil spills are treated as a widespread problem that poses a great threat to any ecosystem. Following first response actions, bioremediation has emerged as the best strategy for combating oil spills and can be enhanced by the following two complementary approaches: bioaugmentation and biostimulation. Bioaugmentation is one of the most controversial issues of bioremediation. Studies that compare the relative performance of bioaugmentation and biostimulation suggest that nutrient addition alone has a greater effect on oil biodegradation than the addition of microbial products because the survival and degradation ability of microbes introduced to a contaminated site are highly dependent on environmental conditions. Microbial populations grown in rich media under laboratory conditions become stressed when exposed to field conditions in which nutrient concentrations are substantially lower. There is increasing evidence that the best approach to overcoming these barriers is the use of microorganisms from the polluted area, an approach proposed as autochthonous bioaugmentation (ABA) and defined as a bioaugmentation technology that exclusively uses microorganisms indigenous to the sites (soil, sand, and water) slated for decontamination. In this work, we examined the effectiveness of strategies combining autochthonous bioaugmentation with biostimulation for successful remediation of polluted marine environments. Seawater was collected from a pristine area (Agios Onoufrios Beach, Chania) and was placed in a bioreactor with 1% v/v crude oil to facilitate the adaptation of the indigenous microorganism population. The pre-adapted consortium and the indigenous population were tested in combination with inorganic or lipophilic nutrients in the presence (or absence) of biosurfactants (rhamnolipids) during 90-day long experiments. Chemical analysis (gas chromatography-mass spectrometry) of petroleum hydrocarbons confirmed the results of previous work demonstrating that the biodegradation processes were enhanced by the addition of lipophilic fertilizers (uric acid and lecithin) in combination with biosurfactants (rhamnolipids), resulting in increased removal of petroleum hydrocarbons as well as reduction of the lag phase within 15 days of treatment. Considering this outcome and examining the results, the use of biostimulation additives in combination with naturally pre-adapted hydrocarbon-degrading consortia (bioaugmentation) has proved to be an effective treatment and is a promising strategy that could be applied specifically when an oil spill approaches near a shore line and an immediate hydrocarbon degradation effort is needed.

Enhanced ex situ bioremediation of crude oil contaminated beach sand by supplementation with nutrients and rhamnolipids

Mediterranean coastal regions are particularly exposed to oil pollution due to extensive industrialization, urbanization and transport of crude and refined oil to and from refineries. Bioremediation of contaminated beach sand through landfarming is both simple and cost-effective to implement compared to other treatment technologies. The purpose of the present study was to investigate the effect of alternative nutrients on biodegradation of crude oil contaminated beach sand in an effort to reduce the time required for bioremediation employing only indigenous hydrocarbon degraders. A natural sandy soil was collected from Agios Onoufrios beach (Chania, Greece) and was contaminated with weathered crude oil. The indigenous microbial population in the contaminated sand was tested alone (control treatment) or in combination with inorganic nutrients (KNO3 and K2HPO4) to investigate their effects on oil biodegradation rates. In addition, the ability of biosurfactants (rhamnolipids), in the presence of organic nutrients (uric acid and lecithin), to further stimulate biodegradation was investigated in laboratory microcosms over a 45-day period. Biodegradation was tracked by GC/MS analysis of aliphatic and polycyclic aromatic hydrocarbons components and the measured concentrations were corrected for abiotic removal by hopane normalizations. It was found that the saturated fraction of the residual oil is degraded more extensively than the aromatic fraction and the bacterial growth after an incubation period of approximately 3 weeks was much greater from the bacterial growth in the control. The results show that the treatments with inorganic or organic nutrients are equally effective over almost 30 days where C12-C35n-alkanes were degraded more than 97% and polyaromatic hydrocarbons with two or three rings were degraded more than 95% within 45 days. The results clearly show that the addition of nutrients to contaminated beach sand significantly enhanced the activity of indigenous microorganisms, as well as the removal of total recoverable petroleum hydrocarbons (TRPH) over a 45-day study period.

Allochthonous bioaugmentation in ex situ treatment of crude oil-polluted sediments in the presence of an effective degrading indigenous microbiome

Oil-polluted sediment bioremediation depends on both physicochemical and biological parameters, but the effect of the latter cannot be evaluated without the optimization of the former. We aimed in optimizing the physicochemical parameters related to biodegradation by applying an ex-situ landfarming set-up combined with biostimulation to oil-polluted sediment, in order to determine the added effect of bioaugmentation by four allochthonous oil-degrading bacterial consortia in relation to the degradation efficiency of the indigenous community. We monitored hydrocarbon degradation, sediment ecotoxicity and hydrolytic activity, bacterial population sizes and bacterial community dynamics, characterizing the dominant taxa through time and at each treatment. We observed no significant differences in total degradation, but increased ecotoxicity between the different treatments receiving both biostimulation and bioaugmentation and the biostimulated-only control. Moreover, the added allochthonous bacteria quickly perished and were rarely detected, their addition inducing minimal shifts in community structure although it altered the distribution of the residual hydrocarbons in two treatments. Therefore, we concluded that biodegradation was mostly performed by the autochthonous populations while bioaugmentation, in contrast to biostimulation, did not enhance the remediation process. Our results indicate that when environmental conditions are optimized, the indigenous microbiome at a polluted site will likely outperform any allochthonous consortium.

Microcosm evaluation of autochthonous bioaugmentation to combat marine oil spills

Oil spills can be disastrous to any ecosystem. Bioremediation through bioaugmentation (addition of oil-degrading bacteria) and biostimulation (addition of nutrients N&P) options can be a promising strategy for combating oil spills following first response actions. However, bioaugmentation is one of the most controversial issues of bioremediation since nutrient addition alone has a greater effect on oil biodegradation than the addition of microbial products that are highly dependent on environmental conditions. There is increasing evidence that the best way to overcome the above barriers is to use microorganisms from the polluted area, an approach proposed as autochthonous bioaugmentation (ABA) and defined as the bioaugmentation technology that uses exclusively microorganisms indigenous to the sites (soil, sand, and water) to be decontaminated. In this study, we examined the effectiveness of an ABA strategy for the successful remediation of polluted marine environments. A consortium was enriched from seawater samples taken from Elefsina Gulf near the Hellenic Petroleum Refinery, a site exposed to chronic crude oil pollution. Pre-adapted consortium was tested alone or in combination with inorganic nutrients in the presence (or not) of biosurfactants (rhamnolipids) in 30-day experiments. Treatment with fertilizers in the presence of biosurfactants exhibited the highest alkane and PAH degradation and showed highest growth over a period of almost 15 days. Considering the above, the use of biostimulation additives in combination with naturally pre-adapted hydrocarbon degrading consortia has proved to be a very effective treatment and it is a promising strategy in the future especially when combined with lipophilic fertilizers instead of inorganic nutrients. Such an approach becomes more pertinent when the oil spill approaches near the shoreline and immediate hydrocarbon degradation is needed.

Introduction to Food Waste Treament: The 14001 Standards

The aim of this chapter is to introduce the main ideas concerning treatment of foodwaste in the spirit of the standards and directives of the EU and to examine theenvironmental performance of the food industry as it seeks to comply with the ISO14001 standards. How is the food scientist and=or engineer going to comply withthese requirements? Basic tools in the hands of the food scientist and=or engineer isthe knowledge of physical and physicochemical properties of the foods, the massand energy balances, the application of these principles to the unit operations, andunit processes taken place in the industry or activity of concern. This enables thefood scientist and=or engineer to predict the quantities and compositions of thevarious streams, especially those of the effluents. Then the environmental behaviorof the organization could be obtained. From this point and on the remediationtactics and strategies could be followed. Life cycle analysis is important becausenot only the inputs and outputs of a given food industry or in general food activity isof importance for the environment but also the pre- and posthistory. Usually theinput to an organization is the output of another organization and vice versa.Environmental aspects of a food endeavor is not a onetime concern. In the begin-ning, we ensure compliance with the environmental laws and regulations. Follow upis also a requirement set by the EU directives.