Adriano Pinto Mariano

Laboratory study on the bioremediation of diesel oil contaminated soil from a petrol station

The purpose of the present study was to investigate possible methods to enhance the rate of aerobic biodegradation of hydrocarbons (ex-situ treatments). In this work, the bioremediation processes were applied to a sandy soil with a high level of contamination originated from the leakage of a diesel oil underground storage tank at a petrol station. Laboratory scale experiments (Bartha biometer flasks) were used to evaluate the biodegradation of the diesel oil. Enhancement of biodegradation was carried out through biostimulation (addition of nitrogen and phosphorus solutions or Tween 80 surfactant) and bioaugmentation (bacterial consortium isolated from a landfarming system). To investigate interactions between optimizing factors, and to find the right combination of these agents, the study was based on full factorial experimental design. Efficiency of biodegradation was simultaneously measured by two methods: respirometric (microbial CO2 production) and gas chromatography. Acute toxicity tests with Daphnia similis were applied for examination of the efficiency of the processes in terms of the generation of less toxic products. Results showed that all bioremediation strategies enhanced the natural bioremediation of the contaminated soil and the best results were obtained when treatments had nutritional amendment. Respirometric data indicated a maximum hydrocarbon mineralization of 19.8%, obtained through the combination of the three agents, with a total petroleum hydrocarbons (TPH) removal of 45.5% in 55 days of treatment. At the end of the experiments, two predominant bacteria species were isolated and identified (Staphylococcus hominis and Kocuria palustris).

Butanol production in a first-generation Brazilian sugarcane biorefinery: Technical aspects and economics of greenfield projects

The techno-economics of greenfield projects of a first-generation sugarcane biorefinery aimed to produce ethanol, sugar, power, and n-butanol was conducted taking into account different butanol fermentation technologies (regular microorganism and mutant strain with improved butanol yield) and market scenarios (chemicals and automotive fuel). The complete sugarcane biorefinery with the batch acetone-butanol-ethanol (ABE) fermentation process was simulated using Aspen Plus®. The biorefinery was designed to process 2 million tonne sugarcane per year and utilize 25%, 50%, and 25% of the available sugarcane juice to produce sugar, ethanol, and butanol, respectively. The investment on a biorefinery with butanol production showed to be more attractive [14.8% IRR, P(IRR>12%)=0.99] than the conventional 50:50 (ethanol:sugar) annexed plant [13.3% IRR, P(IRR>12%)=0.80] only in the case butanol is produced by an improved microorganism and traded as a chemical.

Biosurfactant synthesis by Pseudomonas aeruginosa LBI isolated from a hydrocarbon-contaminated site

Aims:  Pseudomonas aeruginosa LBI (Industrial Biotechnology Laboratory) was isolated from hydrocarbon‐contaminated soil as a potential producer of biosurfactant and evaluated for hydrocarbon biodegradation. The emulsifying power and stability of the product was assessed in the laboratory, simulating water contamination with benzene, toluene, kerosene, diesel oil and crude oil at various concentrations.

Utilization of pentoses from sugarcane biomass: Techno-economics of biogas vs. butanol production

This paper presents the techno-economics of greenfield projects of an integrated first and second-generation sugarcane biorefinery in which pentose sugars obtained from sugarcane biomass are used either for biogas (consumed internally in the power boiler) or n-butanol production via the ABE batch fermentation process. The complete sugarcane biorefinery was simulated using Aspen Plus®. Although the pentoses stream available in the sugarcane biorefinery gives room for a relatively small biobutanol plant (7.1-12 thousand tonnes per year), the introduction of butanol and acetone to the product portfolio of the biorefinery increased and diversified its revenues. Whereas the IRR of the investment on a biorefinery with biogas production is 11.3%, IRR varied between 13.1% and 15.2% in the butanol production option, depending on technology (regular or engineered microorganism with improved butanol yield and pentoses conversion) and target market (chemicals or automotive fuels). Additional discussions include the effects of energy-efficient technologies for butanol processing on the profitability of the biorefinery.

Biodegradability of commercial and weathered diesel oils

This work aimed to evaluate the capability of different microorganisms to degrade commercial diesel oil in comparison to a weathered diesel oil collected from the groundwater at a petrol station. Two microbiological methods were used for the biodegradability assessment: the technique based on the redox indicator 2,6 - dichlorophenol indophenol (DCPIP) and soil respirometric experiments using biometer flasks. In the former we tested the bacterial cultures Staphylococcus hominis, Kocuria palustris, Pseudomonas aeruginosa LBI, Ochrobactrum anthropi and Bacillus cereus, a commercial inoculum, consortia obtained from soil and groundwater contaminated with hydrocarbons and a consortium from an uncontaminated area. In the respirometric experiments it was evaluated the capability of the native microorganisms present in the soil from a petrol station to biodegrade the diesel oils. The redox indicator experiments showed that only the consortia, even that from an uncontaminated area, were able to biodegrade the weathered diesel. In 48 days, the removal of the total petroleum hydrocarbons (TPH) in the respirometric experiments was approximately 2.5 times greater when the commercial diesel oil was used. This difference was caused by the consumption of labile hydrocarbons, present in greater quantities in the commercial diesel oil, as demonstrated by gas chromatographic analyses. Thus, results indicate that biodegradability studies that do not consider the weathering effect of the pollutants may over estimate biodegradation rates and when the bioaugmentation is necessary, the best strategy would be that one based on injection of consortia, because even cultures with recognised capability of biodegrading hydrocarbons may fail when applied isolated.

An alternative process for butanol production: Continuous flash fermentation

The objective of this work is to introduce and demonstrate the technical feasibility of the continuous flash fermentation for the production of butanol. The evaluation was carried out through mathematical modeling and computer simulation which is a good approach in such a process development stage. The process consists of three interconnected units, as follows: the fermentor, the cell retention system (tangential microfiltration) and the vacuum flash vessel (responsible for the continuous recovery of butanol from the broth). The efficiency of this process was experimentally validated for the ethanol fermentation, whose main results are also shown. With the proposed design the concentration of butanol in the fermentor was lowered from 11.3 to 7.8 g/l, which represented a significant reduction in the inhibitory effect. As a result, the final concentration of butanol was 28.2 g/l for a broth with 140 g/l of glucose. Solvents productivity and yield were, respectively, 11.7 g/l.h and 33.5 % for a sugar conversion of 95.6 %. Positive aspects about the flash fermentation process are the solvents productivity, the use of concentrated sugar solution and the final butanol concentration. The last two features can be responsible for a meaningful reduction in the distillation costs and result in environmental benefits due to lower quantities of wastewater generated by the process.

The use of vinasse as an amendment to ex-situ bioremediation of soil and groundwater contaminated with diesel oil

This work investigated the possibility of using vinasse as an amendment in ex-situ bioremediation processes. Groundwater and soil samples were collected at petrol stations. The soil bioremediation was simulated in Bartha biometer flasks, used to measure the microbial CO2 production, during 48 days, where vinasse was added at a concentration of 33 mL.Kg-1of soil. Biodegradation efficiency was also measured by quantifying the total petroleum hydrocarbons (TPH) by gas chromatography. The groundwater bioremediation was carried out in laboratory experiments simulating aerated (bioreactors) and not aerated (BOD flasks) conditions. In both the cases, the concentration of vinasse was 5 % (v/v) and different physicochemical parameters were evaluated during 20 days. Although an increase in the soil fertility and microbial population were obtained with the vinasse, it demonstrated not to be adequate to enhance the bioremediation efficiency of diesel oil contaminated soils. The addition of the vinasse in the contaminated groundwaters had negative effects on the biodegradation of the hydrocarbons, since vinasse, as a labile carbon source, was preferentially consumed.

Optimization Strategies Based on Sequential Quadratic Programming Applied for a Fermentation Process for Butanol Production

In this work, the mathematical optimization of a continuous flash fermentation process for the production of biobutanol was studied. The process consists of three interconnected units, as follows: fermentor, cell-retention system (tangential microfiltration), and vacuum flash vessel (responsible for the continuous recovery of butanol from the broth). The objective of the optimization was to maximize butanol productivity for a desired substrate conversion. Two strategies were compared for the optimization of the process. In one of them, the process was represented by a deterministic model with kinetic parameters determined experimentally and, in the other, by a statistical model obtained using the factorial design technique combined with simulation. For both strategies, the problem was written as a nonlinear programming problem and was solved with the sequential quadratic programming technique. The results showed that despite the very similar solutions obtained with both strategies, the problems found with the strategy using the deterministic model, such as lack of convergence and high computational time, make the use of the optimization strategy with the statistical model, which showed to be robust and fast, more suitable for the flash fermentation process, being recommended for real-time applications coupling optimization and control.

Use of weathered diesel oil as a low-cost raw material for biosurfactant production

Abstract - This work aimed to investigate the capability of biosurfactant production by Staphylococcus hominis , Kocuria palustris and Pseudomonas aeruginosa LBI, using weathered diesel oil from a long-standing spillage as raw material. The effect of the culture media (Robert or Bushnell-Haas) and of the carbon source (spilled diesel oil or commercial diesel oil) on biosurfactant production was evaluated. Erlenmeyer flasks (250 mL) containing the cell broth were agitated (240 rpm) for 144 h at 27±2 o C. Biosurfactant production was monitored according to the De Nouy ring method using a Kruss K6 tensiometer. Considering the possibility of intracellular storage of biosurfactant in the cell wall of the cultures S. hominis and K. palustris , experiments were also done applying ultrasound as a way to rupture the cells. For the conditions studied, the cultures did not indicate production of biosurfactants. Results obtained with a hydrocarbon biodegradability test based on the redox indicator 2,6-dichlorophenol indophenol showed that only the commercial diesel was biodegraded by the cultures.

Dynamics of a continuous flash fermentation for butanol production

University of Campinas, UNICAMP School of Chemical Engineering, P.O. Box 6066, 13083-970 Campinas SP