Expressão heteróloga e modelagem de uma enzima lipolítica utilizando a abordagem metagenômica

Pós-graduação em Microbiologia Agropecuária - FCAV

Synthesis of polycationic bentonite-ionene complexes and their benzene adsorption capacity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Incêndios de petróleo e pretoquímicos: biorremediação de áreas afetadas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversity of hydrocarbon-degrading Klebsiella strains isolated from hydrocarbon-contaminated estuaries

To investigate the diversity and the catabolic capacity of oil-degrading Klebsiella strains isolated from hydrocarbon-contaminated sediments in Santos-Sao Vicente estuary systems in Brazil. Klebsiella strains obtained from the estuary were characterized using 16S rRNA gene sequencing and BOX-PCR patterns, testing their catabolic capacity to degrade toluene, xylene, naphthalene and nonane, and identifying the catabolic genes present in the oil-degrading strains. Results show that Klebsiella strains were widespread in the estuary. Twenty-one isolates from the Klebsiella genus were obtained; 14 had unique BOX patterns and were further investigated. Among four distinct catabolic genes tested (todC1, ndoB, xylE and alkB1), only the todC1 gene could be amplified in two Klebsiella strains. The biodegradation assay showed that most of the strains had the ability to degrade all of the tested hydrocarbons; however, the strains displayed different efficiencies. The oil-degrading Klebsiella isolates obtained from the estuary were closely related to Klebsiella pneumoniae and Klebsiella ornithinolytica. The isolates demonstrated a substantial degree of catabolic plasticity for hydrocarbon degradation. The results of this study show that several strains from the Klebsiella genus are able to degrade diverse hydrocarbon compounds. These findings indicate that Klebsiella spp. can be an important part of the oil-degrading microbial community in estuarine areas exposed to sewage.

Results from a 15-year study on hydrocarbon concentrations in water and sediment from Admiralty Bay, King George Island, Antarctica

Admiralty Bay on the King George Island hosts the Brazilian, Polish and Peruvian research stations as well as the American and Ecuadorian field stations. Human activities in this region require the use of fossil fuels as an energy source, thereby placing the region at risk of hydrocarbon contamination. Hydrocarbon monitoring was conducted on water and sediment samples from the bay over 15 years. Fluorescence spectroscopy was used for the analysis of total polycyclic aromatic hydrocarbons (PAHs) in seawater samples and gas chromatography with flame ionization and/or mass spectrometric detection was used to analyse individual n-alkanes and PAHs in sediment samples. The results revealed that most sites contaminated by these Compounds are around the Brazilian and Polish research stations due to the intense human activities, mainly during the summer. Moreover, the sediments revealed the presence of hydrocarbons from different sources, suggesting a mixture of the direct input of oil or derivatives and derived from hydrocarbon combustion. A decrease in PAH concentrations occurred following improvement of the sewage treatment facilities at the Brazilian research station, indicating that the contribution from human waste may be significant.

Multi-molecular markers and metals as tracers of organic matter inputs and contamination status from an Environmental Protection Area in the SW Atlantic (Laranjeiras Bay, Brazil)

The sources and concentrations of aliphatic hydrocarbons (AHs) and polycyclic aromatic hydrocarbons (PAHs), faecal and biogenic sterols, and trace metals at 10 sampling sites located in Laranjeiras Bay, a large Environmental Protection Area in the southern Atlantic region of Brazil, were determined to assess the sources of organic matter and the contamination status of estuarine sediments. Organic compounds were determined by GC-FID and GC-MS, and ICP-OES was used to evaluate trace metals. The total AHs concentration ranged from 0.28 to 8.19 mu g g(-1), and n-C-29 and n-C-31 alkanes were predominant, indicating significant inputs from higher terrestrial plants. Unresolved complex mixtures (UCM) were not detected at any site, suggesting that the study area was not significantly contaminated by fossil fuels. The total PAH concentration varied from 3.85 to 89.2 ng g(-1). The ratio between selected PAH isomers showed that combustion of biomass, coal, and petroleum is the rnain source of PAHs in the study area. The concentrations of the faecal sterols coprostanol and epicoprostanol were below the detection limits, suggesting that sewage was not a significant contributor to sedimentary organic matter. The concentrations of the trace metals (As, Cr, Cu, Ni, Pb and Zn) were low, except near sites located at the mouths of rivers that discharge into the study area and near urbanised regions (Paranagua city and the adjoining harbour). In general, the concentrations of PAHs were below the threshold effect concentrations (TEL) levels. Although the As, Cr and Ni concentrations were above the TEL levels, the study area can be considered as preserved from human activities.

Proceedings of the Twentieth Annual Biochemical Engineering Symposium

The 20th Annual Biochemical Engineering Symposium was held at Kansas State University on April 21,1990. The objectives of the symposium were to provide: (i) a forum for informal discussion of biochemical engineering research being conducted at the participating institutions and (ii) an opportunity for students to present and publish their work. Twenty-eight papers presented at the symposium are included in this proceedings. Some of the papers describe the progress of ongoing projects, and others contain the results of completed projects. Only brief summaries are given of the papers that will be published in full elsewhere. The program of the symposium and a list of the participants are included in the proceedings. ContentsCell Separations and Recycle Using an Inclined Settler, Ching-Yuan Lee, Robert H. Davis and Robert A. Sclafani Micromixing and Metabolism in Bioreactors: Characterization of a 14 L Fermenter, K.S. Wenger and E.H. Dunlop Production, Purification, and Hydrolysis Kinetics of Wild-Type and Mutant Glucoamylases from Aspergillus Awamori, Ufuk Bakir, Paul D. Oates, Hsiu-Mei Chen and Peter J. Reilly Dynamic Modeling of the Immune System, Barry Vant-Hull and Dhinakar S. Kompala Dynamic Modeling of Active Transport Across a Biological Cell: A Stochastic Approach, B.C. Shen, S.T. Chou, Y.Y. Chiu and L.T. Fan Electrokinetic Isolation of Bacterial Vesicles and Ribosomes, Debra T.L. Hawker, Robert H. Davis, Paul W. Todd, and Robert Lawson Application of Dynamic Programming for Fermentative Ethanol Production by Zymomonas mobilis, Sheyla L. Rivera and M. Nazmul Karim Biodegradation of PCP by Pseudomonas cepacia, R. Rayavarapu, S.K. Banerji, and R.K. Bajpai Modeling the Bioremediation of Contaminated Soil Aggregates: a Phenomenological Approach, S. Dhawan, L.E. Erickson and L.T. Fan Biospecific Adsorption of Glucoamylase-I from Aspergillus niger on Raw Starch, Bipin K. Dalmia and Zivko L. Nikolov Overexpression in Recombinant Mammalian Cells: Effect on Growth Rate and Genetic Instability, Jeffrey A. Kern and Dhinakar S. Kompala Structured Mathematical Modeling of Xylose Fermentation, A.K. Hilaly, M.N. Karim, I. C. Linden and S. Lastick A New Culture Medium for Carbon-limited Growth of Bacillus thuringiensis, W. -M. Liu and R.K. Bajpai Determination of Sugars and Sugar Alcohols by High Performance Ion Chromatography, T. J. Paskach, H.-P. Lieker, P.J. Reilly, and K. Thielecke Characterization of Poly-Asp Tailed B-Galactosidase, M.Q. Niederauer, C.E. Glatz, l.A. Suominen, C.F. Ford, and M.A. Rougvie Computation of Conformations and Energies of cr-Glucosyl Disaccharides, Jing Zepg, Michael K. Dowd, and Peter J. Reilly Pentachlorophenol Interactions with Soil, Shein-Ming Wei, Shankha K. Banerji, and Rakesh K. Bajpai Oxygen Transfer to Viscous Liquid Media in Three-Phase Fluidized Beds of Floating Bubble Freakers, Y. Kang, L.T. Fan, B.T. Min and S.D. Kim Studies on the Invitro Development of Chick Embryo, A. Venkatraman and T. Panda The Evolution of a Silicone Based Phase-Separated Gravity-Independent Bioreactor, Peter E. Villeneuve and Eric H. Dunlop Biodegradation of Diethyl Phthalate, Guorong Zhang, Kenneth F. Reardon and Vincent G. Murphy Microcosm Treatability of Soil Contaminated with Petroleum Hydrocarbons, P. Tuitemwong, S. Dhawan, B.M. Sly, L.E. Erickson and J.R. Schlup

Composition of organic matter from samples of massive sulfides, bottom sediments, and seawater collected within the Logachev and Broken Spur hydrothermal fields, Mid-Atlantic Ridge

Lipid components of hydrothermal deposits from the unusual field at 14°45'N MAR and from the typical field at 29°N MAR were studied. For the first time mixed nature of organic matter (OM) from hydrothermal sulfide deposits was established with use of biochemical, gas chromatographic, and molecular methods of studies. In composition of OM lipids of phytoplankton, those of chemosynthesis bacteria and non-biogenic synthesis lipids were determined. Specific conditions of localization of sulfide deposits originated from ''black smokers'' (reducing conditions, absence of free oxygen, presence of reduced sulfur preventing OM from decomposition) let biogenic material, including bacterial one, be preserved in sulfide deposits. The hydrothermal system at 14°45'N MAR is characterized by geological, geochemical and thermodynamic conditions allowing abiogenic synthesis of methane and petroleum hydrocarbons. For sulfide deposits at 29°N and other active hydrothermal fields known at MAR, abiogenic synthesis of hydrocarbons occurs in lower scales.

SADA and DESYRE DSSs descriptive classification criteria (main issues related to DSS's)

Brownfield rehabilitation is an essential step for sustainable land-use planning and management in the European Union. In brownfield regeneration processes, the legacy contamination plays a significant role, firstly because of the persistent contaminants in soil or groundwater which extends the existing hazards and risks well into the future; and secondly, problems from historical contamination are often more difficult to manage than contamination caused by new activities. Due to the complexity associated with the management of brownfield site rehabilitation, Decision Support Systems (DSSs) have been developed to support problem holders and stakeholders in the decision-making process encompassing all phases of the rehabilitation. This paper presents a comparative study between two DSSs, namely SADA (Spatial Analysis and Decision Assistance) and DESYRE (Decision Support System for the Requalification of Contaminated Sites), with the main objective of showing the benefits of using DSSs to introduce and process data and then to disseminate results to different stakeholders involved in the decision-making process. For this purpose, a former car manufacturing plant located in the Brasov area, Central Romania, contaminated chiefly by heavy metals and total petroleum hydrocarbons, has been selected as a case study to apply the two examined DSSs. Major results presented here concern the analysis of the functionalities of the two DSSs in order to identify similarities, differences and complementarities and, thus, to provide an indication of the most suitable integration options.

Modelización de un proceso de biorremediación de suelos contaminados con gasoil

El objeto de esta tesis doctoral es encontrar, mediante herramientas estadísticas, un modelo matemático que prediga la evolución temporal de las concentraciones de los hidrocarburos totales del petróleo (TPH) en un proceso de biorremediación de suelos contaminados con gasoleo. Obtenido el modelo matemático realizar las comparaciones pertinentes para determinar la eficiencia de distintos tratamientos, la influencia tanto del suelo como del nivel de concentración. Para cumplir este objetivo se realizó un diseño de experimentos que tomaba en cuenta dos tipos de suelos, dos niveles de concentración de hidrocarburo y seis tratamientos. Se realizaron en total 1824 ensayos en laboratorio repartidos en 8 campañas de campo durante un año de experiencia tanto en parcelas de 4 x 4 m como en microcosmos. Los resultados obtenidos muestran que el proceso de biorremediación se puede ajustar a un comportamiento exponencial, que es una ecuación de cinética de primer orden, y que las comparaciones realizadas han mostrado resultados satisfactorios de la eficiencia de algunos tratamientos. ABSTRACT The purpose of this thesis is using statistical tools find a mathematical model to predict the time evolution of the total petroleum hydrocarbons (TPH) concentrations in the bioremediation process of diesel contaminated soil. Retrieved mathematical model make relevant comparisons to determine the efficiency of different treatments and the influence of both soil and concentration levels. To achieve this goal a design of experiments was developed, it took into account two types of soil, two concentration levels of hydrocarbon and six treatments. There were a total of 1824 laboratory tests spread over 8 field campaigns during a year of experience in both plots of 4 x 4 m in microcosm. The results obtained show that the bioremediation process can be adjusted to an exponential model, it is an equation of kinetic of first order and that the comparisons have shown satisfactory results of the efficiency of some treatments.

A Study of Shallow Groundwater Migration through Anthropogenic Fill along a Seawall

This report provides the findings and opinions of a historical document review, hydraulic balance calculation, and proposed additional study for a property that was historically used as a bulk petroleum storage and distribution facility. The property lies along the base, west, of a heavily vegetated bluff with a tidally influenced body of water west-adjacent to the property. The western portion of the property is bounded by a seawall spanning approximately 3,200 linear feet trending north-south. The seawall’s construction details are not known, save for a 225-foot section of driven sheet pile wall located within the northern portion of the property’s seawall. Due to the presence of petroleum hydrocarbons in soil and groundwater at the property, a cleanup action for the property will likely be overseen by the state regulatory agency. The property is currently undergoing remedial investigation in an effort to identify the lateral and vertical extent in which contaminants at the property have come to be located, also known as the “site” as defined by the Model Toxics Control Act (MTCA). The majority of the property bounded within the seawall area has been characterized; however, the shoreline sediments located immediately west-adjacent of the seawall have not been properly delineated. Identifying the bounds of the site to the west within sediment is pivotal for the purposes of the remedial investigation. Since the west-adjacent shoreline is so extensive, conducting a complete sediment sampling event along the entire shoreline would be cost-prohibitive due to analytical costs and logistical issues at the property. Because of the extensive nature of the shoreline, it would greatly benefit the client and project to focus sampling efforts at areas of greater risk for contaminants along the shoreline by identifying potential preferential pathways for contaminants to migrate off of the property and into adjacent shoreline sediments. The review of historical studies of the property yielded some useful information; however much of the findings included within the historical studies were lacking original raw data, therefore limiting the information obtained. The calculated hydraulic balance for the property yielded a relatively large surplus of recharge to groundwater after precipitation events, reinforcing the concept that contaminant have potentially historically, and currently, been migrating into the adjacent shoreline through preferential pathways along the seawall. Due to the limitation within the historical studies for the property as well as the groundwater recharge identified in the hydraulic balance, an additional study was proposed in an effort to provide additional aquifer characteristics along the seawall, and the ability to observe flow propagation at and proximate to the seawall in two-dimensions through time without the need to piece separate studies together. This proposed study includes a single simultaneous tidal study which comprises select monitoring points along the seawall. This report has identified the need for additional data that can be collected through available avenues for the property based upon the client’s desires and project needs. Ultimately, the proposed additional study is suggested based upon its relatively low capital investment, and ability meet the requirements relevant to the specific project needs and scope. Assuming preferential pathways are identified through the additional study proposed within this report, a representative and cost-effective sediment sampling plan can then be put in place in an effort to define the site.

Estimating the Extent of Soil Contamination During a Redevelopment Excavation

From October 2014 to March 2015, I provided excavation oversight services at a property with substantial environmental concerns. The property in question is located near downtown Seattle and was formerly occupied by the Washington’s first coal gasification plant. The plant operated from 1888 to 1908 and produced coal gas for municipal use. A coal tar like substance with a characteristically high benzene concentration was a byproduct of the coal gasification process and heavily contaminated at or below the surface grade of the plant as shown in previous investigations on the property. Once the plant ceased operation in 1908 the property was left vacant until 1955 when the site was filled in and a service station was built on the property. The main goal of the excavation was not to achieve cleanup on the property, but to properly remove what contaminated soil was encountered during the redevelopment excavation. Areas of concern were identified prior to the commencement of the excavation and an estimation of the extent of contamination on the property was developed. “Hot spots” of contaminated soil associated with the fill placed after 1955 were identified as areas of concern. However, the primary contaminant plume below the property was likely sourced from the coal gasification plant, which operated at an approximate elevation of 20 feet. We planned to constrain the extents of the soil contamination below the property as the redevelopment excavation progressed. As the redevelopment excavation was advanced down to an elevation of approximately 20 feet, soil samples were collected to bound the extents of contamination in the upper portion of the site. The hot spots, known pockets of carcinogenic polycyclic aromatic hydrocarbons (cPAH) located above 20 feet elevation, were excavated as part of the redevelopment excavation. Once a hot spot was excavated, soil samples were collected from the north, south, east, west and bottom sidewalls of the hot spot excavation to check for remaining cPAH. Additionally, four underground storage tanks (USTs) associated with the service station were discovered and subsequently removed. Soil samples were also collected from the resulting UST excavation sidewalls to check for remaining petroleum hydrocarbons. Once the excavation reached its final excavation depth of 20 to 16 feet in elevation, bottom of excavation samples were collected on a 35 foot by 35 foot grid to test for concentrations of contaminants remaining onsite. Once the redevelopment excavation was complete, soils observed from borings drilled for either structural elements, geotechnical wells, or environmental wells were checked for any evidence of contamination using field screening techniques. Evidence of contamination was used to identify areas below the final excavation grade which had been impacted by the operation of the coal gasification plant. Samples collected from the excavation extents of hot spots and USTs show that it was unlikely that any contamination traveled from the post-1955 grade down to the pre-1955 grade. Additionally, the lack of benzene in the bottom of excavation samples suggests that a release from the coal gasification plant occurred below the redevelopment excavation final elevations of 20 to 16 feet. Qualitative data collected from borings for shoring elements and wells indicated that the spatial extent of the subsurface contaminant plume was different than initially estimated. Observations of spoils show that soil contamination extends further to the southwest and not as far to the east and north than originally estimated. Redefining the extent of the soil contamination beneath the property will allow further subsurface investigations to focus on collecting quantitative data in areas that still represent data gaps on the property, and passing over areas that have shown little signs of contamination. This information will help with the formation of a remediation plan should the need to clean up the site arise in the future.

Aplicação de tratamentos eletroquímicos integrados para remediação de solos e águas contaminadas com petróleo e pesticidas

Electrochemical technologies have been proposed as a promising alternative for the treatment of effluents and contaminated soils. Therefore, the objective of this work was to study the treatment of contaminated soils and wastewaters using electrochemical technologies. Thus, the study regarding the scale-up of the electrochemical system with continuous flow treatment of wastewater of the petrochemical industry was investigated using platinum electrodes supported on titanium (Ti / Pt), and boron-doped diamond (BDD). The results clearly showed that under the operating conditions studied and electrocatalytic materials employed, the better removal efficiency was achieved with BDD electrode reducing the chemical oxygen demand (COD) from 2746 mg L-1 to 200 mg L-1 in 5 h consuming 56.2 kWh m-3 . The decontamination of soils and effluents by petrochemical products was evaluated by studying the effects of electrokinetic remediation for removal of total petroleum hydrocarbons (HTP) from contaminated soil with diesel. The efficiency of this process was dependent on the electrolyte used Na2SO4 (96.46%), citric acid (81.36%) and NaOH (68.03%) for 15 days. Furthermore, the effluent after treatment of the soil was treated by electrochemical oxidation, achieving a good elimination of the organic polluting load dissolved. Depending on the physical behavior of wastewater contaminated with oil (emulsified state); atrazine emulsified effluents were investigated. The main characteristics of the effluent produced during the washing of contaminated soil were studied, being dependent on the surfactant dosage used; which determined its electrolytic treatment with BDD. The electrochemical oxidation of emulsified effluent of atrazine was efficient, but the key to the treatment is reducing the size of micelles.

(Table) Content of organic compounds in the bottom sediments of the Volga River estuary

Materials from different spheres of the Earth are ultimately delivered to bottom sediments, which serve as a natural recorder of the functioning of other spheres and originate as a result of the accumulation of their substances. Sedimentary material and species of river-transported elements are subjected to dramatic reworking in marginal filters, where river and sea waters are mixed. These processes are most important for the Caspian Sea, where runoffs of rivers (especially the Volga River) and the intense development and transportation of hydrocarbon fuel by tankers and pipelines (related to the coastal petroleum industry in the Sumgait and Baku ports, Apsheron Peninsula) are potential sources of hydrocarbon pollution. Previously obtained data showed that the total content of hydrocarbon fraction (i.e., the sum of aliphatic hydrocarbons (AHC) and polycyclic aromatic hydrocarbons (PAH)) in bottom sediments varied within 29-1820 µg/g. The content of petroleum hydrocarbons in the northeastern Caspian region varied from 0.052 to 34.09 µg/g with the maximum content in the Tengiz field. The content of six polyarenes in the Volga delta sediments was no more than 40 ng/g. To determine the recent HC pollution of bottom sediments and trends in the functioning of the Volga marginal filter, in summer of 2003 and 2004 we analyzed bottom sediments (58 samples) in the river waterway; Kirovsk channel; Bakhtemir and Ikryanoe branches; tributaries of the Kizan, Chagan, and other rivers; and the Caspian seashore.

Treatment of phenanthrene and benzene using microbial fuel cells operated continuously for possible in situ and ex situ applications

Bioelectrochemical systems could have potential for bioremediation of contaminants either in situ or ex situ. The treatment of a mixture of phenanthrene and benzene using two different tubular microbial fuel cells (MFCs) designed for either in situ and ex situ applications in aqueous systems was investigated over long operational periods (up to 155 days). For in situ deployments, simultaneous removal of the petroleum hydrocarbons (>90% in term of degradation efficiency) and bromate, used as catholyte, (up to 79%) with concomitant biogenic electricity generation (peak power density up to 6.75 mWm−2) were obtained at a hydraulic retention time (HRT) of 10 days. The tubular MFC could be operated successfully at copiotrophic (100 ppm phenanthrene, 2000 ppm benzene at HRT 30 days) and oligotrophic (phenanthrene and benzene, 50 ppb each, HRT 10 days) substrate conditions suggesting its effectiveness and robustness at extreme substrate concentrations in anoxic environments. In the MFC designed for ex situ deployments, optimum MFC performance was obtained at HRT of 30 h giving COD removal and maximum power output of approximately 77% and 6.75 mWm−2 respectively. The MFC exhibited the ability to resist organic shock loadings and could maintain stable MFC performance. Results of this study suggest the potential use of MFC technology for possible in situ/ex situ hydrocarbon-contaminated groundwater treatment or refinery effluents clean-up, even at extreme contaminant level conditions.