Network decontamination is a well known mobile agent problem with many applications. We assume that all nodes of a network are contaminated (e.g., by a virus) and a set of agents is deployed to decontaminate them. An agent passing by a node decontaminates it, however a decontaminated node can be recontaminated if any of its neighbours is contaminated. In the vast literature a variety of models are considered and different assumptions are made on the power of the agents. In this thesis we study variation of the decontamination problem in mesh and tori topologies, under the assumption that when a node is decontaminated, it is immune to recontamination for a predefined amount of time t (called immunity time). After the immunity time is elapsed, recontamination can occur. We focus on three different models: mobile agents (MA), cellular automata (CA), and mobile cellular automata (MCA). The first two models are commonly studied and employed in several other contexts, the third model is introduced in this thesis for the first time. In each model we study the temporal decontamination problem (adapted to the particular setting) under a variety of assumptions on the capabilities of the decontaminating elements (agents for MA and MCA, decontaminating cells for CA). Some of the parameters we consider in this study are: visibility of the active elements, their ability to make copies of themselves, their ability to communicate, and the possibility to remember their past actions (memory). We describe several solutions in the various scenarios and we analyze their complexity. Efficiency is evaluated slightly differently in each model, but essentially the effort is in the minimization of the number of simultaneous decontaminating elements active in the system while performing the decontamination with a given immunity time.

Excavation is a common method to clean up areas with increased levels of metal contamination. Identifying the degree of soil contamination is normally done by a laboratory analysis but it could also be done using a portable X-ray fluorescence spectrometer (XRF) in the field. This project has investigated how well XRF results correlate with the results from the laboratory and how the water ratio affects the results in XRF measurement. The purpose has been to gain an increased knowledge of how soil samples should be handled to obtain a reliable XRF result. Data was compared between lab results and XRF results. Soil samples with different water ratios have been examined to see whether moist affect the XRF results. The results indicate that the reliability of XRF results depends on the metal being studied. The water ratio also affects the result to varying degrees depending on the metal. This study has involved arsenic, lead, nickel, copper and zinc. The main focus is on arsenic, and the results show no significant difference between the two aforementioned methods. There is however a significant difference at some water quotas, but the result correlate very well and can be estimated based on the equation calculated in the results. Results show that a portable XRF is a reliable instrument for dry samples of arsenic, but laboratory analysis should be used to confirm the results at low levels of arsenic close to the applicable limit values due to margin of error when measuring with a portable XRF.

Experiments were performed to investigate the effects of wash treatments, consisting of hot water, 2% lactic, 2% acetic, or 2% levulinic acid, for decontamination of pathogenic bacteria previously inoculated onto meat surfaces, to inhibit growth of pathogenic bacteria inoculated onto previously washed meat surfaces, and on the organoleptic quality of sliced turkey roll and beef trim. Acid washes were no more effective at reducing Escherichia coli O157:H7 on beef plate, Listeria monocytogenes on sliced turkey roll, and Salmonella on pork belly than was water wash. Only lactic acid treatment was more effective than water at reducing Salmonella on chicken skin, but by less than 1 log CFU/cm2. Increasing wash temperatures with 2% levulinic acid did not reduce E. coli O157:H7 on beef plate. Organic acid washes did not protect against growth of L. monocytogenes and E. coli O157:H7. Acetic acid prevented growth of Salmonella, but only on chicken skin. Organic acid spray treatments of sliced turkey roll and beef trim did not affect consumer liking of turkey roll or cooked ground beef patties. Acid treatments had some effect on instrumental color measurements, but these appear to have little practical significance. Overall, washing with 2% organic acid solutions was no more effective at reducing pathogenic bacteria on meat surfaces than washing with water.

This study examines the fundamental properties of Colloidal Gas Aphrons, CGA, relevant to its application in soil and groundwater decontamination. It also presents the results of laboratory experiments on the applicability of CGA in treating soils contaminated with organic chemicals. CGA is a collection of spherical, 10 to 100 micron-sized gas bubbles dispersed in an aqueous surfactant solution with a volumetric gas fraction (quality) of at most 0.74. It is characterized by its quality, bubble size, stability and apparent viscosity. The stability can be described by the half life, the time needed for 50% of its liquid phase to separate by gravitational drainage. CGA is a non-Newtonian fluid. The apparent viscosity is higher than that of its liquid or gas phase and increases with increase in quality. At quality above 0.6, CGA is a pseudoplastic fluid and can be modelled by the power law. In this quality range, apparent viscosity decreases with increase in shear rate. Measured apparent viscosity can be as high as 25 centipoise at low shear rates for a CGA quality of 0.72. But at low quality, CGA is best described by the Bingham fluid model. CGA flow through porous media follows the discontinuous fluid flow model with the liquid phase advancing faster than the gas phase. The presence of CGA bubbles causes considerable reduction of effective mobility compared to water flow. The bubbles act to reduce the effective flow area by first blocking the wider pores and re-directing flow to narrower pores. The tenacity of the bubbles in the presence of adequate surfactant molecules account for its flow characteristics in porous media. CGA is more effective in flushing hydrophobic organics from saturated sand-packed columns compared with surfactant flushing. CGA provides the surface active agent needed to lower the interfacial tension of the non-wetting phase and at the same time creates enough viscous force to mobilize any trapped fluid. Its ability to preferentially block large pores in a heterogeneous media and direct fluid flow to smaller pores also contributes to the demonstrated effectiveness as a soil flushing agent. / Ph. D.

Abstrakt The diploma thesis is focused on the issue of securing serviceability of decontamination sites using the forces and the means available after cancellation of rescue battalions, and on the current position of the Fire Rescue Service. The objective of the thesis was to draw attention to the problem arising from insufficient securing of decontamination after cancellation of the rescue battalions intended for this activity. The output of the analysis of the present state and the state before cancellation of the rescue batallions is the finding that the solution currently used to secure decontamination is less effective. Therefore I have proposed a more effective solution based on maintaining rescue battalions together with the Fire Rescue Service. The raised hypotheses were confirmed through the concise charts that addressed the capabilities of the rescue units and the communication between individual brigades and departments will be verified in the planned exercise ZÓNA 2010 that will take place from 22 to 23 September 2010. In my thesis I made use of the available literature, findings from my bachelor thesis and from exercises ZÓNA 2007 and ZÓNA 2008, observations of experts, and terrain reconnaissance leading to proposing a suitable decontamination site. The thesis includes a proposal of a decontamination site in Hluboká nad Vltavou {--} Zámostí. I believe that there would be justification for submission of this proposal to the competent bodies. I regard my thesis as a good material for assessing the current state of this issue, proposing various solutions, and returning to greater care of citizens, as it used to be in previous year.

Experimental studies into the use of cold atmospheric plasmas for inactivating foodborne pathogens are presented in this thesis. Eliminating the possibility that treatment delivered by a plasma to a population or assemblage of micro-organisms is unevenly distributed is an essential pre-requisite to attempting to interpret inactivation kinetics with a view to elucidating mechanisms of inactivation. A filtration method of depositing cells evenly on the surface of a membrane without cell stacking was developed and used throughout the work described here. Two atmospheric plasma systems were evaluated and each brought about microbial inactivation in a distinct way. A pulsed radio frequency plasma jet operated at 3.47 MHz caused gross morphological changes to L. innocua whereas a low frequency air mesh plasma system operated at a frequency of 24 kHz led to the inactivation of these bacteria without inducing observable structural changes. Changing the operating parameters of the plasma jet system had a significant effect on the composition of the reactive plasma species generated as revealed by changes to the mode of inactivation of bacteria. In addition to inactivating bacteria, the pulsed plasma jet was shown to be highly effective in degrading and removing amyloid aggregates from the surface of mica coupons. Amyloids have widely been used as a non-infectious model for prions, and the results obtained here show potential for the application of gas plasma technology for removing prions from abiotic surfaces in medical and other applications. It has widely been assumed that bacterial envelopes are the principal sites at which reactive plasma species bring about damage to cells. However, changing the composition of the bacterial membranes of E. coli and Listeria innocua by cultivating them at widely different temperatures to induce changes proved not to result in enhanced inactivation. Flow cytometry was also used to provide additional insights into possible mechanisms of inactivation. The following fluorescent dyes were used either singly or in combination; SYTO 13, DiBAC4(3), cFDA and PI. The results obtained with the dyes DiBAC4(3) and PI showed that Gram positive bacteria became depolarised prior to the bacterial membrane becoming compromised, possibly suggesting that the inactivating plasma species are affecting membrane proteins responsible for maintaining the bacterial charge. Differences between the fluorescent dye staining of Gram negative and Gram positive species were obtained using SYTO13 and PI demonstrating that the different membrane structures affect their interaction with the plasma. In additional studies, the air mesh plasma was used to treat multi-drug resistant strains of Methicillin resistant Staphylococcus aureus (MRSA) in an attempt to reverse antibiotic resistance. MRSA PM 64 was shown to reverse its antibiotic resistance to Oxacillin, Kanamycin and Trimethoprim. Culturing the bacteria in a nutrient limited media led to increased resistance towards plasma treatment and maintenance of their high levels of antibiotic resistance.

This thesis presents experimental studies of various non-thermal atmospheric pressure gas discharges generated using short pulsed excitation as an alternative to widely used sinusoidal excitation. Several pulse generators are detailed that provide high voltage pulses ranging from hundreds of microseconds to less than ten nanoseconds in duration. A key enabler to the generation of a stable discharge is a suitably high repetition rate; this prerequisite precludes many conventional pulsed power technologies. Fortunately, recent advances in semiconductor technology have made it possible to construct solid state switches capable of producing high voltage pulses with repetition rates of many kilohertz. Pulsed excitation introduces many opportunities to tailor the applied voltage and consequently enhance the discharge which are not possible with sinusoidal excitation sources. Through detailed electrical and optical analysis it is shown that pulsed excitation is not only more energy efficient than a comparable sinusoidal source but produces a higher flux of excited species that are essential in many applications. When pulse widths are reduced to a sub-microsecond timescale a novel barrier-free mode of operation is observed. It is shown that diffuse large area plasmas are easily produced at kilohertz repetition rates without the usually indispensable dielectric barriers. Experimental results show that a short pulse width prevents the onset of the undesirable glow-to-arc transition thus introducing an added degree of stability. A further benefit of pulsed excitation is the ability to produce gas discharges with a high instantaneous peak power yet low average power consumption, resulting in a high density plasma that exhibits roomtemperature characteristics. Finally, as an acid test to highlight the many benefits of pulsed excitation several real-world applications are considered. It is shown that in all cases pulsed gas discharges provide real benefits compared to their sinusoidal counterparts.

Afin de satisfaire à un cahier des charges toujours plus exigeant, l'industrie spatiale se doit d'innover en permanence, notamment dans le domaine du maintien des performances. De fait, la minimisation de la contamination moléculaire, qui provient du dégazage sous vide des matériaux constitutifs du satellite, permet d'éviter une baisse progressive des performances de ce dernier. En effet, les molécules organiques issues de ce dégazage peuvent se déposer sur les optiques ou les revêtements de contrôle thennique, altérant ainsi leur fonctionnement. De par leur porosité contrôlée, les zéolithes possèdent des propriétés compatibles avec une utilisation en orbite. Le principal obstacle à l'utilisation de ces matériaux concerne leur mise en forme. Contrairement aux poudres, les films zéolithiques permettent d'éviter la génération de poussières et présentent une meilleure durabilité tout le long de la mission en orbite.Le but de ce travail est d'étudier les propriétés d'adsorption de différentes structures zéolithiques vis-à-vis de divers polluants organiques Des films de zéolithes de types BEA et FAU ont été déposés sur des supports en verre et en fibres de carbone, respectivement. Dans les deux cas, des films homogènes, dotés d'une épaisseur compatible avec une utilisation en orbite, ont été obtenus. Les quantités adsorbées par les films ont été comparées à celles des poudres zéolithiques correspondantes. Il ressort de cette étude que les capacités d'adsorption sont légèrement plus faibles pour les films que pour les poudres, en raison de la présence de barrières diffusionnelles dans les films. Cependant, des études de désorption menées sous vide et en température ont établi que les films présentaient de meilleures capacités de rétention que les poudres, confirmant leur applicabilité dans le domaine de la décontamination moléculaire en orbite. / Molecular contamination control has been described as a key parameter for the optimization of on-orbit satellit functioning. In space conditions, the outgassing of organic pollutants jeopardizes the performances of certain instrument by contaminating critical surfaces such as thermal radiators and optical systems. The chemical nature of these pollutant has been investigated by the NASA, which listed the most frequent ones in a table. Several types of outgassed molecule were identified, mainly deriving from plasticizers (phthalates, adipates...). Moreover, elastomers, adhesives and other compounds such as hydrocarbons were also found in important quantities.In order to minimize the molecular contamination, techniques involving pre-flight bakeouts were set up, but rapidly proved to be expensive and time-consuming, as an important number of outgassing cycles was required to efficiently decrease the molecular contaminationThat is why other porous materials had to be investigated, highlighting zeolite relevancy for satellite decontamination.These aluminosilicate materials exhibit an ordered molecular-sized porous network which induces high sorption properties and an outstanding size and shape selectivity. Another topic of interest deals with the shaping of zeolite for their use in space conditions. Unlike powder materials, zeolite films avoid the generation of dust particles in low earth orbit and can be easily embedded in appropriate areas throughout the lifetime of the mission.This work deals with the preparation of zeolite films and the adsorption of organic contaminants onto their surface. Zeolite beta and faujasite films were prepared on glass substrates and carbon fibers, respectively. The sorption properties towards several alkanes and aromatic molecules were investigated and the performances compared to the powders counterparts. It was shown that the films presented slightly lower sorption properties in comparison to powders, due to diffusional barriers. Nevertheless, the properties were satisfactory in respect to the requirements of space industry.

En cas de contamination cutanée radiologique par des composés d’uranium, les seuls traitements actuellement employés consistent en un rinçage de la zone contaminée par de l’eau et des détergents, ou par une solution de sel calcique de l’acide diéthylène-triamine-pentaacétique (Ca-DTPA). Ces derniers ne sont cependant pas efficaces vis-à-vis de l'uranium. De plus, en l'absence de traitement d'urgence, le passage transcutané de ce radionucléide est rapide, et induit une exposition interne après sa distribution dans l’organisme par le biais de la circulation sanguine. Une partie de l'uranium ainsi biodisponible est alors stockée dans les organes cibles que sont principalement les reins et le squelette, où ses effets toxiques se manifestent. C'est pourquoi une formulation topique consistant en une nanoémulsion huile dans eau, incorporant des molécules de calixarène tricarboxylique en tant qu'agent chélatant spécifique de l'uranium, a été initialement développée. Les travaux menés dans le cadre de cette thèse visent à évaluer l'efficacité de décontamination ex vivo et in vivo de ce nouveau traitement d'urgence à la fois sur peaux intactes et sur peaux lésées superficiellement. Pour cela, le modèle d'excoriation a été utilisé. Des modèles de lésions reproductibles ont également été mis en place afin de mimer des incisions par micro-piqûres et microcoupures. Ces études démontrent que la nanoémulsion de calixarène pourrait constituer un traitement de décontamination efficace, moins agressif que l’emploi de l’eau savonneuse actuellement employée. Sa potentielle toxicité cutanée a également été évaluée in vitro par l'utilisation d'épiderme humain reconstitué, combinée à trois différents tests de toxicité (MTT, LDH et IL-1-α). Dans le cadre de ces études, il a ainsi été démontré que la nanoémulsion de calixarène n’induit pas de toxicité cutanée, même après un temps de contact prolongé jusqu'à 24 h. / In case of radiological skin contamination by uranium compounds, the only treatments currently available consist in rinsing the contaminated skin area with water and detergent, or with a calcium salt of diethylene triamine pentaacetic acid (Ca-DTPA) solution. However, these procedures are not specific and no efficient treatment for cutaneous contamination due to uranium exists. In the absence of such treatments, uranium diffusion through the skin is fast, inducing an internal exposure after its distribution inside the body through the bloodstream. One part of the bioavalaible uranium is uptaken in target organs which are the kidneys and the skeleton, where its toxic effects occur. Therefore a topical formulation consisting of an oil-in-water nanoemulsion incorporating a tricarboxylic calixarene molecule, as a specific chelating agent for uranium, was previously developed. The work achieved in this thesis aimed at evaluating the ex vivo and in vivo decontamination efficiency of this new emergency treatment on intact and superficially wounded skin. For this purpose, skin excoriation model was used. Reproducible models of superficial wounds consisting of micro-cuts and micro-punctures were also developed in order to evaluate the efficiency of the nanoemulsion on physical wounds such as incisions. These studies showed that the calixarene nanoemulsion could be an efficient decontaminant treatment, less aggressive than using the current treatment: soaped water. Its potential cutaneous toxicity was evaluated on in vitro reconstructed human epidermis using three different toxicity tests (MTT, LDH and IL-1-α). These studies demonstrated that the calixarene nanoemulsion did not induce skin toxicity even after 24 h of exposure time.

Dada a importância do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT), a necessidade de uma eficiente caracterização bacteriológica dos focos como ponto fundamental do sistema de vigilância e as dificuldades encontradas pelos laboratórios quanto aos métodos de isolamento de Mycobacterium bovis fizeram crescer o interesse do meio científico por estudos, sobretudo moleculares, de isolados M. bovis. Para a realização dessas técnicas moleculares, é necessária abundância de massa bacilar, obtida através da manutenção dos isolados em laboratório e repiques em meios de cultura. Entretanto o crescimento fastidioso do M. bovis em meios de cultura traz grandes dificuldades para essas operações. Assim sendo, o presente estudo teve por objetivos: 1º) Comparar três métodos de descontaminação para homogeneizados de órgãos, etapa que precede a semeadura em meios de cultura, onde 60 amostras de tecidos com lesões granulomatosas, provenientes de abatedouros bovinos do Estado de São Paulo, foram colhidas e imersas em solução saturada de Borato de Sódio e transportadas para o Laboratório de Zoonoses Bacterianas do VPS-FMVZ-USP, onde foram processadas até 60 dias após a colheita. Essas amostras foram submetidas a três métodos de descontaminação: Básico (NaOH 4%), Ácido (H2SO4 12%) e 1- Hexadecylpyridinium chloride a 1,5% (HPC) e o quarto método foi representado pela simples diluição com solução salina (controle). Os resultados foram submetidos à comparação de proporções, pelo teste de &#967;², na qual verificou-se que o método HPC foi o que apresentou menor proporção de contaminação (3%) e maior proporção de sucesso para isolamento de BAAR (40%). 2º) Comparar três diferentes meios criopreservates para M. bovis, foram utilizados 16 isolados identificados pela técnica de spoligotyping. Cada um desses isolados foi solubilizado em três meios (solução salina, 7H9 original e 7H9 modificado), e armazenado em três diferentes temperaturas (-20ºC, -80ºC e -196ºC), sendo descongelado em três diferentes tempos (45, 90 e 120 dias de congelamento). Antes do congelamento e após o descongelamento foram feitos cultivos quantitativos em meios de Stonebrink Leslie. Os porcentuais de redução de Unidades Formadoras de Colônias (UFC) nas diferentes condições foram calculados e comparados entre si através de métodos paramétricos e não-paramétricos. Os resultados obtidos foram: na análise da variável tempo, em 90 dias de congelamento foi observada uma maior proporção de perda de M. bovis, quando comparado ao tempo 120 dias (p=0,0002); na análise da variável temperatura, foi observada uma diferença estatística significativa entre as proporções de perda média nas temperaturas de -20ºC e -80ºC (p<0,05); na análise da variável meio, foi observada uma diferença significativa (p=0,044) entre os meios A e C, para 45 dias de congelamento e -20ºC de temperatura de criopreservação. Embora as medianas dos porcentuais de perdas de UFC terem sido sempre inferiores a 4,2%, permitiram sugerir que o melhor protocolo de criopreservação de isolados de M. bovis é solubilizá-los em 7H9 modificado e mantê-los à temperatura de -20ºC / In the context of the National Program of Control and Eradication of Brucellosis and Tuberculosis (PNCEBT), the necessity of an efficient bacteriologic characterization of the infected herds as a cornerstone of the monitoring system and the difficulties faced by the laboratories regarding the methods for Mycobacterium bovis isolation led to a growing interest for scientific studies, especially molecular, of M. bovis isolates. To use these molecular techniques it is necessary to have an abundant bacillary mass, obtained through the maintenance of isolates in laboratory and replication in culture media. However the fastidious growth of M. bovis in culture media brings out great difficulties for these activities. Thus, the present study has the following objectives: First, to compare three decontamination methods for organ homogenates, phase that precedes the sowing in culture media, 60 samples of tissues with granulomatosis injuries, proceeding from bovine slaughterhouses in the State of São Paulo, were obtained, immersed in sodium borato saturated solution and transported to the Laboratório de Zoonoses Bacterianas of the VPS-FMVZ-USP, where they were processed up to 60 days after the sampling. These samples were submitted to three methods of decontamination: Basic NaOH 4%, Acid (H2SO4 12%) and 1- Hexadecylpyridinium chloride (HPC) 1.5% and a simple dilution with saline solution (control method). The results were analysed by means of the &chi test to compare proportions, and it was verified that HPC method presented the smallest proportion of contamination (3%) and the greatest proportion of success for M. bovis isolation (40%). Second, to compare three different cryopreservation media for M. bovis, 16 isolates identified by the technique of spoligotyping were used. Each one of these isolates was solubilized in three media (original saline solution, 7H9 and 7H9 modified), and stored in three different temperatures (-20ºC, -80ºC and -196ordm;C), and defrosted in three different time periods (45, 90 and 120 days of freezing). Before the freezing and after the unfreezing, quantitative cultivations in Stonebrink Leslie media were carried out. The proportions of Colony-Forming Units (CFU) loss in the different conditions were calculated and compared with one another through parametric and non-parametric methods. The results obtained were: in the analysis of the variable time, at 90 days of freezing a bigger proportion of CFU loss was observed when compared to 120 days (p=0,0002); in the analysis of the variable temperature, a statistically significant difference was observed between the average proportions of CFU loss for the temperatures of -20ºC and -80ºC (p<0,05); in the analysis of the variable media, a significant difference was observed (p=0,044) between the media A and C, for 45 days of freezing and -20ºC of cryopreservation temperature. Althougth the medium ones of the proportion of losses of CFU to always have been inferior 4,2%, had allowed to suggest that the best protocol for cryopreservation of M. bovis isolates is to solubilize them in 7H9 modified medium and to keep them at a temperature of -20ºC