The theoretical part of the thesis is divided in the five main chapters. First chapter generally defines the term ?contamination?, which is closely related to the decontamination. Second chapter is focused on the microbial contamination, defines infectious agents, the mechanisms of its spread and points out microbes most likely to be misused as a biological weapon. Third chapter of the theoretical part addresses the decontamination itself. It defines the decontamination in general aiming for microbial decontamination, i.e. the biological agent surface and environment removal. Two basic groups of decontamination e.g. physical and chemical procedures are mentioned in this chapter. Fourth chapter presents valid standards describing various types of examinations for the desinfecting preparations efficacy evaluation. The last chapter of the theoretical part covers large-scale facilities, defines them and describes experimental facilities at the Department of large-scale testing in the National institute for nuclear, chemical and biological protection in Kamenna. The aim of the thesis is to create own methodology of the disinfactant efficiency testing with emphasis on the real condition simulation. The decontamination was carried out by means of following methods: aerosol generator method, decontamination by evaporation, manual sprayer and engine sprayer respectively. For the analysis we chose these disinfactants: Chirosan? Plus, Incidin OxyDes, Kohrsolin? FF, Korsolex? basic, Persteril? 36 and Sanosil? Super 25 Ag. The disinfectant efficiency was tested in 30, 60 and 120 m3 facilities. The tested B-agents were: Bacillus anthracis, Bacillus atrophaeus, Escherichia coli, Staphylococcus aureus and Streptococcus pneumoniae. For the decontamination efficiency verification we used a modified surface assay, a modified suspension assay and an agarose assay. The following issues were of concern: We found out that, based on the analysis of different techniques of decontamination, the large-scale facility decontamination is feasible. The most effective method of decontamination appears the manual and engine spraying with the latter more convenient for spaces over the 30 m3 in the matter of time. The decontamination by evaporation was inefficient for the large-scale facility decontamination. Experiments revealed that method based on aerosol distribution is convenient for large-scale facility decontamination. However, the nature of decontaminating preparation has to be considered. All disinfactants tested have shown good bactericidal activity. Nevertheless, only Korsolex? basic a Persteril? 36 have shown sufficient sporicidal activity with equal effect. The disadvantage of the preparation Persteril? 36 is its significant corrosive activity, on the other hand is inexpensive. The preparation Korsolex? basic contains anticorrosive compounds, but is more expensive than Persteril? 36. Our results demonstrated that the most convenient method for the decomtamination of facilities up to the 30 m3 is the manual aerosol sprayer. For facilities of larger volume the engine sprayer decontamination is more convenient. From the preparations tested, only Korsolex? basic a Persteril? 36 are, with respect to advantages and disadvantages, usable in the Department of large-scale testing in the National institute for nuclear, chemical and biological protection in Kamenna.

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This thesis consists of three main parts. In the first part, the general procedures and principles of decontamination, the decontamination substances and mixtures, as well as the technique that is used for decontamination, have been described. Next, the description of the decontamination of the first responders, as this part of the decontamination process has been given; it has been elaborated and checked in practice. In the second part, there is the drafted plan of the mass extemporaneous decontamination of people in the selected public facility, according to the general methods of decontamination. Next, there is the drafted documentation for resolving an extraordinary event: an accident with subsequent leakage of a dangerous chemical substance, resulting in the need to carry out the mass decontamination of people in selected public facilities. In the last part, I have discussed and compared the advantages and possible disadvantages of the selected object of for the implementation of mass decontamination of people. There is also a suggested way of how to carry out this decontamination in this object in the most efficient way. The processing of this documentation has clearly confirmed my hypothesis. The mass decontamination of people in the selected location, in this case specifically in the Domazlice Hostel and Swimming Pool, can be feasibly performed with an improvised method. After giving the specific supplemented data (the names of the personnel, the telephone numbers, places of storage material etc.), the suggested documentation is usable for the object in practice. The documentation can be also easily adapted to other similar public facilities.

O objetivo do trabalho foi avaliar a capacidade de cepas de bactérias ácido-láticas (BAL) em remover a aflatoxina M1 (AFM1) em solução tampão fosfato salina (TFS) e em amostras de leite. Nos ensaios com TFS, verificou-se a influência do tempo de contato (15 min. ou 24 horas) entre as células de sete cepas de BAL e AFM1, as diferenças entre a eficiência de remoção das bactérias viáveis e inviabilizadas termicamente, e a estabilidade do complexo BAL/AFM1 formado. As três cepas de BAL com maior percentual (> 33%) de remoção da AFM1 nos ensaios com TFS foram re-avaliadas utilizando-se leite UHT (ultra-high-temperature) desnatado artificialmente contaminado com AFM1. Para isso, foram utilizadas somente células inviabilizadas termicamente, verificando-se o efeito da temperatura (4ºC ou 37ºC) sobre a capacidade de remoção da toxina por 15 minutos. A remoção média da AFM1 pelas cepas de BAL em TFS variou entre 5,60±0,45 e 45,67±1,65% (n=3), sendo que as células inviáveis obtiveram percentuais de remoção de AFM1 significativamente maiores que as células viáveis, em ambos os tempos de contato analisados (15 min. ou 24 horas), não havendo diferença significativa entre os tempos. Observou-se que o complexo BAL/AFM1 obtido nos ensaios com TFS é instável, pois 40,57±4,66 a 87,37±1,82% da AFM1 retida pela bactéria foram recuperados em solução após a lavagem do complexo com TFS. As três cepas de BAL com maior percentual de remoção da AFM1 em TFS (Lactobacillus rhamnosus, Lactobacillus delbrueckii spp. bulgaricus e Bifidobacterium lactis) não apresentaram diferenças significativas nos ensaios com leite UHT a 37ºC. Somente B. lactis apresentou maior capacidade de remover a AFM1 do leite UHT a 4ºC. Os resultados demonstraram que a remoção de AFM1 empregando-se as BAL em alimentos é viável para reduzir as concentrações da toxina a níveis seguros. Entretanto, estudos adicionais são necessários a fim de investigar os mecanismos envolvidos na remoção da toxina pelas BAL com vistas à aplicação em indústrias de alimentos. / The purpose of this study was to evaluate the ability of strains of lactic acid bacteria (LAB) to remove aflatoxin M1 (AFM1) in phosphate buffer saline (PBS) and in milk samples. In the assays with PBS, the influence of contact time (15 min. or 24 hours) between the cells of seven LAB strains and AFM1 was evaluated, as well as the differences between the removal efficiency of viable and non-viable (heat-killed) bacteria, and the stability of AFM1/LAB complex produced. The three LAB strains with the highest percentage (> 33%) of AFM1 removal in the tests with PBS were reevaluated using UHT (ultra-high-temperature) skimmed milk spiked with AFM1. For these assays, only non-viable bacterial cells were used for checking the effect of temperature (4ºC or 37ºC) on the toxin removal capacity during 15 min. The mean AFM1 removal by LAB strains in PBS ranged from 5.60±0.45 and 45.67±1.65% (n=3). Non-viable cells showed AFM1 removal percentages significantly higher than viable cells in both contact times (15 min. or 24 hours), although there were not significant differences between these contact times. The AFM1/LAB complex resulted from the tests with PBS was unstable, as 40.57±4.66 to 87.37±1.82% of AFM1 retained by the bacteria were recovered in solution after washing the complex with PBS. The three LAB strains with the highest percentage of AFM1 removal in the PBS assays (Lactobacillus rhamnosus, Lactobacillus delbrueckii spp. bulgaricus and Bifidobacterium lactis) showed no significant differences in the UHT skimmed milk assays at 37ºC. Only B. lactis had greater ability to remove AFM1 in UHT milk at 4ºC. The results demonstrated that the removal of AFM1 by using LAB in foods is viable to reduce the toxin concentrations until safe levels. However, additional studies are needed to investigate the mechanisms involved in the toxin removal by LAB aiming its application in food industries.

O trabalho teve por finalidade avaliar o efeito da adição de cepa de bactéria ácido-lática e biomassa de Saccharomyces cerevisiae resultante da fermentação alcoólica de cerveja em rações sobre o desempenho de frangos de corte intoxicados com aflatoxina B1 (AFB1). Para este fim, selecionou-se a cepa bacteriana (Lactobacillus rhamnosus, LYO) que apresentou maior crescimento celular médio, sendo seca posteriormente. O resíduo da fermentação alcoólica (RFC) contendo S. cerevisiae foi obtido em microcervejaria, seco e triturado. Preliminarmente, foram realizados ensaios in vitro para avaliação da capacidade de remoção da AFB1 pela LYO (inviabilizada por tratamento térmico a 121ºC por 15 minutos, em solução ou seca por atomização ou por liofilização) e RFC em solução tampão (pH 3,0 e 6,0) a 25ºC. O experimento in vivo foi conduzido utilizando-se 200 pintos machos de um dia de idade, provenientes de matrizes comerciais de frangos de corte. As aves foram distribuídas aleatoriamente em 8 tratamentos com 25 aves cada, em arranjo fatorial 2 x 2 x 2, correspondendo a dois níveis (0 e 2.000 µg/kg) de incorporação de AFB1 às rações e dois níveis (0 e 1%) de incorporação de LYO (seca por atomização) e RFC. As aves receberam as rações experimentais do 1º ao 21º dia de vida. Semanalmente, avaliou-se o ganho de peso (GP), consumo de ração (CR) e a conversão alimentar (CAL), e ao término do experimento, determinou-se a viabilidade das aves sobreviventes comparado ao total. Análises de bioquímica sérica aos 14 e 21 dias das aves incluíram a determinação de proteína total (PT), glicose (GLI), albumina (ALB), globulina (GLO), cálcio (CA), ácido úrico (AU) e as enzimas aspartato-amino-transferase (AST) e gama-glutamil-transferase (GGT). Ao final do período de intoxicação, todas as aves foram sacrificadas e pesadas, determinando-se o peso relativo do fígado, rins e bursa de Fabricius, além das análises histopatológicas desses órgãos. Nos ensaios in vitro, LYO em solução conseguiu remover a AFB1 em 46,0% e 35,8% em pH 3,0 e 6,0, respectivamente, enquanto que RFC mostrou-se capaz de adsorver a AFB1 em 50,5% e 48,5% nos respectivos valores de pH. No entanto, LYO atomizada perdeu completamente sua capacidade de adsorção, enquanto que LYO liofilizada conseguiu manter a sua habilidade em adsorver a AFB1 em 36,6% em pH 3,0 e 27,2% em pH 6,0. No estudo in vivo, os tratamentos com LYO isolada ou associada com RFC não apresentaram efeitos satisfatórios sobre as variáveis estudadas nos frangos de corte, exceto pelo aumento no CA, ALB, PT e GLO. O RFC resultou em uma maior amenização dos efeitos nos índices de produtividade e bioquímica sérica, como aumento do GP, ALB, PT, GLO e CA, além de diminuição das alterações histopatológicas, em comparação ao grupo controle. Os experimentos demonstraram que o RFC apresenta potencial para uso como aditivo anti-aflatoxinas para frangos de corte, e que novos estudos são necessários para viabilizar a produção em escala de LYO liofilizada para uso na alimentação animal. / The study aimed to evaluate the effect of adding strain of lactic acid bacteria and biomass of Saccharomyces cerevisiae from the beer alcoholic fermentation in diets on performance of broilers intoxicated with aflatoxin B1 (AFB1). For this purpose, the bacterial strain (Lactobacillus rhamnosus, LYO), which presented higher cell growth, was selected and subsequently dried. The residue of alcoholic fermentation (RAF) containing S. cerevisiae was obtained in microbrewery, dried and ground. Preliminarily, it was accomplished in vitro tests to evaluate the capacity of AFB1 removal by LYO (non-viable by heat treatment at 121ºC for 15 minutes, in solution or dried by spray-drying or lyophilization) and RAF in buffer solution (pH 3.0 and 6.0) at 25ºC. The in vivo test was conducted using 200 one day old male chicks, from arrays of commercial broilers. The birds were randomly assigned to 8 treatments with 25 birds each in a factorial 2 x 2 x 2, corresponding to two levels (0 and 2,000 µg/kg) of AFB1 incorporation to feed and two levels (0 and 1%) of incorporation of LYO (spray-dried) and RAF. The chicks received the experimental diets from 1st to 21st days of life. Weight gain (WG), feed intake (FI) and calculation of feed conversion (FC) were assessed weekly and at the end of the experiment, it was determined the viability of the surviving broilers compared to the total. Serum biochemical analysis at days 14 and 21 included the determination of total protein (TP), glucose (GLU), albumin (ALB), globulin (GLO), calcium (CA), uric acid (UA) and the enzymes aspartate-aminotransferase (AST) and gamma-glutamyl-transferase (GGT). At the end of the intoxication period, broilers of all treatments were sacrificed and weighed, determining the relative weights of liver, kidneys and bursa of Fabricius, beside the histopathological analysis of these organs. in vitro tests showed that LYO in solution could bind 46.0% and 35.8% of AFB1 at pH 3.0 and 6.0, respectively, while RFC was able to bind 50.5% and 48.5% of AFB1 the respective pH values. However, atomized LYO completely lost its adsorption capacity, whereas lyophilized LYO could maintain its ability to bind AFB1 by 36.6% at pH 3.0 and 27.2% at pH 6.0. in vivo study results demonstrated that the treatments with LYO, isolated or associated with RFC, showed no satisfactory effect on the variables analyzed in broilers, except for an increase in CA, ALB, PT and GLO. The use of RFC resulted in greater mitigation of effects on performance and serum biochemistry parameters, such as increased GP, ALB, PT, GLO and CA, besides the reduction of histopathological changes, compared to the control group. The experiments demonstrated that RFC has potential for use as anti-aflatoxin additive for broilers, and further studies are necessary to enable the scale production of LYO lyophilized for use in animal feed.

Orientadores: Francisco Humberto Nociti Junior, Marcio Zaffalon Casati / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-05T21:22:40Z (GMT). No. of bitstreams: 1 Goncalves_PatriciaFurtado_D.pdf: 1584484 bytes, checksum: 320d77faa174f7b65746904c2c6166c6 (MD5) Previous issue date: 2006 / Resumo: Tendo sido demonstrada a possibilidade de preservação do cemento como forma alternativa de tratamento da doença periodontal, o presente estudo teve como objetivo avaliar o efeito do cemento dental na regeneração periodontal. Foram utilizados nove cães, sem raça definida, que receberam ligaduras de fio de algodão nos terceiros pré-molares inferiores, bilateralmente, para produzir lesões de bifurcação classe III. Após o levantamento de retalhos mucoperiosteais, os defeitos foram aleatoriamente escolhidos para receber um dos seguintes tratamentos: (A) remoção do tecido de granulação e uma cuidadosa raspagem e alisamento radicular, utilizando curetas e brocas, objetivando a remoção dos depósitos duros e moles e do cemento radicular, e polimento radicular com taças de borracha e pasta profilática; (B) somente a remoção do tecido de granulação e polimento radicular com taças de borracha e pasta profilática, objetivando a preservação do cemento radicular. Em ambos os grupos, os defeitos foram tratados com o princípio da Regeneração Tecidual Guiada (membranas reabsorvíveis - RESOLUT XT®) e os retalhos foram posicionados coronariamente e suturados. Após quatro meses os animais foram sacrificados para uma análise histomorfométrica. No grupo B, houve uma maior extensão (3,59±1,67 e 6,20±2,26mm para os grupos A e B, respectivamente; p=0,004) e espessura (18,89±9,47 e 52,29±22,48µm para os grupos A e B, respectivamente; p=0,001) de novo cemento, uma maior extensão de novo osso (1,86±1,76 e 4,62±3,01mm para os grupos A e B, respectivamente; p=0,002), além de uma maior extensão de regeneração periodontal, quando comparado ao grupo A (p<0,05 ¿ Wilcoxon). Observou-se ainda, no grupo B, uma menor formação de tecido epitelial/conjuntivo sobre a superfície radicular (2,77±0,79 e 1,10±1,48mm para os grupos A e B, respectivamente; p=0,009), além de uma maior porcentagem de preenchimento da área do defeito (80,95±17,46 e 94,50±10,27 para os grupos A e B, respectivamente; p<0,05), com uma maior proporção de novo osso e uma menor proporção de tecido epitelial/conjuntivo (p<0,05). Dentro dos limites deste estudo, conclui-se que o cemento dental pode modular a regeneração periodontal / Abstract: The aim of the present study was to histometrically evaluate, in dogs, the effect of root cementum on periodontal regeneration. Nine mongrel dogs were used to obtain bilateral chronic class III furcation defects by placing cotton ligatures around both third mandibular premolars. The teeth were randomly assigned to receive one of the following treatments: (A) Scaling and root planning, by means of hand and rotatory instruments, in order to remove soft and hard deposits as well as all root cementum; (B) Removal of only soft microbial deposits, by polishing the root surface with rubber cups and polishing paste, aiming at maximum root cementum preservation. Both groups were treated with Guided Tissue Regeneration (GTR), with the use of resorbable poliglycolic-lactic-acid membranes (RESOLUT XT®) covering the buccal e lingual aspects of the furcation defect, and the flaps coronally positioned. Four months after the treatment, the animals were sacrificed and the histological sections obtained. Data analysis showed that a superior length (3.59±1.67 vs. 6.20±2.26mm; p=0.004) and a thicker layer (18.89±9.47 vs. 52.29 ±22.48µm; p=0.001) of new cementum were observed for group B (p<0.05). Additionally, group B presented and a superior length of new bone (1.86±1.76 vs. 4.62±3.01mm; p= 0.002) and less soft tissue along the root surface (2.77±0.79 vs. 1.10±1.48mm; p=0.009), and a larger proportion of periodontal regeneration when compared to control (p<0.05). A larger area of new bone (p=0.018) and a smaller area of soft tissue (p=0.024) were also observed for group B (p<0.05 - Wilcoxon). Within the limits of this study, it can be concluded that root cementum may modulate periodontal regeneration / Doutorado / Periodontia / Doutor em Clínica Odontológica