Orientadores: Brenda Paula Figueiredo de Almeida Gomes, Rogerio de Castilho Jacinto / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-13T00:24:34Z (GMT). No. of bitstreams: 1 Montagner_Francisco_M.pdf: 40440080 bytes, checksum: 91c7e1c3b65d68cb2d0c5c34049c40ea (MD5) Previous issue date: 2009 / Resumo: Os objetivos do presente estudo foram analisar a microbiota endodôntica em dentes que se apresentavam com necrose pulpar e abscesso periapical; investigar a presença de enterococos, fungos e enterobactérias na saliva e nos canais radiculares; e, determinar in vitro a suscetibilidade antimicrobiana de bactérias anaeróbias estritas isoladas com método de E-test e produção de _-Lactamases. Coletas microbiológicas de 30 canais radiculares e 30 amostras de saliva dos mesmos pacientes foram processadas e identificadas empregando-se métodos de cultura microbiana e testes bioquímicos. Análise estatística foi realizada através dos testes Qui-quadrado de Pearson ou de Fisher. Dos 159 microrganismos isolados nos canais radiculares, predominaram anaeróbios estritos e bactérias Gram-positivas, sendo Peptostreptococcus micros a espécie mais freqüentemente isolada. Fungos, enterobactérias e enterococos foram pouco isolados dos canais radiculares, sendo mais freqüentemente observados em amostras de saliva. Associações de espécies e sinais e sintomas de origem endodôntica foram encontrados. Penicilina G, amoxicilina, amoxicilina + ácido clavulânico, metronidazol foram bastante efetivos frente aos microrganismos testados. Todas as cepas de P. buccae, P. intermedia/nigrescens, P. disiens, P. micros e P. propionicum foram sensíveis à amoxicilina e ao metronidazol. F. nucleatum demonstrou ser bastante resistente aos antibióticos lactâmicos. Observou-se proporção variável de cepas capazes de produzir lactamases. Os resultados sugerem que a infecção endodôntica primária associada a casos sintomáticos é mista, com predomínio de anaeróbios estritos. A presença de fungos, enterococos e enterobactérias não foi significativa tanto nos canais radiculares quanto nas amostras de saliva. Os antibióticos testados mostraram-se efetivos, no entanto resistência microbiana foi detectada em taxas variáveis de acordo com a espécie estudada. / Abstract: The aim of the present study was to analyze the endodontic microbiota of teeth with pulp necrosis and spontaneous pain; detect the presence of enterococci, fungi and enteric bacteria in saliva and root canals; and, to determine, in vitro the antimicrobial sensitivity of anaerobic isolates by E-test and _-lactamasis production. Thirty samples were collected from both root canals and saliva, and processed using strict microbiological techniques. Statistical analysis was performed with Persons X2 test or Fisher's Exact Test, as appropriated. A hundred and fifty nine strains were isolated from the root canals, with a predominance of strict anaerobes and gram-positive bacteria, and Peptostreptococcus micros was the most frequently isolated species. Fungi, enterobacteria and enterococci were seldom found in root canals. There were no Enterobacteria in both places. Positive associations between specific species and signs and symptoms of endodontic origin were present. Penicillin G, amoxicillin, amoxicilin + clavulanate, and metronidazole were effective against the tested microorganisms. All strains of P. buccae, P. intermedia/nigrescens, P. disiens, P. micros e P. propionicum were susceptible to amoxicillin and metronidazole. About 12% to 33% of the isolates were able to produce beta-lactamasis. The present results suggested that the primary endodontic infection in symptomatic teeth was mixed, and was predominantly formed by strict anaerobic strains. Fungi, enterococci and enterobacteria were not significantly found in root canals and saliva. The tested antibiotics were effective, however resistance was detected in several clinical isolates. / Mestrado / Endodontia / Mestre em Clínica Odontológica

Orientador: Francisco Jose de Souza Filho / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-13T16:43:23Z (GMT). No. of bitstreams: 1 Rabang_HelenaRosaCampos_D.pdf: 2819542 bytes, checksum: 75016e34f1b77318e9fd8f990bdad508 (MD5) Previous issue date: 2009 / Resumo: Os objetivos deste trabalho foram: investigar microbiologicamente, por método de cultura e pelo método do Checkerboard DNA-DNA hybridization, canais radiculares antes (C1) e após preparo químico-mecânico (C2); e, analisar radiograficamente, lesões periapicais induzidas em dentes de cães, avaliando a possível relação entre comprimento de dente e tamanho de lesão periapical. Foram utilizados 21 pré-molares inferiores de quatro cães. Os dentes foram acessados, as polpas removidas e permaneceram expostos ao meio oral por 120 dias. Após este período, as imagens das reações periapicais induzidas foram avaliadas pelo software Imagelab 2.4, mensurando área e perímetro. O preparo químico-mecânico foi procedido, sendo os dentes divididos em três diferentes grupos, de acordo com a substância química-auxiliar testada: G1- NaCl 0,09%; G2- Clorexidina 2% gel; e G3- NaOCl 5,25% + EDTA 17%. Em cada uma das amostras foi investigada a microbiota por método de Cultura que propicia crescimento de bactérias anaeróbias estritas; e, a presença de até 40 espécies bacterianas foi investigada pela técnica de Checkerboard DNA-DNA hybridization. As médias das áreas das imagens radiográficas das lesões foram de 0,06 cm2, para os segundos pré-molares, 0,12 cm2 para os terceiros pré-molares e 0,18 cm2 para os quartos pré-molares. Houve associação entre os dentes de maior comprimento com as maiores áreas de lesão periapical. Em C1 foram isoladas 60 bactérias cultiváveis, pertencendo a 32 diferentes espécies, variando de 0 a 9 espécies por canal, sendo 70% anaeróbios facultativos e 18% anaeróbios estritos, com predominio de Gram-positivos (73%). Os gêneros bacterianos mais freqüentemente isolados foram: Streptococcus, Staphylococcus, Neisseria, Propionibacterium, Actinomyces e Prevotella. Pelo Checkerboard foram detectadas 158 bactérias, pertencendo a 29 diferentes espécies, variando de 0 a 19 espécies por canal, sendo 67% anaeróbios estritos, com predomínio de Gram-negativos (67%). Os microrganismos mais frequentemente encontrados foram Campylobacter gracilis, Capnocytophaga sputigena, Leptotrichia bucallis, Prevotela intermedia, Streptococcus gordonii e Gemella morbilorum. Em C2 todos os canais apresentaram ausência de crescimento microbiano pela Cultura, porém, pelo Checkerboard foram detectadas 33 bactérias, pertencentes a 9 diferentes gêneros e 11 diferentes espécies, variando de 1 a 8 por canal, em 10 dos 21 dentes analisados. Gemella morbilorum, Capnocytophaga sputigena, Leptotrichia bucallis e Fusobacterium nucleatum sp. Polymorphum (não detectada em C1), foram detectadas com maior freqüência após o preparo químico mecânico. No G1 houve uma redução 89% de células bacterianas. A utilização de clorexidina gel 2% (G2) e do NaOCl 5,25 (G3) permitiram redução de 97% e 99%, respectivamente, não diferindo entre si, ao nível de significância de 5%. Concluíu-se que, a microbiota de canais radiculares de dentes de cães com lesão periapical induzida, por exposição ao meio oral, após 120 dias, apresenta grande número de espécies; que o preparo químico mecânico promoveu uma grande redução de microrganismos; e que os resultados sugerem existência de relação positiva entre comprimento de dente e tamanho de lesão periapical / Abstract: The objectives of this study were: to investigate, by Culture and by Checkerboard DNA-DNA hybridization, microorganisms from infected root canals before (S1) and after chemomechanical preparation (S2), and analyse radiographically induced periapical lesions in dog's teeth, for a possible relation between tooth length and lesion size. Twenty one lower premolars of four mongrel dogs were used. Their chambers were accessed and their pulps removed and left open to the oral environment for the total period of 120 days. The radiographs were evaluated by Imagelab 2.4 software to measure the area and perimeter images of the induced periapical lesions. Chemomechanical preparation was proceded, the teeth were assigned to three groups, according to the substance tested: G1- NaCl 0.09%; G2- chlorhexidine gel 2%; G3-. NaOCl 5.25% + EDTA 17%. Methodology for handling, culture and incubation for growth of strict anaerobe species was used; and, 40 bacterial species were investigated in each one of the samples by Checkerboard DNA-DNA hybridization. Measurements of the radiographic image data of the lesions were 0.06 cm2 for the second premolars, 0.12 cm2 for the third premolars and 0.18 cm2 for the fourth premolars. There was a positive association between tooth length and size of periapical lesion. In S1, a total of 60 cultivable isolates were recovered from 32 different species, ranging from 0 to 9 per canal. Facultative anaerobe species comprised 70% of the samples and 18% were strict anaerobic species with one microbiota predominantly Gram-positive (73%). The most frequent genera recovered from the canals were: Streptococcus, Staphylococcus, Neisseria, Propionibacterium, Actinomyces and Prevotella. One hundred and fifty eight bacteria were detected from 29 different species, ranging from 0 to 19 per canal. Strict anaerobe species comprised 67% of the samples with one microbiota predominantly Gram-negative (67%). The most frequent species detected by Checkerboard from the canals (S1) were: Campylobacter gracilis, Capnocitophaga sputigena, Leptotrichia bucallis, Prevotela intermedia, Streptococcus gordonii e Gemella morbilorum. In S2, no microbial growth was observed by culture. However, by checkerboard, 33 bacteria were detected, from 9 different genera and 11 differet species, ranging from 1 to 8 per canal, in 10 of the 21 investigated teeth. Gemella morbilorum, Capnocytophaga sputigena, Leptotricchia bucallis e Fusobacterium nucleatum sp. Polymorphun (not detected in S1), were detected with higher frequency after chemomechanical preparation. There was 89% of bacteria reduction in G1. Use of chlorhexidine gel 2% (G2) and NaOCl 5.25% (G3) allow bacteria reduction of 97% and 99%, respectively, with no statistical difference, with 5% significance. In conclusion, the root canal microbiota of dog teeth with induced periapical lesion, by exposure to the oral environment, after 120 days presents a great number of different species; results suggest positive relationship between the tooth length and size of the periapical lesion. / Doutorado / Endodontia / Doutor em Clínica Odontológica

Orientador: Lucia Regina Durrant / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-15T18:41:33Z (GMT). No. of bitstreams: 1 Dias_ElisangelaFrancisconGuimaro_D.pdf: 1126460 bytes, checksum: 218340ed688851cd46f637d8f8b3cd16 (MD5) Previous issue date: 2010 / Resumo: Azocorantes são compostos aromáticos com um ou mais grupos azo (-N=N-). São os maiores e a mais importante classe de corantes sintéticos usados em aplicações industriais. Eles são considerados compostos xenobióticos recalcitrantes aos processos de biodegradação, e a presença destes azocorantes nos ecossistemas aquáticos é a causa de sérios problemas ambientais e relacionados com a saúde. Neste trabalho, a habilidade em degradar azocorantes de 62 linhagens bacterianas previamente isoladas de efluente industrial foi investigada. A seleção das linhagens foi realizada através de testes de descoloração visual em meio líquido com azocorantes suplementados com diferentes fontes de carbono. O processo de descoloração foi realizado sob condições microaerofílicas ou estáticas até que nenhuma cor fosse observada, seguido de agitação para promover a biodegradação dos metabólitos produzidos. A descoloração e a biodegradação dos azocorantes bem como dos metabólitos produzidos foram monitoradas por análises de UV-vis, Carbono Orgânico Total (COT), Espectroscopia no Infravermelho com Transformadas de Fourier (FTIV) e Cromatografía Líquida de Alta Eficiência e Espectrometria de Massa (CLAE-EM). A atividade de enzimas oxidativas (peroxidase, lacase and tirosinase) foram analisadas para verificar se estas estavam envolvidas no metabolismo de biodegradação dos azocorantes. Análises de toxicidade foram realizadas antes e após a degradação dos azocorantes utilizando o organismo teste Daphnia magna.As aminas aromáticas geradas da biodegradação dos azocorantes foram testadas com o propósito de obter a polimerização. A enzima lacase de Myceliophthora Thermophila foi usada para catalizar reações de acoplamento entre as aminas aromáticas produzidas. As análises de UV-vis mostraram que os corantes foram descoloridos (80%) em condições microaerofílicas ou estáticas. Não houve nenhuma mudança significativa na cor, no estágio aeróbio seguinte. O tempo de descoloração mostrou relação com o meio de cultura utilizado e com a estrutura química dos corantes. Os corantes monoazo foram descoloridos entre 8 a 120 hs. Os diazo e os triazo foram descoloridos após 120 e 168 hs. As linhagens bacterianas descoloriram os corantes somente quando o meio foi suplementado com glicose e piruvato ou extrato de levedura. Na ausência destes compostos, as culturas foram incapazes de descolorir, indicando um requerimento obrigatório de uma fonte suplementar de carbono para alcançar a descoloração. Resultados mostraram que, quando o meio foi incubado em condições microaerofílicas ou estáticas a redução no COT (Carbono Orgânico Total) foi menor do que em condições aeróbias, onde 70% de redução foi observado. A presença de altas concentrações de aminas aromáticas em condições microaerofílicas ou estáticas confirma que houve redução das ligações azo. Porém, houve a confirmação da oxidação destas aminas no estágio aeróbio, indicando que um proceso oxidativo foi responsável pela biodegradação dos metabólitos. Foi observada atividade de tirosinase para na linhagem de Brevibacterium sp, sugerindo o papel desta enzima no processo de descoloração dos azocorantes, não sendo observada atividade de lacase e peroxidase. Nas análises de Espectroscopia no Infravermelho com Transformadas de Fourier (FTIV) após condições microaerofílicas ou estáticas foram observadas bandas em regiões atribuidas a grupamentos amina. Estas bandas desapareceram no estágio aeróbio e foram observadas novas bandas nas regiões associadas com ácido carboxílicos e íons NH3+, confirmando mineralização parcial dos produtos de degradação dos azocorantes, bem como dos metabólitos do meio de cultura. Os metabólitos produzidos pela biodegradação do azocorante RR 198 foram analisados por CLAEEM, para tentar identificar alguns metabólitos desconhecidos. Entre os possíveis compostos produzidos da biodegradação do RR198, 4-cloro-N-o-toluil-1,3,5-triazina-2-amino; sódio 4-aminonaftaleno-2-sulfonado e 3,6 -dimetil-7-(o-toluildiazenil) naftaleno-1-amino, tiveram razoável semelhança com os metabólitos aromáticos encontrados na amostra. Após condições aeróbias, a intensidade dos íons presentes nestes metabólitos foram reduzidos. Estes resultados também foram confirmados pelas análises de FTIV e poderia ser explicado pela diminuição dos compostos aromáticos gerados nas condições microaerofílicas ou estáticas. Foi observado que após um longo período de tempo, a lacase catalizou a polimerização das aminas aromáticas presentes nas soluções descoloridas. Os produtos gerados precipitaram e adquiriram cor, como confirmado pelas análises de UV-Vis. Os tamanhos das partículas foram significativamente maiores após o tratamento com lacase, como mostra as análises de Espectroscopia de Correlação de Fótons. Todas as linhagens bacterianas usadas neste estudo foram capazes de descolorir e degradar os azocorantes em condições microaerofílicas ou estáticas. Em condições aeróbias, ocorreu parcial mineralização dos produtos de degradação dos azocorantes, bem como dos metabólitos do meio, como confirmado para o organismo teste Daphnia magna e Carbono Orgânico Total (COT). Após o estudo, estas bactérias foram identificadas através de análises de sequência de rDNA 16S como Staphylococcus arlettae, Klebsiella sp, Microbacterium sp, Leucobacter albus e Brevibacterium sp / Abstract: Azo dyes, which are aromatic compounds with one or more azo (-N=N-) groups, are the most important and largest class of synthetic dyes used in commercial applications. They are considered as xenobiotic compounds that are very recalcitrant to biodegradation processes. The presence of these dyes in the aqueous ecosystem are a cause of serious environmental and health concerns. In this work, the ability of 62 bacterial strains previously isolated from an industrial activated sludge process treating effluent containing azo dyes was investigated. The selection was undertaken, through visual decolorization, in liquid media with azo dyes supplemented with diferent carbon sources. Decolorization process was performed under microaerophilic or static conditions until no color was observed. The medium was then aerated to promote the biodegradation of the metabolites produced. The azo dyes decolorization and biodegradation and the aromatic amines produced were monitored by UV-Vis, Total Organic Carbon (TOC), Fourier Transformed Infra Red (FTIR) and High Performence Liquid Chromatography (HPLC- MS). Activity of the oxidoreductase enzymes (peroxidase, laccase and tyrosinase) was evaluated in cultures of the bacterial isolates. Acute toxicity tests with Daphnia magna (Crustacea, Cladocera) were carried out after and before microaerobic or static and aerobic conditions. The aromatic amines generated from the biodegradation of the azo dyes were tested for their ability to undergoing polymerization using a laccase from M. thermophila to catalyze the coupling reactions of the aromatic amines. The decolorization time showed a relationship with the culture medium and chemical structure of the dyes. The monoazo dyes were decolourized within 8 to 120 h. The diazo and triazo were decolourized required 120 to168 h, approximately. UV-Vis analysis showed complete decolorization (>80%) in the microaerophilic or static conditions. No significant color changes were detected in the following aerobic stage. However the bacterial strains could only decolourize the dyes effectively when the medium was supplemented with glucose and pyruvate or yeast extract. In the absence of these compounds, the cultures were unable to decolorize the dyes, thus indicating an obligate requirement for a supplementary carbon source for dye decolorization. When the medium was incubated under microaerophilic conditions, the reduction in TOC was low even after 7 days of incubation. Conversely, a significant increase in TOC reduction (>70%) was observed in the aerobic stage. The reduction of azo bonds is known to yield the production of high concentrations of amines in the microaerophilic or static stage therefore confirmed the azo bond was reduced. Therefore the oxidation of these aromatic amines was confirmed by the absence of amine in the aerobic stage indicating that an oxidative process was responsible by metabololite biodegradation. Tyrosinase activity was observed for Brevibacterium sp, suggesting the role of this enzyme in the decolorization process, but no-activity was observed for laccase and peroxydase. In the Fourier Transformed Infra Red (FTIR) analysis after microaerophilic or static decolorization, new bands were observed in region attributed to amine groups. These bands disappeared in the aerobic stage and a new broad region associated with carboxylic acid and NH3+ ions were observed. However, in the aerobic stage the partial mineralization of the dye degradation products and of the medium metabolites was confirmed. The decolorization products of the RR198 dye were analyzed by High Performence Liquid Chromatography (HPLC- MS) for tentative identification of the unknown metabolites tentating identifield compounds included, 4-chloro-N-o-tolyl-1,3,5-triazin-2-amine; sodium 4-aminonaphthalene-2-sulfonate and 3,6-dimethyl-7-(o-tolyldiazenyl) naphthalen-1-amine. After aerobic conditions the intensity of these metabolites was reduced. These results were also confirmed by FTIR and could be explained by degradation of these aromatics coumpounds previously generated in microaerophylic or static stage. After an extended period of time, laccase catalyzed polymerization of the aromatic amines in the destained solutions. The products generated precipitated spontaneously from the solution and acquired some color as confirmed by the UV-Vis analysis. The particle size was also significantly higher after laccase treatment as show by Photon Correlation Spectroscopy analysis (PCS). The bacterial strains used in this study were able to totally destain the azo dyes under microaerophilic or static condition. In the aerobic stage, partial mineralization of the dye decolorization products as well as of the medium metabolites was also confirmed by toxicity testing and TOC measurements. The strains were identified by 16S rDNA gene sequence analysis as Staphylococcus arlettae, Klebsiella sp, Microbacterium sp, Leucobacter albus e Brevibacterium sp. / Doutorado / Doutor em Ciência de Alimentos

Orientador: Lucia Regina Durrant / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-03T18:02:39Z (GMT). No. of bitstreams: 1 CuadrosOrellana_Sara_D.pdf: 8786475 bytes, checksum: 37002d082a1fea19d88bc66b7dced99e (MD5) Previous issue date: 2003 / Resumo: O metabolismo de compostos aromáticos depende de fatores físico-químicos como temperatura, pH e salinidade, e é bem conhecido e caracterizado em microrganismos mesófilos e em condições ambientais moderadas. No entanto, pouco se conhece sobre o metabolismo desses compostos em ambientes hipersalinos. este trabalho, cinco ambientes hipersalinos foram analisados quanto à presença de arqueas halofílicas capazes de metabolizar compostos aromáticos: Salar de Uyuni (Bolívia), salinas de Cahuil (Chile), salinas de Cabo Rojo (Porto Rico), sabkhas da região do Golfo Pérsico (Arábia Saudita) e Mar Morto (Israel e Jordânia). A estratégia empregada para o enriquecimento e isolamento de arqueas halofílicas capazes de crescer em presença de compostos aromáticos foi bem sucedida. Foram isoladas 12 linhagens capazes de crescer em presença de 1 ,2--benzoantraceno (2 mM) e 44 linhagens capazes de crescer em presença de ácido p-hidroxibenzóico (10 mM) como única fonte de carbono e energia. O isolado MM 17, proveniente de uma amostra de água do Mar Morto, apresentou o melhor resultado de crescimento e foi capaz de degradar completamente os ácidos benzóico (10 mM) e p-hidroxibenzóico (10 mM) após 200 h de cultivo. As análises bioquímica e genética dos isolados, juntamente com a análise dos perfis de lipídeos polares, indicaram que as linhagens estudadas pertencem a pelo menos dois gêneros: Haloferax e Halobacterium. Foi determinada a secreção de uma proteína de alto peso molecular por Haloferax volcanii em resposta à presença de 2 mM 1 ,2-benzoantraceno no meio de cultivo. Com o objetivo de estudar o mecanismo de secreção protéica em haloarqueas, o qual não é completamente entendido_ foram realizadas a clonagem molecular dos genes secD e secY de Haloferax volcanii, a expressão heteróloga em Escherichia coli e a purificação dos produtos gênicos, além de uma tentativa preliminar de obtenção de anticorpos policlonais através da imunização de coelhos / Abstract: The metabolism of aromatic compounds depends on physical-chemical factors such as temperature, pH and salinity, and is well known and characterized in mesophilic microorganisms under mild environmental conditions. Little is known, however, about the metabolism of these compounds in highly saline environments. Here, five hypersaline sites were tested for the presence of halophilic archaea able to metabolize aromatic compounds: the Uyuni Salt Marsh (Bolivia), the crystallizer ponds in Cahuil (Chile), the crystallizer ponds in Cabo Rojo (Puerto Rico), the sabkhas in the Persian Gulf (Saudi Arabia) and the Dead Sea (Israel and Jordan). The strategy used for the enrichment and isolation of halophilic archaea able to grow in aromatic compounds was successful. Twelve strains able to grow in 1,2-benzoantracene (2 mM) and 44 strains able to grow in p-hydroxybenzoic acid (10 mM) as the sole carbon and energy source were isolated. Strain MM17, isolated from a Dead Sea water sample, showed the best growth and was able to degrade benzoic (10 mM) and p-hydroxybenzoic (10 mM) acids afier 200 h of cultivation. Biochemical and genetic analyses of the isolates, together with the analysis of polar lipid profiles, indicate that the strains belong to at least two different genera: Haloferax and Halobacterium. The secretion of a high molecular weight protein by Haloferax volcanii following cultivation in 2 mM 1,2-benzoantracene was observed. To study the mechanism of protein secretion in halophilic archaea, a process that is not completely understood, preliminary studies were conducted, which included cloning of secO and secY genes of Haloferax volcanii, their heterologous expression in Escherichia coli and the purification of the gene products. In addition, a preliminary attempt to obtain polyclonal antibodies through rabbit immunization was made / Doutorado / Doutor em Ciência de Alimentos

No description available.

The ability to mount an efficient immune response should be an important life-history trait as parasitism can impact upon an individual's fecundity and survival prospects, and hence its fitness. However, immune function is likely to be costly as resources must be divided between many important traits. Whilst many studies have examined host resistance to particular parasite types, fewer have considered general immune responses. Studies that have considered general immune responses tend to do so in vertebrate models. However, the complexity of the vertebrate immune system makes the examination of evolutionary aspects of immune function difficult. Using larvae of the genus Spodoptera (Lepidoptera: Noctuidae) as a model system, this study examines' genetic and phenotypic aspects of innate immunity. The aims were to assess the levels of additive genetic variation maintained in immune traits, to consider possible costs that could maintain this variation, and to assess the role of phenotypic plasticity in ameliorating those costs. A key finding of this study was that high levels of additive genetic variation were maintained in all of the measured Immune traits. Analysis of the genetic correlations between traits revealed potential trade-offs within the immune system and between immune components and body condition. In addition, it was shown that larvae living at high densities invest more in immune function than those living in solitary conditions, suggesting that larvae can minimise the costs of immune function by employing them only when the risk of pathogenesis is high.

M. Tech. / Background: Waterborne disease contributes significantly to the total global disease burden. Populations in rural areas of South Africa depend on untreated waters for consumption and sanitation. Contamination of public water supplies by harmful bacteria such as pathogenic E. coli poses a major risk for public health. Ingestion of these pathogenic microorganisms present in the contaminated and untreated waters could cause infection, leading to systemic inflammatory responses manifested by the production of various proinflammatory cytokines. To date, there is no human system test available to detect whether water, following ingestion, would cause disease (i.e. whether the water is infectious). The current water testing methods only test for the presence of indicator organisms, such as faecal coliforms, total coliforms, and Escherichia coli. A reliable in-vitro bioassay that could assess whether the water would cause an inflammatory response was investigated in this study. Objectives: Pro-inflammatory cytokines and whole-blood have been used in similar studies to detect the inflammatory responses following exposure to specific stimulants such as dust, lipopolysaccharide (LPS), E. coli and various others. It has been reported that larger numbers of these contaminants induced higher levels of pro-inflammatory cytokine expression. This implies that the pro-inflammatory cytokine expression could be used as a marker of infection since, inflammation occurs in response to infection. Successful infection is thus necessary for inflammation to occur, and high levels of pro-inflammatory cytokine expression confirm that infection has occurred. Thus if pro-inflammatory cytokines could serve as indicators for infection, these cytokines could be used as indicators for bacterial pathogenicity of water.

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Good quality of drinking water is the foundation of a functioning society. Many Swedish municipals investigates ways to protect water supplies in convention areas. The purpose of this study was to investigate the risks of microbiological contamination of drinking water by private sewers and geothermal heat pumps. The purpose was also to investigate the risks of contamination associated with safety distances from energy wells and private sewers. Water samples were taken from private groundwater wells in the convention area of Hertsölandet in Luleå municipal, to test for microbiological contamination. To investigate safety distances, maps were made to locate private sewers, private groundwater wells and energy wells in the area. The result indicated that the groundwater wells was not contaminated by wastewater but that individual weaknesses in the groundwater wells contributed to microorganisms in the drinking water. However, too few samples were taken to provide a reliable result. The samples were taken at spring time and it should possibly had been more representative to take the samples after the summer season, when the use of water and production of wastewater has increased. Contaminations associated with safety distances to geothermal heat pumps and other groundwater wells could not be detected. The results shown that several groundwater wells were located too close to sewers according to safety distance. It is possible that the risks will increase as more people moves permanently to this area.

>Magister Scientiae - MSc / Extreme weather events in combination with geographical changes in groundwater utilization, groundwater availability, aquifer recharge, and ultimately changes in the quality of water resources, are expected in the future. As a consequence of changing weather patterns and urbanization the demand for groundwater is likely to increase in certain areas. We know that most waterborne pathogenic health epidemics are associated with contamination of farm water and wastewater. There is however limited understanding of the nature and extent of chemical, physical and biological processes that control the fate and transport of the microorganisms in primary and secondary aquifers. In this thesis, transport results are reported, where E. coli and PDR1 were selected as the biological tracers transported through a primary aquifer at the University of the Western Cape. In conjunction with the microbes salt and Rhodamine (chemical tracers) were injected to compare their fate and transport mechanism in the primary aquifer medium. A series of controlled Darcy experiments under laboratory and field conditions were conducted. Each provided a different data and information. The results from laboratory studies were used to improve design of the field studies. In both cases, the data collected provided information on fate and transport of microbes in groundwater. The field design phase of the experiment was an up-scaling of the laboratory phase of this project. The amount of chemical tracers injected into the aquifer was increased in proportion to the size of the research site. Tracer tests using chemical and microbial tracers were conducted simultaneously. Results of laboratory tests demonstrate a 5 times slower transport of microbes, compared to tests with salts during the laboratory phase. The salts at field scale show a breakthrough occurring after 2 days whereas the microbes –did not break through during the 28 days of the observation period. A new borehole was drilled closer to the pumping borehole to eliminate distance or travel time, but this had no effect on field results for the microbes. / National Research Foundation