Syfte: Syftet med denna studie var att studera huruvida algen Ascophyllum Nodosum (AN) utövar någon effekt på proteasaktivitet i oralt plack, samt om effekten finns i algen från början eller om man måste inta den oralt för att få en systemisk effekt.En förhöjd proteasaktivitet har förknippats med gingivit och parodontit. Vi förväntar oss en minskad proteasaktivitet hos försökspersonerna, och därmed en minskad risk för gingivit och parodontit, efter intag av AN under en månads tid. Material och metod: Ett in vitro med en pilotstudie, och ett in vivo försök utfördes. I in vitro försöket användes pulveriserat och upplöst Ascophyllum Nodosum. I in vitro studien deltog 5 personer i åldersgruppen 20 till 30 år. Plackprover på försökspersonerna togs före och efter intag av algen i 4 veckor. Under båda försökstillfällena fick försökspersonerna inte ha borstat tänderna på 12h innan försöket. Resultat: Vid kombinering av pilotstudien och in vitro studien ses ingen signifikant skillnad gällande Ascophyllum Nodosums proteasaktivitet i pulveriserad form. Våra resultat erhållna från in vivo studien visar att det har skett en ökad proteasaktivitet i placket hos försökspersonerna efter en månads intag av Ascophyllum Nodosum. Konklusion: Denna studie visar på en tendens till en ökad proteasaktivitet orsakad av Ascophyllum nodosum. Studien har inte undersökt vilka proteaser som påverkats. På grund av komplexiteten i den orala miljön och de många olika typerna av proteaser, behöver fler studier utföras för att studera de exakta effekterna på den orala miljön. / Aims: The purpose of this study was to investigate if the alga Ascophyllum Nodosum (AN) exerts any effect on protease activity in plaque, if such an effect is present in the algae from the beginning or if it has to be taken orally to exert a systemic effect.Increased protease activity has been associated with gingivitis and periodontitis. We expected a reduced protease activity, and thus a potentially reduced risk for gingivitis and periodontitis, after ingestion of AN for a month.Materials and methods: One in vitro trial with a pilot study, and one in vivo trial was carried out. In the in vitro trial pulverized and dissolved AN was used to make a solution, tested for protease activity.In the in vitro study 5 subjects, aged 20 to 30 years, participated. Plaque samples were taken before and after ingestion of the algae for 4 weeks. Subjects were instructed not to brush their teeth 12 h before sampling.Results: When combining the results from the pilot and in vitro studies, no AN protease activity could be detected. Our in vivo results showed an increased protease activity in the plaque after a month of AN intake.Conclusion: This study indicates a tendency to an increased protease activity caused by Ascophyllum Nodosum. However, the study did not examine which protease was affected. Because of the complexity in the oral environment and the many different types of protease, more studies need to be executed to study the exact effects of AN on the oral environment.

Fungal infections are one of the leading causes of death in humans, causing infections that range from mild superficial infections to severe, and life-threatening invasive infections that affect the bloodstream and vital organs. Invasive fungal infections have a high mortality rate, leading to approximately 1.5 million deaths annually. The most common pathogens responsible for these infections are Candida, Cryptococcus, and Aspergillus. Currently, treatment options for invasive fungal infections are limited to three main classes of antifungal drugs: azoles, polyenes, and echinocandins. The emergence of new fungal species, such as Candida auris, which displays high resistance and mortality rates (30-60%), has further complicated treatment efforts. Thus, there is a critical need for new therapeutic strategies to combat these life-threatening pathogens. C. auris isolates have demonstrated significant resistance, especially to azoles (fluconazole) and polyenes (amphotericin B, AmB). To address this, we screened approximately 2,600 FDA-approved drugs and clinical compounds to identify agents capable of inhibiting C. auris growth and enhancing or restoring the antifungal activity of existing antifungals. This screening revealed that HIV protease inhibitors, such as lopinavir, atazanavir, saquinavir and ritonavir, significantly enhanced the antifungal activity of azoles (fluconazole, voriconazole, itraconazole, and posaconazole) and polyene (AmB). Mechanistic studies showed that the HIV protease inhibitors inhibited the fungal efflux pump, and interfered with glucose utilization, leading to reduced ATP levels in C. auris. Moreover, HIV protease inhibitors, in combination with AmB, were able to inhibit the virulence factors of Candida species. Furthermore, HIV protease inhibitors, in combination with itraconazole or posaconazole, resulted in a significant reduction of the C. auris burden in mice kidneys. Two additional drugs, darapladib and rilapladib, phospholipase A2 inhibitors, were identified as potent inhibitors of C. auris. Darapladib and rilapladib demonstrated superior killing kinetics compared to itraconazole. Interestingly, C. auris did not develop any detectable resistance to both drugs at sub-inhibitory concentration over 16-passages. Mechanistic studies revealed that darapladib increased the plasma membrane permeability and caused DNA leakage, likely due to a direct interaction with ergosterol, as suggested by competition assays with exogenous ergosterol. We further validated the broad-spectrum, fungicidal, and potent activity of darapladib in combination with AmB, showing significant synergy against multidrug-resistant fungal pathogens, including C. auris, Aspergillus fumigatus and mucormycosis-associated species. Additionally, darapladib demonstrated a superior safety profile compared to AmB, exhibiting lower affinity for cholesterol, and no toxicity in combination with AmB to kidney cells. Finally, rilapladib (at 2× MIC) inhibited the C. auris burden by 1.5 Log10, whereas darapladib (at 2× MIC) achieved complete eradication of the C. auris burden in an in vivo C. elegans model. / Doctor of Philosophy / Fungi are a group of living organisms that are neither plants nor animals. While Some fungi are beneficial, others can cause serious diseases in humans. Candida auris is a human fungal pathogen that causes serious infections with mortality rate up to 60%. It has become a growing threat to global public health. This pathogen is particularly concerning due to its high virulence, resistance to multiple antifungal drugs, and ability to rapidly spread, often leading to hospital outbreaks. In our current research, we have identified several compounds that either inhibit the growth of C. auris and/or enhance the efficacy of existing antifungals, such as azoles and amphotericin B. We have identified HIV protease inhibitors (lopinavir, atazanavir, saquinavir, and ritonavir) that were able to enhance the antifungal activities of both azoles (fluconazole, itraconazole, voriconazole, and posaconazole) and polyene (amphotericin B) against multidrug-resistant C. auris. The combination of HIV protease inhibitors with amphotericin B inhibited the virulence factors of Candida species, including biofilm and hyphae formation. Two drugs darapladib and rilapladib were found to not only inhibit the growth of C. auris alone but also enhance the antifungal activity of amphotericin B against all medically important fungal pathogens tested. Additionally, we have explored the mechanisms by which these compounds act on the pathogen. Finally, we have confirmed the effectiveness of these drugs in live models, including mice and worms.

Xylitol har påvisats förebygga karies som orsakas av Streptococcus mutans, en viktig kariespatogen. Syfte: Syftet med denna litteraturstudie var att beskriva xylitolens påverkan på Streptococcus mutans. Metod: Studiedesignen utgjordes av en allmän litteraturstudie som sammanställer resultatet av tidigare forskning inom området. Sökningen genomfördes i databaserna Dentistry of Oral Sciences Source, Medline och Cinahl. Denna litteraturstudie grundas på vetenskapliga invitrostudier med kvantitativ ansats som valdes utifrån inklusions- och exklusionskriterier samt relevanta sökordskombinationer. En urvalsprocess av artiklar utfördes för att säkerställa en systematisk, noggrann samt reliabel presentation av urvalet. Endast artiklar av högkvalitet inkluderades i denna litteraturstudie. Resultat: Resultatet i denna litteraturstudie baserades på en sammanställning av femton vetenskapliga artiklar, som indikerade att xylitol har en inhiberande inverkan på Streptococcus mutans livsduglighet, tillväxt och biofilmbildning, acidogenitet, uppbyggnad, polysackariders kvantitet samt genuttryck. Slutsats: Sammantaget tyder resultatet på att xylitol medför antibakteriell effekt på Streptococcus mutans, genom att hämma dess metaboliska aktiviteter samt virulensfaktorer. / Xylitol has been shown to prevent dental caries caused by Streptococcus mutans, a major caries pathogen. Aim: To describe the impact of xylitol on Streptococcus mutans. Method: The study design included a general literature study summarizing the results of previous research in the chosen field. The research was conducted in the databases Dentistry of Oral Sciences Source, Medline and Cinahl. This literature study is based on scientific invitro studies with a quantitative approach that were selected based on inclusion and exclusion criteria and relevant keyword combination. An article selection process was carried out to ensure a systematic, accurate and reliable presentation of the sample. High-quality articles were included in this study. Results: The result of this literature study were based on fifteen scientific articles, which indicated that xylitol has an inhibitory effect on viability, growth and biofilm formation, acidogenicity, structure, polysaccharide amount and gene expression of Streptococcus mutans. Conclusion: The result of this literature study suggest that xylitol has an antibacterial effect on Streptococcus mutans by inhibiting its metabolic activities and virulence factors.

Salmonella enterica sérovar Typhi est une bactérie à Gram négatif qui cause la maladie systémique nommée fièvre typhoïde. Cette maladie affecte environ 9 millions de personnes par année et se propage par la voie fécale-orale par ingestion d’aliments ou d’eau contaminés. Ainsi, la fièvre typhoïde est particulièrement problématique dans les pays ayant des systèmes d’assainissement peu efficaces. Elle peut être traitée par des antibiotiques, mais comme pour de nombreuses bactéries, la résistance est de plus en plus commune. À la suite d’une infection aiguë, environ 5% des patients deviennent porteurs chroniques asymptomatiques grâce à la formation de biofilms dans la vésicule biliaire et excrètent la bactérie dans leurs selles continuellement. Étant le seul réservoir connu de S. Typhi, cet état constitue une source importante de persistance et de propagation de la maladie. L’état de porteur est intraitable par antibiotiques en raison de la haute tolérance aux stress des biofilms et une chirurgie pour enlever la vésicule biliaire est souvent la solution la plus efficace. Il est connu que les biofilms de Salmonella contiennent principalement de la cellulose, des fimbriae curli, de l'acide colanique et des protéines BapA. Cependant, S. Typhi a accumulé de nombreux pseudogènes au cours de son évolution humain-spécifique, dont certains sont impliqués dans la biosynthèse de composants communs de biofilm chez Salmonella, comme la cellulose et l'acide colanique. Il est également proposé que l'expression de curli soit dysfonctionnelle chez S. Typhi. La production de biofilms chez cette souche n'a jamais été caractérisée dans des conditions in vitro optimisées. Par conséquent, nous supposons que les biofilms de S. Typhi soient uniques dans leur production et leur composition. Cette étude caractérise la composition et la structure d’un biofilm de S. Typhi in vitro. Nous déterminons qu’aucun composant de biofilm communément identifié chez Salmonella ne joue un rôle majeur dans les biofilms de S. Typhi. Nous identifions un rôle important pour l’ADN extracellulaire et l’intégrité des LPS. / Salmonella enterica serovar Typhi is a Gram-negative rod-shaped bacterium that causes the systemic disease of typhoid fever. This disease affects about 9 million people per year and is spread through the fecal-oral route by ingestion of contaminated food or water. Thus, typhoid fever is particularly problematic in countries with poor sanitation systems. It is currently treatable by antibiotics but as with many other bacteria, resistance is becoming more and more common. Following acute infection, ~5% of patients become chronic asymptomatic carriers through biofilm formation in the gallbladder and continuously shed the bacteria in their feces. Being the only known reservoir of S. Typhi, this is an important source of persistence of the disease in endemic areas and propagation to new areas. The carrier state is untreatable by antibiotics due to the high stress tolerance of biofilms. Gallbladder-removal surgery is often the most efficient solution. Salmonella biofilms are known to primarily contain cellulose, curli fimbriae, colanic acid and BapA proteins. However, S. Typhi evolved to be human-specific over time and has accumulated many pseudogenes in the process, some of which are genes in the biosynthetic pathways of biofilm-related components of Salmonella, such as cellulose and colanic acid. It is also proposed that curli expression is dysfunctional in S. Typhi. Biofilm production in this strain has never been characterized in optimized in vitro conditions. Therefore, we hypothesized that S. Typhi biofilms are unique in their production and composition. This study characterizes S. Typhi biofilm composition and structure in vitro. We determine that no biofilm components commonly identified in other bacteria play a major role in S. Typhi biofilms. We identify an important role for extracellular DNA and lipopolysaccharide-layer integrity.

Der Einfluss des Biofilms, der Inkrustation, der Fließgeschwindigkeit und der Wasserqualität auf die Freisetzung von PAK wurde in Batch-, Drehkolbenreaktorversuchen und an einem Rohrleitungssystem untersucht. Chlorung von tauchgeteerten Edelstahlcoupons mit Biofilm führten zu einer fast vollständigen Ablösung des Biofilms und einer Zunahme der Freisetzung von Fluoren, Phenanthren und Fluoranthen. Die PAK-Freisetzung von tauchgeteerten Edelstahlcou-pons in einer Lösung von Trinkwasser mit 0,1% Natriumazid als Hemmstoff der Atmungskette, der den Biofilm nicht ablöst, führten zu einer Zunahme aller untersuchten PAK in der gleichen Größenordnung wie ohne Bio-film. In chloriertem Teerleachat wurden unter der teilweisen Anwendung einer einfachen Methode der kontinu-ierlichen Extraktion folgende PAK-Derivate analysiert: Chloracenaphthen, 1-Chlorpyren, 9-Fluorenon, Anthron. Da Reaktionsprodukte von Phenanthren und Fluoranthen nicht gefunden werden konnten, wird eine Weiterreaktion zu 9-Fluorenon vorgeschlagen, die durch die Pechoberfläche katalysiert wird. Instationäre Korrosion und nicht vorhandener aerober Abbau sind die Gründe für eine PAK-Zunahme während Stagnationphasen. / The influence of biofilm, encrustation, flow velocity and water quality on the release of PAHs was investigated in batch, continuously flown reactor experiments and in a pipe rig system. Chlorination of coal-tar coated stainless steel slides with biofilm resulted in an almost complete removal of the biofilm and an increase of the release of fluorene, phenanthrene and fluoranthene. PAH release of coal-tar coated stainless steel slides exposed to a solution of drinking water with 0,1% sodium azide as an inhibitor of the respiratory chain which did not remove the biofilm led to an increase of all observed PAHs up to the same magnitude as without biofilm. In chlorinated coal-tar leachate the following derivatives were detected while partly utilizing an easy method for a continuously extrac-tion: chloroacenaphthene, 1-chloropyrene, 9-fluorenone, anthrone. While reaction products of phenanthrene and fluoranthene could not be found a further reaction to 9-fluorenone catalysed by the coal-tar surface is proposed. Instationary corrosion and no aerobic degradation are the reasons for PAH increase during stagnation periods.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / A Escherichia coli enteroagregativa (EAEC) é um patotipo emergente e heterogêneo que causa a diarréia aguda ou persistente em indivíduos de diferentes faixas etárias e em pacientes imunocomprometidos. Além disso, EAEC é um dos principais agentes etiológicos da diarréia dos viajantes. O padrão de aderência agregativa de EAEC está associado ao plasmídeo de aderência agregativa (pAA). Genes presentes no plasmídeo e no cromossomo codificam proteínas envolvidas na secreção extracelular de fatores de virulência na superfície ou diretamente na célula hospedeira. A capacidade de produção de muco e biofilme, elaboração de toxinas, aderência e indução de inflamação intensa na mucosa intestinal são importantes características da patogenicidade de EAEC. Nesse estudo, determinamos o perfil genotípico de genes do sistema de secreção Tipo V (SST5) e sistema de secreção Tipo VI (SST6) em cepas de EAEC. Os genes do SST5 ocorreram com mais frequência que os genes do SST6. A presença de pelo menos um gene do SST5 foi detectada em 79% das cepas, enquanto que os genes relacionados ao SST6 foram detectados em apenas 42% das cepas analisadas. A produção de biofilme foi observada em teste quantitativo e verificamos que 67% das cepas produziram biofilme. No teste qualitativo, o tipo de biofilme que predomina é o biofilme moderado (11 cepas), seguido do biofilme forte (9 cepas) e do biofilme discreto (4 cepas). A presença ou ausência de genes do SST5 e SST6 não parece interferir com a capacidade de produção de biofilme, nem com o tipo de biofilme formado. Em ensaios de citotoxicidade, apenas 25% das cepas EAEC (sobrenadante) causaram redução significativa na viabilidade de células T84 avaliada pelo teste de redução com MTT. Nossos resultados mostram que as cepas EAEC isoladas de crianças com diarréia aguda ou de grupo controle são invasoras para células T84. Ao compararmos a capacidade invasora das cepas clinicas e controle, observamos que a média do índice de internalização obtido nas 15 cepas do grupo clinico foi de 5,7% 1,7 e para as 9 cepas do grupo controle foi de 2.4 % 0,7; entretanto essa diferença observada não foi estatisticamente significativa. Não foi possível correlacionar o perfil genotípico dos genes do SST5 e SST6 com o perfil fenotípico analisado (formação de biofilme, citotoxicidade e invasão).O que pode ser atribuído a heterogeneidade genotípica e fenotípica, uma característica relevante de cepas EAEC. / Enteroaggregative Escherichia coli (EAEC) is an emerging and heterogeneous pathotype that causes acute or persistent diarrhea in individuals of different age groups and HIV-positive patients. In addition, EAEC is a major etiologic agent of travelers' diarrhea. The pattern of aggregative adherence of EAEC is associated with the aggregative adherence plasmid (pAA). Genes present in the plasmid and chromosome encode proteins involved in extracellular secretion of virulence factors on the surface or directly into the host cell. The production of mucus, biofilm and toxins; and induction of intense inflammation in the intestinal mucosa and mucosal adherence are major features of EAEC pathogenesis. In this study, we determined the genotypic profile of type V secretion system (T5SS) and type VI secretion system (T6SS) genes in EAEC strains. T5SS genes occurred more frequently than T6SS genes. The presence of at least one T5SS gene was detected in 79% of the strains, while genes related with T6SS were detected in only 42% of the strains tested. The biofilm production was observed in quantitative test, we have found that 67% of strains produced biofilm. In the qualitative test, we detected three distinctive patterns of biofilm formation: moderate (11 strains), strong (9 strains) and discrete biofilm (4 strains). The presence or absence of T5SS and T6SS genes do not interfere with the ability to produce biofilm, neither with biofilm pattern. In cytotoxicity assays, only 25% of EAEC strains (supernatant)resulted in significant reduction of T84 cells activity as evaluated by MTT reduction test. Our results show that EAEC strains isolated from children with acute diarrhea or the control group are able to invade T84 cells. When comparing the invasive ability of clinical and control strains, we observed that the average rate of internalization obtained in 15 clinical strains was 5.7% 1.7, for the nine strains of control group was 2.4% 0. 7, however this difference was not statistically significant. It was not possible to correlate the genotypic profile of T5SS and T6SS genes with the phenotypic profile analysed (biofilm formation, cytotoxicity and invasion), which can be attributed to genotypic and phenotypic heterogeneity, an important characteristic of EAEC strains

Conselho Nacional de Desenvolvimento Científico e Tecnológico / A Escherichia coli enteroagregativa (EAEC) é um patotipo emergente e heterogêneo que causa a diarréia aguda ou persistente em indivíduos de diferentes faixas etárias e em pacientes imunocomprometidos. Além disso, EAEC é um dos principais agentes etiológicos da diarréia dos viajantes. O padrão de aderência agregativa de EAEC está associado ao plasmídeo de aderência agregativa (pAA). Genes presentes no plasmídeo e no cromossomo codificam proteínas envolvidas na secreção extracelular de fatores de virulência na superfície ou diretamente na célula hospedeira. A capacidade de produção de muco e biofilme, elaboração de toxinas, aderência e indução de inflamação intensa na mucosa intestinal são importantes características da patogenicidade de EAEC. Nesse estudo, determinamos o perfil genotípico de genes do sistema de secreção Tipo V (SST5) e sistema de secreção Tipo VI (SST6) em cepas de EAEC. Os genes do SST5 ocorreram com mais frequência que os genes do SST6. A presença de pelo menos um gene do SST5 foi detectada em 79% das cepas, enquanto que os genes relacionados ao SST6 foram detectados em apenas 42% das cepas analisadas. A produção de biofilme foi observada em teste quantitativo e verificamos que 67% das cepas produziram biofilme. No teste qualitativo, o tipo de biofilme que predomina é o biofilme moderado (11 cepas), seguido do biofilme forte (9 cepas) e do biofilme discreto (4 cepas). A presença ou ausência de genes do SST5 e SST6 não parece interferir com a capacidade de produção de biofilme, nem com o tipo de biofilme formado. Em ensaios de citotoxicidade, apenas 25% das cepas EAEC (sobrenadante) causaram redução significativa na viabilidade de células T84 avaliada pelo teste de redução com MTT. Nossos resultados mostram que as cepas EAEC isoladas de crianças com diarréia aguda ou de grupo controle são invasoras para células T84. Ao compararmos a capacidade invasora das cepas clinicas e controle, observamos que a média do índice de internalização obtido nas 15 cepas do grupo clinico foi de 5,7% 1,7 e para as 9 cepas do grupo controle foi de 2.4 % 0,7; entretanto essa diferença observada não foi estatisticamente significativa. Não foi possível correlacionar o perfil genotípico dos genes do SST5 e SST6 com o perfil fenotípico analisado (formação de biofilme, citotoxicidade e invasão).O que pode ser atribuído a heterogeneidade genotípica e fenotípica, uma característica relevante de cepas EAEC. / Enteroaggregative Escherichia coli (EAEC) is an emerging and heterogeneous pathotype that causes acute or persistent diarrhea in individuals of different age groups and HIV-positive patients. In addition, EAEC is a major etiologic agent of travelers' diarrhea. The pattern of aggregative adherence of EAEC is associated with the aggregative adherence plasmid (pAA). Genes present in the plasmid and chromosome encode proteins involved in extracellular secretion of virulence factors on the surface or directly into the host cell. The production of mucus, biofilm and toxins; and induction of intense inflammation in the intestinal mucosa and mucosal adherence are major features of EAEC pathogenesis. In this study, we determined the genotypic profile of type V secretion system (T5SS) and type VI secretion system (T6SS) genes in EAEC strains. T5SS genes occurred more frequently than T6SS genes. The presence of at least one T5SS gene was detected in 79% of the strains, while genes related with T6SS were detected in only 42% of the strains tested. The biofilm production was observed in quantitative test, we have found that 67% of strains produced biofilm. In the qualitative test, we detected three distinctive patterns of biofilm formation: moderate (11 strains), strong (9 strains) and discrete biofilm (4 strains). The presence or absence of T5SS and T6SS genes do not interfere with the ability to produce biofilm, neither with biofilm pattern. In cytotoxicity assays, only 25% of EAEC strains (supernatant)resulted in significant reduction of T84 cells activity as evaluated by MTT reduction test. Our results show that EAEC strains isolated from children with acute diarrhea or the control group are able to invade T84 cells. When comparing the invasive ability of clinical and control strains, we observed that the average rate of internalization obtained in 15 clinical strains was 5.7% 1.7, for the nine strains of control group was 2.4% 0. 7, however this difference was not statistically significant. It was not possible to correlate the genotypic profile of T5SS and T6SS genes with the phenotypic profile analysed (biofilm formation, cytotoxicity and invasion), which can be attributed to genotypic and phenotypic heterogeneity, an important characteristic of EAEC strains

Der Einfluss des Biofilms, der Inkrustation, der Fließgeschwindigkeit und der Wasserqualität auf die Freisetzung von PAK wurde in Batch-, Drehkolbenreaktorversuchen und an einem Rohrleitungssystem untersucht. Chlorung von tauchgeteerten Edelstahlcoupons mit Biofilm führten zu einer fast vollständigen Ablösung des Biofilms und einer Zunahme der Freisetzung von Fluoren, Phenanthren und Fluoranthen. Die PAK-Freisetzung von tauchgeteerten Edelstahlcou-pons in einer Lösung von Trinkwasser mit 0,1% Natriumazid als Hemmstoff der Atmungskette, der den Biofilm nicht ablöst, führten zu einer Zunahme aller untersuchten PAK in der gleichen Größenordnung wie ohne Bio-film. In chloriertem Teerleachat wurden unter der teilweisen Anwendung einer einfachen Methode der kontinu-ierlichen Extraktion folgende PAK-Derivate analysiert: Chloracenaphthen, 1-Chlorpyren, 9-Fluorenon, Anthron. Da Reaktionsprodukte von Phenanthren und Fluoranthen nicht gefunden werden konnten, wird eine Weiterreaktion zu 9-Fluorenon vorgeschlagen, die durch die Pechoberfläche katalysiert wird. Instationäre Korrosion und nicht vorhandener aerober Abbau sind die Gründe für eine PAK-Zunahme während Stagnationphasen. / The influence of biofilm, encrustation, flow velocity and water quality on the release of PAHs was investigated in batch, continuously flown reactor experiments and in a pipe rig system. Chlorination of coal-tar coated stainless steel slides with biofilm resulted in an almost complete removal of the biofilm and an increase of the release of fluorene, phenanthrene and fluoranthene. PAH release of coal-tar coated stainless steel slides exposed to a solution of drinking water with 0,1% sodium azide as an inhibitor of the respiratory chain which did not remove the biofilm led to an increase of all observed PAHs up to the same magnitude as without biofilm. In chlorinated coal-tar leachate the following derivatives were detected while partly utilizing an easy method for a continuously extrac-tion: chloroacenaphthene, 1-chloropyrene, 9-fluorenone, anthrone. While reaction products of phenanthrene and fluoranthene could not be found a further reaction to 9-fluorenone catalysed by the coal-tar surface is proposed. Instationary corrosion and no aerobic degradation are the reasons for PAH increase during stagnation periods.

Vid framställning av papper och pappersmassa krävs en stor mängd energi och även stora mängder vatten som med fördel tas från en närliggande sjö eller vattendrag. Avloppsvattnet passerar flera olika reningssteg såsom mekanisk, kemisk och biologisk rening innan det återinförs till recipienten. Vid aerob biologisk rening sker en nedbrytning och oxidation av det organiska materialet som utförs av mikroorganismer. Aeroba mikroorganismer kräver närvaro av syre varvid luft tillsätts till dammen. Luftningen utgör drygt hälften av det externa reningsverkets totala energibehov och försvåras av extraktivämnen som finns i träden. Vid Stora Enso Skoghalls bruk används en luftad damm som biologisk rening. Det finns problem med att få syret att räcka till i hela dammen. Om ett försteg placeras innan dammen skulle detta kunna bryta ned en del av de syreförbrukande ämnena. Förstegets utformning skulle vara en MBBR (Moving Bed Biofilm Reactor) innehållande bärare där termofila bakterier bildar biofilm. Två olika försöksuppställningar byggdes upp i ett laboratorium på Karlstads Universitet innehållande MBBR och luftade dammar. De luftade dammarna skulle likna Skoghalls bruks luftade damm med samma uppehållstid och innehålla samma sorts mikroorganismer. Processvattnet hämtades från Skoghalls bruk och pumpades kontinuerligt in till försöksuppställningarna. Vattnet togs från två olika ställen på bruket och innehöll dels CTMP och dels CTMP blandat med vatten från blekeriet och indunstningen. För utvärderingar av försöken utfördes analyser av COD, TOC, SÄ, SVI, ytspänning, extraktivämnen samt mikroskopering. Resultatet uppvisade en relativt jämn COD-och TOC-reduktion för de luftade dammarna på runt 70 %, vilket är jämförbara värden med Skoghalls bruks luftade damm. För det termofila MBBR-försteget med temperaturen 45 °C uppvisades en reduktionskapacitet på 30 – 40 %. Resultatet vid körning med vatten innehållande endast CTMP påvisades en reduktion på 30 %. Mätning av ytspänning och extraktivämnen ger en indikation på om syresättningen av avloppsvattnet kan bli effektivare. Det visade sig att biofilmen i de termofila stegen är mycket känslig och relativt enkel att slå ut om förhållandena för de termofila bakterierna inte är optimala eller om inkommande vatten innehåller någon giftig substans. Bioslammet i samtliga luftade dammar bestod av både bakterier, protozoer och metazoer. I referensdammen uppvisades stora och kompakta flockar medan det för dammarna som föregicks av termofila MBBR påvisade mindre slam och mindre kompakta flockar. För dammen innehållande endast CTMP påvisades förutom att flockarna var små även flytslam och skumproblem. / The production of pulp and paper requires a large amount of energy and also large amounts of water which, by advantage, can be taken from a nearby lake or river. The wastewater passes through several different purification steps, such as mechanical, chemical and biological treatment before being reintroduced to the recipient. During aerobic biological treatment, a degradation and oxidation of the organic material occurs and is performed by microorganisms which require oxygen. Aeration consumes about half of the external plant's total energy and is aggravated by extractives present in the trees. At Stora Enso Skoghall mill an aerated pond is used as biological treatment, but there are troubles to get the oxygen to last for the entire pond. If a pre-treatment is placed before the pond this would be able to reduce the substances which demands oxygen. The implementation of the pre-treatment would be a MBBR (Moving Bed Biofilm Reactor) containing carriers and where thermophilic bacteria form biofilms. The experimental setups were built up in a laboratory at the University of Karlstad. The aerated ponds would resemble the aerated pond at Skoghall mill, with an identical retention time and contain equivalent microorganisms. The wastewater was continuously pumped into the experimental setups and was received from Skoghall mill and contained only CTMP water and CTMP mixed with water from the bleach plant and evaporation plant. As evaluation of the experiments analyzes of COD, TOC, SS, SVI, surface tension, extractives and microscopy were done. The results showed a relatively uniform COD and TOC reduction for the aerated ponds at around 70 %, which is comparable with the real pond at Skoghall mill. For the thermophilic MBBR with a temperature of 45° C and with probe 10-water, a capacity of reduction at 30 - 40% was noticed. The result with water containing only CTMP showed a reduction of 30%. Measurements of surface tension and extractives show if the concentration of extractives is decreasing in the thermophilic pre-treatments and also in the aerated ponds, which indicates a more effective oxygenation of the water. It was also found that the biofilm in the thermophilic stages are very sensitive and relatively easy to decimate if the conditions for the thermophilic bacteria are not optimal or if the incoming water contains any toxic substance. The sludge in all aerated ponds consisted of bacteria, protozoa and metazoa. The reference pond showed large and dense flocks while for the ponds that were preceded by thermophilic MBBR indicated less sludge and less dense flocks. For pond containing CTMP-water the flocks were small, but in addition, floating sludge was detected and foam problems arose.

Dans cette étude, nous avons isolé et cultivé des bactéries intimement liées aux spores du champignon mycorhizien Glomus irregulare prélevées dans la rhizosphère de plants d’Agrostis stolonifera L. récoltés dans un sol naturel. Le séquençage des 29 morphotypes isolés a révélé la présence de seulement sept taxons bactériens (Variovorax paradoxus, Microbacterium ginsengiosoli, Sphingomonas sp., Bacillus megaterium, B. simplex, B. cereus et Kocuria rhizophila). Des isolats de chacun de ces sept taxons ont ensuite été cultivés in vitro sur le mycélium de G. irregulare afin d’observer par microscopie leur capacité à croitre et à s’attacher au mycélium en absence d’éléments nutritifs autres que ceux fournis par le champignon. Tous les isolats, sauf B. cereus, ont été capables de bien croitre dans le système expérimental et de s’attacher au mycélium en formant des structures ressemblant à des biofilms sur la surface du champignon. Toutefois, B. simplex formait ces structures plus rapidement, soit en 15 jours, alors que les autres isolats les ont formés après 30 jours (K. rhizophila et B. megaterium) ou 45 jours (V. paradoxus, M. ginsengiosoli et Sphingomonas sp.). D’autre part, la technique PCR-DGGE a permis d’analyser la diversité bactérienne associée aux spores. La diversité des taxons associés aux spores de G. irregulare qu’il a été possible d’isoler et de cultiver in vitro a été nettement moindre que celle qui était présente sur la surface des spores, alors que la biodiversité bactérienne totale du sol a été encore beaucoup plus élevée. Les bactéries associées aux champignons mycorhiziens jouent probablement un rôle important dans la capacité des plantes à résister aux stress biotiques et abiotiques auxquels elles sont soumises. / In this study, we isolated and cultivated bacterial cells intimately associated with Glomus irregulare spores in a natural soil Agrostis stolonifera rhizosphere. Sequencing of the 29 morphotypes isolated revealed the presence of only seven bacterial taxa (Variovorax paradoxus, Microbacterium ginsengiosoli, Sphingomonas sp., Bacillus megaterium, B. simplex, B. cereus and Kocuria rhizophila). These seven isolates were cultivated in vitro on the mycelium of G. irregulare to allow microscopic observation of growth and attachment to the mycelium in absence of nutritive sources other than those derived from the fungal mycelium. All isolates but B. cereus were able to grow on the experimental system and to attach to the mycelium to form biofilm-like structures on their surface. However, B. simplex formed these structures more quickly, in 15 days, than the remaining isolates that have formed them only after 30 days (K. rhizophila and B. megaterium) or 45 days (V. paradoxus, M. ginsengiosoli and Sphingomonas sp.). In addition, PCR-DGGE was used to compare bacterial diversity. The bacterial biodiversity associated with spores of G. irregulare that were isolated and cultured in vitro was significantly lower than that present on the spore surface, while total soil bacterial diversity was much higher. The bacteria associated with mycorrhizal fungi probably have an important role in the ability of plants to withstand biotic and abiotic stresses to which they are submitted.