Indiana University-Purdue University Indianapolis (IUPUI) / The innate and adaptive immune responses are critical for host survival. The TNFα/NF-κB signaling pathway is a major regulator of the immune response. The TNFα/NF-κB signaling pathway has also been proposed to play a role in the regulation of hematopoiesis. In the TNFα signaling pathway, full induction of NF-κB (specifically the p65 subunit) dependent transcription is regulated by a co-activator SIMPL. The biological significance of SIMPL in TNFα dependent responses is poorly understood. To study SIMPL in vitro and in vivo in mammalian cells, a knockdown system utilizing shRNA (short hairpin RNA) was used. Analysis of hematopoietic progenitor cells infected with a retrovirus encoding the SIMPL shRNA was used to study the role of SIMPL in hematopoiesis. The ability of progenitor cells lacking SIMPL to grow and differentiate was not compromised. In contrast in the progenitors cells lacking SIMPL, TNFα mediated inhibition of colony formation was significantly enhanced. These growth inhibitory effects of SIMPL were not due to an increase in apoptosis. The enhanced inhibitory affects were specific for TNFα and not found in other common hematopoietic inhibitors (TGF-β1 and IFNγ). Results of this work reveal that SIMPL is a component of the hematopoiesis that is required for TNFα dependent effects upon myeloid progenitors.

Postoperativa sårinfektioner är den tredje mest frekventa anmälda vårdrelaterade infektionen i Sverigeenligt Socialstyrelsen (2006). Operationsrummet är det mest kritiska området för infektionskontroll.Enighet tycks råda kring den bakteriella orsaken för postoperativa sårinfektioner, nämligenStaphylocous aureus samt att den förekommer i den normala hudfloran hos oss människor, är individberoende samt luftburen. Att en betydande del bakteriebärande partiklar kontaminerar såret underoperation har även visats samt att de härstammar från den omgivande luften. En relation mellan haltenluftburna bakterier och infektionsfrekvens har gjorts för ledplastikoperationer av höft- och knäleder.Dock är relationen mellan sårkontaminering och infektionsfrekvensen inte helt fastställd för övrigkirurgi och frågasätts även för ledplastikoperationer. För att minska infektionsfrekvensen idag användsdock en kombination av antibiotikaprofylax, ventilationssystem och kläder. Trots svårigheten i attbedöma halten S.aureus utifrån mängden bakteriebärande partiklar är det trots allt den ända metodenatt mäta ventilationssystemets effektivitet på. Det blir än mer viktigt med att uppnå låga halter av dessai luften i och med ökandet av multiresistenta bakterier.Stor oenighet råder bland dagens experter kring vilken systemvalslösning som är bäst lämpad givetolika förutsättningar och krav på renlighet. Debatten har varit stor främst då det under senaste tidenimplementerats olika ventilationsprinciper inom sjukvården i landet. Debatten grundar sig i om ettlaminärt system med hastighet under 0,4 m/s beter sig omblandande eller faktiskt laminärt samt omkläder har samma effekt beroende på luftföringsprincip. Det efterfrågas även en större ömsesidigförståelse mellan ingenjörer och kirurger för att medvetengöra personalens inverkan på spridning avbakteriebärande partiklar under pågående operation.Mot bakgrund av detta har målet med denna studie varit att sätta ihop en kritiskkunskapssammanställning kring frågor rörande renhet, arbetsmiljö och inomhusklimat ioperationssalar baserat på fallstudier och litteraturstudie inom ämnena. Samt att utifrån dettaidentifiera risker med utföranden, utmaningar, kunskaps- och samverkansbehov i frågorna. Störredelar av studien görs med utgångspunkt från fallstudier utförda genom kvalitativa intervjuer därerfarenheter och information om ämnet samlas in i syfte att kartlägga och exponera sammansattaproblem, utmaningar samt skillnader i utförande och systemlösningar. Författaren ämnar inte utföraegna mätningar eller simuleringar utan studien vilar på tidigare forskning i områden kringspridningsmekanismerna och källor till infektionsspridande partiklar, termisk komfort och luftkvaliteti operationssalsmiljö samt empirin från fallstudier och kvalitativa intervjuer.Redskapen som använts har varit en litteraturstudie i kombination med kvalitativa fallstudier därintervjuer har skett med tolv respondenter från olika yrkesgrupper som alla till olika grad varitdelaktiga vid beslut/design/utförande av operationssalarna och systemlösningarna. Intervjuerna harvarit halvstrukturerade och analyserats utifrån ett pragmatiskt synsätt. Studien i sin helhet har utförtsmed ett abduktivt förhållningssett där befintliga teorier inom ämnet har jämförts med ny empiri ochsammanställts i en syntes.Resultatet har utmynnat i följande: För att minska risken för postoperativa sårinfektioner krävs båderutiner i form av när och hur antibiotikan appliceras, personlig hygien och kläder i samspel medventilationstekniken men även funktionen av att upprätthålla olika barriärer mellan infektionskälla ochpatient. För att upprätthålla detta krävs ett samspel mellan ventilationstekniken, byggtekniken och denmäskliga faktorn. Det handlar om att begränsa förekomsten av bakteriebärande partiklar i salen ocheffektivt transportera bort och spä ut de som oundvikligt förekommer samt begränsa dess livsduglighetöver tid. För patientsäkerheten innebär det även i förlängningen att erbjuda både personal och patient2 (100)en god arbetsmiljö och termiskt klimat för att undvika kirurgiska misstag och nedkylning (hypotermi)av patient.Av de två ventilationsprinciperna behandlade i denna studie, omblandande och laminär strömningvisas det att båda kan uppnå godkända nivåer på halter luftburna bakterier under pågående operation.Systemen skiljer i luftföringsprincip men bygger båda till olika grad på riktad strömning ochutspädning. Utspädningsprincipen tycks inte till fullo kunna bedöma det laminära systemets förmågaatt reducera halten bakteriebärande partiklar likväl antar dess strömning ändå en omblandandekaraktär då det är känsligare för hinder. Det antyds att systemet upprätthåller en viss strömningseffektmed undertempererad luft men anses även på grund av detta i samband med stora flöden utsättapatienten för större risk för hypotermi i jämförelse med omblandande systemet. Gemensamt för debåda är beroendet av rätta rutiner hos personal, en upprättad tryckdifferens mellan utrymmen ochskyddande klädsel för att minska spridning från angränsande utrymmen och oss människor.Beslutsprocessen gällande val av system kräver mer oberoende forskning, tydligare och adekvatajämförelser samt en försiktighet vid analys av tidigare studier kring ventilationssystemens inverkan påpostoperativa sårinfektioner. En enskilds parameters inverkan på infektionsfrekvensen kan idag intebedömmas. Med denna vetskap är det även av stor vikt med samarbete och förståelse mellan ingenjöreroch vårdverksamheter för att upplysa varandra om operationsverksamhetens behov ochsystemlösningarnas begränsningar samt de förutsättningar som ställs för att en låg infektionsfrekvensska uppnås.

(ENGLISH) Hematopoiesis is dependent on the actions of hematopoietic stem cells (HSCs). This process is tightly controlled through a complex array of extrinsic and intrinsic factors. Even though the hematopoiesis seems to be well conserved across the disparate vertebrate animals, erythroid and thrombocytic differentiation have changed during the evolution of mammals. Specifically, adult mammalian red blood cells have the unique feature of being enucleated, and mammalian thrombocytes are not individual cells, but fragments of megakaryocytes, instead. It is likely that these enhancements provided a survival advantage to early mammalian species; however, they also bring up the question of evolutionary origin of these cells that studied using zebrafish (Danio rerio) model. First, it was necessary to generate a toolbox of a recombinant cytokines and optimized culture media that allowed us to manipulate zebrafish hematopoietic cells ex vivo in liquid and clonal cultures. Interestingly, teleost species underwent an extra duplication event during their evolution and as a result, two copies (paralogs) of some of the genes are present in zebrafish. This was also the case for majority of the cytokines from our toolbox and here, we provide functional characterization of these paralogs. Strikingly, our results...

Une des complications importantes d’un traitement intensif de chimio/radio-thérapie est l’aplasie de la moelle osseuse qui peut persister longtemps même après une greffe de cellules souches. Le PBI-1402 est un petit lipide qui a été associé à la diminution de l’apoptose des neutrophiles induite par des agents cytotoxiques. Nos travaux ont démontré que la culture in vitro de progéniteurs hématopoiétiques humains en présence de PBI-1402 induit une augmentation significative du nombre de progéniteurs érythroides (PEryth) (p<0,05). En évaluant la sensibilité des PEryth à l’érythropoietine (Epo), nous avons démontré que le PBI-1402 n’a pas d’effet sensibilisateur et que les cellules répondent de façon similaire aux cellules contrôles. De plus, la combinaison de l’Epo et du « stem cell factor » avec le PBI-1402 permet de prolonger et d’augmenter l’activation d’ERK1/2 (p<0,05), un important signal mitogène. Cet effet est associé à une inhibition de l’activation de la phosphatase MKP-1 dans les cellules exposées au PBI-1402. Nous démontrons aussi la capacité du PBI-1402 à amplifier la prolifération des PEryth et sa capacité à réduire la durée et l’intensité de l’anémie dans un modèle in vivo murin. Des souris ayant reçu une dose létale d’irradiation et subi une transplantation syngénique de moelle osseuse, ont été traitées oralement avec le PBI-1402 pendant 14 jours. Ces souris démontrent une réduction significative de l’anémie post-transplantation versus les souris contrôle (p<0,05). De plus, la moelle osseuse des souris traitées au PBI-1402 présente un nombre de BFU-E et CFU-E plus élevé comparativement au contrôle. Ces résultats démontrent donc le potentiel du PBI-1402 à réduire l’anémie post-transplantation et accélérer la reconstitution érythroïde. / One of the most important complications of intensive radiotherapy or chemotherapy is cytopenia, which can persist for significant amount of time even after stem cell transplantation. PBI-1402, a small lipid, was previously shown to be associated with decreased neutrophil apoptosis caused by cytotoxic agents. Our work has shown that day primary human hematopoietic cell in vitro culture in the presence of PBI-1402 resulted in an increased number of erythroid progenitors (p<0,05). Dose-response experiments evaluating sensitivity to erythropoietin (Epo) of cells exposed to PBI-1402 indicated that PBI-1402 did not have a sensitizing effect and that both treated and control cells respond similarly to Epo. In addition, PBI-1402, used in combination with stem cell factor (SCF) and Epo, enhanced and prolonged ERK1/2 phosphorylation (p<0.05), a signalling pathway important for erythroid progenitor cell proliferation. This effect was associated with a decrease of the phosphatase MKP-1 activation in PBI-1402 exposed cells. This translated into and increased proliferation of erythroid progenitors as well as a reduced duration and level of anemia in an in vivo murine transplantation model. Lethally irradiated mice that received syngeneic stem cell transplantation were treated orally with PBI-1402 for 14 days. These mice demonstrated a significant reduction in post-transplantation anemia in a dose dependent manner compared to control (vehicle)(p<0.05). Moreover, PBI-1402-treated mice harboured significantly higher numbers of BFU-E and CFU-E in bone marrow compared to control (p<0.05). These results demonstrate that PBI-1402 treatment significantly reduced transplantation-induced anemia with concomitant acceleration in erythroid recovery.

Introdução: O leite pode ser considerado um dos alimentos mais completos por apresentar alto teor de proteínas e sais minerais, porém, também é considerado excelente meio de cultura para microrganismos. Objetivos: Identificar o número de UFC de Staphylococcus spp. e Klebsiella pneumoniae, cultivados isoladamente ou em associação, em leite integral estéril, a 6 oC, 27 oC e 37 oC e descrever a curva de morte térmica, quando submetidas ao processo de pasteurização lenta isoladamente ou em associação. Material e Métodos: Avaliação através da contagem das UFC, do comportamento de Staphylococcus spp. e K. pneumoniae, isoladas de tanque de refrigeração de leite, submetidas a temperaturas que simulam o leite em condições de refrigeração (6 °C), condições ambientais (25 °C) e na temperatura ideal de crescimento de patógenos mesófilos (36°C). Avaliação da sensibilidade de Staphylococcus spp. e K. pneumoniae isoladas e em associações ao processo de pasteurização lenta. Resultados: Na escala 1 de Mac Farland a média de UFC de K. pneumoniae foi maior que a de Staphylococcus spp. A 6 °C as bactérias no leite, isoladas ou em associação crescem na mesma velocidade. A 25 °C a K. pneumoniae cresce mais que o Staphylococcus spp. A 25 °C K. pneumoniae associada ao Staphylococcus spp. cresce mais do que quando encontra-se isolada. A 36 °C K. pneumoniae associada ao Staphylococcus spp. cresce mais do que quando encontra-se isolada e ainda mais que a 25 °C. A pasteurização lenta foi efetiva, pois reduziu em no mínimo 90% as UFC no leite após 30 minutos a 65 °C. Conclusão: a medida que aumenta a temperatura, até 36 °C, a Klebsiella pneumoniae apresenta crescimento superior ao Staphylococcus spp., a contaminação de leite por Klebsiella pneumoniae (contaminação ambiental) irá influenciar mais na qualidade do produto final, comparando-se à contaminação por Staphylococcus spp., oriundo de mastite e a interação de microrganismos altera a morte dos mesmos, recomendando-se que novos estudos sejam realizados para que se entenda melhor esse processo. / Introduction: The milk can be considered one of the most complete food for its high contents of protein and minerals, but also it is considered an excellent medium of culture for microorganisms. Objective: To identify the number of the Colony Forming Units of Staphylococcus spp. and Klebsiella pneumoniae, in integral sterile milk, at different temperatures (6ºC, 27ºC e 37ºC) and to describe a thermal curve death, when submitted to the process of slow pasteurization isolated or in association. Materials and methods: Evaluation by counting of the colony forming units and the behavior of the Staphylococcus spp. and Klebsiella pneumoniae, isolated and in association occurred in the cooling tank, submitted to temperatures that simulate the conditions of milk in cooling (6ºC), environmental condition (25ºC) and the optimal temperature for the growth of microorganisms (36ºC). Evaluation of the sensibility of Staphylococcus spp. and Klebsiella pneumoniae isolated and in association during the pasteurization process. Results: In Scale 1 of Mac Farland the average of the colony form units of the K. pneumoniae was greater than the colony of Staphylococcus spp. At 6ºC the microorganisms in the milk isolated or in association grew at the same speed. At 25ºC K. pneumoniae associated with Staphylococcus spp. grew more than when they were isolated. At 36ºC K. pneumoniae associated with Staphylococcus spp. grew more then when it was isolated and more than at 25ºC. So we can conclude that the slow pasteurization was effective, because it has reduced at least 90% of the colony form unity in the milk after 30 minutes at 65ºC. Conclusion: By raising the temperature up to 36ºC, a Klebsiella pneumoniae had a superior growth when compared with Staphylococcus spp.; the contamination of milk by Klebsiella pneumoniae (environmental contamination) influences the most the quality of the final product when compared to the contamination by Staphylococcus spp. from mastitis, we can also conclude that the interaction of microorganisms modify the death rate of them. New studies and researches are recommended to a deeper and better comprehension of this process.

Introdução: O leite pode ser considerado um dos alimentos mais completos por apresentar alto teor de proteínas e sais minerais, porém, também é considerado excelente meio de cultura para microrganismos. Objetivos: Identificar o número de UFC de Staphylococcus spp. e Klebsiella pneumoniae, cultivados isoladamente ou em associação, em leite integral estéril, a 6 oC, 27 oC e 37 oC e descrever a curva de morte térmica, quando submetidas ao processo de pasteurização lenta isoladamente ou em associação. Material e Métodos: Avaliação através da contagem das UFC, do comportamento de Staphylococcus spp. e K. pneumoniae, isoladas de tanque de refrigeração de leite, submetidas a temperaturas que simulam o leite em condições de refrigeração (6 °C), condições ambientais (25 °C) e na temperatura ideal de crescimento de patógenos mesófilos (36°C). Avaliação da sensibilidade de Staphylococcus spp. e K. pneumoniae isoladas e em associações ao processo de pasteurização lenta. Resultados: Na escala 1 de Mac Farland a média de UFC de K. pneumoniae foi maior que a de Staphylococcus spp. A 6 °C as bactérias no leite, isoladas ou em associação crescem na mesma velocidade. A 25 °C a K. pneumoniae cresce mais que o Staphylococcus spp. A 25 °C K. pneumoniae associada ao Staphylococcus spp. cresce mais do que quando encontra-se isolada. A 36 °C K. pneumoniae associada ao Staphylococcus spp. cresce mais do que quando encontra-se isolada e ainda mais que a 25 °C. A pasteurização lenta foi efetiva, pois reduziu em no mínimo 90% as UFC no leite após 30 minutos a 65 °C. Conclusão: a medida que aumenta a temperatura, até 36 °C, a Klebsiella pneumoniae apresenta crescimento superior ao Staphylococcus spp., a contaminação de leite por Klebsiella pneumoniae (contaminação ambiental) irá influenciar mais na qualidade do produto final, comparando-se à contaminação por Staphylococcus spp., oriundo de mastite e a interação de microrganismos altera a morte dos mesmos, recomendando-se que novos estudos sejam realizados para que se entenda melhor esse processo. / Introduction: The milk can be considered one of the most complete food for its high contents of protein and minerals, but also it is considered an excellent medium of culture for microorganisms. Objective: To identify the number of the Colony Forming Units of Staphylococcus spp. and Klebsiella pneumoniae, in integral sterile milk, at different temperatures (6ºC, 27ºC e 37ºC) and to describe a thermal curve death, when submitted to the process of slow pasteurization isolated or in association. Materials and methods: Evaluation by counting of the colony forming units and the behavior of the Staphylococcus spp. and Klebsiella pneumoniae, isolated and in association occurred in the cooling tank, submitted to temperatures that simulate the conditions of milk in cooling (6ºC), environmental condition (25ºC) and the optimal temperature for the growth of microorganisms (36ºC). Evaluation of the sensibility of Staphylococcus spp. and Klebsiella pneumoniae isolated and in association during the pasteurization process. Results: In Scale 1 of Mac Farland the average of the colony form units of the K. pneumoniae was greater than the colony of Staphylococcus spp. At 6ºC the microorganisms in the milk isolated or in association grew at the same speed. At 25ºC K. pneumoniae associated with Staphylococcus spp. grew more than when they were isolated. At 36ºC K. pneumoniae associated with Staphylococcus spp. grew more then when it was isolated and more than at 25ºC. So we can conclude that the slow pasteurization was effective, because it has reduced at least 90% of the colony form unity in the milk after 30 minutes at 65ºC. Conclusion: By raising the temperature up to 36ºC, a Klebsiella pneumoniae had a superior growth when compared with Staphylococcus spp.; the contamination of milk by Klebsiella pneumoniae (environmental contamination) influences the most the quality of the final product when compared to the contamination by Staphylococcus spp. from mastitis, we can also conclude that the interaction of microorganisms modify the death rate of them. New studies and researches are recommended to a deeper and better comprehension of this process.

Une des complications importantes d’un traitement intensif de chimio/radio-thérapie est l’aplasie de la moelle osseuse qui peut persister longtemps même après une greffe de cellules souches. Le PBI-1402 est un petit lipide qui a été associé à la diminution de l’apoptose des neutrophiles induite par des agents cytotoxiques. Nos travaux ont démontré que la culture in vitro de progéniteurs hématopoiétiques humains en présence de PBI-1402 induit une augmentation significative du nombre de progéniteurs érythroides (PEryth) (p<0,05). En évaluant la sensibilité des PEryth à l’érythropoietine (Epo), nous avons démontré que le PBI-1402 n’a pas d’effet sensibilisateur et que les cellules répondent de façon similaire aux cellules contrôles. De plus, la combinaison de l’Epo et du « stem cell factor » avec le PBI-1402 permet de prolonger et d’augmenter l’activation d’ERK1/2 (p<0,05), un important signal mitogène. Cet effet est associé à une inhibition de l’activation de la phosphatase MKP-1 dans les cellules exposées au PBI-1402. Nous démontrons aussi la capacité du PBI-1402 à amplifier la prolifération des PEryth et sa capacité à réduire la durée et l’intensité de l’anémie dans un modèle in vivo murin. Des souris ayant reçu une dose létale d’irradiation et subi une transplantation syngénique de moelle osseuse, ont été traitées oralement avec le PBI-1402 pendant 14 jours. Ces souris démontrent une réduction significative de l’anémie post-transplantation versus les souris contrôle (p<0,05). De plus, la moelle osseuse des souris traitées au PBI-1402 présente un nombre de BFU-E et CFU-E plus élevé comparativement au contrôle. Ces résultats démontrent donc le potentiel du PBI-1402 à réduire l’anémie post-transplantation et accélérer la reconstitution érythroïde. / One of the most important complications of intensive radiotherapy or chemotherapy is cytopenia, which can persist for significant amount of time even after stem cell transplantation. PBI-1402, a small lipid, was previously shown to be associated with decreased neutrophil apoptosis caused by cytotoxic agents. Our work has shown that day primary human hematopoietic cell in vitro culture in the presence of PBI-1402 resulted in an increased number of erythroid progenitors (p<0,05). Dose-response experiments evaluating sensitivity to erythropoietin (Epo) of cells exposed to PBI-1402 indicated that PBI-1402 did not have a sensitizing effect and that both treated and control cells respond similarly to Epo. In addition, PBI-1402, used in combination with stem cell factor (SCF) and Epo, enhanced and prolonged ERK1/2 phosphorylation (p<0.05), a signalling pathway important for erythroid progenitor cell proliferation. This effect was associated with a decrease of the phosphatase MKP-1 activation in PBI-1402 exposed cells. This translated into and increased proliferation of erythroid progenitors as well as a reduced duration and level of anemia in an in vivo murine transplantation model. Lethally irradiated mice that received syngeneic stem cell transplantation were treated orally with PBI-1402 for 14 days. These mice demonstrated a significant reduction in post-transplantation anemia in a dose dependent manner compared to control (vehicle)(p<0.05). Moreover, PBI-1402-treated mice harboured significantly higher numbers of BFU-E and CFU-E in bone marrow compared to control (p<0.05). These results demonstrate that PBI-1402 treatment significantly reduced transplantation-induced anemia with concomitant acceleration in erythroid recovery.

Magister Chirurgiae Dentium - MChD (Oral Medicine and Periodontics) / The aim of this study was to assess whether a mouthwash containing carbohydrate derived fulvic acid, is effective in reducing the salivary microbial count pre-operatively. Endeavours have been made to reduce the risk of infection, bacteraemia and cross-contamination during dental procedures by the application of topical antimicrobial agents. To date chlorhexidine is the most widely evaluated and efficacious agent against oral biofilms but there have been reports of adverse effects ranging from contact dermatitis to severe anaphylactic shock. A new mouth rinse containing carbohydrate derived fulvic acid are reported to have broad spectrum antimicrobial activity against specific oral microbes and Candida albicans with no side effects.

This diploma thesis deals with microbiological quality of hot water in a hospital in Cesky Krumlov as well as hot water piping condition with respect to Legionella. It also deals with arrangements for improving the quality of hot water in this hospital. The subject and target of this thesis are the findings of the mentioned parameters based on own observations, technical documentation and analysis of warm water performed by an accredited laboratory. The hospital in Cesky Krumlov, the construction of which began in 1909, was built in 1911 as the 'Nemocnice Císaře Františka Josefa I.' As the time went by, new buildings were built up. The hospital was renamed to 'Všeobecná veřejná nemocnice' in 1942. In the following years the hospital expanded until the current condition. Water quality and presence of Legionella is a world-wide issue, which we must fight against. The thesis is divided into two parts. First, theoretical part, which includes both general and specific knowledge about Legionella, its health risks, the environment in which it occurs, possibilities of eliminating, suitability of the material for hot water pipes, gained in professional literature and a part of the Czech legislation dealing with public health protection, the law 'No. 258/2004 Coll.' in actual version and quality of drinking water and hot water. In the second, empirical part, I focus on the situation of the hospital in Cesky Krumlov from 2009 till 2015, concerning newly built boiler room and the whole distribution system of hot water regarding Legionella. Particularly I focused on materials used for ecologisation of the gas boiler room and materials used for hot water pipes and actual state of water distribution in individual buildings of this health care facility. I mentioned the analysis of the water samples performed by an accredited laboratory with corresponding technical documentation, which concerns the water distribution system, provided by the medical facility. I used secondary data analysis to evaluate these documents. Regarding the results from 2009 to 2015 the microbiological quality of warm water doesn't meet the limits defined in the Attachment No. 3 of the above mentioned law Order 252/2004 Coll. Limit value for colony forming units of Legionella is set to maximum of 100 CFU. In most of the samples this limit was exceeded by series of units up to hundred thousands of units. The last results in 2015 prove improvement of the unfavorable situation. In that year the analysis was tested three times. First in February, where the CFU units were 'only' in hundreds, with the top value of 550 CFU in the Postacute Care ward. Second analysis was performed in June that same year. Results were acceptable. The highest value was 64 CFU. Last analysis in the hospital was performed in December. Increased concentration of Legionella was again in the Surgery building. There were established of 1400 colony forming units. There were 300 CFU in the Postacute Care ward and the limit value of 100 CFU was in the ward of Internal medicine. The answers to research questions 'How to prevent reproduction of Legionella peumophylis in the water distribution system of the hospital in Cesky Krumlov?' and 'How to ensure a sufficient amount of quality hot water in the medical facility?' are included at the end of the stated recommendations. A risk factor causing the fast reproduction of Legionella is the old water distribution system in most of the buildings and low water temperature of water leaving the boiler, which is 55°C. All mentioned results and evaluations are included in this thesis as well as attached pictures, which show the environment and situation within the Health Care Institutions.

This diploma theses focuses on comparison of natural and intentionally inducated supressiveness of environment induced by application of entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, Isaria fumosorosea a Lecanicillium muscarium. In tests were evaluated the in vitro parameters as well as the effectiveness in vivo biotests on insect host larva Tenebrio molitor. The species of entomopatoghenic fungi were applied in suspensions, single strains and also in combination of two strains. In the in vitro conditions the possibilities of objective evaluation of the supresivity level were tested by using the CFU test (Colony Forming Units) on three different nutrient media (PDA, PDA + A , PDA + D), as one of the basic evaluation parameters. Further the germination tests were evaluated according to GI (Germination Index), determination of radial growth (comparison of median cultures) and interaction of strain suspensions on nutrient media PDA. In the in vivo biotests were watched the epizooties from suspensions of these entomopathogenic fungi on insect larva Tenebrio molitor in competitive test of strains according to FDI (Fungl development index) evaluation scala. Chosen larva covered by fully sporulating mycelium from epizootie were further evaluated in CFU test. The result were dominant strain/s on the larva from applied suspensions.