Orientador: Maria Luiza Moretti / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-05T09:55:21Z (GMT). No. of bitstreams: 1 Bratfich_OrlandoJose_D.pdf: 1128343 bytes, checksum: 5a4d981849f245636703326e9af0c8ad (MD5) Previous issue date: 2005 / Resumo: Isolados clínicos de Staphylococcus epidermidis provenientes de infecções de corrente sangüínea do hospital de clínicas HC ¿ UNICAMP, Campinas foram analisados. Os isolados foram caracterizados por análises fenotípicas e genotípicas. Os testes de susceptibilidade aos antimicrobianos foram realizados para comparar os métodos de disco difusão, concentração inibitória mínima e diluição em Agar, para determinar a relevância clínica da resistência aos antimicrobianos. Foram analisados a presença do gene mecA por PCR, produção de biofilme, genotipagem por PFGE com enzima SmaI. Dados epidemiológicos dos pacientes foram analisados para detectar a importância clínica dessas cepas. Para oxacilina 82 isolados foram resistentes, sendo que 78 (95,12%) apresentaram reação de PCR positiva para o gene mecA, dois isolados apresentaram-se como hiperprodutores de ß-lactamases. Para teicoplanina os testes de CIM apresentaram 52 isolados sensíveis, 35 isolados intermediários e 13 isolados resistentes e, quando comparado com os testes de disco difusão foram observadas discordâncias em 43% dos isolados, onde o teste de disco difusão não foi capaz de identificar isolados intermediários e resistentes (P < 0,001). Para vancomicina um único isolado apresentou resultado intermediário com CIM de 8,0 µg/mL. Isolados vancomicina resistentes não foram observados. Analises dos dados epidemiológicos, através de regressão logística, as variáveis estatisticamente significantes foram neutropenia (P = 0,002) e quimioterapia (P = 0,015). As análises genotípicas por PFGE demonstraram a disseminação clonal de S. epidermidis no ambiente hospitalar e verificou-se que testes dilucionais são necessários quando se estudam glicopeptideos. A análise filogenética demonstrou grande estabilidade e adaptabilidade genética do gene mecA responsável pela resistência a oxacilina / Abstract: Clinical strains of Staphylococcus epidermidis taken from bloodstream infection from UNICAMP (teaching hospital in Campinas São Paulo State, Brazil) were analyzed. The strains were characterized by phenotypic and genotypic analysis. The antimicrobial susceptibility tests were made to compare the disk diffusion tests, MIC, and agar-salt screening-test to determine the relevant clinical antimicrobial resistance. There were also made PCR assays for mecA detection, biofilm production, genotypic by SmaI-digested and pulsed-field gel electrophoresis. Patients epidemiological data were employed to determine the major clinical importance of these strains. For oxacillin 82 strains were resistant, and 78 (95.12%) had positive PCR assay for mecA gene, and two strains presented characteristics of ß-lactamases hyperproduction. In the MIC tests for teicoplanin, 52 strains were susceptible; 35 strains intermediate and 13 strains were resistant, and when compared to the results obtained by disk diffusion tests, MIC, were observed discordant results in 43% of the strains where the disk diffusion test was not able to identify strains intermediate and resistant (P < 0.001). MIC test had only one intermediate of 8.0 µg/mL for vancomycin. Strains vancomycin-resistant were not observed. The variable data statistically significant in the logistic regression analysis related to the treatment of the BSI by S. epidermidis were: neutropenia (P = 0.002) and chemotherapy (P = 0.015). The genotyping analysis of isolates demonstrated a clonal distribution of S. epidermidis in the hospital environment and it was checked that dilution tests are necessary when glycopeptides are being studied.. The phylogenetic analysis showed great genetic stability and adaptability of the mecA gene responsible for the oxacillin resistance / Doutorado / Ciencias Basicas / Doutor em Clínica Médica

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-04-03T17:25:23Z No. of bitstreams: 1 Oliveira, Hanna Lara da Cruz Dinéas de [Dissertação, 2013].pdf: 1766254 bytes, checksum: 30cf8e6b52cfd32b623ecf94d3cee01b (MD5) / Made available in DSpace on 2017-04-03T17:25:23Z (GMT). No. of bitstreams: 1 Oliveira, Hanna Lara da Cruz Dinéas de [Dissertação, 2013].pdf: 1766254 bytes, checksum: 30cf8e6b52cfd32b623ecf94d3cee01b (MD5) / Os estafilococos coagulase negativos emergem como importantes patógenos nosocomiais, frequentemente isolados em bacteremias humanas, fato intimamente relacionado com o aumento do uso de dispositivos médicos ao longo dos últimos anos. Sua capacidade de aderir a superfícies poliméricas e produzir biofilmes constitui o principal fator de virulência associado a estes dispositivos. A fim de minimizar esta possibilidade, antissépticos são largamente utilizados para desinfecção de pele e mucosas na rotina hospitalar, no entanto, tem-se observado uma diminuição da susceptibilidade bacteriana a estes agentes. A diminuição da susceptibilidade dos microrganismos a antissépticos, associados a sua capacidade de formar biofilmes tem elevado a morbidade, mortalidade, período de internação e os custos relativos ao cuidado com a saúde. No estudo aqui apresentado, foram analisadas 81 cepas de Staphylococcus epidermidis resistentes a meticilina (MRSE) e 55 cepas de Staphylococcus haemolyticus resistentes a meticilina (MRSHa), todas resistentes a meticilina, isoladas de pacientes internados em hospital Universitário localizado na cidade de Niterói-RJ nos anos de 2008 e 2010. A determinação do perfil de susceptibilidade aos biocidas triclosan, clorexidina e cloreto de benzalcônio mostrou que frente ao triclosan, as amostras apresentaram as menores taxas de susceptibilidade bacteriana. Papaína não apresentou atividade antibacteriana. Entretanto, foi capaz de reduzir expressivamente a formação de biofilme (p<0,06) em ambas as espécies, mostrando-se a mais eficiente entre os produtos analisados. Foi capaz também de desintegrar biofilmes maduros formados por Staphylococcus epidermidis. Os biocidas não levaram à redução significante de biofilmes, exceto a clorexidina, que foi capaz de reduzir a formação do biofilme pelas MRSE. Foi possível verificar que cepas de MRSHa tratadas com cloreto de benzalcônio têm 40% menos probabilidade de formar biofilme se comparada às tratadas com triclosan e clorexidina. A análise estatística nos mostrou que a expressão do gene qacA/B influenciou significativamente a formação de biofilme e confirmou a relação existente entre os valores de concentração mínima inibitória (CMI) e a formação de biofilme (p< 0,001) / Coagulase negative staphylococci have emerged as important nosocomial pathogens, commonly isolated in human bacteremia, a fact closely related to the increased use of medical devices over the last few years. Its ability to adhere to polymer surfaces and produce biofilms is the main virulence factor associated with these devices. To minimize this possibility, antiseptics are widely used for disinfection of skin and mucosa in routine hospital, however, there has been an increase in bacterial resistance to these agents. Decreased susceptibility of microorganisms to antiseptics, associated with its ability to form biofilms has high morbidity, mortality, length of stay and costs related to health care. In the study presented here, we analyzed 81 strains of methicillin-resistant Staphylococcus epidermidis (MRSE) and 55 strains of methicillin-resistant Staphylococcus haemolyticus (MRSHa), all meticillin- resistant, isolated from patients hospitalized at University Hospital in the city of Niterói, Rio de Janeiro in 2008 and 2010. The determination of the susceptibility profile of biocides triclosan, chlorhexidine and benzalkonium chloride demonstrated that front of triclosan, the strains have the biggest rates of bacterial susceptibility. Papain showed no antibacterial activity. However, it was able to significantly reduce biofilm formation (p <0.06) in both species, being the most efficient among the analyzed products. It was also able to degrade established biofilms formed by Staphylococcus epidermidis. Biocides showed no significant reduction of biofilms, except chlorhexidine, which was able to reduce biofilm formation by MRSE. We noticed that strains MRSHa treated with benzalkonium chloride are 40% less likely to form biofilm compared to those treated with chlorhexidine and triclosan. Statistical analysis showed that the gene expression qacA/B was significantly influential to biofilm formation and confirmed positive relationship between the values of MIC and biofilm formation (p <0.001)

Bacterial contamination of platelet concentrates (PCs) poses the highest transfusion-associated infectious risk with skin flora, such as Staphylococcus epidermidis and Staphylococcus capitis, being the predominant contaminants. These bacteria are able to form surface-attached aggregates or biofilms, which are present in the skin of healthy blood donors and can subsequently be isolated from contaminated PCs. Disinfection of the venipuncture area before donation with a combination of 2% chlorhexidine-gluconate and 70% isopropanol is used at Canadian Blood Services. However, not all bacteria are eliminated during skin disinfection since contaminated PCs are still captured during routine PC screening. In this thesis, the ability of biofilm-forming S. epidermidis and S. capitis to resist the currently used disinfectants was explored. It was demonstrated that although a combination of chlorhexidine and isopropanol has a bactericidal effect, it is unable to completely eradicate skin flora biofilms. Several countries have implemented Pathogen Inactivation Technologies (PITs) as a measure to help control transfusing bacterially-contaminated PCs by exposing PC units to ultra violet light. However, no investigations have been done to evaluate the ability of PITs against bacterial biofilms, which was one of the objectives of this thesis. Data revealed that the efficacy of a currently used PIT, the Mirasol® system, is similar for S. epidermidis present in PCs produced from whole blood inoculated with biofilm or non-biofilm cells. However, treatment effectiveness was strain dependent. In conclusion, further investigation to improve donor skin disinfection and PITs should be considered. Surveillance at Canadian Blood Services shows that contamination rates in single-donor apheresis PCs (Aph-PCs) is generally higher than in four-donor buffy coat platelet pools (BC-PCs). This study investigated whether the BC-PC production method contributes to this observation as BC-PCs are derived from WB that is left to rest overnight while Aph-PCs are collected directly from the donor. Data showed that WB hold during BC-PC production does not have a broad-spectrum bactericidal effect and therefore other factors contribute to low rates of contamination in BC-PCs. The work presented in this thesis provides an insight to bacterial residence and persistence during blood product manufacturing and makes suggestions for PC safety improvements.

The presented master thesis deals with the monitoring of the influence of polysaccharides which are used as an additive in the cosmetic products, on the monitored types of bacteria which are part of the skin microbiome. And it also deals with the study the effect of polysaccharides as prebiotics on selected species of bacteria that are part of the skin microbiome. Two polysaccharides were selected to determine the effects on the skin microbiome: Nanomoist and PoLevan S. The first part of the thesis focuses on the literature search which deals with skin anatomy, skin diseases and skin microbiome and its function. The experimental part is focused on monitoring changes in the quantity of selected microorganisms of the skin microbiome, before and after application of polysaccharides to the skin using qPCR. Staphylococcus epidermidis, Cutibacterium acnes, Escherichia coli and Micrococcus luteus were monitored. The PCR products were detected by agarose gel electrophoresis. The bacterium Staphylococcus epidermidis was detected on the skin to the greatest extent, especially after the application of the polysaccharides Nanomoist and PoLevan S. Thus, a positive effect of both polysaccharides on the growth of this bacterium was found.

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Hudens mikrobiom utgör ett skydd mot patogener och domineras framförallt av grampositiva bakterier som Staphylococcus, där S. epidermidis är den mest förekommande arten. På huden är S. epidermidis en kommensal art och har en aktiv roll i att begränsa tillväxten och koloniseringen av patogener som S. aureus. S. epidermidis är en viktig art i att bibehålla stabiliteten av hudens mikrobiom och en reduktion av S. epidermidis innebär ökade risker för dysbios. S. aureus är en vanlig orsak till hudinfektioner och har upptäckts finnas i en hög utsträckning på patienter med atopisk dermatit samt hos diabetiker med risk för att utveckla fotsår. Syftet med denna studie var att optimera qPCR och undersöka om metoden kunde användas för att uppskatta förhållandet mellan S. aureus och S. epidermidis, vilket skulle kunna identifiera patienter med risk för hudsjukdomar. Tre primerpar undersöktes. Två var specifika mot S. aureus och en var specifik mot S. epidermidis. Flera tester genomfördes för att optimera qPCR och specificiteten av primerparen. Den annealingtemperatur och primerkoncentration som gav mest adekvata resultat användes vid den relativa kvantifieringen. Den utvecklade metoden kunde uppskatta förhållandet mellan S. aureus och S. epidermidis vid de lägre koncentrationerna (80–280 kopior/μl) bättre än vid de högre koncentrationerna (80 000–280 000 kopior/μl), men vidare optimering av metoden behövs.

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Tiefe sternale Wundheilungsstörungen nach medianer Sternotomie treten mit einer Inzidenz 0,4 – 5 % selten auf. Dennoch stellen sie mit einer Mortalität von 10 – 47 % ernst zu nehmende Komplikationen in einem multimorbiden Patientengut dar. Mikrobiologisch dominieren vorwiegend Erreger des grampositiven Spektrums. Mischinfektionen und multiresistente Erreger verstärken zudem die mikrobiologische Komplexität. Auf Grundlage dessen sind tiefe sternale Wundinfektionen im klinischen Alltag nur schwer kontrollierbar und stellen damit eine große Herausforderung dar. In dem Zeitraum von Mai 2012 bis Mai 2013 wurden 52 Patienten mit tiefen sternalen Wundheilungsstörungen im Universitätsklinikum Leipzig und im Herzzentrum Leipzig behandelt. Die sternalen Wundinfektionen traten hierbei als postoperative Komplikationen nach herzchirurgischen Eingriffen auf. Das einheitliche Behandlungskonzept bestand aus einem radikalen chirurgischen Wunddébridement und einer lappenplastischen Brustwandrekonstruktion unter simultaner resistogrammgerechter Antibiotikatherapie. Im Rahmen des Wunddébridements wurden drei mikrobiologische Abstriche nach standardisiertem Schema entnommen (präoperativ, intraoperativ vor Débridement, intraoperativ nach Débridement). Die Datenauswertung erfolgte retrospektiv anhand der Patientenakten und klinikinternen Datenbanken. Von besonderem Interesse waren patientenspezifische Parameter wie Alter, Geschlecht und das Vorhandensein von Komorbiditäten. Ebenso waren die mikrobiologischen Abstriche, die Methode der plastischen Rekonstruktion sowie Komplikationen im stationären Verlauf von großer Relevanz. Ziel dieser retrospektiven Arbeit war es, das vorherrschende bakteriologische Keimspektrum zu analysieren und das Patientenkollektiv im Hinblick auf Risikofaktoren für sternale Wundheilungsstörungen zu untersuchen. Ebenso wurde die Effektivität des angewandten Therapiekonzeptes unserer Klinik im Patientenkollektiv geprüft. Wir konnten nachweisen, dass Staphylokokkus epidermidis in 64,5 % der Fälle der dominierende Erreger im intraoperativen Abstrich war. Infektionen bedingt durch multiresistente Erreger kamen nur bei 12,9 % der Patienten vor. Intraoperative Wundbesiedelungen mit MRSA waren in der vorliegenden Arbeit nicht nachweisbar. Zudem konnten wir die ernst zu nehmende Pathogenität des Staphylokokkus epidermidis belegen. Im Vergleich zur Kontrollgruppe entwickelten Patienten mit Nachweis von Staphylokokkus epidermidis deutlich schneller sternale Wundheilungsstörungen nach der kardiochirurgischen Operation (47,4 Tage vs. 71,5 Tage). Ebenso resultierten mehr Revisionseingriffe (55,5 % vs. 27,3 %) und hierdurch längere Hospitalisierungszeiten (65,6 T age vs. 37,4 T age). Die antibiotische Behandlung des Staphylokokkus epidermidis ist daher bei diesem Patientengut im Therapiealgorithmus neben einem radikalen Débridement essentiell. Nur in der Kombination kann das Infektionsgeschehen effektiv beherrscht und der Patient aus dieser lebensbedrohlichen Situation geführt werden.

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No description available.