Class of 2013 Abstract / Specific Aims: The purpose of this study was to assess risk factors that are associated with inpatient charges and mortality rates with methicilin-resistant Staphylococcus aureu (MRSA), Aspergillus, and Candida in children with leukemia. The hypotheses is MRSA and opportunistic fungal infections are associated with higher inpatient charges and worse outcomes relative to those without. Children with leukemia are especially at risk due to underdeveloped and compromised immune systems. The rational is that identifying the risk factors that affect mortality and inpatient charges of these infections should add to the current knowledge of treating and preventing these infections in immune compromised patients. Methods: Retrospective cohort study using the Agency for Healthcare Research Quality (AHRQ) KIDS 2009 database. Inclusion criteria was defined as 17 years of age or younger and a ICD-9 code for an active infection with MRSA, Aspergillus, or Candida. Regression analysis's were performed to identify factors that had an impact on mortality, length of stay, and direct patient charges. Main Results: Values that were significant (p≤0.05) for predicting an increase in mortality were Age (year), MRSA, Aspergillus, Candida, and Deyo-Charlson comorbidity scores. Significant predictors of increased Inpatient Charges (p≤0.05) was Age (year), MRSA, Aspergillus, Candida, Female Sex, Deyo-Charlson comorbidity scores, Urban Hospitals, and the Southern Region. Significant predictors of Increased Length of Stay (p≤0.05) MRSA, Aspergillus, Candida, Female Sex, Deyo-Charlson, Urban Hospitals, Teaching Hospitals, and the Southern Region. Conclusion: Although uncommon in leukemia cases involving pediatrics or young adults, statistically significant and large risks of higher mortality, length of stay, and inpatient charges were noted in cases involving MRSA, aspergillus, and candida. Increasing Deyo-Charlson comorbidities scores were also consistently important predictors for poor outcomes in these leukemia patients and, with certain outcomes, increasing age and female sex.

The present thesis focuses on nosocomial infections and their causes. The theoretical part summarizes information on especially the sources of these infections, the way of their transfer and the most frequent infection agents. Clostridium difficile is studied in more detail. The illness caused by this pathogen leads to the complications especially with in-patients with a previous antibiotic therapy and immuno-compromised individuals. And this is the problem the present thesis is also concerned with. The thesis also divides nosocomial infections by their clinical manifestations and contains basic anti-epidemic measures and surveillance methods. Its empirical part focuses on the description of an occurrence of select nosocomial infections in Strakonice Hospital, a.s., especially an occurrence of the Clostridium difficile pathogen and methicillin-resistant Staphylococcus aureus (MRSA).

O objetivo foi avaliar a ação do ácido indol-3-acético (AIA) na ausência e presença da peroxidase de raiz forte (HRP) sobre a viabilidade de cepas certificadas de Staphylococcus aureus ATCC 6538 (produtora de biofilme), ATCC 14458 (portadora do gene seb), ATCC 43300 (portadora do gene mecA) e ATCC 29213 (controle). Foi determinada a concentração inibitória mínima (CIM) e, após, escolheu-se uma concentração subinitória para avaliar a capacidade de formação de biofilme, atividade de enzimas antioxidantes, integridade de DNA, integridade de membrana para todas as cepas e avaliação da expressão do gene seb da cepa ATCC 14458. A CIM observada foi de 40 mM de AIA para cepa ATCC 6538, 60 mM para ATCC 43300,50 mM para ATCC 14458 e ATCC 29213. No entanto, houve diferença significativa (P<0,05), na presença de HRP, somente para as cepas ATCC 14458 e 43300. A concentração subinibitória de AIA utilizada foi de 30 mM para ATCC 6538 e ATCC 14458, e de 40 mM para ATCC 43300 e ATCC 29213.mM. A capacidade de formação de biofilme pelo S. aureus(biofilme positivo) não foi alterada pela presença de AIA ou AIA/HRP. A atividade específica de catalase e superóxido dismutase foi aumentada pela ação de AIA/HRP (P<0,05) para todas as cepas, porém, não houve diferença (P>0,05) entre AIA e AIA/HRP. O DNA manteve-se íntegro, porém, houve diminuição na integridade de membrana (P<0,05) para todas as cepas cultivadas na concentração subinibitória de AIA, sendo este efeito potencializado na presença de HRP somente para ATCC 43300. Houve redução na expressão do gene seb da cepa ATCC 14458 (P<0,05). Conclui-se que o AIA, associado à HRP apresenta efeito bacteriostático em cepas de Staphylococcus aureus portadoras de diferentes fatores de virulência. / The objective was to evaluate the action of indole-3-acetic acid (IAA) in the absence and presence of horseradish peroxidase (HRP) on the viability of certified strains of Staphylococcus aureus ATCC 6538 (producer of biofilm), ATCC 14458 (carrying the seb gene), ATCC 43300 (carrying the mecA gene) and ATCC29213 (control). Concentration was determined minimum inhibitory (CIM) and, after, we chose a concentration subinitória to assess the ability of biofilm formation, activity of antioxidant enzymes, integrity DNA, membrane integrity for all strains and evaluation of gene expression seb of strain ATCC 14458. The MIC for the strains ATCC 6538 was 40 mM IAA , for the strains ATCC 14458 and 29213 was 50 mM and for the strain ATCC 43300 was 60 mM IAA, however, significant differences (P<0.05) compared to the addition of HRP to the culture medium, was found only for the doses of MIC of the strains ATCC14458 and 43300. The concentration subinibitória IAA used was 30 mM for ATCC 6538 and ATCC 14458, and 40 mM to ATCC 43300 and ATCC 29213.mM for ATCC 6538 and ATCC 14458, and 50 mM to ATCC 43300 and ATCC 29213.The ability of biofilm formation by S. aureus(biofilm-positive) was not altered by the presence of IAA or IAA/HRP in the culture medium. The activity of catalase and superoxide dismutase was influenced by the action of IAA and IAA/HRP (P<0.05), but there was no difference (P≥0.05) between IAA and IAA/HRP. The DNA remained intact, however, there were changes (P<0.05) in the membrane integrity of microorganisms to sub-inhibitory concentrations of IAA and IAA/HRP, as well as the expression of the sebgene was lower (P<0.05) in microorganisms treated with IAA and IAA/HRP. It is concluded that the IAA has bacteriostatic effect on S. aureus carrying different virulence factors.

Made available in DSpace on 2016-04-28T20:17:24Z (GMT). No. of bitstreams: 1 2007- Lidiane de Castro Soares.pdf: 1336181 bytes, checksum: 47680a74331a705e5ecd649c1639aa3f (MD5) Previous issue date: 2007-02-28 / Funda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Coagulasenegative staphylococci (SCN) take part of normal microbiota. Although it has been considered saprophytic, nowadays there is a concern about its pathogenic potential.Nevertheless, all advance in SCN identification assays, these microorganisms are continually neglected in laboratorial routine of infectious diseases, because of the wide range of species. In spite of the difficulties, it is necessary to make an appropriated species identification in order to differentiate the potential pathogenic agents and to determine its antimicrobial susceptibility pattern. Resistance in SCN is related to the presence of mecA gene, which expresses a heterogeneous pattern that can be detected through diverse phenotypics tests. In this study, 72 SCN isolates obtained from external ear conducts of dogs, bovine mastitis and human nosocomial infections were evaluated. Staphylococcus xylosus was the most prevalent microorganism in animal and S. cohnii subsp.urealyticus in human sample. The antimicrobial resistance patterns were evaluated through disc diffusion test, and a high level of resistance to penicillin and ampicillin were detected. The most efficient antibiotics evaluated were gentamicin, vancomicin and the association ampicillin+sulbactam. Oxacillin resistance was phenotypically detected by modified disc diffusion test, agar screen, broth microdilution and agar dilution. Presence of mecA gene where detected in 5,55% of the isolates by Polimerase Chain Reaction (PCR). Correlation with the detection of mecA gene was used as gold standard for evaluation of sensitivity and specificity of phenotypical assays. The low number of mecA positives isolates did not allowed the association between cefoxitin and oxacillin resistance as a test to detect the presence this gene. The broth microdilution and agar dilution tests presented the best accuracy in phenotypic detection of the oxacillin resistance. / Os estafilococos coagulasenegativos (ECN) fazem parte da microbiota normal da pele e apesar de terem sido considerados sapr?fitas por muito tempo, o seu significado cl?nico como agente etiol?gico tem aumentado com o passar dos anos. No entanto, apesar de todo avan?o nas t?cnicas de identifica??o dos ECN e do conhecimento destes como agentes etiol?gicos em diversos processos infecciosos, estes microrganismos muitas vezes s?o negligenciados na rotina laboratorial, devido a enorme diversidade de esp?cies encontradas. A identifica??o das esp?cies de ECN, embora de dif?cil realiza??o para a maioria dos laborat?rios cl?nicos, ? necess?ria para diferenciar o potencial patog?nico e o perfil de resist?ncia de cada isolado. A resist?ncia ? oxacilina em ECN ? mediada pelo gene mecA e usualmente heterog?nea, sendo detectada por v?rios m?todos fenot?picos. Neste estudo, foram avaliados 72 isolados de ECN provenientes de amostras do conduto auditivo de c?es da ra?a Beagles, de mastite bovina e de infec??es humanas. Staphylococcus xylosus foi o microrganismo mais isolado, nas amostras animais, e S. cohnii subsp.urealyticus em humanos, dentro de uma ampla gama de esp?cies identificadas. O perfil de resist?ncia aos antimicrobianos em isolados de animais e humanas foi avaliado atrav?s da t?cnica de difus?o em disco, na qual detectouse um elevado n?vel de resist?ncia ? penicilina e ampicilina. A gentamicina, vancomicina e associa??o ampicilina+sulbactam foram eficientes frente aos isolados testados. A avalia??o da resist?ncia ? oxacilina foi realizada atrav?s da difus?o em disco modificada, ?gar screen, microdilui??o em caldo e em ?gar. A presen?a do gene mecA foi determinada pelo m?todo da Rea??o em Cadeia de Polimerase (PCR), sendo 5,55% dos isolados mecA positivos. Os resultados fenot?picos de resist?ncia a cefoxitina e a oxacilina foram correlacionados com a detec??o do gene mecA, que foi utilizado como padr?o ouro para avalia??o da sensibilidade e especificidade das t?cnicas utilizadas. O reduzido n?mero de isolados mecA + n?o permitiu estabelecer o grau de correla??o entre a cefoxitina e a oxacilina como m?todo de predi??o do gene, embora ambas tenha apresentado o mesmo percentual de resist?ncia. O teste fenot?pico que apresentou melhor acur?cia na detec??o da resist?ncia ? oxacilina foi a microdilui??o em caldo.

Made available in DSpace on 2014-06-11T19:27:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-28Bitstream added on 2014-06-13T20:35:47Z : No. of bitstreams: 1 maluta_rp_me_jabo.pdf: 175883 bytes, checksum: 21d3132e7d1e30684a695722f9886e2d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo desse estudo foi isolar cepas de estafilococos resistentes a meticilina (MRS) de humanos e cães e verificar os seus respectivos perfis de suscetibilidade a diversos antimicrobianos em um hospital veterinário de ensino no Brasil. Foram analisados espécimes de 50 pessoas e de 50 cães para isolamento de estafilococos que foram identificados através de características morfotintoral, reações bioquímicas e resistência a antimicrobianos. Foi verificado o perfil de suscetibilidade a oxacilina, penicilina, ciprofloxacina, gentamicina, clindamicina, eritromicina, sulfametoxazol+trimetoprim e vancomicina. Não foram isolados Staphylococcus aureus resistentes a meticilina (MRSA) e Staphylococcus intermedius resistentes a meticilina (MRSI). Foram isoladas duas cepas de estafilococos coagulase-negativas resistentes a meticilina (MRCoNS) de cães (4%) e 18 cepas de MRCoNS de seres humanos (36%). Todas as cepas de estafilococos isoladas foram sensíveis a vancomicina. O isolamento de MRCoNS merece atenção devido à hipótese que estes podem transferir o gene mecA para estafilococos coagulase-positivos. A ausência de cepas de MRSA resistentes a vancomicina é importante por ser este um antibiótico que pode ser usado como alternativa extrema. Programas de vigilância de MRS devem ser estimulados em unidades de saúde veterinárias. / The aim of this study was to isolate methicillin-resistant staphylococci (MRS) strains from humans and dogs and verify their susceptibility profile to several antimicrobials in a veterinary teaching hospital in Brazil. Specimens from 50 people and 50 dogs were analyzed to the isolation of staphylococci. They were identified trough gram staining, biochemical reactions and resistance to antimicrobials. Susceptibility profile to oxacillin, penicillin, ciprofloxacin, gentamicin, clindamycin, erythromycin, trimethoprim + sulfamethoxazole and vancomycin was verified. It was not isolated any strain of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus intermedius (MRSI). Two strains of methicillin-resistant coagulasenegative staphylococci (MRCoNS) were isolated from dogs (4%) and 18 strains were isolated from humans (36%). MRCoNS isolation is important because there is the hypothesis that mecA transferring from MRCoNS to coagulase-positive staphylococci could occur. The absence of MRSA strains resistant to vancomycin is important because this antimicrobial could be used as an important alternative against MRSA infections. Surveillance programs aiming MRS should be stimulated in veterinary health units.

Orientador: Fernado Antônio de Ávila / Banca: José Moacir Marin / Banca: Luiz Florencio Franco Margatho / Resumo: O objetivo desse estudo foi isolar cepas de estafilococos resistentes a meticilina (MRS) de humanos e cães e verificar os seus respectivos perfis de suscetibilidade a diversos antimicrobianos em um hospital veterinário de ensino no Brasil. Foram analisados espécimes de 50 pessoas e de 50 cães para isolamento de estafilococos que foram identificados através de características morfotintoral, reações bioquímicas e resistência a antimicrobianos. Foi verificado o perfil de suscetibilidade a oxacilina, penicilina, ciprofloxacina, gentamicina, clindamicina, eritromicina, sulfametoxazol+trimetoprim e vancomicina. Não foram isolados Staphylococcus aureus resistentes a meticilina (MRSA) e Staphylococcus intermedius resistentes a meticilina (MRSI). Foram isoladas duas cepas de estafilococos coagulase-negativas resistentes a meticilina (MRCoNS) de cães (4%) e 18 cepas de MRCoNS de seres humanos (36%). Todas as cepas de estafilococos isoladas foram sensíveis a vancomicina. O isolamento de MRCoNS merece atenção devido à hipótese que estes podem transferir o gene mecA para estafilococos coagulase-positivos. A ausência de cepas de MRSA resistentes a vancomicina é importante por ser este um antibiótico que pode ser usado como alternativa extrema. Programas de vigilância de MRS devem ser estimulados em unidades de saúde veterinárias. / Abstract: The aim of this study was to isolate methicillin-resistant staphylococci (MRS) strains from humans and dogs and verify their susceptibility profile to several antimicrobials in a veterinary teaching hospital in Brazil. Specimens from 50 people and 50 dogs were analyzed to the isolation of staphylococci. They were identified trough gram staining, biochemical reactions and resistance to antimicrobials. Susceptibility profile to oxacillin, penicillin, ciprofloxacin, gentamicin, clindamycin, erythromycin, trimethoprim + sulfamethoxazole and vancomycin was verified. It was not isolated any strain of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus intermedius (MRSI). Two strains of methicillin-resistant coagulasenegative staphylococci (MRCoNS) were isolated from dogs (4%) and 18 strains were isolated from humans (36%). MRCoNS isolation is important because there is the hypothesis that mecA transferring from MRCoNS to coagulase-positive staphylococci could occur. The absence of MRSA strains resistant to vancomycin is important because this antimicrobial could be used as an important alternative against MRSA infections. Surveillance programs aiming MRS should be stimulated in veterinary health units. / Mestre

O objetivo foi avaliar a ação do ácido indol-3-acético (AIA) na ausência e presença da peroxidase de raiz forte (HRP) sobre a viabilidade de cepas certificadas de Staphylococcus aureus ATCC 6538 (produtora de biofilme), ATCC 14458 (portadora do gene seb), ATCC 43300 (portadora do gene mecA) e ATCC 29213 (controle). Foi determinada a concentração inibitória mínima (CIM) e, após, escolheu-se uma concentração subinitória para avaliar a capacidade de formação de biofilme, atividade de enzimas antioxidantes, integridade de DNA, integridade de membrana para todas as cepas e avaliação da expressão do gene seb da cepa ATCC 14458. A CIM observada foi de 40 mM de AIA para cepa ATCC 6538, 60 mM para ATCC 43300,50 mM para ATCC 14458 e ATCC 29213. No entanto, houve diferença significativa (P<0,05), na presença de HRP, somente para as cepas ATCC 14458 e 43300. A concentração subinibitória de AIA utilizada foi de 30 mM para ATCC 6538 e ATCC 14458, e de 40 mM para ATCC 43300 e ATCC 29213.mM. A capacidade de formação de biofilme pelo S. aureus(biofilme positivo) não foi alterada pela presença de AIA ou AIA/HRP. A atividade específica de catalase e superóxido dismutase foi aumentada pela ação de AIA/HRP (P<0,05) para todas as cepas, porém, não houve diferença (P>0,05) entre AIA e AIA/HRP. O DNA manteve-se íntegro, porém, houve diminuição na integridade de membrana (P<0,05) para todas as cepas cultivadas na concentração subinibitória de AIA, sendo este efeito potencializado na presença de HRP somente para ATCC 43300. Houve redução na expressão do gene seb da cepa ATCC 14458 (P<0,05). Conclui-se que o AIA, associado à HRP apresenta efeito bacteriostático em cepas de Staphylococcus aureus portadoras de diferentes fatores de virulência. / The objective was to evaluate the action of indole-3-acetic acid (IAA) in the absence and presence of horseradish peroxidase (HRP) on the viability of certified strains of Staphylococcus aureus ATCC 6538 (producer of biofilm), ATCC 14458 (carrying the seb gene), ATCC 43300 (carrying the mecA gene) and ATCC29213 (control). Concentration was determined minimum inhibitory (CIM) and, after, we chose a concentration subinitória to assess the ability of biofilm formation, activity of antioxidant enzymes, integrity DNA, membrane integrity for all strains and evaluation of gene expression seb of strain ATCC 14458. The MIC for the strains ATCC 6538 was 40 mM IAA , for the strains ATCC 14458 and 29213 was 50 mM and for the strain ATCC 43300 was 60 mM IAA, however, significant differences (P<0.05) compared to the addition of HRP to the culture medium, was found only for the doses of MIC of the strains ATCC14458 and 43300. The concentration subinibitória IAA used was 30 mM for ATCC 6538 and ATCC 14458, and 40 mM to ATCC 43300 and ATCC 29213.mM for ATCC 6538 and ATCC 14458, and 50 mM to ATCC 43300 and ATCC 29213.The ability of biofilm formation by S. aureus(biofilm-positive) was not altered by the presence of IAA or IAA/HRP in the culture medium. The activity of catalase and superoxide dismutase was influenced by the action of IAA and IAA/HRP (P<0.05), but there was no difference (P≥0.05) between IAA and IAA/HRP. The DNA remained intact, however, there were changes (P<0.05) in the membrane integrity of microorganisms to sub-inhibitory concentrations of IAA and IAA/HRP, as well as the expression of the sebgene was lower (P<0.05) in microorganisms treated with IAA and IAA/HRP. It is concluded that the IAA has bacteriostatic effect on S. aureus carrying different virulence factors.