Made available in DSpace on 2016-12-23T13:55:59Z (GMT). No. of bitstreams: 1 Dissertacao mestrado Nazareth Klein.pdf: 9490018 bytes, checksum: a860548b52d0cb66494dd6a26c4306df (MD5) Previous issue date: 2006-10-23 / Este estudo foi conduzido para se avaliar a prevalência e a distribuição das espécies de Candida spp., isoladas de diversos materiais biológicos, obtidos de pacientes internados no Hospital Universitário Cassiano Antonio de Moraes (HUCAM), Vitória-ES, durante o período de janeiro de 2003 a dezembro 2005, e descrever o perfil de suscetibilidade de Candida não-albicans a drogas antifúngicas. A dentificação da espécie Candida albicans foi realizada no meio cromogênico CHROMagar-Candida® e confirmada pela indução positiva de tubo germinativo e/ou clamidoconídeo. A identificação das espécies não-albicans foi realizada pelos seus padrões morfológicos e bioquímicos e pelo sistema comercial API 20 C AUX. O perfil de suscetibilidade às drogas antifúngicas anfotericina B, itraconazol e fluconazol foi estabelecido pelo método de referência, microdiluição em caldo, de acordo com o documento M27-A2 do CLSI, 2002. Estudo comparativo foi realizado, empregandose o método Etest® para as cepas de Candida tropicalis, sendo incluída nesta etapa, a droga voriconazol. Os resultados mostraram 268 isolados de Candida spp., no geral, um predomínio de Candida não-albicans (54%), incluindo C. tropicalis (26%), C. parapsilosis (14%), C. glabrata (10%), C. Krusei (2%), C. guilliermondii (1.5%) e C. lusitaniae (0.5%). Ocorreu uma marcante transição na prevalência de Candida não-albicans, durante os três anos de estudo, oscilando na freqüência de 39% dos isolamentos em 2003 para 74% em 2005. As espécies não-albicans foram mais isoladas de urina (47%), seguido de sangue (39%) e cateter (8%). As maiores ocorrências foram na Unidade de Tratamento Intensivo Geral e Clínica Médica, com 23% em cada setor. A resistência para as espécies não-albicans ao fluconazol foi de 4% e, de suscetível dose-dependente (SDD) ao fluconazol e ao itraconazol foi de 2% e 3%, respectivamente (metodologia CSLI). Com esta metodologia, 98% das cepas de C. tropicalis foram sensíveis ao fluconazol e ao itraconazol, mas 2% foram SDD para estas drogas. Com o método Etest®, 2% dessas cepas foram resistentes (R) a ambas as drogas e 19% foram SDD ao itraconazol. Nenhum isolado de Candida não-albicans foi resistente a anfotericina B ou ao voriconazol. Todos os isolados de Candida albicans foram sensíveis às três drogas analisadas. Em conclusão: a) Houve uma marcante tendência para o isolamento de C. não-albicans no HUCAM, com predomínio de C. tropicalis. b) Observou-se, reduzida suscetibilidade ao fluconazol e ao itraconazol apenas para as espécies C. tropicalis, C. krusei e C. glabrata. c) O método Etest®, usando o meio de ágar Casitone, apresentou ótima concordância com o método de referência para as drogas anfotericina B (100%) e fluconazol (96%), no entanto, para o itraconazol, essa concordância foi de 62%. / This study was carried out to evaluate the prevalence and distribution of the species of Candida spp. Isolated from several biological materials obtained from patients interned at the Hospital Universitário Cassiano Antonio de Moraes (HUCAM), Vitória-ES, during the period of January 2003 to December 2005, and to describe the profile of susceptibility of Candida non-albicans to antifungal drugs. The identification of the species of Candida albicans was made into the chromogenic medium CHROMagar-Candida® and confirmed through the positive induction of a germinative tube and/or clamidoconídeo. The identification of the species nonalbicans was made through of morphologic and biochemical patterns and through the commercial system API 20 C AUX. The profile of susceptibility to the antifungal drugs amphotencin B itraconazole and fluconazole was established by the reference method, broth microdilution, according to the document M27-A2 from CLSI, 2002. A comparative study was conducted, using the method Etest® for the strains of Candida tropicalis being included at this stage, the drug voriconazole. The results showed that 268 isolates of Candida spp. were obtained during the period of the study and, in general, a predominance was observed in the isolation of Candida nonalbicans (54%) at HUCAM, including C. tropicalis (26%), C. parapsilosis (14%), C. glabrata (10%), C. krusei (2%), C. guilliermondii (1.5%) and C. lusitaniae (0.5%). There was a relevant transition in the prevalence of Candida non-albicans during the three years of study, oscillating in the frequency from 39% of the isolations in 2003 to 74% in 2005. The non-albicans species were more isolated from urine (47%), followed by blood (39%) and catheter (8%). The biggest occurrences were noticed in patients interned at the General Intensive Care Unit and Medical Clinic with a rate of 23% in each one. The resistance rate noticed for the non-albicans species to fluconazol was of 4% and, of susceptible dose-dependent (SDD) to fluconazole and to itraconazole was of 2% and 3% respectively (methodology CSLI). With the use of this methodology, it was observed that 98% of the strains of C.tropicalis were sensitive to fluconazole and to itraconazole, but 2% were SDD to these drugs. When the method Etest® was used, 2% of these samples were resistant to both drugs and 19% were SDD to itraconazole. No isolated of Candida non-albicans was resistant to amphotericin B or to voriconazole. All the isolates of Candida albicans were sensitive to the three drugs analyzed. With this study it was concluded that there was a striking tendency to the isolation of C. non-albicans, with the predominance of C. tropicalis. Reduced susceptibility to fluconazole and itraconazole was also noticed only in the species C. tropicalis, C. krusei and C. glabrata, reinforcing the concern about a high number of isolations of these species non-albicans at HUCAM. The Etest® method using the medium of agar Casitone, showed an excellent concordance with the reference method to the drugs amphotericin B (100%) and fluconazole (96%). However, for the itraconazole this agreement was of 62%.

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Introdução: Pneumonias causadas por Staphylococcus aureus meticilina resistentes (Methicillin resistant Staphylococcus aureus - MRSA) estão associadas com uma alta mortalidade em pacientes internados em Centros de Terapia Intensiva (CTI). Não é incomum que ocorra falha no tratamento de pneumonia por MRSA com vancomicina. Alguns estudos têm mostrado que há uma relação entre mortalidade em infecções por MRSA e concentrações inibitórias mínimas (CIM) elevadas para vancomicina por Etest®, apesar dos isolados serem sensíveis à vancomicina (CIM ≤2μg/mL), conforme o Clinical Laboratory Standards Institute (CLSI). Objetivos: O objetivo deste estudo foi avaliar as CIMs para vancomicina em pacientes com diagnóstico de pneumonia por MRSA, descrever a relação entre a CIM da vancomicina obtido por Etest® e microdiluição em caldo com mortalidade em 30 dias em pacientes com pneumonia por MRSA internados em um hospital terciário, acadêmico, no sul do Brasil. Métodos: Estudo prospectivo de coorte. Foram incluídos todos os pacientes com pneumonia adquirida no hospital (hospital adquired pneumonia - HAP) e pneumonia associada à ventilação (ventilator associated pneumonia - VAP) por MRSA entre Junho de 2009 e Dezembro de 2011. A CIM da vancomicina do primeiro isolado do trato respiratório foi determinado por Etest® e microdiluição em caldo. As variáveis selecionadas para serem analisadas incluíram idade, sexo, comorbidades, dosagem sérica de vancomicina, escore APACHE II (Acute Physiological and Chronic Health Evaluation II) e a presença do accessory gene regulator II (agr II). O desfecho principal foi morte em 30 dias. Resultados: Foram incluídos 85 pacientes. A mortalidade em 30 dias foi de 42,4%. Em sessenta e oito pacientes (80,1%) as CIMs da vancomicina obtidos por Etest® foram menores ou iguais a 1,0μg/mL, enquanto todas as CIMs (N=85) obtidas por microdiluição em caldo foram ≤1,0μg/mL. Não houve correlação entre as CIMs por Etest® e microdiluição em caldo com morte em 30 dias. Quarenta e seis isolados de MRSA (54,1%) foram positivos para o locus agr II. A média da vancomicina sérica foi 20,7μg/mL (12,0 – 29,0) e a média do escore APACHE II foi 23,0 (17,5 – 29,0). O escore APACHE II foi relacionado com mortalidade em 30 dias. Conclusões: As CIMs da vancomicina obtidas por Etest® e microdiluição em caldo não foram associadas com altos níveis de mortalidade em pacientes com pneumonia grave por MRSA. / Background: Pneumonias due to methicillin resistant Staphylococcus aureus (MRSA) are associated with significant mortality in Intensive Care Units (ICU) patients. Vancomycin treatment failure in MRSA pneumonia patients is not uncommon. Some reports found a link between high mortality in MRSA infections and higher vancomycin minimum inhibitory concentrations (MICs) by Etest®, despite the susceptibility of the isolates (MIC ≤2,0μg/mL), according to the Clinical Laboratory Standards Institute (CLSI). Objectives: The purpose of this study was evaluate vancomycin MICs in patients with MRSA pneumonia, describe the relationship between vancomycin MIC generated by Etest® and broth microdilution with 30-day mortality in MRSA pneumonia patients. Methods: We conducted a prospective cohort study. All patients with MRSA hospital acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP) admitted in ICU between June 2009 and December 2011 were included. Vancomycin MIC for the first isolate of the respiratory tract was determined by Etest® and broth microdilution. The variables selected for the analysis included age, sex, comorbid conditions, vancomycin serum trough concentration, APACHE II (Acute Physiological and Chronic Health Evaluation II) score and the presence of the accessory gene regulator II (agr II). The primary outcome was mortality at 30-day. Results: Eithy five patients were included. All case mortality at the 30-day was 42.4%. Sixty eight of the patients (80.1%) of the vancomycin MICs generated by Etest® were equal or below 1.0μg/mL. While all MICs generated by broth microdilution were ≤1.0μg/mL. MICs generated by Etest® and broth microdilution were not associated with 30-day mortality. Forty six isolates of MRSA (54.1%) from patients were positive for agr II. The median serum vancomycin was 20.7μg/mL (12.0 – 29.0) and the median APACHE II score was 23.0 (17.5 – 29.0). The APACHE II score was related to 30-day mortality. Conclusions: Vancomycin MICs by Etest® and broth microdilution were not associated with higher mortality leves in patients with severe MRSA pneumonia.

Introdução: Pneumonias causadas por Staphylococcus aureus meticilina resistentes (Methicillin resistant Staphylococcus aureus - MRSA) estão associadas com uma alta mortalidade em pacientes internados em Centros de Terapia Intensiva (CTI). Não é incomum que ocorra falha no tratamento de pneumonia por MRSA com vancomicina. Alguns estudos têm mostrado que há uma relação entre mortalidade em infecções por MRSA e concentrações inibitórias mínimas (CIM) elevadas para vancomicina por Etest®, apesar dos isolados serem sensíveis à vancomicina (CIM ≤2μg/mL), conforme o Clinical Laboratory Standards Institute (CLSI). Objetivos: O objetivo deste estudo foi avaliar as CIMs para vancomicina em pacientes com diagnóstico de pneumonia por MRSA, descrever a relação entre a CIM da vancomicina obtido por Etest® e microdiluição em caldo com mortalidade em 30 dias em pacientes com pneumonia por MRSA internados em um hospital terciário, acadêmico, no sul do Brasil. Métodos: Estudo prospectivo de coorte. Foram incluídos todos os pacientes com pneumonia adquirida no hospital (hospital adquired pneumonia - HAP) e pneumonia associada à ventilação (ventilator associated pneumonia - VAP) por MRSA entre Junho de 2009 e Dezembro de 2011. A CIM da vancomicina do primeiro isolado do trato respiratório foi determinado por Etest® e microdiluição em caldo. As variáveis selecionadas para serem analisadas incluíram idade, sexo, comorbidades, dosagem sérica de vancomicina, escore APACHE II (Acute Physiological and Chronic Health Evaluation II) e a presença do accessory gene regulator II (agr II). O desfecho principal foi morte em 30 dias. Resultados: Foram incluídos 85 pacientes. A mortalidade em 30 dias foi de 42,4%. Em sessenta e oito pacientes (80,1%) as CIMs da vancomicina obtidos por Etest® foram menores ou iguais a 1,0μg/mL, enquanto todas as CIMs (N=85) obtidas por microdiluição em caldo foram ≤1,0μg/mL. Não houve correlação entre as CIMs por Etest® e microdiluição em caldo com morte em 30 dias. Quarenta e seis isolados de MRSA (54,1%) foram positivos para o locus agr II. A média da vancomicina sérica foi 20,7μg/mL (12,0 – 29,0) e a média do escore APACHE II foi 23,0 (17,5 – 29,0). O escore APACHE II foi relacionado com mortalidade em 30 dias. Conclusões: As CIMs da vancomicina obtidas por Etest® e microdiluição em caldo não foram associadas com altos níveis de mortalidade em pacientes com pneumonia grave por MRSA. / Background: Pneumonias due to methicillin resistant Staphylococcus aureus (MRSA) are associated with significant mortality in Intensive Care Units (ICU) patients. Vancomycin treatment failure in MRSA pneumonia patients is not uncommon. Some reports found a link between high mortality in MRSA infections and higher vancomycin minimum inhibitory concentrations (MICs) by Etest®, despite the susceptibility of the isolates (MIC ≤2,0μg/mL), according to the Clinical Laboratory Standards Institute (CLSI). Objectives: The purpose of this study was evaluate vancomycin MICs in patients with MRSA pneumonia, describe the relationship between vancomycin MIC generated by Etest® and broth microdilution with 30-day mortality in MRSA pneumonia patients. Methods: We conducted a prospective cohort study. All patients with MRSA hospital acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP) admitted in ICU between June 2009 and December 2011 were included. Vancomycin MIC for the first isolate of the respiratory tract was determined by Etest® and broth microdilution. The variables selected for the analysis included age, sex, comorbid conditions, vancomycin serum trough concentration, APACHE II (Acute Physiological and Chronic Health Evaluation II) score and the presence of the accessory gene regulator II (agr II). The primary outcome was mortality at 30-day. Results: Eithy five patients were included. All case mortality at the 30-day was 42.4%. Sixty eight of the patients (80.1%) of the vancomycin MICs generated by Etest® were equal or below 1.0μg/mL. While all MICs generated by broth microdilution were ≤1.0μg/mL. MICs generated by Etest® and broth microdilution were not associated with 30-day mortality. Forty six isolates of MRSA (54.1%) from patients were positive for agr II. The median serum vancomycin was 20.7μg/mL (12.0 – 29.0) and the median APACHE II score was 23.0 (17.5 – 29.0). The APACHE II score was related to 30-day mortality. Conclusions: Vancomycin MICs by Etest® and broth microdilution were not associated with higher mortality leves in patients with severe MRSA pneumonia.

Introdução: Pneumonias causadas por Staphylococcus aureus meticilina resistentes (Methicillin resistant Staphylococcus aureus - MRSA) estão associadas com uma alta mortalidade em pacientes internados em Centros de Terapia Intensiva (CTI). Não é incomum que ocorra falha no tratamento de pneumonia por MRSA com vancomicina. Alguns estudos têm mostrado que há uma relação entre mortalidade em infecções por MRSA e concentrações inibitórias mínimas (CIM) elevadas para vancomicina por Etest®, apesar dos isolados serem sensíveis à vancomicina (CIM ≤2μg/mL), conforme o Clinical Laboratory Standards Institute (CLSI). Objetivos: O objetivo deste estudo foi avaliar as CIMs para vancomicina em pacientes com diagnóstico de pneumonia por MRSA, descrever a relação entre a CIM da vancomicina obtido por Etest® e microdiluição em caldo com mortalidade em 30 dias em pacientes com pneumonia por MRSA internados em um hospital terciário, acadêmico, no sul do Brasil. Métodos: Estudo prospectivo de coorte. Foram incluídos todos os pacientes com pneumonia adquirida no hospital (hospital adquired pneumonia - HAP) e pneumonia associada à ventilação (ventilator associated pneumonia - VAP) por MRSA entre Junho de 2009 e Dezembro de 2011. A CIM da vancomicina do primeiro isolado do trato respiratório foi determinado por Etest® e microdiluição em caldo. As variáveis selecionadas para serem analisadas incluíram idade, sexo, comorbidades, dosagem sérica de vancomicina, escore APACHE II (Acute Physiological and Chronic Health Evaluation II) e a presença do accessory gene regulator II (agr II). O desfecho principal foi morte em 30 dias. Resultados: Foram incluídos 85 pacientes. A mortalidade em 30 dias foi de 42,4%. Em sessenta e oito pacientes (80,1%) as CIMs da vancomicina obtidos por Etest® foram menores ou iguais a 1,0μg/mL, enquanto todas as CIMs (N=85) obtidas por microdiluição em caldo foram ≤1,0μg/mL. Não houve correlação entre as CIMs por Etest® e microdiluição em caldo com morte em 30 dias. Quarenta e seis isolados de MRSA (54,1%) foram positivos para o locus agr II. A média da vancomicina sérica foi 20,7μg/mL (12,0 – 29,0) e a média do escore APACHE II foi 23,0 (17,5 – 29,0). O escore APACHE II foi relacionado com mortalidade em 30 dias. Conclusões: As CIMs da vancomicina obtidas por Etest® e microdiluição em caldo não foram associadas com altos níveis de mortalidade em pacientes com pneumonia grave por MRSA. / Background: Pneumonias due to methicillin resistant Staphylococcus aureus (MRSA) are associated with significant mortality in Intensive Care Units (ICU) patients. Vancomycin treatment failure in MRSA pneumonia patients is not uncommon. Some reports found a link between high mortality in MRSA infections and higher vancomycin minimum inhibitory concentrations (MICs) by Etest®, despite the susceptibility of the isolates (MIC ≤2,0μg/mL), according to the Clinical Laboratory Standards Institute (CLSI). Objectives: The purpose of this study was evaluate vancomycin MICs in patients with MRSA pneumonia, describe the relationship between vancomycin MIC generated by Etest® and broth microdilution with 30-day mortality in MRSA pneumonia patients. Methods: We conducted a prospective cohort study. All patients with MRSA hospital acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP) admitted in ICU between June 2009 and December 2011 were included. Vancomycin MIC for the first isolate of the respiratory tract was determined by Etest® and broth microdilution. The variables selected for the analysis included age, sex, comorbid conditions, vancomycin serum trough concentration, APACHE II (Acute Physiological and Chronic Health Evaluation II) score and the presence of the accessory gene regulator II (agr II). The primary outcome was mortality at 30-day. Results: Eithy five patients were included. All case mortality at the 30-day was 42.4%. Sixty eight of the patients (80.1%) of the vancomycin MICs generated by Etest® were equal or below 1.0μg/mL. While all MICs generated by broth microdilution were ≤1.0μg/mL. MICs generated by Etest® and broth microdilution were not associated with 30-day mortality. Forty six isolates of MRSA (54.1%) from patients were positive for agr II. The median serum vancomycin was 20.7μg/mL (12.0 – 29.0) and the median APACHE II score was 23.0 (17.5 – 29.0). The APACHE II score was related to 30-day mortality. Conclusions: Vancomycin MICs by Etest® and broth microdilution were not associated with higher mortality leves in patients with severe MRSA pneumonia.

Antifungal drug resistance is a growing problem. Several mechanisms contribute to the development of resistance to antifungal agents. In South Africa, little is known about the antifungal susceptibility of local yeast isolates. The aim of this study was therefore to determine the antifungal susceptibility profile of local Candida species and Cryptococcus neoformans isolates and the molecular mechanisms of resistance to different antifungal agents in Pretoria. A total of 250 yeast isolates were collected from the diagnostic laboratory in the Department of Medical Microbiology at the University of Pretoria-National Health Laboratory Services. The isolates were subcultured on Sabouraud dextrose agar media for purity of yeast colonies. Identification to species level was performed using biochemical techniques. The antifungal susceptibility of 87 isolates was determined by the Etest for three azole antifungals (fluconazole, posaconazole and voriconazole), amphotericin B and caspofungin. Clinical breakpoint susceptibility was determined according to the CLSI clinical breakpoint reference methods. Polymerase chain reaction was performed on C. albicans isolates to amplify the ERG11 gene and the FKS1 gene. Sequencing was done for the amplification products and the sequence data were analysed by the CLC genome workbench software. Among the 250 isolates collected, Candida species accounted for 82.8% and C. neoformans accounted for 17.2% of the isolates. C. albicans was the most commonly isolated (76.8% of Candida species), of which 30% were resistant to caspofungin. Fluconazole resistance was detected in 56.7% of C. parapsilosis isolates, the highest fluconazole resistance among Candida species. Cross-resistance was found between fluconazole and voriconazole. Resistance to posaconazole was detected in 66.7% of C. glabrata isolates, whilst all other Candida species and C. neoformans isolates were fully susceptible. Furthermore, C. neoformans var. gattii isolates were less susceptible to azole antifungal agents than C. neoformans var. neoformans isolates. Molecular alterations are one of the mechanisms of resistance to azole and caspofungin antifungal agents. The amino acid substitutions D116E, K128T and V437I were detected in the ERG11 gene of two azole susceptible isolates. The new amino acid substitution E517Q was detected in the ERG11 gene of a resistant isolate. The S642L substitution was detected in the FKS1 gene of all the isolates that were caspofungin resistant and caspofungin susceptible. C. albicans was the most commonly isolated yeast species in Pretoria. Cross-resistance was detected between fluconazole and voriconazole. Therefore, these two agents are not a good alternative to use in the treatment of resistant isolates. With Candida species and C. neoformans isolates, there was less resistance to posaconazole than to fluconazole and voriconazole. The identification of the two varieties of C. neoformans is important in order to establish the differences in their antifungal susceptibility. Resistance to azole and caspofungin antifungal agents existed without the previously described molecular alterations in the ERG11 and FKS1 genes of resistant isolates. Further studies are required to explain the role of new amino acid substitutions, as well as the involvement of other mechanisms in resistance to antifungal drugs. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Medical Microbiology / unrestricted

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Made available in DSpace on 2014-06-11T19:22:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-08-15Bitstream added on 2014-06-13T18:48:56Z : No. of bitstreams: 1 rodrigues_lo_me_arafcf.pdf: 364674 bytes, checksum: 25ad80987f7d2eb4d8bf537154a09922 (MD5) / Universidade Estadual Paulista (UNESP) / A produção de beta-lactamase de espectro estendido (ESBL) em membros da família Enterobacteriaceae pode conferir resistência a cefalosporinas de amploespectro, aztreonam e penicilinas. Devido a esse fenômeno, a detecção exata dos produtores de ESBL é essencial para a seleção apropriada da antibioticoterapia. Para detectar a produção de beta-lactamase de espectro estendido (ESBL) em bacilos Gram-negativos, foi usado um teste de triagem com os discos de aztreonam (ATM), ceftazidima (CAZ), cefotaxima (CTX) e ceftriaxona (CRO) sobre 300 cepas, das quais trinta e cinco eram suspeitas da presença de ESBL. A produção de ESBL foi demonstrada por três métodos fenotípicos confirmatórios de fácil utilização. Os três testes fenotípicos para confirmar a produção de ESBL incluíram o teste do sinergismo (double disk), E-test? ESBL e disco combinado. Os discos utilizados no teste do sinergismo e do disco combinado foram: aztreonam (30?g-ATM), cefotaxima (30?g-CTX), ceftazidima (30?g-CAZ), cefpodoxima (10?g-CPD) ceftriaxone (30?g-CRO) e amoxicilina+ácido clavulânico(30?g-AMC), cefotaxima+ácido clavulânico (30?g-10?g), ceftazidima+ácido clavulânico (30?g- 10?g), cefpodoxima+ácido clavulânico (10?g-1? g). Para E-test foram utilizadas fitas contendo as cefalosporinas: ceftazidima versus ceftazidima/ácido clavulânico; cefotaxima versus cefotaxima/ácido clavulânico. Os testes fenotípicos confirmaram a presença de ESBL em cinco cepas de enterobactérias (1,66%). Todos os métodos são de fácil execução, contudo o método do Etest requer experiência para interpretar os resultados. Os três testes oferecem uma solução viável para confirmar a produção de ESBL no laboratório clínico. / The production of extended spectrum beta-lactamase (ESBL) in the members of the family Enterobacteriaceae can check resistance to cephalosporins of extended-spectrum, aztreonam and penicilins. Due to this phenomenon, the exact detection of the producers of ESBL are essential for the appropriate selection of antimicrobial therapy. To detect the production of extended spectrum beta-lactamase (ESBL) in Gram-negative bacilli, a test of screening was used with the discs of aztreonam (ATM), ceftazidime (CAZ), cefotaxime (CTX) e ceftriaxone (CRO) in 300 strains, of which thirty-five were suspicious of the presence of ESBL. The production of ESBL was demonstrated by three phenotypic methods confirmed of easy utilization. The three phenotypic tests to confirm the production of ESBL included the test of sinergy (double disk), E-test? ESBL and combination disk. The disks used on the test sinergy and the combination disk were: aztreonam (30?g-ATM), cefotaxime (30?g-CTX), ceftazidime (30?g-CAZ), cefpodoxime (10?g-CPD) ceftriaxone (30?g- CRO) e amoxicillin+clavulanic acid (30?g-AMC), cefotaxime+clavulanic acid (30?g- 10? g), ceftazidime+clavulanic acid (30?g-10? g), cefpodoxime+clavulanic acid (10?g- 1? g). For E-test, were utilized strips containing the cephalosporins: ceftazidime and ceftazidime/clavulanic acid; cefotaxime and cefotaxime/clavulanic acid. The phenotypic tests confirmed the presence of ESBL in five strains Enterobacteriaceae (1,66%). All of the methods are of easy execution; however, the method of Etest requires experiment to interpret the results. The three tests offer a viable solution to confirm the production of ESBL on a clinic laboratory.

Orientador: Maria de Lourdes Ribeiro de Souza da Cunha / Resumo: A alta frequência de Estafilococos Coagulase-Negativa (CoNS) na pele de indivíduos saudáveis e em doenças associadas ao sangue, associados à seleção de cepas resistentes devido a uso indiscriminado de antimicrobianos, tornou mais estreitos os limites entre o ambiente hospitalar e o comunitário quanto à distribuição de cepas. Objetivou-se, com este estudo, caracterizar isolados de CoNS de origem hospitalar e comunitária da cidade de Botucatu-SP quanto ao perfil clonal, analisar os aspectos de resistência à oxacilina pela aferição de metodologia de detecção, e investigar os determinantes de heterorresistência à vancomicina nessas cepas. As espécies estudadas incluíram S. epidermidis, S. haemolyticus, S. warneri, S. hominis, S. lugdunensis, S. capitis, S. saprophyticus, S. pasteuri, S. simulans e S. xylosus. O teste de disco-difusão (TDD) com discos de oxacilina e cefoxitina, fitas de Etest impregnadas com oxacilina e pesquisa do gene mecA por PCR em tempo real foram realizadas. A triagem em ágar com 6 e 8 µg/ml de vancomicina, microdiluição em caldo para aferição da Concentração Inibirtória Mínima (MIC), microscopia eletrônica de transmissão para verificar espessamento da parede celular e alterações fenotípicas por testes bioquímicos foram realizadas. O perfil clonal foi determinado por PFGE (Pulsed-Field Gel Eletrophoresis) e para clones de S. epidermidis, o MLST (Multilocus Sequence Typing). S. epidermidis apresentou alta diversidade clonal, mas presença de clusters no ambien... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The high frequency of Coagulase-Negative Staphylococci (CoNS) on the skin of healthy individuals and in bloodstream infections, together with the selection of resistant strains, has narrowed the boundaries between the hospital and the community environment for the distribution of strains. This study aimed to characterize CoNS isolated from clinical and colonization specimens of patients and individuals from Botucatu-SP, to compare their clonal profile, to analyze the determination of oxacillin resistance by the evaluation of the methodology of detection, and to investigate the determinants of reduced susceptibility to vancomycin in those strains. CoNS species included S. epidermidis, S. haemolyticus, S. warneri, S. hominis, S. lugdunensis, S. capitis, S. saprophyticus, S. pasteuri, S. simulans and S. xylosus. The disc diffusion test (DDT) using oxacillin and cefoxitin discs was employed, Etest strips impregnated with oxacillin and mecA gene detection by real-time PCR were used. An agar screening with 6 and 8 µg/ml of vancomycin, the broth microdiluition method for the Minimal Inhibitory Concentration (MIC), the transmission eletronic microscopy for evaluation of cellwall thickening and phenotypic modifications by biochemical tests were performed. Clonal profile was determined by PFGE (Pulsed-Field Gel Eletrophoresis) and, for S. epidermidis clones, MLST (Multilocus Sequence Typing). S. epidermidis presented high clonal diversity, despite some clusters circulating within hospi... (Complete abstract click electronic access below) / Doutor