Epidemiological characterization of resistance and PCR typing of shigella flexneri and shigella sonnei strains isolated from bacillary dysentery cases in Southeast Brazil

Orientador: Wanderley Dias da SilveiraTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: Bactérias do ¿gênero¿ Shigella spp. apresentam-se como bacilos gram-negativos, anaeróbios facultativos, imóveis, não formam esporos e pertencem à família Enterobacteriaceae. De acordo com testes de aglutinação com anti-soros específicos, estas bactérias são classificadas em quatro sorogrupos: Sorogrupo A (Shigella dysenteriae), Sorogrupo B (Shigella flexneri), Sorogrupo C (Shigella boydii) e Sorogrupo D (Shigella sonnei). Estas bactérias são responsáveis pela Shiguelose ou Disenteria Bacilar enfermidade endêmica que anualmente acomete milhões de pessoas em todo o mundo, sendo que mais de 70% de todos os casos ocorrem em crianças de 1 até 5 anos de idade, possuindo grande importância epidemiológica devido à alta morbi-mortalidade. Os principais determinantes de patogenicidade neste grupo bacteriano são: o plasmídio de alto peso molecular, que determina o fenótipo invasivo desta espécie; genes cromossômicos, que regulam a expressão dos genes de virulência no plasmídio e a produção de uma exotoxina que atua destruindo a barreira de células epiteliais. No Brasil, até então, não foram encontrados trabalhos publicados que comparem as diferentes amostras bacterianas de Shigella spp. isoladas de casos de Shiguelose, relacionando suas características biológicas e estrutura clonal. Sendo assim, neste trabalho, estudamos as características biológicas (sorotipagem, perfil de resistência a antimicrobianos, adesão e invasão, análise do perfil de DNA plasmidial) de diferentes amostras de Shigella spp. relacionando-as através de técnicas de Biologia Molecular (ERIC-PCR, REPPCR e DRE-PCR) permitindo, assim, determinar a clonalidade epidemiológica destas. As amostras de Shigella spp. foram isoladas de diferentes surtos, de diversas cidades das regiões de Campinas e de São João da Boa Vista, e pertencem à coleção do Instituto Adolfo Lutz de CampinasAbstract: Shigella spp are gram-negative, anaerobic facultative, non-motile, and non-sporulated bacilli of the Enterobacteriaceae family, responsible for ¿Shigellosis¿ or the Bacillary Dysentery (BD) disease, an important cause of worldwide morbidity and mortality. The pathogenic determinants of Shigella spp include high molecular weight plasmids responsible for the bacterial invasive capacity, as well as chromosomal genes encoding for different pathogenic, factors such as exotoxins that destroy epithelial host cells. Little is known about the antibiotic resistance profiles and the population structure of Shigella species isolated from humans in Brazil. In this work, we have studied the antibiotic resistance profiles and the clonal structure of Shigella strains isolated from humans in different cities located in the region of Campinas, a city in the state of São Paulo, Brazil. We have also related the antibiotic resistance of these strains with the bacterial clonal groups determined by the molecular techniques ERIC, REP, and DRE-PCR. Our data indicate that many strains of S. flexneri and S. sonnei are multiresistant, and our results also support the circulation of specific clones among the cities. These data indicate that the human sanitary conditions in the cities analyzed herein should be improved.DoutoradoMicrobiologiaDoutor em Genetica e Biologia Molecula

Micro-leakage at the implant-abutment interface with different tightening torques in vitro

OBJECTIVES: This study evaluated the microleakage at the implant/abutment interface of external hexagon (eH) implants and abutments with different amounts of bacteria and tightening torques. MATERIAL AND METHODS: A bacterial suspension was prepared to inoculate the implants. The first phase of this study used nine EH implants and abutments that were divided into three groups with different amounts of bacterial suspension (n=3): V0.5: 0.5 µL; V1.0: 1.0 µL e V1.5: 1.5 µL, and tightened to the manufacturer's recommended torque. The second phase of this experiment used 27 assemblies that were similar to those used in the first phase. These samples were inoculated with 0.5 µL of bacterial suspension and divided into three groups (n=9). T10: 10 Ncm; T20: 20 Ncm and T32: 32 Ncm. The samples were evaluated according to the turbidity of the broth every 24 hours for 14 days, and the bacteria viability was tested after that period. The statistical evaluation was conducted by Kruskal-Wallis testing (

F42 : epidemiologia em amostras de Escherichia coli de origem suina e sua associação com enterotoxinas

Orientador : Domingos da Silva LeiteDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaMestrad

Micro-leakage at the implant-abutment interface with different tightening torques in vitro

OBJECTIVES: This study evaluated the microleakage at the implant/abutment interface of external hexagon (eH) implants and abutments with different amounts of bacteria and tightening torques. MATERIAL AND METHODS: A bacterial suspension was prepared to inoculate the implants. The first phase of this study used nine EH implants and abutments that were divided into three groups with different amounts of bacterial suspension (n=3): V0.5: 0.5 µL; V1.0: 1.0 µL e V1.5: 1.5 µL, and tightened to the manufacturer's recommended torque. The second phase of this experiment used 27 assemblies that were similar to those used in the first phase. These samples were inoculated with 0.5 µL of bacterial suspension and divided into three groups (n=9). T10: 10 Ncm; T20: 20 Ncm and T32: 32 Ncm. The samples were evaluated according to the turbidity of the broth every 24 hours for 14 days, and the bacteria viability was tested after that period. The statistical evaluation was conducted by Kruskal-Wallis testing (p<.05). RESULTS: During the first phase, groups V1.0 and V1.5 was presented with bacterial contamination in all samples after 24 h. During the second phase, two samples from group T10 and one from T20 presented positive results for bacterial contamination. Different amounts of bacterial solution led to overflow and contamination during the first 24 h of the experiment. The tightening torques did not statistically affect the microleakage in the assemblies. However, the group that was tightened to 32 Ncm torque did not show any bacterial contamination. CONCLUSION: After 14 days of experimentation, the bacteria were proven to remain viable inside the implant internal cavity

Bacterial microleakage at the implant-abutment interface in Morse taper implants

AIM: To evaluate the microleakage at the implant-abutment (I-A) interface of Morse tapered implants inoculated with different volumes of bacterial suspension.METHODS: Morse tapered I-A sets were selected and divided in two groups depending on the type of abutment: passing screw (PS) and solid (S), and then subdivided into four subgroups (n=6) according to the suspension volume: PS1: 0.1 µL; PS3: 0.3 µL; PS5: 0.5 µL; PS7: 0.7 µL; S1: 0.1 µL; S3: 0.>3 µL; S5: 0.5 µL and S7: 0.7 µL. A control test was performed to verify the presence of external contamination during the inoculation and the implants were incubated for microbiological analysis. The microleakage was evaluated every 24 h for 7 days by the clarity of solution. After this period, the implants were disassembled for confirmation of bacterial viability.RESULTS: All the specimens with 0.7 µL and one sample of S5 presented turbidity in the control test indicating external contamination, and were excluded from the study. After 7 days of observation, none of the specimens presented positive results for microleakage and the bacterial viability was confirmed in all specimens. The 0.1 µL and 0.3 µL volumes did not present bacterial microleakage, meaning that these volumes may be inadequate for analysis.CONCLUSIONS: None of the sets evaluated showed bacterial microleakage at the I-A interface and the volume of 0.7 µL exceeded the internal capacity of the implants

Bacterial microleakage at the implant-abutment interface in Morse taper implants

evaluate the microleakage at the implant-abutment (I-A) interface of Morse tapered implants inoculated with different volumes of bacterial suspension. Methods: Morse tapered I-A sets were selected and divided in two groups depending on the type of abutment: passing screw (PS) and solid (S), and then subdivided into four subgroups (n=6) according to the suspension volume: PS1: 0.1 µL; PS3: 0.3 µL; PS5: 0.5 µL; PS7: 0.7 µL; S1: 0.1 µL; S3: 0.3 µL; S5: 0.5 µL and S7: 0.7 µL. A control test was performed to verify the presence of external contamination during the inoculation and the implants were incubated for microbiological analysis. The microleakage was evaluated every 24 h for 7 days by the clarity of solution. After this period, the implants were disassembled for confirmation of bacterial viability. Results: All the specimens with 0.7 µL and one sample of S5 presented turbidity in the control test indicating external contamination, and were excluded from the study. After 7 days of observation, none of the specimens presented positive results for microleakage and the bacterial viability was confirmed in all specimens. The 0.1 µL and 0.3 µL volumes did not present bacterial microleakage, meaning that these volumes may be inadequate for analysis. Conclusions: None of the sets evaluated showed bacterial microleakage at the I-A interface and the volume of 0.7 µL exceeded the internal capacity of the implants

FREQUENCY OF Candida SPECIES IN A TERTIARY CARE HOSPITAL IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL

Infections by Candida species are a high-impact problem in public health due to their wide incidence in hospitalized patients. The goal of this study was to evaluate frequency, susceptibility to antifungals, and genetic polymorphism of Candida species isolated from clinical specimens of hospitalized patients. The Candida isolates included in this study were obtained from blood cultures, abdominal fluids, and central venous catheters (CVC) of hospitalized patients at the Clinical Hospital of the Federal University of Uberlândia during the period of July 2010 - June 2011. Susceptibility tests were conducted by the broth microdilution method. The RAPD-PCR tests used employed initiator oligonucleotides OPA09, OPB11, and OPE06. Of the 63 Candida isolates, 18 (28.5%) were C. albicans, 20 (31.7%) were C. parapsilosis complex species, 14 (22.2%) C. tropicalis, four (6.4%) C. glabrata, four (6.4%) C. krusei, two (3.3%) C. kefyr, and one (1.6%) C. lusitaniae. In vitro resistance to amphotericin B was observed in 12.7% of isolates. In vitro resistance to azoles was not detected, except for C. krusei. The two primers, OPA09 and OPB11, were able to distinguish different species. Isolates of C. albicans and C. parapsilosis complex species presented six and five clusters, respectively, with the OPA09 marker by RAPD-PCR, showing the genetic variability of the isolates of those species. It was concluded that members of the C. parapsilosis complex were the most frequent species found, and most isolates were susceptible to the antifungals amphotericin B, flucozanole, and itraconazole. High genetic polymorphisms were observed for isolates of C. albicans and C. parapsilosis complex species, mainly with the OPA09 marker.As infecções causadas por espécies de Candida são problema de grande impacto para a saúde pública, devido à alta incidência em pacientes hospitalizados e como causa de mortalidade. O presente estudo teve como objetivo avaliar a frequência de Candida spp. isoladas de pacientes hospitalizados, assim como a sensibilidade aos antifúngicos e o polimorfismo genético por RAPD-PCR. Os microrganismos incluíram isolados de hemocultura, líquido abdominal e ponta de cateter venoso central de pacientes internados no Hospital de Clínicas da Universidade Federal de Uberlândia, região do Triângulo Mineiro, Minas Gerais, Brasil, no período de julho de 2010-junho de 2011. Os testes de sensibilidade aos antifúngicos foram realizados por microdiluição em caldo e na análise por RAPD-PCR foram utilizados os oligonucleotídeos OPA09, OPB11, e OPE06. Dos 63 isolados, 18 (28,5%) foram C. albicans, 20 (31,7%) C. parapsilosis, 14 (22,2%) C. tropicalis, quatro (6,4%) C. glabrata, quatro (6,4%) C. krusei, dois (3,3%) C. kefyr, e um (1,6%) C. lusitaniae. Resistência in-vitro à anfotericina B foi observada em 12,7% dos isolados. Não foi observada resistência in-vitro aos azólicos, exceto para os isolados de C. krusei. Os oligonucleotídeos OPA09 e OPB11 possibilitaram distinguir diferentes espécies. Isolados de C. albicans apresentaram seis clusters e o complexo C. parapsilosis, cinco clusters, com o iniciador OPA09, por RAPD-PCR, mostrando a variabilidade genética daquelas espécies. Conclui-se que o complexo C. parapsilosis foi a espécie mais frequente, e a maioria dos isolados foi sensível in vitro aos antifúngicos testados. Alto polimorfismo genético foi observado para os isolados de C. albicans e complexo C. parapsilosis, principalmente com o oligonucleotídeo OPA09