El género Enterococcus es conocido por ser de origen fecal o intestinal, pero tiene una amplia distribución en la naturaleza y se le puede encontrar en suelos, aguas, plantas y en productos alimenticios, siendo capaz de sobrevivir en medios poco enriquecidos. Los estudios reportados sobre estos microorganismos generalmente inciden en su aspecto clínico y su resistencia a antibióticos, y algunos se ubican en un contexto ambiental evaluando métodos para su detección o enumeración para uso en aguas recreacionales. Está aumentando la importancia de este microorganismo como agente causal de infecciones adquiridas en hospitales, pero el interés de estudio en este género radica en su alta resistencia natural a múltiples antimicrobianos y a su capacidad de adquirir y transferir dicha resistencia. Se sabe que Enterococcus es un microorganismo introducido al ecosistema marino debido a la contaminación de éste ambiente con desechos orgánicos, pero son pocos los reportes sobre estudios de resistencia antimicrobiana de éste género provenientes de muestras de agua de mar, siendo necesario este tipo de investigación que nos permita conocer la importancia de estos microorganismos en estos ambientes. / The genus Enterococcus is recognized as being of fecal origin but have a wide distribution in nature, they can be found in soil, water, plants and food products, being able to survive in low-enriched media. Studies on these microorganisms usually affect their appearance and clinical resistance to antibiotics, and there are some who are placed in an environmental context, evaluating methods of detection or enumeration in waters for recreational use. It is increasing the importance of this microorganism as a causative agent of infections acquired in hospitals, but the interest in this kind of study lies in its high natural resistance to multiple antimicrobials and their ability to acquire and transfer the resistance. Despite that Enterococcus is a microorganism introduced to the marine ecosystem by contamination with organic wastes, there are few reports on studies of antimicrobial resistance of the Enterococcus genus water samples from the sea, being necessary to this type of research that allows us to know the importance of these microorganisms in these environments.

El objetivo del estudio fue determinar el efecto in vitro de la medicación intraconducto hidróxido de calcio con omeprazol frente al crecimiento bacteriano del Enterococcus faecalis. El diseño de estudio fue experimental. Los medicamentos hidróxido de calcio y omeprazol fueron diluidos, obteniéndose las concentraciones requeridas. Posteriormente, se colocó 9 ml de cada uno en placas petri, agregando 1 ml del inóculo; procediéndose a la siembra. No se observó Unidades Formadoras de Colonias (UFC), por lo que se evidencia que el efecto in vitro del hidróxido de calcio, así como la asociación de hidróxido de calcio con omeprazol inhiben el crecimiento de Enterococcus faecalis. Se realizó una prueba binomial donde los eventos esperados fueron que haya o no crecimiento bacteriano. La significación estadística fue del 5%. El estudio concluyó que la asociación in vitro de hidróxido de calcio con omeprazol, inhibió el crecimiento bacteriano del Enterococcus faecalis, sin evidenciarse potencialización con el uso del inhibidor de la bomba de protones.

Universiẗat, Diss.--Jena, 2008.

Thesis (M.S.)--University of Puerto Rico, Mayagüez Campus, 2008. / Tables. Printout. Includes bibliographical references (leaves 64-70).

Thesis (Ph. D.)--Rutgers University, 2007. / "Graduate Program in Food Science." Includes bibliographical references.

Thesis (Ph.D.)--McMaster University, 2004. / Advisor: G.D. Wright. Includes bibliographical references. Also available via World Wide Web.

Orientador: Idomeo Bonetti Filho / Banca: Milton Carlos Kuga / Banca: Rodrigo Ricci Vivan / Resumo: O presente estudo avaliou a atividade antimicrobiana, escoamento, pH e a liberação de cálcio do MTA Fillapex (MTAF) (G1) ou MTA Fillapex com 10% (em massa) de hidróxido de cálcio (HC) (MTAF10) (G2), comparados com o AH Plus (AP) (G3) e o Sealapex (SE) (G4). A atividade antimicrobiana foi realizada através de teste de difusão radial sobre Enterococos faecalis (29212). O escoamento foi realizado de acordo com a norma ISO 6876:2001. Os cimentos foram inseridos em tubos de polietileno e imersos em recipiente com água deionizada. Após 24 horas, 7, 14 e 28 dias os valores do pH e cálcio liberado foram mensurados. Os valores obtidos na liberação de cálcio foram analisados através dos testes de Kruskal Wallis e Dunn e a atividade antimicrobiana, pH e escoamento foram analisados através dos testes de ANOVA e Tukey (p=0,05). A atividade antimicrobiana foi similar entre os cimentos (p>0,05). G1 e G2 apresentaram respectivamente o maior e menor escoamento que os demais grupos (p<0,05). G2 e G4 apresentaram pH e liberação de cálcio maior que G3 (p<0,05), em todos os períodos. G1 apresentou maior pH que G3 (p<0,05), exceto no período de 7 dias (p>0,05). G4 apresentou maior pH do que G1 e G2, mas o cálcio liberado foi similar (p>0,05). G3 apresentou menor liberação de cálcio entre todos os grupos (p<0,05). A adição de 10% de HC no MTAF não alterou o pH e liberação de cálcio do cimento e reduziu o escoamento, porém fora das padronizações técnicas. / Abstract: This study evaluated the antimicrobial activity, flow, pH and calcium release of MTA Fillapex (MTAF) (G1) or MTA Fillapex plus 10% in weight of calcium hydroxide powder (CH) (MTAF10) (G2), compared to AH Plus (AP) (G3) and Sealapex (SE) (G4). The flow test was performed according to ISO 6876:2001 requirements. The sealers were placed into plastic tubes and immersed in deionized water. After 24 hours, 7, 14 and 28 days the water of each tube was removed and tested to evaluate the pH values and the level of released calcium. Calcium release values were analyzed statistically by Kruskal Wallis and Dunn tests and antimicrobial activity, pH values and flow were analyzed by ANOVA and Tukey tests (α = 5%). The antimicrobial activity was similar among all groups (p>0.05). G1 presented higher flow among all sealers (p<0.05). The addition of 10% CH into MTAF reduced the flow (p<0.05), but in discordance with ISO requirements. G2 and G4 presented pH values and calcium release higher than G3 (p<0.05), in all periods. G1 presented pH value higher than G3 (p<0.05), except in 7 days period (p>0.05). G4 presented higher pH values than G1 and G2, but the calcium release was similar for all periods (p>0.05). G3 presented lower calcium release among all groups (p<0.05). The addition of 10% calcium hydroxide in MTA Fillapex caused reduction in flow and no negative interference in pH and/or calcium release. The flow obtained does not follow ISO requirements. / Mestre

Enterococcus species are integral members of the gastrointestinal microfloral of humans, animals, birds, as well as insects. Their presence in water and food has been greatly associated with faecal contamination. This study was aimed at evaluating the incidence of Enterococcus species in cow dung and environmental water sources in three commercial dairy farms. In addition, their antibiotic profiles were determined as well as resistance and virulence genes. Furthermore, the genetic relatedness of the isolates was determined by molecular typing method (RAPD PCR). Three hundred and thirty four water and faecal samples consisting of 117, 116 and 101 were collected from Seven Star Middle Drift and Fort Hare Dairy trusts respectively. Of the 334 samples collected, 289 were of faecal origin and 45 from water sources within the farms. All samples were screened for enterococci using culture base growth media and molecular methods targeting the tuf gene. Speciation was done using species-specific primers and the incidences of various species within the farms determined. Furthermore resistance to antibiotics and multidrug-resistant phenotypes were established using the disk diffusion method. Genes coding for virulence and resistance were also determined. From the samples collected, 313 (289 faecal and 24 water) presumptive enteroccocci were isolated, 305 of 313 (97.45 percent) were confirmed as Enterococcus of which 239 of 305 (78.38 percent) were identified as E. hirae, 15 of 305 (4.92 percent) as E. faecium, 12/305 (3.93 percent) as E. durans, 6 of 305 (1.97 percent) as E. faecalis and 33 of 305 (10.82 percent) were unidentified. Out of the five virulence genes that were targeted in the study only gelE (71.80 percent of 219/305) and ace (27.2 percent 83/305) were present in the isolates. Phenotypic resistance to antibiotics was observed is in all twelve antibiotics tested with multidrug resistance phenotypes detected in some enterococcal isolates most predominant in Seven Star and Middledrift dairy trust. Finally RAPD profiles of the isolates showed high relatedness between the strains from water and cow dung sources in all three commercial dairy farms suggesting possible contamination from cow dung to the water sources or vice versa.

As bactérias do gênero Enterococcus spp. estão amplamente distribuídas na natureza, atuando como patógenos oportunistas e freqüentemente causam infecções em pacientes imunocomprometidos. Uma característica destes microrganismos é a resistência intrínseca a muitos dos antimicrobianos utilizados habitualmente no tratamento de infecções por cocos Gram positivos. Por outro lado, os Enterococos têm adquirido diferentes determinantes genéticos que conferem resistência a vários antimicrobianos. Este trabalho tem como objetivo determinar a freqüência da resistência a níveis elevados de aminoglicosídeos (gentamicina), tetraciclina, bem como avaliar a diversidade genética das amostras, pesquisando os genes envolvidos na resistência, além de detectar o fator de virulência gelatinase em isolados clínicos e alimentares de Enterococcus spp. Os testes de suscetibilidade antimicrobiana dos 52 isolados de alimentos revelaram que 7,7% (n=4) apresentaram resistência somente à tetraciclina, 5,8% (n=3) foram resistentes somente a gentamicina e 28,9% (n=15) apresentaram resistência a ambos antimicrobianos. Nos 40 isolados clínicos, 42,5% (n=17) foram resistentes a tetraciclina, 20% (n=8) resistentes a gentamicina e 17,5% (n=7) foram resistentes a ambos antimicrobianos. A pesquisa de genes de resistência em isolados alimentares revelou que 90% (n=17) dos microrganismos resistentes a tetraciclina apresentaram o gen tet(M), e 100% das amostras resistentes a gentamicina apresentaram o gene aac(6’)-le-aph(2”)-Ia. Entretanto, 30% dos microrganismos sensíveis apresentaram algum dos genes de resistência. Nos isolados clínicos 95,8% (n=23) das amostras resistentes a tetraciclina apresentaram o gene tet(M) e 100% dos Enterococos resistentes a gentamicina apresentaram o gen aac(6’)-le-aph(2”)-Ia. No entanto, dos isolados que se mostraram sensíveis, 37,5% apresentaram algum dos genes de resistência analisados. Ainda, foi verificado que 78% (n=49) dos microrganismos isolados de alimentos hidrolisaram a gelatina, e somente 50% (n=20) dos isolados clínicos apresentaram a mesma atividade. / Bacteria of the Enterococcus spp. genus are widely distributed in the natural environment, Enterococci are opportunistic pathogens, and often are the causal agents of infections in immunosuppressed patients. One of the remarkable characteristics of these microorganisms is the intrinsic resistance they offer against several of the antimicrobial agents routinely prescribed in the treatment of Gram-positive cocci. On the other hand, enterococci have been proved to acquire different genetic markers that lend resistance to a variety of other antimicrobial ,searching the involved genes in the resistance, besides detecting the virulence factor gelatinase in isolated of Enterococcus spp. from foods and humans samples. This study aimed to determine the frequency of antimicrobial resistance against high concentrations of aminoglycosides (gentamicin), tetracycline, as well as assess the genetic diversity of microbial strains isolated from foods and humans. Antibiotic susceptibility tests of Enterococcus spp. isolated from food showed that of the 52 isolates, 7,7% (n=4) were tetracycline resistant, 5,8% (n=3) were resistant only the gentamicin and 28,9% (n=15) was resistant to both antibiotics. Already in 40 clinical samples 42,5% (n=17) of the samples were tetracycline resistant, 20% (n=8) were gentamicin resistant and 17,5% (n=7) were resistant to both antimicrobials. The research of genes of resistance in isolates from food disclosed that in 90% (n=17) of tetraciclyn resistant Enterococcus had the tet(M) gen, and 100% gentamicin resistant samples had aac(6')-le-aph(2")-Ia gene and 30% of the sensible microorganism had presented some of the resistance genes. Already in clinical isolates 95,8% (n=23) tetraciclyn resistant samples contained the tet(M) gene and 100% of the gentamicin resistant enterococcus had presented gen aac(6')-le-aph(2")-Ia; however 37,5% enterococcus sensible had presented some of the genes of resistance in research. In this work, also it was verified that 78% (n=49) of the foods isolates produced gelatinase, and 50% (n=20) of the clinical isolates demonstrated the capacity of degradation of the gelatin substrate.

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