Presencia de infección de sitio quirúrgico en procedimientos gineco-obstétricos en un hospital de II nivel de atención, Honduras 2017-2018 / Presence of surgical site infection in obstetric-gynecological procedures in a level II care hospital, Honduras 2017-2018

OBJETIVO: Identificar la presencia de infección de sitio quirúrgico y factores de riesgo en pacientes sometidas a cirugías Gineco-Obstétricas de forma programada o de urgencia en un hospital de II nivel de atención en Honduras. METODOLOGÍA: Estudio observacional, descriptivo, retrospectivo; recopilando 226 fichas del registro de infección de sitio quirúrgico recuperadas de los expedientes clínicos brindados por el servicio de estadística del Hospital Mario Catarino Rivas. Captando pacientes sometidas a cirugías Gineco-Obstétricas, durante el 2017 y 2018. RESULTADOS: 99 fichas cumplieron los criterios de inclusión, reportando una edad de 24 años [RIQ, 19,0 - 30,0], peso 82,0 kg [RIQ, 51,7 - 98,25], talla 154 cm [150,0 - 158,0] y el IMC de 25,8 ± 3,6 kg/m2. Un 9,1% presento antecedentes de inmunosupresión. 5,1% presento ISQ. El 55.6% de las cirugías se realizó el mismo día de ingreso del paciente. El tiempo entre la profilaxis antibiótica y el comienzo de la intervención quirúrgica es de 60 minutos [RIQ, 40,0 - 160,0]. La duración de los procedimientos quirúrgicos son de 45 minutos [RIQ, 35,0 - 55,0]. Los microrganismos aislados en los cultivos fueron Cocos gram positivos (2/5), Enterococcus faecalis (1/5), Klebsiella pneumoniae (2/5). CONCLUSIÓN: La presencia de ISQ en cirugías Gineco-Obstétricas del HMCR es del 5.1%, identificando los siguientes factores de riesgo: edad extrema, obesidad, diabetes mellitus, estado inmunitario (VIH), profilaxis antibiótica (temprana); por último, la técnica y el tiempo quirúrgico.

OBJECTIVE: To identify the presence of surgical site infection and risk factors in patients undergoing Gynecological-Obstetric surgeries on a scheduled or emergency basis in a 2nd level of care hospital in Honduras. METHODOLOGY: Observational, descriptive, retrospective study, compiling 226 data sheets of the surgical site infection record recovered from the clinical records provided by the statistics service of the "Hospital Mario Catarino Rivas". Recruiting patients undergoing Gynecological-Obstetric surgeries, during 2017 and 2018. RESULTS: 99 tabs met the inclusion criteria, reporting an age of 24 [RIQ, 19.0 - 30.0], weight 82.0 kg [RIQ, 51.7 - 98.25], size 154 cm [150.0 - 158.0] and BMI of 25.8 ± 3.6 kg/m2. 9.1% have a history of immunosuppression. 5.1% present ISQ. 55.6% of surgeries were performed on the same day as the patient's admission. The time between antibiotic prophylaxis and the onset of surgery 60 minutes [RIQ, 40.0 - 160.0]. Duration of surgical procedures 45 minutes [RIQ, 35.0 - 55.0]. Isolated micro-morphisms in crops were Cocos gram positives (2/5), Enterococcus faecalis (1/5), Klebsiella pneumoniae (2/5). CONCLUSION: The presence of ISQ in HMCR Gynecological-Obstetric surgeries is 5.1%, identifying the following risk factors: extreme age, obesity, diabetes mellitus, immune status (HIV), early antibiotic prophylaxis; finally, technique and surgical time.

Eficacia del instrumento XP-endo Finisher y del sistema EndoActivator en la reducción/eliminación del biofilm bacteriano: un ensayo ex vivo / Ability of the XP-endo Finisher instrument and the EndoActivator system to reduce/eliminate of the bacterial biofilm: an ex vivo assay

Objetivos: Comparar ex vivo la eficacia del instrumento XP-endo Finisher y del sistema EndoActivator en la reducción/eliminación del biofilm microbiano en conductos radiculares infectados. Materiales y métodos: Se utilizaron 23 premolares inferiores humanos extraídos cuya longitud fue estandarizada en 17 mm. Todos los conductos se prepararon con el sistema WaveOne Gold Medium (#35.06). Los dientes se esterilizaron, se inocularon con Enterococcus faecalis y se separaron en dos grupos experimentales de 10 piezas cada uno. De los 3 dientes remanentes, 1 fue utilizado como control positivo y 2, como controles negativos. En el grupo 1, las soluciones irrigantes se agitaron con XP-endo Finisher. En el grupo 2, se utilizó EndoActivator. Se tomaron muestras antes de la contaminación, luego de esta y después de la agitación de los irrigantes mediante conos de papel estériles. La carga microbiana fue sembrada en agar sangre y los conos se cultivaron en caldo tripteína de soja. La remoción de la carga microbiana se determinó por la presencia o ausencia de turbiedad del medio. Las unidades formadoras de colonias (UFC) remanentes se cuantificaron y los resultados se categorizaron como R1 (≤10 UFC) o R2 (>10 UFC). Los datos fueron analizados mediante la prueba de Fisher. Resultados: No hubo diferencias significativas entre XP-endo Finisher y EndoActivator (P>0,05). El número de usos no influyó sobre la capacidad operativa de ambos instrumentos (AU)

Aim: To compare ex vivo the effectiveness of the XP-endo Finisher and the EndoActivator in biofilm reduction/ removal from infected root canals. Materials and methods: Twenty three extracted human single-rooted lower premolars were selected and standardised to 17 mm in length. All the canals were prepared with WaveOne Gold Medium reciprocating files (#35.06). The teeth were autoclaved and inoculated with Enterococcus faecalis. The infected teeth were then assigned to 2 experimental groups of 10 teeth each according to the final irrigation/agitation protocol. Of the three remaining teeth, one was used as a positive control, and the other two were used as negative controls. In Group 1 the irrigating solutions were agitated with XP-endo Finisher while in Group 2 the EndoActivator was used. All root canals were sampled before and after contamination, and again after irrigant agitation with sterile paper points. The microbial load was spread on blood agar plates and the paper points were cultured in sterile trypticase soy broth. The removal of the microbial load was determined by visual observation of the turbidity of the media and by quantification of the number of colony-forming units (UFC). The results were categorized as R1 (≤10 UFC) or R2 (>10 UFC). Data were analysed by the Fisher's exact test at P<0.05. Results: No significant differences was found between XP-endo Finisher and EndoActivator (P>0.05) regarding their effectiveness in the reduction/removal of the microbial biofilm. The number of uses of both instruments did not affect their operative performance (AU) Conclusion: XPF and EA were both equally effective for microbial biofilm reduction/removal from ex vivo infected root canals (AU)

Decontamination of Gutta-percha Cones employed in Endodontics / Descontaminação de cones de guta-percha empregados em endodontia

ABSTRACT The guttapercha cones used in endodontic treatment are produced in aseptic conditions and their composition includes zinc oxide, which is responsible for antibacterial activity. However, there is the possibility of microbial contamination by manipulation, aerosol or during storage. Although several chemical agents have been tested for their decontamination, there is no consensus on the best disinfection protocol to be used. The aim of this study was to evaluate the decontamination of guttapercha cones contaminated with the bacteria Enterococcus faecalis, by using chlorhexidine digluconate (CHX) and sodium hypochlorite (NaClO) at different concentrations for short exposure times. For this purpose, guttapercha cones (size 40) were selected at random from a sealed box and immersed for 1 min in a microbial suspension. Then they were immersed in specific Petri dishes for different groups containing: CHX 2%, NaClO 1% or NaClO 2.5% for 30 s or 1 min, and subsequently placed in tubes containing BHI broth. After incubating the tubes for 48 h, it was observed that 1% and 2.5% NaClO and 2% CHX were effective for decontaminating the cones at those exposure time intervals. Microbial growth was detected in one of the replicates of the group with CHX applied for 30 s. To prevent the possibility of failures at this stage, the exposure time of guttapercha cones to the decontaminating agent should not be reduced.

RESUMO Os cones de gutapercha utilizados no tratamento endodôntico são produzidos em condições assépticas e possuem óxido de zinco em sua composição, responsável pela atividade antibac te riana. No entanto, existe a possibilidade de contaminação microbiana por manipulação, aerossol ou seu armazenamento. Embora vários agentes químicos já tenham sido testados para sua descontaminação, não há consenso sobre o melhor proto colo de desinfecção a ser usado. Nosso objetivo foi avaliar a descontaminação de cones de gutapercha contaminados com a bactéria Enterococcus faecalis, utilizando digluconato de clorexidina (CHX) e hipoclorito de sódio (NaClO) em diferentes concentrações e tempos de exposição curtos. Para esse fim, 40 cones de gutapercha foram selecionados aleatoriamente, de uma caixa selada e imersos por 1 min em uma suspensão microbiana. Em seguida, foram imersos em placas de Petri específicas para diferentes grupos contendo: CHX 2%, NaClO 1% ou 2,5%, nos tempos de exposição de 30s e 1min e subseqüentemente imersos em tubos contendo caldo BHI. Após incubação dos tubos por 48 h, observouse que NaClO 1% e 2,5% e CHX 2% foram eficazes para a descontaminação dos cones nesses intervalos de tempo de exposição. Em uma das réplicas do grupo com CHX aplicado por 30s foi detectado crescimento microbiano. O tempo de exposição dos cones de gutapercha ao agente de desconta minação não deve ser reduzido para evitar a possibilidade de falhas nesse estágio.

Investigação da população de Enterococcus faecalis em pacientes com pólipo e câncer colorretal em comparação a indivíduos saudáveis / Investigation of enterococcus faecalis population in patients with polyp and colorectal cancer in comparison of healthy individuals

ABSTRACT BACKGROUND: Colorectal cancer is one of the most commonly diagnosed cancers around the world. One of the factors involved in the development of colorectal cancer is the changes in the normal flora of the intestine. OBJECTIVE: In this study, the mean copy number of Enterococcus faecalis in people with polyps and people with colorectal cancer has been evaluated in comparison with healthy controls. METHODS: In this study, 25 patients with colorectal cancer and 28 patients with intestinal polyps were selected and stool specimens were taken. In addition, 24 healthy individuals were selected as control group. Extraction of bacterial DNA from the stool sample were performed. The molecular methods of PCR for confirmation of standard strain and absolute Real Time PCR (qRT-PCR) method were used to evaluate the number of Enterococcus faecalis in the studied groups. RESULTS: The results of this study indicate that the mean copy number of Enterococcus faecalis in patients with colorectal cancer was 11.2x109 per gram of stool, and in patients with polyps was 9.4x108 per gram of stool. In healthy people, this number was 9x108 per gram of stool. There was a significant difference between the implicit copy numbers in the three groups. (P<0.05). CONCLUSION: Enterococcus faecalis in faecal flora of people with colorectal cancer was significantly higher than those with polyps and healthy people. This could potentially signify the ability of this bacterium to induce colorectal cancer. More studies are needed to prove this theory.

RESUMO CONTEXTO: O câncer colorretal é um dos cânceres mais comumente diagnosticados em todo o mundo. Um dos fatores envolvidos no desenvolvimento do câncer colorretal é a mudança na flora normal do intestino. OBJETIVO: O número médio de cópias de Enterococcus faecalis em pessoas com pólipos e pessoas com câncer colorretal foram avaliados em comparação com controles saudáveis. MÉTODOS: Neste estudo, 25 pacientes com câncer colorretal e 28 pacientes com pólipos intestinais foram selecionados e amostras de fezes foram adquiridas. Além disso, 24 indivíduos saudáveis foram selecionados como grupo controle. A extração do DNA bacteriano da amostra coletada foi executada. Os métodos moleculares de PCR para confirmação da cepa padrão e o método absoluto de PCR em tempo real (qRT-PCR) foram utilizados para avaliar o número de Enterococcus faecalis nos grupos estudados. RESULTADOS: Os resultados deste estudo indicam que o número médio de cópias de Enterococcus faecalis em pacientes com câncer colorretal foi de 11,2x109 por grama de fezes, e em pacientes com pólipos foi de 9,4x108 por grama de fezes. Em pessoas saudáveis, este número foi de 9x108 por grama de fezes. Houve diferença significativa entre os números de cópia implícita nos três grupos. (P<0,05). CONCLUSÃO: Enterococcus faecalis na flora fecal de pessoas com câncer colorretal foi significativamente maior do que aqueles com pólipos e pessoas saudáveis. Isto poderia potencialmente significar a capacidade desta bactéria para induzir o câncer colorretal. Mais estudos são necessários para provar esta teoria.

Effectiveness of XP-Endo Finisher in the reduction of bacterial load in oval-shaped root canals

Abstract: This study investigated the effectiveness of XP-Endo Finisher (XPF) associated with XP-Endo Shaper (XPS) or Reciproc Blue (RB) files in reducing bacterial load in oval-shaped root canals (RC) during chemomechanical preparation (CMP) using 0.9% saline solution (NaCl) or 2.5% sodium hypochlorite (NaOCl). Eighty mandibular incisors with single oval-shaped RC were contaminated with Enterococcus faecalis. The teeth were randomly assigned to eight experimental groups (n = 10) according to the CMP, as follows: G1: XPS, G2: XPS + XPF, G3: RB, and G4: RB + XPF. CMP was performed with NaCl or NaOCl. The reduction of bacterial load was assessed by colony-forming unit count before (S1) and after (S2) CMP. Data normality was verified by using Shapiro-Wilk test. ANOVA, Tukey's test, and Bonferroni post-hoc test were used at a 5% significance level. Culturable bacteria were present in all S1 samples (p>0.05). All instrumentation techniques were effective in reducing bacterial load, irrespective of the irrigating solution (p < 0.05). With the use of NaCl, RB was more effective than XPS (p = 0.035). With the use of NaOCl, XPS and RB presented similar effectiveness (p = 0.779). XPF enhanced the bacterial reduction of both systems tested (p < 0.05). The use of NaOCl improved the CMP, irrespective of the instrumentation technique used (p < 0.05). In conclusion, XPS and RB files are effective in reducing bacterial levels in oval-shaped RC. The use of XPF as a method of agitation of the irrigating solution improved the cleaning efficiency of both file systems tested. Mechanical preparation performed with saline solution decreased culturable bacteria from the root canal, but antimicrobial substances such as NaOCl should be used to achieve a significantly better disinfection.

Evaluation of the waveone gold and one shape new generation in reducing enterococcus faecalis from root canal

Abstract The aim of this study was to evaluate the WaveOne Gold and One Shape New Generation systems regarding the bacterial removal from root canals infected with Enterococcus faecalis by comparing them to the conventional WaveOne and One Shape systems. Forty-eight distobuccal root canals of maxillary molars sterilized with ethylene oxide were infected with E. faecalis for 21 days, and then root canal initial bacterial sample was collected with paper cones and plated on M-enterococcus agar. The specimens were randomly divided into 4 groups according to the instrumentation: WaveOne Gold, One Shape New Generation, WaveOne and One Shape. After instrumentation, samples were collected with use of scraping and paper cones at immediate and 7 days after instrumentation. The bacterial reduction was calculated and then made intragroup analysis by Friedman test and intergroup analysis by Kruskal-Wallis with Dunn's post-hoc test, all at 5% significance. All techniques significantly reduced the number of bacteria in the root canal (p<0.05). WaveOne Gold and One Shape New Generation promoted higher bacterial reduction than WaveOne and One Shape systems (p<0.05), but no significant difference was found between WaveOne Gold and One Shape New Generation or between WaveOne and One Shape (p>0.05). Novel single-file systems promote better bacterial removal than the conventional single-file systems.

Resumo A proposta deste estudo foi avaliar os sistemas WaveOne Gold e One Shape New Generation em relação à remoção bacteriana de canais infectados com Enterococcus faecalis, comparando-os com seus sistemas convencionais WaveOne e One Shape. Quarenta e oito canais disto vestibulares de molares superiores esterilizados em óxido de etileno foram contaminados com E. faecalis por 21 dias, e então acoleta bacteriana inicial foi feita com cone de papel e plaqueadas em M-enterococcus agar. Os espécimes foram aleatoriamente divididos em quarto grupos de acordo com a instrumentação: WaveOne Gold, One Shape New Generation, WaveOne e One Shape. Após instrumentação, amostras foram coletadas utilizando limagem e cones de papel imediatamente e 7 dias após o preparo. A redução bacteriana foi calculada e então feita análise intra grupos com teste de Friedman, e entre grupos utilizando Kruskal-Wallis e teste de Dunn, todos a 5% de significância. Todas as técnicas reduziram significantemente o número de bactérias do canal radicular (p<0.05). WaveOne Gold e One Shape New Generation promoveram maior redução bacteriana que WaveOne e One Shape (p<0.05), mas nenhuma diferença significante foi encontrada entre WaveOne Gold e One Shape New Generation ou entre WaveOne e One Shape (p>0.05). Novos sistemas de lima-única promovem melhor remoção bacteriana que seus sistemas convencionais.

Molecular Typing of Enterococcus faecalis from Persistent Endodontic Infections / Tipificación Molecular de Enterococcus faecalis Provenientes de Infecciones Endodónticas Persistentes

ABSTRACT: Molecular techniques that provide valuable information about the epidemiology of oral strains. The purpose of this study was to determine the genetic relatedness of 83 Enterococcus faecalis strains isolated from treated root canals. These strains were obtained from patients who were treated for persistent endodontic infections. E. faecalis isolates were molecular typed by Pulsed Field Gel Electrophoresis using Smal. Ten clonal groups and 13 pulse types with 38.7 % similarity for the less related strains were identified. Genetic heterogeneity among strains from different patients and a high level of genetic homogeneity among intrapatient strains were observed. Therefore, restriction endonuclease fingerprinting of genomic DNA from E. faecalis strains confirmed the polyclonality of the isolates obtained from the root canals of patients diagnosed with persistent endodontic infections, compared with other reports. These results provide additional data for a better understanding of the epidemiological aspects of root canal infections by E. faecalis.

RESUMEN: Las técnicas moleculares proporcionan información valiosa sobre la epidemiología de aislados orales. El propósito de este estudio fue determinar la relación genética de 83 cepas de Enterococcus faecalis aisladas de conductos radiculares tratados. Estas cepas se obtuvieron de pacientes que fueron tratados por infecciones endodónticas persistentes. Los aislados de E. faecalis se tipificaron molecularmente por electroforesis en gel de campo pulsado usando Smal. Se identificaron diez grupos clonales y 13 pulsotipos con un 38,7 % de similitud para las cepas menos relacionadas. Se observó heterogeneidad genética entre las cepas de diferentes pacientes y un alto nivel de homogeneidad genética entre las cepas intrapacientes. Por lo tanto, la toma de huellas dactilares a traves de restricción de ADN genómico de cepas de E. faecalis confirmó la policlonalidad de los aislados obtenidos de los conductos radiculares de pacientes diagnosticados con infecciones endodónticas persistentes, en comparación con otros informes. Estos resultados proporcionan datos adicionales para una mejor comprensión de los aspectos epidemiológicos de las infecciones del conducto radicular por E. faecalis.

Instrument Design May Influence Bacterial Reduction During Root Canal Preparation

Abstract The aim of this study was to evaluate the bacterial reduction promoted by ProTaper Next and Twisted File by comparing to ProTaper Universal and manual technique. Sixty distobuccal root canals of maxillary molars sterilized with ethylene oxide were contaminated with Enterococcus faecalis broth culture. After incubation for 21 days, bacterial samples were collected and cultured on m-Enterococcus agar plates. The root canals were divided into 4 groups, according to the system used for instrumentation: ProTaper Next, Twisted File, ProTaper Universal, and crown down manual technique. Other 8 uncontaminated root canals were control asepsis. Bacterial samples were collected immediately and 7 days after instrumentation. The bacterial reduction was calculated and then made intragroup analysis by paired t-test and intergroup analysis by ANOVA and Tukey tests, all at 5% significance. All techniques significantly reduced the bacterial number in the root canal (p<0.05). ProTaper Next and Twisted File resulted in more bacterial reduction than ProTaper Universal and manual technique (p<0.05). ProTaper Next and Twisted File were similar (p>0.05). It can be concluded that ProTaper Next and Twisted File promote a higher bacterial reduction than Protaper Universal and manual technique.

Resumo A proposta deste estudo foi avaliar a redução bacteriana promovida pelos sistemas ProTaper Next e Twisted File comparando-os ao sistema ProTaper Universal e técnica manual. Sessenta raízes disto vestibulares de molares superiores esterilizadas com óxido de etileno foram contaminadas com Enterococcus faecalis. Após incubação por 21 dias, amostras foram coletas e cultivadas em m-Enterococcus agar. Os canais foram divididos em 4 grupos de acordo com o sistema de instrumentação: ProTaper Next, Twisted File, ProTaper Universal, e técnica manual crown-down. Outros 8 canais não contaminados foram utilizados como controle de assepsia. Amostra bacteriana foi coletada imediatamente após o prepare e aos 7 dias. A redução bacteriana foi calculada, e então feita a análise intergrupos utilizando teste t-pareado, e análise entre grupos utilizando os testes ANOVA e Tukey, todos com 5% de significância. Todas as técnicas reduziram significantemente o número de bactérias no canal radicular (p<0.05). ProTaper Next e Twisted File resultaram em maior redução bacteriana que ProTaper Universal e técnica manual (p<0.05). ProTaper Next e Twisted File foram semelhantes (p>0.05). Pode-se concluir que ProTaper Next e Twisted File promovem maior redução bacteriana que Protaper Universal e técnica manual.

Operatoria dental y endodoncia 3: capacidad de sellado marginal de un sistema adhesivo utilizado con o sin el complemento de clorhexidina / Operative dentistry and endodontics 3: marginal sealing properties of an adhesive system used with or without chlorhexidine addition

Objetivo: la biodegradación hidrolítica y enzimática de la interfaz resina/dentina constituye uno de los principales motivos del fracaso a distancia de las restauraciones con resinas compuestas. El objetivo de este ensayo ex vivo fue investigar si el uso de gluconato de clorhexidina al 2,0 por ciento, aplicado previamente al adhesivo, puede mejorar la capacidad de sellado de la interfaz resina/dentina en restauraciones de clase I, frente a la penetración bacteriana. Materiales y métodos: se obtuvieron probetas de dentina en forma de discos de 3 mm de altura a partir de 24 terceros molares humanos sanos, extraídos y utilizados dentro de un período de 30 días posextracción. En cada probeta se preparó una cavidad de clase I de 2 x 4 mm con una profundidad de 3 mm, uniendo de esta forma las superficies superior e inferior. Los especímenes se separaron en dos grupos de 10 probetas cada uno (n=10). En el grupo 1, las cavidades fueron tratadas con ácido fosfórico al 35 por ciento y restauradas mediante el sistema adhesivo Klepp Microhybrid Resin System (Klepp-Raysan Int, Estados Unidos. En el grupo 2, las cavidades se restauraron de manera similar a las del grupo 1, pero, previamente a la aplicación del adhesivo, las paredes dentinarias se trataron con gluconato de clorhexidina al 2,0 por ciento. Dos de las cuatro probetas restantes se utilizaron en el grupo 1 como controles positivos (n=1) y negativos (n=1) y las otras dos fueron empleadas de la misma forma que en el grupo 2. Todos los especímenes fueron termociclados a 5-55ºC (1000 ciclos de 20 segundos de duración cada uno) y tratados mediante un proceso de simulación de la agresión hídrica en el medio oral. Finalmente, se los sometió a un ensayo de filtración bacteriana (Enterococcus faecalis) durante 60 días. Los datos registrados se analizaron con la prueba de Log-Rank, el test de supervivencia de Kaplan Meyer y la prueba exacta de Fischer,c on un nivel de significación de p<0,05.

Actividad in vitro de ampicilina-ceftriaxona frente a aislamientos de Enterococcus faecalis recuperados de infecciones invasivas / In vitro activity of ampicillin-ceftriaxone against Enterococcus faecalis isolates recovered from invasive infections

Se evaluó la actividad in vitro de la asociación entre ampicilina y ceftriaxona frente a 30 aislamientos de Enterococcus faecalis obtenidos de infecciones invasivas de pacientes atendidos en el Hospital de Clínicas José de San Martín, Ciudad Autónoma de Buenos Aires. Las sinergias entre ampicilina y ceftriaxona se determinaron mediante la técnica de dilución en caldo Müeller-Hinton con el agregado de diferentes concentraciones subinhibitorias de ceftriaxona o sin este. La asociación fue sinérgica en 22/30 aislamientos. En 14/30 aislamientos la asociación disminuyó los valores de concentración inhibitoria mínima (CIM) y de concentración bactericida mínima (CBM); en 6/30 se observó solamente una disminución de la CIM, mientras que en 2 solo se determinó una reducción de la CBM. La actividad bactericida de la asociación fue mayor a bajas concentraciones de ampicilina (menor de 1µg/ml). Se demostró la sinergia in vitro entre ampicilina-ceftriaxona; se confirmó así la utilidad de esta asociación en el tratamiento de infecciones severas causadas por E. faecalis

In vitro activity of the combination of ampicillin- ceftriaxone against 30 Enterococcus faecalis isolates recovered from invasive infections in patients admitted to Hospital de Clínicas José de San Martin in the city of Buenos Aires was assessed. Ampicillin- ceftriaxone synergies were determined by microdilution in Müeller-Hinton (MH) broth with and without subinhibitory concentrations of ceftriaxone. Synergy was detected in 22/30 isolates. A decrease in both minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) was observed in 14/30 isolates, whereas in 6/30 isolates the decrease was observed in the MIC value and only in the MBC value in the 2 remaining isolates. The bactericidal activity of the combination showed to be higher at low concentrations of ampicillin (< 1µg/ml). We detected in vitro synergy using the ampicillin-ceftriaxone combination and thus, its efficacy was confirmed in the treatment of severe infections by E. faecalis

Capacidad de sellado coronario de dos cementos endodónticos a base de óxido de cinc y eugenol: un estudio ex vivo / Coronal sealing ability of two zinc-oxide and eugenol-based endodontic sealers: an ex vivo study

Objetivo: comparar, mediante un modelo de filtración bacteriana, la capacidad de sellado coronario de obturaciones realizadas con conos de gutapercha complementados con Kleppmetthasona (KMS; Klepp/Raysan, Buenos Aires, Argentina); y Pulp Canal Sealer EWT (PCS; Sybron-Endo, Glendora, Estados Unidos). Materiales y métodos: se utilizaron 24 (n=24) premolares humanos sanos extraídos. En todos los casos, los conductos radiculares fueron preparados por medio de instrumentos rotatorios ProTaper Universal (Dentsply/Tulsa Dental, Tulsa, OK, Estados Unidos) hasta un instrumento #F3. Se distribuyeron 20 de los dientes en dos grupos experimentales (Grupo I y Grupo II) de 10 (n=10) especímenes cada uno. En cada grupo se agregaron un especimen de control positivo (n=1) y un especimen de control negativo (n=1). En el grupo 1, los conductos se obturaron con un cono de gutapercha ProTaper de conicidad correspondiente al instrumento #F3 y se complementó mediante la condensación lateral de conos accesorios. El sellador utilizado fue KMS, a base de óxido de cinc y eugenol que incluye un corticoide en su composición. En el Grupo 2 se realizó el mismo procedimiento, pero se utilizó PCS, un sellador a base de óxido de cinc y eugenol convencional. Para cada especimen, el proceso de filtración bacteriana se realizó por medio de un sistema individual de doble cámara, cada uno de las cuales contenía el medio de cultivo. En todos los casos, el medio de la cámara superior se inoculó con un cultivo de Enterococcus faecalis y el sistema se incubó durante 60 días. En la cámara inferior, la filtración bacteriana se controló diariamente hasta la finalización del experimento. Los datos registrados se analizaron con la prueba de supervivencia de Kaplan-Meyer yla prueba de Fischer, con un nivel de significación de P<0.05. Resultados: todos los controles positivos demostraron filtración bacteriana dentro de las primeras 48 horas. No hubo filtración en ninguno de los controles negativos.

Effect of Passive Ultrasonic Irrigation on Enterococcus faecalis from Root Canals: An Ex Vivo Study

Endodontic irrigation aims to clean and disinfect the root canal system. Passive ultrasonic irrigation (PUI) is based on the use of an ultrasound-activated instrument into the root canal filled with irrigant. The aim of this study was to evaluate, ex vivo, the effectiveness of PUI in eliminating Enterococcus faecalis from root canals. Seventy-five extracted human single-root teeth were used. After root canal preparation, specimens were inoculated with E. faecalis and incubated at 37 °C for 21 days. Specimens were distributed into five groups (n=15), according to the irrigation method: PUI + saline solution (PUI/SS); PUI + 1% NaOCl (PUI/NaOCl); conventional needle irrigation (CNI) + saline solution (CNI/SS); CNI + 1% NaOCl (CNI/NaOCl); No irrigation (control). Microbiological samples were collected at three time points: initial (21 days after inoculation), post-irrigation (immediately after irrigation), and final (7 days after irrigation). Data were obtained in CFU mL^-1 and subjected to analysis by ANOVA and Tukey's tests at 5% significance level. The post-irrigation samples did not demonstrate statistical difference between PUI/SS and CNI/SS nor between PUI/NaOCl and CNI/NaOCl (p>0.05), but PUI/NaOCl and CNI/NaOCl had lower CFU mL^-1 number than the other groups (p>0.05). Statistically significant difference was observed between the initial and post-irrigation samples and between the post-irrigation and final samples (p<0.05) in all groups, except in the control. The final samples of all groups presented bacterial counts similar to the initial samples. PUI or CNI with 1% NaOCl contribute to disinfection, but are unable to eradicate E. faecalis from the root canal system.

A irrigação endodôntica visa à limpeza e desinfecção do sistema de canais radiculares. A irrigação ultrassônica passiva (IUP) baseia-se na utilização de um instrumento ultrassônico ativado no interior do canal radicular preenchido com a solução irrigadora. O objetivo deste estudo foi avaliar, ex vivo, a eficácia da IUP na eliminação de Enterococcus faecalis dos canais radiculares. Foram usados 75 dentes humanos unirradiculares extraídos. Após o preparo biomecânico dos canais radiculares, foram inoculados com E. faecalis e incubados a 37 °C por 21 dias. Os espécimes foram distribuídos em cinco grupos (n=15), de acordo com o método de irrigação: IUP + solução salina (IUP/SS); IUP + NaOCl a 1% (IUP/NaOCl); irrigação manual convencional (IMC) + solução salina (IMC/sol); IMC + NaOCl a 1% (IMC/NaOCl); Sem irrigação (controle). Foram realizadas três coletas microbiológicas: inicial (21 dias após a inoculação), pós-irrigação (imediatamente após a irrigação), e final (7 dias após a irrigação). Os dados foram obtidos em UFC mL^-1 e submetidos aos testes de ANOVA e Tukey. A coleta pós-irrigação não demonstrou diferença estatisticamente significante entre IUP/SS e IMC/SS nem entre IUP/NaOCl e IMC/NaOCl (p>0,05), mas IUP/NaOCl e IMC/NaOCl apresentaram menor número de UFC mL^-1 comparado aos outros grupos (p>0,05). Diferença estatisticamente significante foi observada entre as coletas iniciais e pós-irrigação e entre a pós-irrigação e a coleta final (p<0,05) em todos os grupos, exceto no controle. Na coleta final, todos os grupos apresentaram contagem bacteriana similar à coleta inicial. IUP ou IMC com NaOCl a 1% contribuem para a desinfecção, mas não eliminam E. faecalis do sistema de canais radiculares.

Clonal diversity and antimicrobial resistance of Enterococcus faecalis isolated from endodontic infections

Background Enterococcus faecalis is considered to be one of most prevalent species in the oral cavity, particularly in endodontic infections. The aim of the present study was to investigate the prevalence of E. faecalis in dental root canals, clonal diversity by restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD-PCR) analysis, and the antibiotic susceptibility of E. faecalis isolates. Results Among the bacterial strains isolated from dental root canal specimens (n = 82), E. faecalis was determined to have the highest prevalence followed by Streptococcus viridians, Leuconostoc mesenteroides, Staphylococcus aureus, Streptococcus mitis, and Pediococcus pentosaceus. Cluster analysis of RAPD-PCR and RFLP patterns of the E. faecalis isolates discriminated five and six different genotypes, respectively. Among the tested strains, 43%, 52% and 5% were susceptible, intermediate resistant, and resistant to erythromycin, respectively. In addition, one strain (E-12) was intermediate resistant to linezolid, and one isolate (E-16) was resistant to tetracycline. Interestingly, many of the intermediate resistant/resistant strains were grouped in clusters 5 and 6, according RAPD and to RFLP, respectively. Conclusions E. faecalis demonstrated the highest prevalence in the tested dental root canal specimens collected from Saudi patients and were grouped into five to six different genotypes. Different levels of antimicrobial susceptibility were observed in the tested E. faecalis strains, which clearly indicated that although bacterial strains may be similar, point mutations can result in extreme susceptibility or resistance to various antibiotics. This phenomenon is a cause for concern for clinicians in the treatment of dental infections caused by E. faecalis.

Apical extrusion of Enterococcus faecalis using three different rotary instrumentation techniques: An in vitro study.

Aims: To compare the apical extrusion of Enterococcus faecalis after instrumentation with three different Ni-Ti rotary instruments- An in vitro study. Settings and Design: In vitro study Methods and Material: Forty freshly extracted mandibular premolars were mounted in bacteria collection apparatus and root canals were contaminated with a suspension of Enterococcus faecalis. The contaminated teeth were divided into 4 groups of 10 teeth each according to rotary system used for instrumentation: Group1: Hyflex files, Group 2: GTX files, Group 3: Protaper files and Group 4: control group (no instrumentation). Bacteria extruded after preparations were collected into vials and microbiological samples were incubated in BHI broth for 24 hrs. The colony forming units were determined for each sample. Statistical Analysis Used: Statistical analysis was done using one way ANOVA followed by post hoc independent “t” test. Results: GTX files extruded least amount of bacteria followed by Hyflex files. Maximum extrusion of E. faecalis was seen in rotary Protaper group. Conclusion: Least amount of extrusion was seen with GTX files followed by Hyflex files and then rotary Protaper system.

Enterococcus faecalis fiofilm: formation and development in vitro observed by scanning electron microscopy

Se realizó la detección de biofilm de Enterococcus faecalis aislados de conductos radiculares, desarro llándolos en microcubetas realizando tinción con Cristal Violeta 10 por ciento, elución con alcohol y tres procedimientos: sin fijar, fijación con calor y fija ción con formaldehido 10 por ciento. Se evaluó el biofilm formado realizando la lectura con lector de microplaca Versamax-Microplate-Reader (USA). Se incubaron 20 porciones radiculares estériles en el medio TS caldo con E. faecalis (108) durante 48 hs, 4, 7, 14 y 30 días.Transcurrido cada tiempo experimental se procesaron y observaron al microscopio electrónico de barrido (MEB). Se evidenció significativamente mayor formación de biofilm en microplacas cuando se realizó fijación con formaldehido, que al fijar con calor y sin fijar (ANOVA p<0,0001). Al MEB seobservó crecimiento del E. faecalis en todos los tiempos y desarrollo de biofilm a partir de 14 días de incubación. La fijación con formaldehido 10% fue la técnica más apropiada para detectar biofilm de E. faecalis desarrollado enmicroplacas. Mediante el MEB se confirmó la formación de biofilm después de 14 días de incubación...

Espermocultivo: crecimiento bacteriano del eyaculado y su relación con los parámetros seminales / Spermoculture: bacterial growth of the ejaculate and its relationship with the seminal parameters

ANTECEDENTES: En el semen, algunos microorganismos pueden encontrar las condiciones óptimas para sobrevivir, ocasionando daños a los espermatozoides y desencadenando procesos de infertilidad o infecciones del tracto reproductivo. Entender el papel de los microorganismos aislados en el semen, contribuye a mejorar el diagnóstico de casos de infertilidad donde la única causa aparente son los procesos infecciosos. OBJETIVO: Describir y correlacionar los parámetros seminales y el crecimiento bacteriano del eyaculado. MÉTODOS: Identificación de los microorganismos aislados en 43 espermocultivos-clínicos y 28 espermocultivos-investigación. Se realizó conteo de las unidades formadoras de colonia a los espermocultivos-investigación y análisis de las características espermáticas. Resultados: Se obtuvo crecimiento bacteriano en 14 (32,6%) de los espermocultivos-clínicos y 15 (53,6%) de los espermocultivos-investigación. Los microorganismos aislados fueron Enterococcus faecalis, Escherichia coli, Morganella morganii, Staphylococcus spp coagulasa negativo, Klebsiella pneumoniae y microbiota mixta. En este estudio se observó abundante crecimiento de cocos aerobios. Finalmente, no se encontró asociación entre la disminución en la calidad de los parámetros seminales y los microorganismos. CONCLUSIONES: La presencia de bacterias en el semen no afecta la calidad seminal.

BACKKGROUND: Microorganisms can find the optimal conditions for survival in semen, causing damage to the spermatozoa and triggering processes of infertility or reproductive tract infections. Therefore, understanding the role of the microorganisms present in semen can help to improve the diagnosis of infertility cases where the only apparent cause is infectious processes. OBJECTIVE: To describe and correlate semen parameters and bacterial growth in ejaculate. METHODS: Identification of microorganisms isolated in 43 clinical spermocultures and 28 research spermocultures. We assessed colony-forming unit counts and sperm characteristics of research spermocultures. In addition, semen parameters were evaluated in each ejaculate. RESULTS: Bacterial growth was obtained in 14 (32.6%) of the 43 clinical spermocultures and 15 (53.6%) of the 28 research spermocultures. The isolated microorganisms were Enterococcus faecalis, Escherichia coli, Morganella morganii, Staphylococcus coagulase negative, Klebsiella pneumoniae and mixed microbiota. Finally, in this study a large growth of aerobic cocci was observed. We did not find association between the decline in the quality of semen parameters and microorganisms. CONCLUSION: The presence of bacteria in semen does not affect semen quality.

Capacidad de sellado de tres agentes cementantes para restauraciones coronarias indirectas / Sealing ability of three luting agents used for cementation of indirect coronal restorations

Objetivo: comparar ex vivo, por medio de un modelo de filtración bacteriana, la capacidad de sellado de un ionómero vítreo modificado con resinas, de un ionómero vítreo convencional y de un cemento de fosfato de cinc, utilizados para la cementación de coronas coladas (AU). Materiales y métodos: un total de 30 premolares humanos sanos, extraídos, fueron preparados de forma estandarizada para recibir una corona metálica con terminación en chamfer sobre dentina. Se realizaron impresiones, se vaciaron en densita tipo IV, se enceraron y se colaron en aleación de oro tipo III. Luego fueron separadas al azar en tres grupos de 10 (n=10) especímenes cada uno. Las coronas del grupo 1 fueron cementadas con Rely-X Luting Plus (3M/ESPE); y las del grupo 2, con Ketac-Cem (3m/ESPE); y las del grupo 3 (control), con el cemento de fosfato de cinc Harvard Cement (Harvard Debtal International GmbH, Hoppengarten, Alemania). Todos los especímenes fueron termociclados. Posteriormente, la superficie metálica oclusal fue reducida 1 mm hasta exponer la dentina, y las muestras se esterilizaron en autoclave. El proceso de filtración se realizó por medio de un sistema de doble cámara con medio de cultivo. El medio de la cámara superior se inoculó con E. faecalis y el sistema fue incubado durante 60 días. En la cámara inferior, la filtración bacteriana se controló diariamente. Los datos registrados fueron analizados por medio de las pruebas de Kaplan-Meyer y de Fischer, con un nivel de significación de P<0,05. Resultados: el tiempo de supervivencia para Rely-X Luting Plus no pudo ser estimado, ya que fue superior a los 60 días, tiempo máximo de duración de la experiencia. Para Ketac-Cem fue de 60 días, con un intervalo de confianza (95 por ciento) de 29,6-90,4. Y para Harvard Cement fue de 40 días, con un intervalo de confianza de 32,6-47,4. Las diferencias entre los grupos fueron estadísticamente significativas. Conclusión: Rely-X Plus demostró una capacidad de sellado superior.

Trombosis de vena cava inferior y endocarditis en un paciente prematuro: Caso clínico / Inferior vena cava thrombosis and endocarditis in a premature patient: Case report

Thrombosis of the inferior vena cava represents a significant percentage of all venous thrombosis that take place during the neonatal period, generally associated with risk factors such as the use of central venous catheter. The incidence of bacterial endocarditis in preterm infants is low. Objectives: To characterize the case of a preterm neonate with both conditions and to detail the disease changing spectrum in the neonatal population and its therapeutic possibilities. Case report: Premature newborn, 31 + 5 weeks of gestation who presented Enterococcus faecalis bacteremia, developed progressive thrombosis of the inferior vena cava and right atrium secondary to the use of umbilical venous catheter, with subsequent diagnosis of endocarditis. He was treated with anticoagulation with subcutaneous low molecular weight heparin. Given a favorable evolution, it was decided to continue the anticoagulation therapy for 4-6 weeks, and at the time of discharge, aspirin treatment was given. Conclusions: The preterm infant with infective endocarditis and intracardiac thrombus presents an interesting management dilemma. Treatment should be individualized according to the clinical evolution and safety profile of thrombolytic and/or anticoagulant agents. Availability and advantages of low molecular weight heparin have led to its use as an alternative treatment in neonates and infants with deep venous thrombosis.

La trombosis de la vena cava inferior corresponde a un porcentaje importante de las trombosis venosas en la etapa neonatal, generalmente asociado a factores de riesgo como el uso de catéter venoso central. La incidencia de endocarditis bacteriana en recién nacidos prematuros es baja. Objetivos: Caracterizar el caso de un neonato pretérmino en que se asociaron ambas patologías y detallar el espectro cambiante de esta enfermedad en la población neonatal y sus posibilidades terapéuticas. Caso clínico: Recién nacido prematuro de 31 + 5 semanas de gestación, que presentó bacteriemia por Enterococo faecalis, evolucionó con trombosis progresiva de la vena cava inferior y aurícula derecha secundaria al uso de catéter venoso umbilical, con posterior diagnóstico de endocarditis. Se manejó con anticoagulación con heparina de bajo peso molecular en forma subcutánea, dada evolución favorable, se decidió continuar manejo médico con terapia anticoagulante por 4-6 semanas, y al alta manejo con aspirina. Conclusiones: El neonato pretérmino con endocarditis infecciosa y trombo intracardiaco presentan un dilema interesante de manejo, por lo cual se debe individualizar el tratamiento según la evolución clínica y el perfil de seguridad de los agentes trombolíticos y/o anticoagulantes. La disponibilidad y las ventajas de la heparina de bajo peso molecular ha dado lugar a su uso como una alternativa de tratamiento en neonatos y niños con trombosis venosa profunda.

Evaluation of photodynamic therapy using a diode laser and different photosensitizers against Enterococcus faecalis

Photodynamic therapy (PDT) has been proven to be effectivein disinfecting root canals. The aim of this present study wasto evaluate the effects of PDT on the viability of Enterococcusfaecalis using methylene blue (MB) and malachite green(MG) as photosensitizers. Solutions containing E. faecalis(ATCC 29212) were prepared and harvested by centrifugationto obtain cell suspensions, which were mixed with MBand MG. Samples were individually irradiated by the diodelaser at a distance of 1mm for 30, 60, or 120 seconds. Colonyformingunits (CFU) were determined for each treatment.PDT for 60 and 120 seconds with MG reduced E. faecalis viabilitysignificantly. Similar results were obtained when MBwas used as photosensitizer. PDT using MB and MG haveantibacterial effect against E. faecalis, showing potential tobe used as an adjunctive antimicrobial procedure in endodontictherapy...

Sensibilidad antimicrobiana in vitro en aislamientos de Enterococcus faecalis y Enterococcus faecium obtenidos de pacientes hospitalizados / In vitro antimicrobial susceptibility in Enterococcus faecalis and Enterococcus faecium isolated from hospitalized patients

Introducción. Actualmente se considera a Enterococcus spp. como uno de los agentes de infección hospitalaria más importantes, siendo su resistencia a los antibióticos un problema importante en los centros de salud. Objetivos. Caracterizar la resistencia antimicrobiana en 50 cepas de Enterococcus spp. aisladas de muestras clínicas de pacientes hospitalizados . Materiales y métodos. Se llevó a cabo un estudio de tipo descriptivo observacional de corte transversal en 50 aislamientos clínicos de estas especies microbianas. Se trabajó un aislamiento por paciente. La identificación y la sensibilidad a los antibióticos se realizaron por métodos automatizados y convencionales. El análisis fenotípico de los mecanismos de resistencia a glucopéptidos se hizo según las recomendaciones de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. Resultados. De 50 aislamientos, 30 (60,0 %) y 20 (40,0 %) pertenecían a las especies de Enterococcus faecalis y Enterococcus faecium, respectivamente. La resistencia global expresada por este género fue de 38/50 (76,0 %) para ampicilina; 33/50 (66,0 %) para gentamicina de alto nivel; 34/50 (68,0 %) para estreptomicina de alto nivel; 26/50 (52,0 %) para ciprofloxacina; 4/50 (8,0 %) para linezolid; 17/50 (34,0 %) para teicoplanina; 25/50 (50,0 %) para vancomicina; 31/50 (62,0 %) para minociclina; 34/50 (68,0 %) para tetraciclina y 9/50 (18,0 %) para nitrofurantoina. Frente a los glucopéptidos, 25/50 (50,0 %) y 10/50 (20,0 %) de los aislamientos presentaron los mecanismos Van A y Van B, respectivamente. Conclusiones. Podemos concluir que la mayoría de las veces, las cepas aisladas en el Hospital Hermanos Ameijeiras mostraron porcentajes de resistencia por encima de lo reportado en la literatura científica consultada. El alto porcentaje de cepas con resistencia a la vancomicina podría influir en la aparición de otros gérmenes Gram positivos con resistencia a este fármaco. Se reporta por primera vez en un hospital cubano resistencia de E. faecium a linezolid.

Introduction: Enterococcus spp is currently considered as one of the most important nosocomial pathogens . The antibiotic resistance of this group of bacteria is a particularly important problem in health centers. Objective: To characterize the antibiotic resistance of 50 Enterococcus spp strains isolated from hospitalized patients clinical samples. Materials and methods: We conducted a cross-sectional descriptive observational study in 50 clinical isolates of Enterococcus spp. Only one isolate per patient was analyzed . The identification and antibiotic susceptibility were studied by conventional and automated methods . The phenotypic analysis of glycopeptide resistance mechanisms was performed as recommended by the Spanish Society of Clinical Microbiology and Infectious Diseases . Results: Of 50 isolates obtained from clinical samples, 30 ( 60.0%) belonged to Enterococcus faecalis and 20 (40.0 %) to Enterococcus faecium . The global resistance expressed by this genre was as follows: Ampicillin, 38/50 ( 76.0%); high-level gentamicin, 33/50 ( 66.0%); high-level streptomycin, 34/50 (68.0 %) ; ciprofloxacin, 26/50 (52.0 %); linezolid, 4/50 (8.0 %); teicoplanin, 17/50 ( 34.0%); vancomycin, 25/50 (50.0 %); minocycline, 31/50 ( 62.0%); tetracycline, 34/50 (68.0 %); nitrofurantoin, 9/50 ( 18.0%). As regards glycopeptides, 25/50 (50.0%) showed a Van A mechanism and 10/50 (20.0 %) a Van B mechanism . Conclusions: The isolates obtained at Hospital Hermanos Ameijeiras showed higher resistance rates than those reported in the consulted literature. The high percentage of vancomycin-resistant strains might have influenced the development of other Gram-positive bacteria resistant to this drug. This is the first report on Enterococcus faecium resistant to linezolid in a Cuban hospital.