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.

Development of Chlorhexidine Digluconate and Lippia sidoides Essential Oil Loaded in Microemulsion for Disinfection of Dental Root Canals: Substantivity Profile and Antimicrobial Activity.

The dental intracanal disinfection is crucial to achieve the success of endodontic treatment, avoiding the maintenance of endodontic infections. Chlorhexidine digluconate can act as an irrigating agent for it. However, it can cause tissue irritation in high concentrations. Therefore, combinations with other antimicrobial agents and more efficient therapeutic alternatives are studied, which make it possible to administer drugs more safely and with minimal adverse effects. Thus, the objective of this study was the development of a microemulsion containing chlorhexidine digluconate and essential oil of Lippia sidoides to be used for disinfection of dental root canals and to evaluate its profile of substantivity and antimicrobial activity. The microemulsions were obtained through phase diagrams, using the spontaneous formation method. We completed a physical-chemical characterization and evaluate the stability of the microemulsions, in addition to the substantivity profile in a bovine root dentin model, and in vitro antibacterial effect on Enterococcus faecalis. A method for quantifying chlorhexidine was developed using UV-Vis spectroscopy. The microemulsions showed acid pH, conductivity above 1.3 µScm-1, and dispersion index similar to water. The microemulsions showed antimicrobial inhibition halos similar to the commercial gel conventionally used, but with four times more substantivity to dentinal tissues. Microemulsions were obtained as a therapeutic alternative to formulations available on the market, presenting themselves as a system with great potential for the administration of drugs for disinfection of root canals.

Trend of clinical vancomycin-resistant enterococci isolated in a regional Italian hospital from 2001 to 2018.

A retrospective study of the epidemiology of vancomycin-resistant enterococci (VRE) in a regional hospital of central Italy in 2001-2018 demonstrated an increased VRE prevalence since 2016. A total of 113 VRE isolates, 89 E. faecium (VREfm) and 24 E. faecalis (VREfs), were collected in the study period. All strains showed high-level resistance to vancomycin; 107 also showed teicoplanin resistance. Altogether, 84 VREfm and 20 VREfs carried vanA, whereas 5 VREfm and 1 VREfs carried vanB. MLST analysis documented that 89 VREfm isolates mainly belonged to ST78, ST80, and ST117. Most strains were isolated from 2001 to 2007, ST78 being the predominant clone. VREfm re-emerged in 2016 with a prevalence of the ST80 lineage. Most VREfs were isolated from 2001 to 2006; although they belonged to 7 different STs, there was a prevalence of ST88 and ST6. Notably, ST88 was sporadically recovered throughout the study period. The increasing rate of VREfm isolation from 2016 to 2018 may be related to the influx of new successful clones and to the renewed and widespread use of vancomycin. Improved infection control measures in hospital wards should be adopted to limit the spread of new epidemic VRE strains.

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.

The gutta-percha 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 gutta-percha 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, gutta-percha 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 gutta-percha cones to the decontaminating agent should not be reduced.

Os cones de guta-percha utilizados no tratamento endodôntico são produzidos em condições assépticas e possuem óxido de zinco em sua composição, responsável pela atividade antibac-teriana. 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 protocolo de desinfecção a ser usado. Nosso objetivo foi avaliar a descontaminação de cones de guta-percha 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 guta-percha 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, observou-se 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 guta-percha ao agente de descontaminação não deve ser reduzido para evitar a possibilidade de falhas nesse estágio.

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.

The polytetrafluoroethylene (PTFE) channel model of cyclic-buildup biofilm and traditional biofilm: The impact of friction, and detergent on cleaning and subsequent high-level disinfection.

OBJECTIVE: To evaluate the efficacy of detergent and friction on removal of traditional biofilm and cyclic-buildup biofilm (CBB) from polytetrafluoroethylene (PTFE) channels and to evaluate the efficacy of glutaraldehyde to kill residual bacteria after cleaning. METHODS: PTFE channels were exposed to artificial test soil containing 108 CFU/mL of Pseudomonas aeruginosa and Enterococcus faecalis, followed by full cleaning and high-level disinfection (HLD) for five repeated rounds to establish CBB. For traditional biofilm, the HLD step was omitted. Cleaning with enzymatic and alkaline detergents, bristle brush, and Pull Thru channel cleaner were compared to a water flush only. Carbohydrate, protein, viable count, adenosine triphosphate (ATP) levels were analyzed and atomic force microscopy (AFM) was performed. RESULTS: In the absence of friction, cleaning of traditional biofilm and CBB was not effective compared to the positive control (Dunn-Bonferroni tests; P > .05) regardless of the detergent used. ATP, protein, and carbohydrate analyses were unable to detect traditional biofilm or CBB. The AFM analysis showed that fixation resulted in CBB being smoother and more compact than traditional biofilm. CONCLUSION: Friction during the cleaning process was a critical parameter regardless of the detergent used for removal of either traditional biofilm or CBB. Glutaraldehyde effectively killed the remaining microorganisms regardless of the cleaning method used.

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

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.

Assessment of vancomycin resistance transfer among enterococci of clinical importance in milk matrix.

Dissemination of vancomycin resistance in enterococci has been associated with horizontal transfer of mobile genetic elements. Aim of the study was to evaluate if milk matrix is a suitable environment to support transferability of vancomycin resistance (vanA) gene from clinical vancomycin-resistant Enterococcus faecium to vancomycin-sensitive Enterococcus faecalis. Enterococci strains were firstly screened for the presence of cpd (inducible sex pheromone determinant) gene, vanA and tetL genes (vancomycin and tetracycline resistance markers, respectively) and the gelE (extracellular metalloendopeptidase) gene to define the mating pairs. Based on these selection markers, we investigated the transferability of eight plasmid-borne vanA harbored by E. faecium (vanA+, cpd-, tetL- and gelE-) into two E. faecalis (vanA-, cpd+, tetL + and gelE+) recipient strains in milk matrix. The strains were mated in a 1:1 ratio in 7% reconstituted milk and incubated at 37 °C. Transconjugants emerged from all 16 matings within 2 h of incubation and were evidenced by dual antibiotic resistance (vancomycin and tetracycline). The vancomycin-resistance of trasconjugants was maintained even after ten subsequent passages on nonselective medium. Transconjugants were positive for vanA, tetL and gelE genes. This study indicates milk matrix as suitable environment to support gene exchange between Enterococcus species.

INVESTIGATION OF ENTEROCOCCUS FAECALIS POPULATION IN PATIENTS WITH POLYP AND COLORECTAL CANCER IN COMPARISON OF HEALTHY INDIVIDUALS.

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.

High Prevalence of the aac(6')-Ie-aph(2'')-Ia Gene in Hospital Isolates of Enterococcus faecalis Co-Resistant to Gentamicin and Penicillin.

Objectives: This study aimed to characterize the molecular mechanism of resistance to gentamicin among penicillin-resistant, ampicillin-susceptible Enterococcus faecalis (PRASEF) isolates by investigating the presence of the aac(6')-Ie-aph(2'')-Ia gene. The co-resistance to antimicrobials of other classes was also evaluated. Results: Among the 151 isolates evaluated, 70 were PRASEF and 81 were penicillin-susceptible and ampicillin-susceptible E. faecalis (PSASEF). No ß-lactamase producing isolate was detected. Eighty-three (55.0%) and 35 (23.2%) out of the 151 E. faecalis isolates showed high-level gentamicin resistance (HLGR) and high-level streptomycin resistance (HLSR) phenotypes. However, a significantly higher rate of PRASEF (88.6%) showed HLGR phenotype in comparison with PSASEF (23.5%) (p < 0.01). Conversely, a significantly lower rate of PRASEF (14.3%) showing HLSR was observed in comparison with PSASEF (30.9%) (p = 0.02). The prevalence of isolates displaying multidrug resistance (MDR) phenotype was significantly higher (p < 0.01) in the group of PRASEF (81.4%) than in PSASEF (18.6%). The majority of PSASEF (61.9%) and PRASEF (90.3%) isolates showing HLGR phenotype was harboring the aac(6')-Ie-aph(2'')-Ia gene, which encodes a bifunctional enzyme that inactivates all aminoglycosides except streptomycin. Conclusion: The aac(6')-Ie-aph(2'')-Ia gene was prevalent among the Brazilian PRASEF isolates that usually exhibit co-resistance to gentamicin and to multiple other drugs.

Bordetella trematum infection: case report and review of previous cases.

BACKGROUND: Bordetella trematum is an infrequent Gram-negative coccobacillus, with a reservoir, pathogenesis, a life cycle and a virulence level which has been poorly elucidated and understood. Related information is scarce due to the low frequency of isolates, so it is important to add data to the literature about this microorganism. CASE PRESENTATION: We report a case of a 74-year-old female, who was referred to the hospital, presenting with ulcer and necrosis in both legs. Therapy with piperacillin-tazobactam was started and peripheral artery revascularization was performed. During the surgery, a tissue fragment was collected, where Bordetella trematum, Stenotrophomonas maltophilia, and Enterococcus faecalis were isolated. After surgery, the intubated patient was transferred to the intensive care unit (ICU), using vasoactive drugs through a central venous catheter. Piperacillin-tazobactam was replaced by meropenem, with vancomycin prescribed for 14 days. Four days later, levofloxacin was added for 24 days, aiming at the isolation of S. maltophilia from the ulcer tissue. The necrotic ulcers evolved without further complications, and the patient's clinical condition improved, leading to temporary withdrawal of vasoactive drugs and extubation. Ultimately, however, the patient's general condition worsened, and she died 58 days after hospital admission. CONCLUSIONS: Despite being a rare finding, B. trematum is typically associated with the clinical manifestation of disorders that predispose to ulcer development, which can be infected by microorganisms. The combination of antibiotic therapy and surgical debridement plays a key role in preventing systemic infections. Monitoring the appearance of new cases of B. trematum is essential, since it can be an emerging microorganism. Isolating and defining the clinical relevance of unusual bacteria yields a more accurate perspective in the development of new diagnostic tools and allows for assessment of proper antimicrobial therapy.

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.

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.

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.

Vancomycin-resistant enterococci isolates colonizing and infecting haematology patients: clonality, and virulence and resistance profile.

BACKGROUND: Vancomycin-resistant enterococci (VRE) are an important agent of colonization and infection in haematology patients. However, the role of virulence on VRE colonization and infection is controversial. AIM: To characterize the lineage, virulence and resistance profile of VRE infection and colonization isolates; as well as their impact on outcome of haematology patients using a regression logistic model. METHODS: Eighty-six isolates (80 Enterococcus faecium and six E. faecalis) from 76 patients were evaluated. Polymerase chain reaction for resistance and virulence genes, and pulsed-field gel electrophoresis and whole genome sequencing of the major clusters, were performed. Bivariate and multivariate analyses were carried out to evaluate the role of virulence genes on outcome. FINDINGS: All isolates harboured the vanA gene. Regarding the virulence genes, 96.5% of isolates were positive for esp, 69.8% for gelE and asa1 genes. VRE infection isolates were more virulent than colonization isolates and harboured more often the gelE gene (P = 0.008). Infections caused by VRE carrying asa1 gene resulted more frequently in death (P = 0.004), but only the predominant clone remained as protector in the multivariate model. The E. faecium strains were assigned to seven STs (ST78, ST412, ST478, ST792, ST896, ST987, ST963) that belonged to CC17. The E. faecalis sequenced belonged to ST9 (CC9). CONCLUSION: E. faecium was predominant, and infection isolates were more virulent than colonization isolates and harboured more often the gene gelE. Infections caused by VRE carrying the asa1 gene appeared to be associated with a fatal outcome.

Instrument Design May Influence Bacterial Reduction During Root Canal Preparation.

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.

ZAAPS Program results for 2015: an activity and spectrum analysis of linezolid using clinical isolates from medical centres in 32 countries.

OBJECTIVES: To report the linezolid in vitro activity obtained during the 2015 ZAAPS Program. METHODS: In total, 7587 organisms causing documented infections were consecutively collected in 65 centres in 32 ex-USA countries. Broth microdilution susceptibility testing was performed. Isolates displaying linezolid MIC results of ≥ 4 mg/L were molecularly characterized. RESULTS: Linezolid inhibited >99.9% of Staphylococcus aureus at ≤ 2 mg/L, with MIC50 results of 1 mg/L, regardless of methicillin resistance. A similar linezolid MIC50 result (0.5 mg/L) was observed for CoNS, with the vast majority of isolates (99.7%) also inhibited at ≤ 2 mg/L. Three CoNS (linezolid MIC, 16-64 mg/L) from Italy were found to contain alterations in the 23S rRNA and/or L3/L4 ribosomal proteins. One isolate also harboured cfr. Linezolid exhibited consistent modal MIC and MIC50 results (1 mg/L) for enterococci regardless of species or vancomycin resistance. One Enterococcus faecalis (linezolid MIC, 8 mg/L) from Galway, Ireland, carried optrA. One Enterococcus faecium (linezolid MIC, 16 mg/L) from Italy contained a G2576T mutation in the 23S rRNA. All Streptococcus pneumoniae, viridans group streptococci and ß-haemolytic streptococci were inhibited by linezolid at ≤ 2, ≤ 2 and ≤ 1 mg/L, respectively, with equivalent MIC90 results (1 mg/L for all groups). CONCLUSIONS: These results document the continued long-term and stable in vitro potency of linezolid and a limited number of isolates with decreased susceptibility to linezolid (MIC, ≥4 mg/L). The latter isolates showed primarily mutations in the 23S rRNA gene and/or L3/L4 proteins, with plasmid-mediated resistance (cfr and optrA) also present, albeit at a low prevalence.