Infección por Clostridium difficile: descripción de las cepas NAP1/027 y de otros serotipos en un centro de alta complejidad de Cali, Colombia, 2012-2015 / Clostridium difficile infection: Description of NAP1/027 and non NAP1/027 strains in a high complexity center in Cali, Colombia, 2012-2015

Resumen Introducción. Clostridium difficile ocasiona infecciones hospitalarias que resultan en altas tasas de morbilidad y mortalidad. La cepa NAP1/027 se ha asociado con una mayor producción de toxinas y con una mayor gravedad, lo que aumenta la carga de la enfermedad. Objetivo. Describir la epidemiología de las infecciones asociadas con C. difficile y las características de la cepa NAP1/027. Materiales y métodos. Se hizo un estudio observacional basado en la revisión de las historias clínicas de los pacientes con muestras de heces positivas para C. difficile identificadas mediante la prueba Xpert™ entre el 2012 y el 2015 en un hospital de alta complejidad. La gravedad de la enfermedad se evaluó con el índice ATLAS. Resultados. Se incluyeron 42 casos de pacientes infectados, 9 de los cuales fueron positivos para la cepa NAP1/027. El uso de antibióticos antes de la infección durante más de siete días fue más frecuente en los casos de pacientes con muestras negativas para NAP1/027. En la mitad de los pacientes, la duración de la diarrea fue mayor de cinco días y no hubo diferencias según el tipo de cepa (p>0,05). Los casos de pacientes positivos para la cepa NAP1/027 se caracterizaron por presentar deposiciones fétidas y sanguinolentas. La gravedad de la infección fue similar entre los grupos. Conclusión. Se comprobó la circulación de la cepa NAP1/027, pero su presencia no supuso diferencias clínicas significativas con respecto a otras cepas, lo cual podría deberse al limitado número de pacientes en este estudio. Sin embargo, su presencia debe alertar a los médicos y a las instituciones de salud, dada su frecuente asociación con la gravedad de la infección y la mortalidad.

Abstract Introduction: Clostridium difficile causes nosocomial infections leading to high morbidity and mortality. The NAP1/027 strain is associated with a higher toxin production and disease severity, which increases the load of the disease. Objective: To describe the epidemiology of the infections associated with C. difficile and the characteristics related to the NAP1/027 strain. Materials and methods: This was an observational study based on the revision of clinical registries of patients with fecal samples that were positive for C. difficile identified by the Xpert test™ between 2012 and 2015 in a high complexity institution. The severity of the disease was evaluated by means of the ATLAS score. Results: We included 42 infected cases, 9 of which were positive for the NAP1/027strain. The use of antibiotics previous to the infection for more than seven days was more frequent in patients with negative results for NAP1/027. The duration of diarrhea in half of the patients was longer than five days and there were no differences according to the type of strain (p>0.05). Positive cases for the NAP1/027 strain were characterized by presenting fetid and bloody stools. The severity of the infection was similar between the groups. Conclusions: In Colombia, the NAP1/027 strain circulates without significant clinical differences, which could be due to the limited number of patients. Nevertheless, the existence of NAP1/027 should alert physicians and health institutions because of its high association with severity and mortality.

Manejo actualizado de la diarrea asociada a Clostridium difficile / Updated management of diarrhea associated with Clostridium difficile

The management of Clostridium difficile (CD) infection has changed in recent years. The latest clinical guidelines and systematic reviews suggest the use of vancomycin orally as the first line of treatment regardless the severity of the crisis (main difference compared to previous recommendations), this is due to changes in its epidemiology, the decrease in effectiveness and the increase of recurrences with the use of metronidazole, particularly in severe crisis. In addition, the use of new agents such as fidaxomicin has been approved. Fulminant crisis require an aggressive management combining oral treatment, enemas and intravenous therapy in addition to a collaborative management with the surgery team. With respect to recurrences, the use of vancomycin in pulses and with extended therapy schemes is suggested; fecal microbiota transplantation (FMT) is also an attractive therapy for patients with multiple recurrences. The following is a summary of the latest recommendations and available evidence regarding the management of CD infection in the most frequent situations, both in first crisis and in its recurrences.

El manejo de la infección por Clostridium difficile (CD) ha tenido modificaciones los últimos años. Las últimas guías clínicas y revisiones sistemáticas sugieren el uso de vancomicina vía oral como primera línea de tratamiento independiente de la severidad de la crisis (diferencia principal con recomendaciones previas), esto debido a cambios en su epidemiología, la disminución de la efectividad y al aumento de las recurrencias con el uso de metronidazol, particularmente en crisis severas. Además, han sido aprobados el uso de nuevos agentes como la fidaxomicina. Las crisis de carácter fulminante requieren un manejo agresivo combinando terapia oral, vía enemas e intravenosa, además de un manejo en conjunto con el equipo de cirugía. Respecto a las recurrencias se sugiere el uso de vancomicina en pulsos y con esquemas de terapia extendida siendo además, el trasplante de microbiota fecal (FMT) una terapia atractiva para pacientes con múltiples recurrencias. A continuación se resumen las últimas recomendaciones y evidencia disponible respecto del manejo de la infección por CD en las situaciones más frecuentes, tanto en la primera crisis como en sus recurrencias.

Detection of vancomycin-resistant enterococci (VRE) in stool specimens submitted for Clostridium difficile toxin testing

Abstract The aim of this study was to determine the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and efficacy of screening stools submitted for C. difficile toxin assay for prevalence of VRE. Between April 2012 and February 2014, 158 stool samples submitted for C. difficile toxin to the Marmara University Microbiology Laboratory, were included in the study. Stool samples were analyzed by enzyme immuno assay test; VIDAS (bioMerieux, France) for Toxin A&B. Samples were inoculated on chromID VRE (bioMerieux, France) and incubated 24 h at 37 °C. Manuel tests and API20 STREP (bioMerieux, France) test were used to identify the Enterococci species. After the species identification, vancomycin and teicoplanin MIC's were performed by E test and molecular resistance genes for vanA vs vanB were detected by polymerase chain reaction (PCR). Of the 158 stool samples, 88 were toxin positive. The prevalence of VRE was 17%(n:19) in toxin positives however, 11.4% in toxin negatives(n:70). All VRE isolates were identified as Enterococcus faecium. These results were evaluated according to Fischer's exact chi-square test and p value between VRE colonization and C. difficile toxin positivity was detected 0.047 (p < 0.05). PPV and NPV were 79% and 47% respectively. In our study, the presence of VRE in C. difficile toxin positives is statistically significant compared with toxin negatives (p < 0.05). Screening for VRE is both additional cost and work load for the laboratories. Therefore VRE screening among C. difficile toxin positive samples, will be cost effective for determination of high risk patients in the hospitals especially for developing countries.

Clostridium difficile infections in HIV-positive patients with diarrhoea.

Background. Patients with HIV/AIDS are at a high risk of being infected with toxin-producing strains of Clostridium difficile (C. difficile) because of frequent hospitalization, exposure to antibiotics and antibiotic prophylaxis for opportunistic infections. There are little data from India on the prevalence of C. difficile infection in such patients. Methods. We assessed the occurrence of C. difficile infections in HIV-positive patients with diarrhoea by looking for the presence of its toxin as well as by culturing. Enzyme immunoassay (EIA, Premier toxins A and B; Meridian Diagnostic Inc.) was used to detect toxin from 237 fresh stool samples collected from HIV-positive patients with diarrhoea. Culture was done on cycloserine–cefoxitin–fructose agar and brain– heart infusion agar. Results. C. difficile was found in 12 of 237 (5.1%, 95% CI 2.64%–8.68%) HIV-positive patients with diarrhoea (9 patients were positive by EIA and 3 by culture). The presence of C. difficile in patients who had received antiretroviral therapy (7/66 [10.6%]) was significantly higher (p<0.016) compared with those who had not (5/171 [3%]). Of the 12 patients positive for C. difficile, 7 were on antiretroviral therapy for a mean (SD) of 34.4 months with mean CD4+ count of 186 (98.81) cells/cmm and 5 patients were anti-retroviral-naïve with mean CD4+ count of 181 (68.7) cells/cmm. All the 12 patients were on antibiotics for previous 2 months and 4 of 12 had been hospitalized in the previous 30 days. Conclusion. C. difficile infections occurred more frequently in patients who had received antiretroviral therapy. Our study population had a lower frequency of C. difficile infections compared to previous studies.

Comparison of Supplemented Brucella Agar and Modified Clostridium difficile Agar for Antimicrobial Susceptibility Testing of Clostridium difficile

BACKGROUND: Antimicrobial susceptibility testing (AST) of Clostridium difficile is increasingly important because of the rise in resistant strains. The standard medium for the AST of C. difficile is supplemented Brucella agar (sBA), but we found that the growth of C. difficile on sBA was not optimal. Because active growth is critical for reliable AST, we developed a new, modified C. difficile (mCD) agar. C. difficile grew better on mCD agar than on sBA. METHODS: C. difficile isolates were collected from patients with healthcare-associated diarrhea. sBA medium was prepared according to the CLSI guidelines. Homemade mCD agar containing taurocholate, L-cysteine hydrochloride, and 7% horse blood was used. For 171 C. difficile isolates, we compared the agar dilution AST results from mCD agar with those from sBA. RESULTS: No significant differences were observed in the 50% minimal inhibitory concentration (MIC50) and 90% minimal inhibitory concentration (MIC90) of clindamycin (CLI), metronidazole (MTZ), moxifloxacin (MXF), piperacillin-tazobactam (PTZ), and rifaximin (RIX), but the values for vancomycin (VAN) were two-fold higher on mCD agar than on sBA. The MICs of CLI, MXF, and RIX were in 100% agreement within two-fold dilutions, but for MTZ, VAN, and PTZ, 13.7%, 0.6%, and 3.1% of the isolates, respectively, were outside the acceptable range. CONCLUSIONS: The MIC ranges, MIC50 and MIC90, were acceptable when AST was performed on mCD agar. Thus, mCD agar could be used as a substitute medium for the AST of C. difficile.

Effect of biotherapeutics on antitoxin IgG in experimentally induced Clostridium difficile infection.

Purpose: Recurrent diarrhoea after successful treatment of primary Clostridium difficile associated disease (CDAD) occurs due to bowel flora alterations and failure to mount an effective antibody response. Apart from antibiotics, risk factors include immunosuppressive and acid-suppressive drug administration. Biotherapeutics such as probiotic and epidermal growth factor (EGF) may offer potential effective therapy for CDAD. Materials and Methods: The effect of biotherapeutics in mounting an antibody response against C. difficile toxins was studied in BALB/c mice challenged with C. difficile after pre-treatment with ampicillin, lansoprazole or cyclosporin. Sera from sacrificed animals were estimated for antitoxin IgG by enzyme linked immunosorbent assay. Results: Antitoxin IgG was significantly higher (P<0.05) in C. difficile challenged groups compared to unchallenged controls, but insignificant (P>0.05) in animals in which C. difficile was given after pre-treatment with cyclosporin compared to those without any pre-treatment, or pre-treatment with antibiotic or lansoprazole. In inter-subgroup comparisons also significant anomaly in production of antitoxin IgG was found. The antitoxin IgG levels were raised in animals administered C. difficile after pre-treatment with ampicillin, but lower in animals administered cyclosporin. High levels of antitoxin IgG were also found in the serum samples of animals receiving lansoprazole and C. difficile. Conclusions: Probiotics showed their beneficial effect by boosting the immune response as seen by production of antitoxin IgG. Oral administration of EGF did not affect the immune response to C. difficile toxins as significant increase was not observed in the serum antitoxin IgG levels in any of the groups investigated.

Recent changes in Clostridium difficile infection / Recentes mudanças da infecção por Clostridium difficile

Clostridium difficile is the main cause of nosocomial diarrhea. Diarrhea associated with C. difficile has increased incidence, morbidity, and mortality in the last few years. The major related risk factors include use of antibiotics, elderly patients and prolonged hospital stay. Many patients receive combinations of antibiotics or multiple antibiotics, which represents the main risk to develop diarrhea associated to C. difficile or its recurrence. Therefore, interventions to improve antibiotic prescribing, as well as compliance with infection control measures can reduce hospital-acquired C. difficile infections. This review addresses the epidemiological changes in C. difficile disease and its treatment.

Clostridium difficile é a principal causa de diarreia hospitalar. A diarreia por C. difficile aumentou sua incidência e sua morbiletalidade nos últimos anos. Os principais fatores de risco relacionados são uso de antibióticos, idosos e permanência hospitalar prolongada. Muitos pacientes recebem combinação de antibióticos ou múltiplos antibióticos, constituindo-se, assim, o principal fator de risco para o desenvolvimento de infecção ou de recorrência de diarreia associada ao C. difficile. Por isso, intervenções que otimizem a prescrição de antibióticos associado à aderência de medidas de controle de infecção podem reduzir aquisição dessa infecção. Assim, esta revisão aborda a mudança da epidemiologia da infecção por C. difficile e seu tratamento.

Treatment of Refractory or Recurrent Clostridium difficile Infection / 대한소화기학회지

The incidence and severity of Clostridium difficile infection (CDI) has increased over the past decades. It is related to the emergence of hypervirulent strains and increased use of antibiotics. The incidence of refractory CDI to standard therapies and the risk for recurrent CDI are also increasing. Current guidelines recommend the first recurrence to be treated with the same agent used for the initial episode. However, data are lacking to support any particular treatment strategy for severe refractory CDI or cases with multiple recurrence. Treatments currently available for CDI are inadequate to prevent recurrence. Widely used method for managing a subsequent recurrence involves tapering followed by pulsed doses of vancomycin. Other potentially effective strategies for recurrent CDI are use of other antibiotics such as fidaxomicin, nitazoxanide, rifaximin, tigecycline, and teicoplanin. There are efforts to recover gut microflora and to optimize immune response to CDI. These include use of probiotics, fecal microbiota transplantation, intravenous immunoglobulin, monoclonal antibodies directed against C. difficile toxins, and active vaccination. However treatment of patients with refractory CDI and those with multiple CDI recurrences is based on limited clinical evidence, and there is an ongoing need for continued research to improve the outcomes these patients.

Clostridium Difficile-Associated Diarrhea: A Review.

Clostridium difficile–associated diarrhea (CDAD) has become a major public health problem. The offending pathogen is acquired by the fecal-oral route from an environmental source or by contact with an infected person or health care worker who serves as a vector. Alteration of the intestinal microflora, frequently by antibiotics, generates a favorable environment results in the proliferation of C. difficile. The pathogen is not invasive but produces two toxins, A and B, that lead to severe inflammation of the colonic mucosa manifested as profound diarrhea, fever, abdominal pain, and leukocytosis. A new hypervirulent strain of C. difficile has become prevalent in the United States, Canada, and the United Kingdom. Identified by pulsed-field gel electrophoresis (PFGE), this strain is called North America PFGE type 1, abbreviated as NAP-1, and characteristically generates large amounts of toxins A and B, in addition to a binary toxin, and is associated with higher morbidity and failure of antibiotic therapy. Mild cases of CDAD may respond to withdrawal of antibiotic therapy, however the majority of patients require C. difficile-specific antimicrobial therapy. The US FDA has approved oral vancomycin for treatment of CDAD, but reluctance to use this antibiotic due to the fear of developing vancomycinresistant organisms in the hospital setting has made metronidazole the recommended first-line therapy for mild to moderate disease. Some newer studies show a high rate of failure, due to infection by NAP-1 or to the presence, in hospitals, of geriatric patients with more concurrent illnesses who may also have been treated with many broad-spectrum antibiotics. The recurrence rate after initial successful treatment can be as high as 20-30%, depending on the initial treatment. A more C. difficile-targeted non-absorbable bacterial RNA polymerase inhibitor, fidaxomicin (also known as OPT-80 and PAR-101), has recently been approved in the US with initial efficacy similar to vancomycin and a lower recurrence rate. Some additional agents that have shown efficacy in humans are nitazoxanide, bacitracin, teicoplanin, and fusidic acid. Rifaximin, polymers that bind C. difficile toxin, monoclonal antibodies to toxins, and preventive measures such as toxoid vaccines are under investigation. Interventions for treatment of recurrences include repeated vancomycin or fidaxomicin courses, probiotics, rifaximin, intravenous immunoglobulin and fecal transplants. Measures for preventing the spread of the pathogen, appropriate diagnostic testing, and treatment may avert morbidity and mortality due to CDAD.

Diagnóstico de diarrea por clostridium difficile: en busca de un enfoque clínico más eficiente / Clinical diagnosis of clostridium difficile diarrhea: searching a more efficient clinical view

Background: The clinical parameters for the suspicion of Clostridium difficile infections, namely the use of antimicrobials and diarrhea, have a low predictive value for the diagnosis. Aim: To search other clinical variables and determine a clinical prediction model for (Clostridium difficile diarrhea. Patients and methods: All patients to whom a Clostridium difficile study was requested, were prospectively studied during 5 months. Clinical variables of these patients were registered. The diagnosis of Clostridium difficile was done using the cytotoxicity test in fibroblast cultures. Results: Ninety two patients were analyzed and in 26, the diagnosis of Clostridium difficile was confirmed. A logistic regression model disclosed an age over 60 years old, the presence of mucus in the stools and a temperature over 37.8 ­C in the previous 24 h, as significant predictors of the infection. The correlation of the model, between the predicted probability and the observed condition, was 81.5 per cent. Conclusions: The presence of the clinical variables identified in this study are associated with a high probability of an infection by Clostridium difficile in patients with diarrhea and the recent use of antimicrobials

Probióticos / Probiotics

Los probióticos son un recurso terapéutico cuya utilidad está demostrada en la prevención y tratamiento de la diarrea aguda y colitis pseudomembranosa asociadas a disbacteriosis provocada por la administración de antibióticos. Otras indicaciones son aún discutibles y requieren de mayores estudios

Probióticos / Probiotics

Los Probióticos son agentes terapéuticos biológicos emergentes, con utilidad probada en diarrea asociada a antibióticos, diarrea en la infancia y del viajero. Hay trabajos recientes que cifran esperanzas en la terapia de la diarrea asociada a C. difficile, en enfermedad inflamatoria intestinal, (tanto en tratamiento agregado a la terapia convencional de la crisis, como en la prevención de rebrotes). Por su excelente tolerancia, disponibilidad y acepatación por la población, debe considerarse dentro del recetario gastroenterológico con interés y expectación

Diarrea asociada a clostridium difficile en un hospital de adultos: estudio descriptivo / Diarrhea associated to clostridium difficile in an adult hospital: a descriptive study

Clostridium difficile es el principal agente causal de diarreas nosocomiales. Para conocer las características epidemiológicas de las infecciones por este agente, se realizó un estudio sobre los casos de diarrea asociados a citotoxina A de C. difficile en deposiciones. Un total de 27 pacientes con 31 eventos fue identificado durante 4 meses de vigilancia pasiva. La mayor parte eran pacientes de sexo femenino (62,9 por ciento) mayores de 65 años (77,7 por ciento) y habían sido sometidos a procedimientos gastrointestinales o recibido lactulosa (59,2 por ciento). Todos los pacientes habían sido expuestos a terapia antimicrobiana, especialmente ciprofloxacina (40,7 por ciento). Todos evolucionaron favorablemente, excepto tres casos que fallecieron durante la hospitalización (11 por ciento), aunque sólo uno de ellos por una causa directamente atribuible a C. difficile. En 7 pacientes se registró un fracaso del tratamiento con metronidazol (25,9 por ciento) y 3 pacientes recayeron durante el seguimiento (11 por ciento). El índice de casos según egresos fue más frecuente en las salas de recuperación neuroquirúrgica y de atención intermedia de medicina

Brote de infección intrahospitalaria por clostridium difficile / Hospital infection outbreaks by clostridium difficile

Un brote de infección nosocomial por clostridium difficile afectó a tres pacientes de sexo femenino admitidas a una misma sala de hospitalización en un plazo no superior a 15 días. Los 3 casos (65 a 84 años) tenían antecedentes de uso de antibióticos (ciprofloxacina en dos de ellas) o habían sufrido procedimientos gastrointestinales (en dos casos). Los 3 casos se presentaron con diarrea acuosa con leucocitos fecales positivos y fueron tratados con metronidazol con buena respuesta. Un caso falleció por causas no claramente relacionadas a una infección por este agente. Las infecciones en el servicio respectivo desaparecieron luego de un programa de capacitación al personal profesional y no profesional y de la aplicación de un aislamiento de contacto. Clotridium difficile es el principal agente de diarreas infecciosas nosocomiales. Las infecciones por esta bacteria pueden presentarse como casos endémicos o en brotes intrahospitalarios bien definidos. La participación de C. difficile como causa de diarrea a nivel hospitalario es explicada por la existencia de pacientes susceptibles y por la alta transmisibilidad de este agente. Diversos factores de riesgo están asociados a la susceptibilidad que presentan algunos pacientes a desarrollar diarreas por este microorganismo esporulado, y ellas incluyen el uso previo de antibióticos, la edad y procedimientos o medicamentos que alteran la microbiota colónica normal. Numerosas publicaciones han sido reportadas sobre brotes nosocomiales provocados por esta bacteria, que han afectado a diferentes tipos de pacientes. El propósito de este trabajo, es comunicar y analizar un brote ligado a este agente, el que se presentó poco después de incorporar en el laboratorio técnicas de detección para esta bacteria

Antibacterial activity of teicoplanin against Clostridium difficile.

The in vitro inhibitory action of teicoplanin, vancomycin, metronidazole and clindamycin against clinical isolates of Clostridium difficile was investigated. Minimum inhibitory concentrations (MICs) were determined using E test. Teicoplanin (MIC range 0.023-0.75 microgram/ml), vancomycin (MIC range 0.5-3 micrograms/ml) and metronidazole (MIC range 0.19-1 microgram/ml) were all very active against the isolates examined. No resistant strains of C. difficile to those three antimicrobial agents were observed, whereas resistance to clindamycin was found in 39.5% of the tested strains. Teicoplanin was about 4-times more potent than vancomycin. It appears to be a more promising antimicrobial for treatment of C. difficile enteric disease.