ABSTRACT
Background: Different strains of invasive Escherichia coli (E. coli), isolated from intestinal mucosa of patients, are related to the pathogenesis of inflammatory bowel diseases (IBD). Aim: To evaluate an association between intracellular E. coli and IBD; its clinical characteristics and use of steroids. Material and Methods: Sixty one patients with Crohn's disease and 83 with ulcerative colitis were studied. To determine the intracellular E. coli content, colonoscopy biopsies of these patients and 29 control subjects were processed using the gentamicin protection assay. Differences in the bacterial content between patient groups were evaluated using Mann-Whitney test, while the association between presence of E. coli with endoscopic activity, location/extension and use of corticosteroid as anti-inflammatory treatment were evaluated with Fisher's exact test or Chi-square test. Results: E. coli strains were detected in 36.1, 39.3 and 10.3% of patients with ulcerative colitis, Crohn's disease and controls, respectively. The number of bacteria per biopsy in Crohn's disease and ulcerative colitis was significantly higher than in controls (p < 0.01 between patients and controls). In ulcerative colitis, significant associations were found between the presence of bacteria and disease location and use of corticosteroids. In Crohn's disease, no association was found. Conclusions: IBD are associated with the presence of intracellular E. coli strains in the intestinal mucosa, suggesting an alteration in the microbiota or loss of integrity of the epithelial barrier. The association of intracellular E. coli with clinical features and the use of corticosteroids in ulcerative colitis suggests that different factors could promote colonization or proliferation of these bacteria.
Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Colitis, Ulcerative/microbiology , Crohn Disease/microbiology , Escherichia coli/isolation & purification , Intestinal Mucosa/microbiology , Reference Values , Colony Count, Microbial , Colitis, Ulcerative/drug therapy , Crohn Disease/drug therapy , Case-Control Studies , Prospective Studies , Adrenal Cortex Hormones/therapeutic use , Statistics, Nonparametric , Anti-Inflammatory Agents/therapeutic useABSTRACT
CONTEXT AND OBJECTIVE: Mycobacterium avium subsp. paratuberculosis (MAP) has attracted the interest of researchers because of similarities between paratuberculosis and Crohn's disease (CD). The aim of this study was to evaluate the frequency of MAP through cultures, histology and polymerase chain reaction (PCR) on intestinal biopsies from Brazilian CD patients. Quantitative real time PCR (qRT-PCR) was performed on positive samples. DESIGN AND SETTING: Analytical cross-sectional study with control group at two federal universities. METHODS: Fresh samples were collected from 25 patients; five with CD, eight with ulcerative colitis (UC) and 12 controls with non-inflammatory bowel disease (nIBD). Formalin-fixed paraffin-embedded (FFPE) samples from 143 patients were also collected: 44 CD, 49 UC and 56 nIBD. RESULTS: None of the fresh samples was positive for MAP. Five FFPE samples (one CD, two UC and two nIBD) and three fresh samples (one in each group) were positive through IS900-PCR. qRT-PCR was performed on these eight samples. Among the FFPE samples, there were 192.12 copies/μl in the CD group, 72.28 copies/μl in UC and 81.43 copies/μl in nIBD. Among the fresh samples, there were 432.99 copies/μl, 167.92 copies/μl and 249.73 copies/μl in the CD, UC and nIBD groups, respectively. The highest bacterial load was in the CD group. CONCLUSION: This study does not provide evidence for a role of MAP in the etiology of CD, although MAP DNA was detected in all three patient groups. This is the first report of MAP presence in human intestinal biopsies in Brazil.
CONTEXTO E OBJETIVO: Mycobaterium avium subsp. paratuberculosis (MAP) tem atraído o interesse de pesquisadores devido às semelhanças entre a paratuberculose e a doença de Crohn (CD). Este estudo objetivou avaliar a frequência de MAP por meio de cultura, histologia e reação da polimerase em cadeia (PCR), em biópsias intestinais de pacientes brasileiros com CD. PCR quantitativa em tempo real (qRT-PCR) foi realizada nas amostras positivas. TIPO DE ESTUDO E LOCAL: Estudo transversal analítico com grupo controle realizado em duas universidades federais. MÉTODOS: Amostras frescas foram coletadas de 25 pacientes; cinco com CD, oito com colite ulcerativa (UC) e 12 controles sem doença inflamatória intestinal (nIBD). Também foram coletadas 149 amostras fixadas em parafina (FFPE): 44 CD, 49 UC e 56 nIBD. RESULTADOS: Nenhuma das amostras frescas foi positiva para MAP. Cinco amostras FFPE (uma CD, duas UC e duas nIBD) e três amostras frescas (uma de cada grupo) foram positivas por IS900-PCR. qRT-PCR foi realizada nessas oito amostras. Nas amostras FFPE, havia 192,12 cópias/μl no grupo CD, 72,28 cópias/μl no UC e 81,43 cópias/μl no nIBD. Nas amostras frescas, havia 432,99 cópias/μl, 167,92 cópias/μl e 249,73 cópias/μl nos grupos CD, UC e nIBD, respectivamente. A maior carga bacteriana foi encontrada no grupo CD. CONCLUSÃO: Este estudo não fornece evidências do papel de MAP na etiologia da CD, embora DNA de MAP tenha sido detectado em pacientes dos três grupos. Este é o primeiro relato da presença de MAP em biópsias intestinais humanas no Brasil.
Subject(s)
Humans , Male , Female , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Inflammatory Bowel Diseases/microbiology , Colitis, Ulcerative/microbiology , Crohn Disease/microbiology , Mycobacterium avium subsp. paratuberculosis/isolation & purification , Case-Control Studies , Polymerase Chain Reaction/methods , Cross-Sectional Studies , Mycobacterium avium subsp. paratuberculosis/pathogenicity , Sequence Analysis, DNA/methodsABSTRACT
Clostridium difficile (CD) infection is increasing in frequency and severity in in-hospital and outpatient clinical settings, with a recurrence that can reach 30% after first episode. The recurrences are usually treated with longer courses of metronidazole or vancomycin. Other treatments have been used, such as probiotics, fidaxomicin, rifaximin, immunoglobulins and monoclonal antibodies against toxins A and B. Fecal microbiota transplantation (FMT) has emerged as a promising strategy in this group of patients, with effectiveness greater than 90%. We present the first case reported in Chile of this therapeutic strategy in a patient with Crohn's disease and recurrent CD infection who presented after the fecal transplantation an Escherichia coli bacteremia, suggesting the need for caution in the use of this strategy. 10 months after the FMT the patient presented a new episode of E. coli bacteremia and two episodes of diarrhea due to CD infection, treated both of them with vancomycin with good clinical response.
La infección por Clostridium difficile (CD) está aumentando en frecuencia y gravedad tanto a nivel intrahospitalario como ambulatorio, con una recurrencia que puede alcanzar hasta 30% después de un primer episodio. Los cuadros recurrentes son generalmente tratados con cursos prolongados de metronidazol y/o vancomicina. Otras terapias han sido sugeridas como el uso de probióticos, fidaxomicina, rifaximina, inmunoglobulina y anticuerpos monoclonales para toxina A y B. El trasplante de microbiota fecal (TMF) ha emergido como una estrategia promisoria en este grupo de pacientes con una efectividad mayor a 90%. Presentamos el primer caso reportado en Chile de esta estrategia terapéutica en un paciente con enfermedad de Crohn y CD recurrente, quien presentó una bacteriemia por Escherichia coli post-TMF, sugiriendo la necesidad de tener precaución con el uso de esta estrategia. El paciente presentó a los 10 meses post-TMF un nuevo episodio de bacteriemia por E. coli y dos episodios de diarrea por CD siendo tratados ambos cuadros con vancomicina con buena respuesta clínica.
Subject(s)
Humans , Male , Middle Aged , Biological Therapy/adverse effects , Clostridioides difficile , Clostridium Infections/therapy , Escherichia coli Infections/etiology , Feces/microbiology , Microbiota , Bacteremia/microbiology , Biological Therapy/methods , Chile , Crohn Disease/microbiology , Recurrence , TransplantationABSTRACT
Background: Colorectal mucosal biopsies occasionally demonstrate the presence of bacteria adherent to the epithelium. This study evaluated the histological and ultrastructural correlates of such bacterial adherence. Materials and Methods: Rectal mucosal biopsies from eight patients in whom histopathological examination of biopsies had earlier demonstrated adherent bacteria were examined by electron microscopy and by bacterial culture. Colorectal biopsies of 69 patients with adherent bacteria detected histologically were retrospectively evaluated for histological changes at sites proximal and distant to adherent bacteria. Results: Escherichia coli of different serogroups were isolated from 7 of 8 rectal biopsies demonstrating bacterial adherence. All isolates showed diffuse or focal adherence to HEp-2 cell monolayers. Ultrastructural changes noted included microvillus damage, pedestal formation, actin web condensation, and protrusions of the apical cytoplasm of epithelial cells into the lumen towards the bacteria. Histological changes noted at light microscopy included reduction in epithelial cell height, focal epithelial cell degeneration, cryptitis and neutrophil infiltration at sites of bacterial adherence whereas these were usually absent at sites distant to adherent bacteria. Bacterial adherence was noted more often in biopsies from Crohn's disease patients than in patients without this diagnosis (P < 0.001). Conclusion: Adherent Escherichia coli in colorectal biopsies were associated with focal epithelial damage and showed an association with Crohn's disease.
Subject(s)
Colon/microbiology , Crohn Disease/microbiology , Biopsy/methods , Colon/microbiology , Escherichia coli/classification , Escherichia coli/isolation & purification , Escherichia coli/pathology , Escherichia coli/ultrastructure , Humans , Intestinal Mucosa/microbiology , PatientsABSTRACT
Paratuberculosis is a chronic intestinal disease of animals caused by Mycobacte-rium avium subsp. paratuberculosis (MAP), which has some pathological features similar to Crohn's disease (CD) in humans. The presence of MAP in food for human consumption and in affected tissues of patients with CD has been detected. Therefore, a causal association between this microorganism and the disease in humans, has been postulated. However, several related studies have failed to confirm this hypothesis and the scientific acceptance of MAP as a zoonotic agent remains controversial. This review presents the main findings related to this issue, contrasting evidences for and against an association between MAP and CD. The need to promote national studies focusing on this area is suggested.
Subject(s)
Animals , Humans , Crohn Disease/microbiology , Mycobacterium avium subsp. paratuberculosis/pathogenicity , Paratuberculosis/transmission , Chile/epidemiology , Food Contamination , ZoonosesABSTRACT
La enfermedad inflamatoria intestinal (EII) es una entidad compleja. Su desarrollo requiere de la interacción entre factores ambientales y la flora gastrointestinal, en un individuo genéticamente susceptible. Nuestro aparato gastrointestinal es un extraordinario, complejo y dinámico modelo de simbiosis o mutualismo con la flora. Los factores ambientales como el tabaco, las infecciones gastrointestinales y los antiinflamatorios no esteroidales juegan probablemente un rol iniciador y/o modificador de la enfermedad. La mucosa intestinal tiene la difícil tarea de limitar la respuesta inflamatoria contra la flora y de mantener la habilidad de generar una respuesta inmune contra los microorganismos patógenos. Esto crea una relación de equilibrio dinámico y frágil que al alterarse cualquiera de sus componentes puede generar un proceso inflamatorio. Existe evidencia que las infecciones pueden tener un rol tanto en el inicio de la enfermedad como en las reagudizaciones de ésta. Es así como las infecciones bacterianas gastrointestinales agudas y la Escherichia coli adherente invasora confieren un riesgo para desarrollar una enfermedad inflamatoria intestinal. Para Mycobacterium avium subespecie paratuberculosis (MAP) sólo se ha establecido una asociación y no un rol patogénico. Por último ha aumentado la incidencia y morbimortalidad de la infección por Clostridium difficile en los pacientes con EII.
Infections in the pathogeny of inflammatory bowel disease Inflammatory bowel disease (IBD) is a complex entity. Its development requires the interaction between environmental factors and gastrointestinal flora in a genetically susceptible subject. Our gastrointestinal tract is an extraordinary, complex and dynamic model of symbiosis or mutualism with the flora. Environmental factors such as tobacco, gastrointestinal infections and non-steroidal anti-inflammatory drugs might play a role of starter and/or modifier of the disease. The intestinal mucosa has the difficult task of limiting the inflammatory response against the flora and of keeping the capability of generating an immune response against pathogenic microorganisms. This creates a dynamic equilibrium relation that is fragile, and that when any of its components is altered, it can cause an inflammatory process. There is evidence that infections can have a role both in the beginning and in the episodes of New-Rebounds of the disease. Therefore, acute gastrointestinal bacterial infections and adherent-invasive Escherichia coli pose a risk of developing an inflammatory bowel disease. In the case of Mycobacterium avium subspecies paratuberculosis (MAP) an association has been established, but not a pathogenic role. The incidence and morbimortality of Clostridium difficile infection has increased in patients with IBD.
Subject(s)
Humans , Inflammatory Bowel Diseases/microbiology , Clostridium Infections/complications , Escherichia coli Infections/complications , Paratuberculosis/complications , Clostridioides difficile/pathogenicity , Colitis, Ulcerative/microbiology , Crohn Disease/microbiology , Escherichia coli/pathogenicity , Mycobacterium avium subsp. paratuberculosis/pathogenicityABSTRACT
El reciente aislamiento de Mycobacterium paratuberculosis de lesiones de pacientes con enfermedad de Crohn (EC) reavivó el interés en el posible origen micobacteriano de esta enfermedad. Sin embargo, las evidencias inmunológicas acumuladas en favor de esta hipótesis son poco sólidas. La respuestaespecífica en la EC podría estar enmascarada por el amplio cruzamiento antigénico que existe entre microbacterias. En este estudio se aplico un procedimiento propuesto para develar la presencia de anticuerpos anti-M. paratuberculosis en la enfermedad de Johne bovina: la absorción de los anticuerpos con reactividad cruzada mediante la preincubación de los sueros con M. phlei. Se cuantificó por enzimoinmunoensayo IgG anti-M. paratuberculosis en 90 muestras de suero de 17 pacientes con EC, 23 con colitis ulcerosa (CU) y 14 con otras patologías intestinales (OP). Muestras de 86 individuos sanos, con tuberculosis, microbacteriosis o micosis sirvieron como control. Se comparó los resultados con los obtenidos con antígenos de M. avium y M. tuberculosis. Se observó una discreta reactividad humoral a micobacterias en pacientes con EC, Cu y OP. Las alteraciones fueron de muy escasa magnitud y frecuencia en relación con las observadas en pacientes con tuberculosis, micobacteriosis y micosis. La absorción de los sueros no develó una presunta respuesta específica en la EC. Tampoco se detectaron elevaciones en los niveles de anticuerpos de pacientes analizados periodicamente. Estos datos no agregan evidencia sólida a la hipótesis del origen micobacteriano de la EC