[Functional biostructure of colonic microbiota (central fermenting area, germinal stock area and separating mucus layer) in healthy subjects and patients with diarrhea treated with Saccharomyces boulardii]. / Biostructure fonctionnelle du microbiote colique(zone de fermentation centrale, zone de réservegerminale et couche de mucus séparatrice) chez lessujets sains et chez les patients atteints de diarrhéetraités par Saccharomyces boulardii.

The colonic content can be compared to a spatially structured high output bioreactor composed of three functionally different regions: a separating mucus layer, a germinal stock area, and a central fermenting area. The stool mirrors this structure and can be used for diagnosis in health and disease. In a first part, we introduce a novel method based on fluorescence in situ hybridization (FISH) of sections of punched-out stool cylinders, which allows quantitatively monitor microbiota in the mucus, the germinal stock and the central fermenting areas. in a second part, we demonstrate the practical implementation of this method, describing the biostructure of stool microbiota in healthy subjects and patients with chronic idiopathic diarrhea treated with Saccharomyces boulardii. Punched stool cylinders from 20 patients with chronic idiopathic diarrhea and 20 healthy controls were investigated using fluorescence in situ hybridization. Seventy-three bacterial groups were evaluated. Fluctuations in assembly of 11 constitutive bacterial groups were monitored weekly for 3 weeks prior to, 3 weeks during, and 3 weeks after oral Saccharomyces boulardii supplementation. Typical findings in healthy subjects were a 5-60 µm mucus separating layer; homogeneous distribution and fluorescence, high concentrations (>10 × 10(10) bacterial/mL) of the three habitual bacterial groups: Bacteroides, Roseburia and Faecalibacterium prausnitzii; and low concentrations of the occasional bacterial groups. The diarrhea could be described in terms of increased separating effort, purging, decontamination, bacterial substitution. Typical findings in diarrhea were: increased thickness of the protective mucus layer, its incorporation in the stool, absolute reduction in concentrations of the habitual bacterial groups, suppression of bacterial metabolism in the central fermenting area (hybridization silence), stratification of the stool structure by watery ingredients, and substitutive increase in the concentrations of occasional bacterial groups. The microbial and clinical symptoms of diarrhea were reversible with Saccharomyces boulardii therapy. The structure-functional analysis of stool microbiota allows to quantitatively monitor colonic malfunction and its response to therapy. Saccharomyces boulardii significantly improves the stool biostructure in patients with chronic idiopathic diarrhea and has no influence on the stool microbiota in healthy subjects.

Mucosal flora in Crohn's disease and ulcerative colitis - an overview.

The intestinal flora harbors varies pathogens. Clostridium perfringens (gas gangrene), Enterococci (endocarditis), Enterobacteriaceae (sepsis), Bacteroides (abscesses) are present in the large intestine of every healthy person in high concentrations. These bacteria are, however, separated from the colonic wall by an impenetrable mucus layer and are tolerated by the host. This separation is disturbed in patients with inflammatory bowel disease (IBD), where bacteria adhere to the mucosa and invade epithelial cells with concomitant inflammatory response. This chronic bowel inflammation can not subside as long as the mucus barrier remains defective. The inflammatory response interferes with the state of tolerance to the intestinal bacteria and leads to characteristic changes in the biostructure of the faecal microbiota. These changes in the biostructure of faecal microbiota are specific for active Crohn's disease and ulcerative colitis (UC) and can be longitudinally monitored. The reason for the defect of the mucus barrier in IBD patients is unclear. Epidemiologic studies indicate a negative role of western lifestyle and foods and document the rise in the incidence of IBD in the industrialized countries during the 20(th) century. In parallel to this, detergents were introduced in households and emulsifiers were increasingly added to food. The cleaning effect of these on the colonic mucus has to be investigated. The present contribution summarizes new data on the biostructure of the intestinal microbiota.

[Biomorphology of the Bacterial Invasion in Chronic pharyngotonsillitis ]. / Biomorphologie der Infektionsherde in rezidivierenden Mandelentzündungen.

BACKGROUND: The study investigates whether relapses of chronicpharyngotonsillitis result from new infections caused by theoro-pharyngeal microbial flora or are reactivations of persistent bacterial infections of the tonsils. METHODS: 90 patients, who were surgically treated for chronicpharyngotonsillitis (age 13 months to 38 years, at least 5 episodes of disease and antibiotic treatment in the past) were included. The surgery was performed in the antibiotic- and symptom-free period (at least 6 weeks after the last exacerbation). Sections of tonsillar tissue were investigated for invasive bacteria using fluorescence in situ hybridization (FISH) with group and species-specific 15/23S RNA based probes. RESULTS: Abundant foci of invasive bacteria were found in 86% of the resected tonsils, despite previous treatment with antibiotica and absent symptoms of ongoing infection. The diffuse infiltration of the tonsils was most predominant in the youger children. Local invasive processes such as abscesses, fissures filled with pus and superficial infiltration of the tonsillar epithelium were more typical for adults. All of the foci were polymicrobial and contained up to 10 different species or groups of bacteria. The local concentrations of invasive bacteria were up to 1012 bacteria/ml. CONCLUSIONS: The chronic pharyngotonsillitis is the result of persistent invasive bacterial infections. The polymicrobial nature of the infectious foci enables them to resist the antibiotic treatment and to exacerbate afterwards. The surgical treatment is unavoidable as long as antibiotic treatment fails to clear the infection.

Spatial organisation of microbiota in quiescent adenoiditis and tonsillitis.

BACKGROUND: The reasons for recurrent adenotonsillitis are poorly understood. METHODS: The in situ composition of microbiota of nasal (5 children, 25 adults) and of hypertrophied adenoid and tonsillar tissue (50 children, 20 adults) was investigated using a broad range of fluorescent oligonucleotide probes targeted to bacterial rRNA. None of the patients had clinical signs of infection at the time of surgery. RESULTS: Multiple foci of ongoing purulent infections were found within hypertrophied adenoid and tonsillar tissue in 83% of patients, including islands and lawns of bacteria adherent to the epithelium, with concomitant marked inflammatory response, fissures filled with bacteria and pus, and diffuse infiltration of the tonsils by bacteria, microabscesses, and macrophages containing phagocytosed microorganisms. Haemophilusinfluenzae mainly diffusely infiltrated the tissue, Streptococcus and Bacteroides were typically found in fissures, and Fusobacteria,Pseudomonas and Burkholderia were exclusively located within adherent bacterial layers and infiltrates. The microbiota were always polymicrobial. CONCLUSIONS: Purulent processes persist during asymptomatic periods of adenotonsillitis. Most bacteria involved in this process are covered by a thick inflammatory infiltrate, are deeply invading, or are located within macrophages. The distribution of the bacteria within tonsils may be responsible for the failure of antibiotic treatment.

Bacterial biofilm within diseased pancreatic and biliary tracts.

BACKGROUND: Bacterial community structures in human pancreatic and biliary tracts were evaluated. METHODS: Gall bladder stones from 153 patients, 20 gall bladder walls, six common duct stones, 52 biliary stents, 21 duodenal biopsies, nine pancreatic duct biopsies, and five bile ducts were investigated using fluorescence in situ hybridisation (FISH) with ribosomal RNA targeted Cy3/Cy5 (carbocyanine) labelled oligonucleotide probes. RESULT: Duodenal, gall bladder, and bile duct walls were free of bacteria. A dense multispecies bacterial biofilm was present within the pancreatic duct of patients with calcific pancreatitis and within biliary stents, irrespective of diagnosis. The concentration, density, and amenability of the biofilm to FISH and DNA staining declined progressively with the grade of stent occlusion. The lowest detectable bacterial concentrations were found by FISH in completely occluded stents and brown/mixed gall stones. Bacteria were not detectable with FISH in cholesterol gall stones. CONCLUSIONS: A wide range of different branches and groups of bacteria participate in the development of biofilms on the surfaces of foreign bodies, such as biliary stents, mixed gall stones, or calcific pancreatic ducts, but not on the surface of pure cholesterol gall stones. Occlusion of stents leads to progressive extinction of the biofilm and mummification of its components. Deposition of cholesterol or other substances within the biofilm matrix may be a novel mechanism of host defence against bacteria present in these biofilms.

Clinical approach to fecal soiling in children.

Fecal soiling is common in childhood and can be caused by stool toileting refusal, fecal incontinence due to organic disease, or encopresis due to functional constipation. Anatomical, neurologic, and inflammatory causes for fecal soiling are ruled out by history and physical examination and, if necessary, by anorectal manometry, barium enema, and rectal biopsy. The initial treatment suggestion for children with stool toileting refusal is to put the child back into pull-ups or diapers. Most children with fecal soiling due to organic disease continue with some degree of incontinence despite optimal medical management. Antegrade enema administration helps those with severe fecal incontinence due to organic causes who do not respond to medical management. Successful treatment of constipation and encopresis requires a combination of medical therapy, nutritional intervention, behavioral intervention, and long-term compliance with laxative use. The combined treatment approach improves the constipation and encopresis in all patients who comply with the treatment program. In some children, cow's milk protein intolerance may be the cause. In them, cow's milk protein needs to be eliminated.

Bilious vomiting in the newborn: rapid diagnosis of intestinal obstruction.

Bilious vomiting in newborns is an urgent condition that requires the immediate involvement of a team of pediatric surgeons and neonatologists for perioperative management. However, initial detection, evaluation and treatment are often performed by nurses, family physicians and general pediatricians. Bilious vomiting, with or without abdominal distention, is an initial sign of intestinal obstruction in newborns. A naso- or orogastric tube should be placed immediately to decompress the stomach. Physical examination should be followed by plain abdominal films. Dilated bowel loops and air-fluid levels suggest surgical obstruction. Contrast radiography may be required. Duodenal atresia, midgut malrotation and volvulus, jejunoileal atresia, meconium ileus and necrotizing enterocolitis are the most common causes of neonatal intestinal obstruction.

Failure to pass meconium: diagnosing neonatal intestinal obstruction.

Timely passage of the first stool is a hallmark of the well-being of the newborn infant. Failure of a full-term newborn to pass meconium in the first 24 hours may signal intestinal obstruction. Lower intestinal obstruction may be associated with disorders such as Hirschsprung's disease, anorectal malformations, meconium plug syndrome, small left colon syndrome, hypoganglionosis, neuronal intestinal dysplasia and megacystis-microcolon-intestinal hypoperistalsis syndrome. Radiologic studies are usually required to make the diagnosis. In addition, specific tests such as pelvic magnetic resonance imaging, anorectal manometry and rectal biopsy are helpful in the evaluation of newborns with failure to pass meconium.

Toilet tales: stool toileting refusal, encopresis, and fecal incontinence.

Defecation difficulties in children present as stool toileting refusal, encopresis (caused by idiopathic or functional constipation), or fecal incontinence (caused by anatomic or neurologic disorders). The appropriate evaluation, treatment, and expected treatment results for children with stool soiling are presented.

Urinary incontinence and urinary tract infection and their resolution with treatment of chronic constipation of childhood.

OBJECTIVES: To evaluate the frequency of urinary incontinence and urinary tract infection in children with chronic constipation and report on the resolution of these with treatment of the underlying constipation. METHODS: We evaluated the frequency of urinary incontinence and urinary tract infection in 234 chronic constipated and encopretic children before, and at least 12 months after, the start of treatment for constipation. RESULTS: Twenty-nine percent complained of daytime urinary incontinence and 34% of nighttime urinary incontinence. Urinary tract infection was present in 11% and was more commonly present in girls than in boys (33% vs 3%). Vesicoureteral reflux was present in four and megacystis in four of the 25 children who had a voiding cystourethrogram because of urinary tract infection. One girl who came in had constipation and acute urinary retention. The treatment for constipation consisted of disimpaction and maintenance treatment, which included the prevention of reaccumulation of stools and reconditioning to normal bowel habits through timed toilet sitting. Follow-up, at least 12 months after start of treatment for constipation, revealed that the constipation was relieved successfully in 52%. Relief of constipation resulted in disappearance of daytime urinary incontinence in 89% and nighttime urinary incontinence in 63% of patients, and disappearance of recurrent urinary tract infections in all patients who had no anatomic abnormality of the urinary tract. CONCLUSION: Urinary symptoms were found in a significant number of children who had functional constipation and encopresis. With treatment of the constipation, most patients became clean and dry and further recurrence of urinary tract infections was prevented.

Fecal incontinence in children.

Functional constipation is the cause of fecal incontinence in 95 percent of affected children, and anatomic or neurologic causes account for up to 5 percent of cases. The history and the physical examination (with emphasis on abdominal, rectal and neurologic examinations) are most helpful in identifying organic disease. In some children, anorectal manometry, a barium enema radiographic examination and a rectal biopsy are necessary to determine the etiology. Most children with fecal incontinence benefit from a strict treatment plan that includes defecation trials, a fiber-rich diet and laxative medications. Surgery followed by medical treatment is required in patients with Hirschsprung's disease and in some patients with anal stenosis or a history of surgical repair of an anorectal malformation.

Biofeedback therapy for defecation disorders.

Biofeedback therapy is a useful adjunct to conventional treatment for many patients with refractory defecation disorders. This article provides an overview regarding the historical evolution of this treatment together with current perspectives regarding the principles and techniques of performing biofeedback therapy and an assessment of its outcome in adults and pediatric patients with defecation disorders.

Balloon defecation as a predictor of outcome in children with functional constipation and encopresis.

OBJECTIVE: To evaluate whether the ability to defecate a rectal balloon might predict 12-month recovery in children with functional constipation and encopresis. METHODS: We evaluated the ability to defecate within 5 minutes a 100 ml waterfilled rectal balloon by 20 healthy children and 139 children with functional constipation and encopresis. RESULTS: All healthy children and only 47% of the patients were able to defecate the balloon. Twelve months after the start of treatment, 51% of patients able to and 34% of patients unable to defecate the balloon had recovered (p < 0.03). Logistic regression revealed that the ability to defecate the balloon and a history of secondary encopresis were related to recovery (p < 0.04). Patients who were unable to defecate the balloon or who did not recover had significantly more impairment in anorectal functions than those who were able to defecate the balloon or who did recover. The ability of the balloon defecation test to predict recovery had a sensitivity of 57%, a specificity of 60%, a positive predictive value of 0.51, and a negative predictive value of 0.66. CONCLUSION: Children with functional constipation and encopresis who were able to defecate the rectal balloon were twice as likely to recover. Even though there was a clinically significant difference in the recovery rates between patients who could and those who could not defecate the balloon, calculation of predictive values showed that the balloon defecation test could not reliably predict recovery.

Encopresis and soiling.

Constipation, encopresis, and fecal incontinence are common problems in children. Constipation can have a variety of causes, such as organic and anatomic causes or intake of medication. Encopresis is the involuntary loss of formed, semiformed, or liquid stool into the child's underwear in the presence of functional (idiopathic) constipation in a child 4 years of age or younger. Fecal incontinence is fecal soiling in the presence of an organic or anatomic lesion, such as Hirschsprung's disease, anal malformation, anal surgery or trauma, meningomyelocele, and some muscle disease. This article reviews the symptoms of functional constipation in young children and the symptoms of functional constipation and encopresis in older children, presents the differential diagnosis of constipation with or without fecal incontinence, describes the evaluation and treatment of these children, and reports on treatment outcome.

Anteriorly located anus: is constipation caused by abnormal location of the anus?

Anteriorly located anus (ALA) is frequently associated with severe constipation accompanied by defecation pain. Between 1988 and 1994, the authors treated 27 children (26 girls, 1 boy; age range, 0 to 11 years) to surgically correct ALA. The operation was performed according to a uniform protocol to longitudinally divide the internal sphincter muscle from the anal skin level to 2 cm above the dentate line on the posterior wall of the anorectum. For anal reconstruction, any of the conventional procedures was employed. Twenty-two of the 27 patients have had follow-up in our clinic for 12 months to 6 years (mean, 2.75 years). Eighteen are completely free of constipation and defecation pain and have regular spontaneous bowel movements. The other four require occasional use of enemas or laxatives. Anal incontinence did not occur in any patient. The results of this study suggest that abnormal function of the internal sphincter is the most likely cause of constipation or defecation pain in patients with ALA and that internal sphincterotomy is the cornerstone of surgical treatment.