Ileostomy Effluent Research Articles

Nutritional management of a malnourished cancer patient with high output ileostomy

High output stoma is a complication that may follow ileostomy formation, with an incidence of 23%. There is no general consensus on the limit of ileostomy production to be defined as high output. However, output of more than 2000 mL/day, can cause fluid and electrolyte imbalance, also malnutrition due to reduced nutrient absorption. Delay in recognition and treatment, especially in cancer patient with high risk of malnutrition, can further deteriorate patient’s nutritional status. A 43-year-old malnourished female with ascending colon cancer underwent tumor resection and ileocolostomy surgery. Starting from the third postoperative day, ileostomy effluent drastically increased to 2700 mL/day, causing severe hyponatremia, hypokalemia, and hypomagnesemia. Risk factors of high output stoma identified were routine prokinetic medication use and unresolved malignancy-related retroperitoneal abscess. Moreover, increment of food intake in the first days after surgery, specifically food high in insoluble fiber, was one of the contributing factors. High output stoma was then resolved by abscess drainage, discontinuation of prokinetic agent, and administration of antimotility agent. Hyponatremia and hypomagnesemia improved with correction, whereas hypokalemia needed longer time to resolve. Enteral nutrition was maintained and increased gradually to prevent further malnutrition. Oral hypotonic fluid intake was limited to 1000 mL/day and isotonic solution consumption was advised. High stoma production due to hypersecretory phase after ileostomy was expected, but thorough management would prevent patient’s deterioration that was caused by the fluid, electrolyte, and nutritional imbalances.

Endogenous small intestinal microbiome determinants of transient colonisation efficiency by bacteria from fermented dairy products: a randomised controlled trial

BackgroundThe effects of fermented food consumption on the small intestine microbiome and its role on host homeostasis are largely uncharacterised as our knowledge on intestinal microbiota relies mainly on faecal samples analysis. We investigated changes in small intestinal microbial composition and functionality, short chain fatty acid (SCFA) profiles, and on gastro-intestinal (GI) permeability in ileostomy subjects upon the consumption of fermented milk products.ResultsWe report the results from a randomised, cross-over, explorative study where 16 ileostomy subjects underwent 3, 2-week intervention periods. In each period, they consumed either milk fermented by Lacticaseibacillus rhamnosus CNCM I-3690, or milk fermented by Streptococcus thermophilus CNCM I-1630 and Lactobacillus delbrueckii subsp. bulgaricus CNCM I-1519, or a chemically acidified milk (placebo) daily. We performed metataxonomic, metatranscriptomic analysis, and SCFA profiling of ileostomy effluents as well as a sugar permeability test to investigate the microbiome impact of these interventions and their potential effect on mucosal barrier function. Consumption of the intervention products impacted the overall small intestinal microbiome composition and functionality, mainly due to the introduction of the product-derived bacteria that reach in several samples 50% of the total microbial community. The interventions did not affect the SCFA levels in ileostoma effluent, or gastro-intestinal permeability and the effects on the endogenous microbial community were negligible. The impact on microbiome composition was highly personalised, and we identified the poorly characterised bacterial family, Peptostreptococcaceae, to be positively associated with a low abundance of the ingested bacteria. Activity profiling of the microbiota revealed that carbon- versus amino acid-derived energy metabolism of the endogenous microbiome could be responsible for the individual-specific intervention effects on the small intestine microbiome composition and function, reflected also on urine microbial metabolites generated through proteolytic fermentation.ConclusionsThe ingested bacteria are the main drivers of the intervention effect on the small intestinal microbiota composition. Their transient abundance level is highly personalised and influenced by the energy metabolism of the ecosystem that is reflected by its microbial composition (http://www.clinicaltrials.gov, ID NCT NCT02920294).3z8GaJs8-4AmNdyX8busiEVideo

High Output Stoma Complicating Nutritional Therapy in Severely Malnourished Patient With Right Ascending Colon Cancer: A Case Report

ObjectivesAscending colon cancer makes up of 27% colorectal cancers, and its incidence has continuously increased. Malnutrition is common in cancer patients, posing as a risk of poor surgical outcome. High output stoma (HOS) is a commonly encountered complication with an incidence of 23%. Not only causes fluid and electrolyte imbalance, it also deteriorates patient's nutritional status, forming a vicious cycle of malnutrition.MethodsA 43-year-old malnourished female with ascending colon cancer presented to the emergency department with signs of bowel obstruction. The patient underwent tumor resection and ileocolostomy surgery. Starting from the third postoperative day, ileostomy effluent drastically increased to 2700 mL/day. Simultaneously, severe hyponatremia, severe hypokalemia, and hypomagnesemia were observed. Intravenous electrolyte correction and antimotility drug were given. It was decided that oral nutrition intake remained to be given. Oral hypotonic fluid intake was limited to 1000 mL/day. Malignancy-related retroperitoneal abscess further complicated patient's condition.ResultsHypersecretory phase was expected in first days after ileostomy surgery. However, HOS, especially with output of more than 2000 mL/day, could cause fluid and electrolyte imbalance. Moreover, HOS could cause malnutrition due to reduced nutrient absorption. Risk factors of HOS identified in this case were prokinetic medication use and unresolved retroperitoneal abscess causing intraabdominal inflammation. Increment of food intake was also observed from prior anorexic period, specifically fruits high in insoluble fiber were consumed. Fruit pulp was found in the stoma effluent, raising concern of reduced bowel absorption capacity. High output stoma was resolved by abscess drainage, discontinuation of prokinetic agent, and administration of antimotility agent. Hyponatremia and hypomagnesemia improved with correction, whereas hypokalemia needed longer time to be resolved. Oral nutrition was maintained and increased gradually to prevent further malnutrition, while stoma production was monitored strictly.ConclusionsThis case report showed that strict evaluation on stoma production and management of HOS in malnourished cancer patients must be done promptly, preventing further deterioration.Funding SourcesNone.

Effects of ileal glucose infusion on enteropancreatic hormone secretion in humans: relationship to glucose absorption

BackgroundsThe distal small intestine plays an important role in regulating the secretion of entero-pancreatic hormones that are critical to the control of glucose metabolism and appetite, but the quantitative contribution of a specific segment to these effects is unknown. PurposesTo determine the effects of 30 cm of the ileum exposed to glucose on the secretion of ghrelin, glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) insulin, C-peptide and glucagon, in relation to glucose absorption in non-diabetic subjects. Basic procedures10 non-diabetic subjects with a loop ileostomy after early-stage rectal cancer resection were studied on 2 days in a double-blind, randomized and crossover fashion, when a catheter was inserted retrogradely 30 cm from the ileostomy for infusion of a glucose solution containing 30 g glucose and 3 g 3-O-methylglucose (as a marker of active glucose absorption), or 0.9% saline, over 60 min. Ghrelin, GIP, GLP-1, insulin, C-peptide, glucagon and ileal glucose absorption (from concentrations of 3-O-methylglucose in serum and glucose in ileostomy effluent) were measured over 180 min. Main findings12.0 ± 1.2 g glucose was absorbed over 180 min. Compared to saline, ileal glucose resulted in minimal increases in blood glucose and plasma insulin and C-peptide, but substantial increases in plasma GLP-1, without affecting ghrelin, GIP or glucagon. The magnitude of the GLP-1 response to glucose was strongly related to the increase in serum 3-O-methylglucose. Principal conclusionsStimulation of the terminal ileum by glucose, even over a short length (30 cm), induces substantial GLP-1 release, coupled primarily to active glucose absorption.Clinical registration:NCT05030376 (ClinicalTrials.gov).

Distinct effects of TU-100 (daikenchuto) on long-lasting dysbiosis in the small intestine in patients with colorectal cancer and inflammatory bowel disease.

The profile of the human small intestinal microbiota remains to be uncovered primarily due to sampling difficulties. Ileostomy provides the intestinal luminal contents as ileostomy effluents (IE) that offer opportunity for performing extensive analyses of nutrients, gastrointestinal fluids, metabolites, and microbiome. In the present study, we evaluated changes in the microbiome, pH, and bacterial short-chain fatty acids (SCFAs) in IE obtained from patients who had undergone ileostomy following surgical resection of colon cancer and inflammatory bowel disease (IBD). We enrolled 11 patients who varied in the duration of ileostomy from 3 days to >5 years after surgery and had no inflammation in the small intestine. The analyses suggested that IE from patients previously having IBD had less diversity and greater intraday and interday fluctuations, and increased pH and decreased levels of propionic acid and acetic acid than those in IE from patients previously having cancer. Furthermore, correlation analysis suggested a possible effect of the intestinal microbiome on luminal pH, presumably via SCFA production. The present study suggested that inflammation in the colon may induce long-term dysbiosis in the small intestine even after removal of diseased parts of the colon. Moreover, pharmaceutical-grade Japanese traditional medicine daikenchuto (TU-100) was found to have beneficial effects on postoperative bowel dysfunction and the human small intestinal microbiota. Taken together, these results suggest the necessity of a direct remedy for dysbiosis and the treatment of gastrointestinal lesions to achieve favorable outcomes for chronic gastrointestinal disorders.

Faecal Calprotectin from Ileostomy Output Is Sensitive and Specific for the Prediction of Small Bowel Inflammation in Patients with Crohn's Disease.

Severe Crohn's disease [CD] can result in extensive bowel resections and need for creation of an ileostomy. Faecal calprotectin [FC] is well studied in CD management, though its role in patients who have an ileostomy is unclear. Our aim is to understand if FC is a useful adjunct to radiographic or endoscopic studies in identifying recurrent CD after surgery in patients with an ileostomy. Between January 1, 2017, and September 30, 2020, we searched the Mayo Clinic electronic medical record retrospectively for adult patients with ICD-10 code for CD, and a surgical history of an ileostomy. Patients were included in the analysis if they had at least one FC measured and a concomitant radiographic imaging and/or endoscopic procedure. An abnormal FC was defined as greater than 60 µg/g. Of 51 patients who met our inclusion criteria, 17 had an FC level >60 µg/g. Of these 17 patients, 14 had imaging and/or an ileoscopy confirming the presence of small bowel inflammation, with a sensitivity of 87.5%. Of the remaining 34 patients with an FC level ≤60 µg/g, 32 patients had imaging and/or ileoscopy demonstrating no small bowel inflammation, with a specificity of 91.4%. FC from an ileostomy effluent had a positive predictive value of 82.3%, a negative predictive value of 94.1% and test diagnostic accuracy of 90.1%. FC from an ileostomy effluent is a highly sensitive and specific test for the assessment and monitoring of small bowel inflammation and disease recurrence in patients with CD.

Perioperative alterations in the intestinal microbiota and functional changes mediate innate immune activation after small bowel transplantation

AimSmall bowel transplantation (SBT) is the only therapy for end-stage short bowel syndrome. However, complicated pathological changes and an increased risk of postoperative infections in the perioperative period are major obstacles to patient survival, but the associated mechanisms remain unclear. MethodsTo explore perioperative alterations in the intestinal microbiota and their functional changes after SBT, 16S rRNA sequencing of ileostomy effluents and plasma analysis were performed pre-SBT and periodically post-SBT. ResultsThe results suggested that the presence of Proteobacteria accelerated bacterial motility and chemotaxis during the first week in post-SBT recipients. Altered gut microbiota impaired intestinal barrier integrity and upregulated 16S rDNA, pathogen-associated molecular pattern (PAMP) and pattern-recognition molecule (PRM) levels in peripheral circulation. Importantly, the levels of neutrophils, monocytes, cytotoxic T lymphocytes, and natural killer cells and the expression of proinflammatory cytokines were increased in the peripheral blood and had potential roles in activating innate immune-mediated inflammatory injury after SBT. ConclusionTogether, our results suggest that altered microbiota and functional changes are probably related to innate immune-mediated inflammatory injury and graft survival after SBT, suggesting that the monitoring and regulation of intestinal microbiota are necessary for SBT patients.

Growth failure and metabolic acidosis due to total body sodium depletion in an infant with an ileostomy.

Sodium is an essential nutrient and inadequate sodium intake and/or excessive sodium losses can result in suboptimal growth. Infants with ileostomies are at significant risk of developing growth failure as a result of excessive sodium loss in their ileostomy effluent. Chronic sodium depletion can also limit the kidney's ability to excrete hydrogen and potassium ions, mimicking electrolyte abnormalities found in type 4 renal tubular acidosis. This report describes an infant with an ileostomy with severe growth failure, hyperkalaemia and metabolic acidosis-all of which promptly resolved with sodium supplementation.

Amylase quantification in the terminal Ileum following formation of an Ileostomy

Amylase is elevated in the foregut and has been used to confirm anastomotic integrity after pancreatic surgery. The physiological activity of pancreatic enzymes in the ileum has been studied in healthy volunteers but not quantitated with the simple and readily available amylase measurements employed with serum tests. We aim to quantitate the levels of amylase in the terminal ileum. This was a prospective, non-randomised, non-blinded, consecutive cohort study conducted at the Royal Brisbane and Women’s Hospital. Consecutive patients undergoing routine surgery with an ileostomy were invited to participate in the study. Ileostomy effluent was collected and analysed daily for the first 5 post-operative days. This validation cohort included 8 males and 3 females, with a mean age of 49 years. Median daily amylase levels ranged from 4470 U/L to 23,000 U/L, with no specimens falling within the laboratory serum reference range of 40 to 130 U/L. Two specimens were not available on day one post-operative due to complete ileus. The sample size of 11 patients is small but was considered sufficient given that 55 effluent specimens were anticipated for analysis. Amylase levels remain highly elevated as the enzyme transits through the length of the small intestine and measured in the terminal ileum, and can be readily quantitated by the existing testing methodology routinely available.

Fermentation Kinetics of Selected Dietary Fibers by Human Small Intestinal Microbiota Depend on the Type of Fiber and Subject.

ScopeAn underexplored topic is the investigation of health effects of dietary fibers via modulation of human small intestine (SI) microbiota. A few previous studies hint at fermentation of some dietary fibers in the distal SI of humans and pigs. Here the potential of human SI microbiota to degrade dietary fibers and produce metabolites in vitro is investigated.Methods and ResultsFructans, galacto‐oligosaccharides, lemon pectins, and isomalto/malto‐polysaccharides are subjected to in vitro batch fermentations inoculated with ileostomy effluent from five subjects. Fiber degradation products, formation of bacterial metabolites, and microbiota composition are determined over time. Galacto‐ and fructo‐oligosaccharides are rapidly utilized by the SI microbiota of all subjects. At 5h of fermentation, 31%–82% of galacto‐oligosaccharides and 29%–89% fructo‐oligosaccharides (degree of polymerization DP4‐8) are utilized. Breakdown of fructo‐oligosaccharides/inulin DP ≥ 10, lemon pectin, and iso‐malto/maltopolysaccharides only started after 7h incubation. Degradation of different fibers result in production of mainly acetate, and changed microbiota composition over time.ConclusionHuman SI microbiota have hydrolytic potential for prebiotic galacto‐ and fructo‐oligosaccharides. In contrast, the higher molecular weight fibers inulin, lemon pectin, and iso‐malto/maltopolysaccharides show slow fermentation rate. Fiber degradation kinetics and microbiota responses are subject dependent, therefore personalized nutritional fiber based strategies are required.

Microbial communities in a dynamic in vitro model for the human ileum resemble the human ileal microbiota.

The important role for the human small intestinal microbiota in health and disease has been widely acknowledged. However, the difficulties encountered in accessing the small intestine in a non-invasive way in healthy subjects have limited the possibilities to study its microbiota. In this study, a dynamic in vitro model that simulates the human ileum was developed, including its microbiota. Ileostomy effluent and fecal inocula were employed to cultivate microbial communities within the in vitro model. Microbial stability was repetitively achieved after 10 days of model operation with bacterial concentrations reaching on average 107 to 108 16S rRNA copy numbers/ml. High diversities similar to those observed in in vivo ileum samples were achieved at steady state using both fecal and ileostomy effluent inocula. Functional stability based on Short Chain Fatty Acid concentrations was reached after 10 days of operation using fecal inocula, but was not reached with ileostomy effluent as inoculum. Principal Components and cluster analysis of the phylogenetic profiles revealed that in vitro samples at steady state clustered closest to two samples obtained from the terminal ileum of healthy individuals, independent of the inoculum used, demonstrating that the in vitro microbiota at steady state resembles that of the human ileum.

Johanson-Blizzard Syndrome: Report of One Case

Genetic causes of exocrine pancreatic insufficiency (EPI) are rare and commonly associated with Cystic fibrosis (CF) or Shwachman Diamond syndrome. Herein we describe a child affected with Johansson Blizard syndrome (JBS), another rare cause of EPI. JBS is an autosomal recessive condition due to mutations in the Ubiquin 3 ligase gene (UBR1). Findings of JBS include craniofacial malformations, EPI, failure to thrive (FTT), hearing loss, developmental delay and anal, genitourinary and other malformations. Case: Our patient was born at 38 weeks via repeat C-section to a 24 year old G2 P1 mother. Pregnancy was complicated by polyhydramnios, hydronephrosis and dilated rectum detected at the 20 week prenatal ultrasound. Growth parameters at birth were normal. She was found to have abdominal distention, imperforate anus, perianal fistula, micro colon, inspissated meconium, persistent cloacae and mild facial dysmorphism. She underwent surgery and had placement of end ileostomy. Testing for CF was negative but steatorrhea was present in the ileostomy effluent. She was started on pancreatic enzymes (Creon 6000 units per feeding), vitamin ADEK and Alimentum formula. At 6 months she had cloacae repair and 2 months later closure of ileostomy. There had been no weight gain till that point and height was at the 3rd percentile. At 19 months, both weight and height were below the 3rdpercentile. She was lost for follow up for one year and was seen again at 31 months of age. Mother reported that the child had been fitted with a cochlear implant and that Creon had been discontinued. Patient continued to have FTT and delayed developmental milestones. She had hypoalbuminemia, anemia, steatorrhea and low fecal pancreatic elastase of < 50 ug/g. Physical exam was significant for plagiocephaly, flat face, elevated palate, up slanting of palpebral fissures and hypoplastic alae nasai. Findings were considered to be consistent with JBS. A baseline genetic evaluation revealed a normal female karyotype and a normal SNP microarray. Preauthorization for UBR1 gene sequencing was obtained and sample sent. A hypercaloric diet and pancreatic enzymes were prescribed as well as iron and vitamin supplementations. At 33 months she had some weight gain but at 36 months her height and weight remained below the 3rd percentile. Hypoalbuminemia has been corrected and hemoglobin levels have improved. Creon supplements were readjusted. Genetic testing for JBS is pending.

Eating marshmallows reduces ileostomy output: a randomized crossover trial.

Anecdotally, many ostomates believe that eating marshmallows can reduce ileostomy effluent. There is a plausible mechanism for this, as the gelatine contained in marshmallows may thicken small bowel fluid, but there is currently no evidence that this is effective. This was a randomized crossover trial. Adult patients with well-established ileostomies were included. Ileostomy output was measured for 1 week during which three marshmallows were consumed three times daily, and for one control week where marshmallows were not eaten. There was a 2-day washout period. Patients were randomly allocated to whether the control or intervention week occurred first. In addition, a questionnaire was administered regarding patient's subjective experience of their ileostomy function. Thirty-one participants were recruited; 28 completed the study. There was a median reduction in ileostomy output volume of 75 ml per day during the study period (P = 0.0054, 95% confidence interval 23.4-678.3) compared with the control week. Twenty of 28 subjects (71%) experienced a reduction in their ileostomy output, two had no change and six reported an increase. During the study period, participants reported fewer ileostomy bag changes (median five per day vs six in the control period, P = 0.0255). Twenty of 28 (71%) reported that the ileostomy effluent was thicker during the study week (P = 0.023). Overall 19 (68%) participants stated they would use marshmallows in the future if they wanted to reduce or thicken their ileostomy output. Eating marshmallows leads to a small but statistically significant reduction in ileostomy output.

TRIGGERED GASTROINTESTINAL RELEASE OF ANTHOCYANINS FROM BILBERRIES (VACCINIUM MYRTILLUS L.)

Bilberries (Vaccinium myrtillus L.) are reported to show preventive activities against inflammatory bowel diseases (IBD), colon cancer or degenerative disease such as Alzheimer’s dementia. These findings are attributed to the antioxidative and anti-inflammatory activities of one of the main ingredients, the anthocyanins. However, anthocyanins are sensitive to environmental conditions, thus their bioavailability in the gastrointestinal tract is an important determinant of their in vivo activity. In the study reported here, the potential benefits of encapsulating an anthocyanin rich bilberry extract (BE) on anthocyanin stability were investigated. Non-encapsulated BE and three different BE loaded microcapsule systems were incubated anaerobically in intestinal fluid, namely ileostomy effluents from volunteers. After exposure to these media released anthocyanins were identified and quantified by HPLC with UV/VIS detection. A constant degradation of anthocyanins from BE was observed to 29.7% of the applied amount within 24 h. In contrast to this, encapsulation of anthocyanins lead to a constant liberation for at least 6 h of about 14.5 to 20.5% of the initial concentration of anthocyanins from the matrix. These results illustrate the ability of encapsulation to provide a constant (triggered) release of anthocyanins into the intestinal system, but in contrast to the non-encapsulated BE lower low amounts being liberated.

Prevalence of Clostridium Difficile in Excluded Colons

Clostridium difficile infection is associated with substantial morbidity and mortality, increased duration of hospitalization, and a marked economic impact. Several case reports and case series have described C. difficile infection in excluded bowels or immediately after reversal of defunctioning ileostomy. The aim of this prospective study is to detect whether the excluded colon is associated with a higher rate of C. difficile colonization than the normal population, which may increase the risk of C. difficile infection. Patients with defunctioning loop ileostomy, undergoing closure of ileostomy to restore bowel continuity, were prospectively recruited. Two stool samples were collected from the ileostomy effluent before closure of ileostomy and two after the procedure including the first bowel movement. All samples were cultured for C. difficile and analyzed for toxins A and B by a Premier EIA test. Demographic data and possible confounding factors were observed and recorded. Twenty-fine adult patients were recruited to this study; five patients were subsequently excluded. Two patients had positive stool cultures for C. difficile in the postoperative samples and another patient developed clinical pseudomembranous colitis with positive toxin. This indicates a possible colonization rate of 3 to 38 per cent (95% confidence interval). Four observed cases out of the 20 subjects taking part in this study would confidently conclude that C. difficile colonization in the excluded colon is 6 to 44 per cent, i.e., higher than the incidence in the healthy adult population, which is 3 per cent. However, the findings of this study prompt larger and well-powered studies to confirm these findings.

The human small intestinal microbiota is driven by rapid uptake and conversion of simple carbohydrates

The human gastrointestinal tract (GI tract) harbors a complex community of microbes. The microbiota composition varies between different locations in the GI tract, but most studies focus on the fecal microbiota, and that inhabiting the colonic mucosa. Consequently, little is known about the microbiota at other parts of the GI tract, which is especially true for the small intestine because of its limited accessibility. Here we deduce an ecological model of the microbiota composition and function in the small intestine, using complementing culture-independent approaches. Phylogenetic microarray analyses demonstrated that microbiota compositions that are typically found in effluent samples from ileostomists (subjects without a colon) can also be encountered in the small intestine of healthy individuals. Phylogenetic mapping of small intestinal metagenome of three different ileostomy effluent samples from a single individual indicated that Streptococcus sp., Escherichia coli, Clostridium sp. and high G+C organisms are most abundant in the small intestine. The compositions of these populations fluctuated in time and correlated to the short-chain fatty acids profiles that were determined in parallel. Comparative functional analysis with fecal metagenomes identified functions that are overrepresented in the small intestine, including simple carbohydrate transport phosphotransferase systems (PTS), central metabolism and biotin production. Moreover, metatranscriptome analysis supported high level in-situ expression of PTS and carbohydrate metabolic genes, especially those belonging to Streptococcus sp. Overall, our findings suggest that rapid uptake and fermentation of available carbohydrates contribute to maintaining the microbiota in the human small intestine.

Microarray Analysis and Barcoded Pyrosequencing Provide Consistent Microbial Profiles Depending on the Source of Human Intestinal Samples

Large-scale and in-depth characterization of the intestinal microbiota necessitates application of high-throughput 16S rRNA gene-based technologies, such as barcoded pyrosequencing and phylogenetic microarray analysis. In this study, the two techniques were compared and contrasted for analysis of the bacterial composition in three fecal and three small intestinal samples from human individuals. As PCR remains a crucial step in sample preparation for both techniques, different forward primers were used for amplification to assess their impact on microbial profiling results. An average of 7,944 pyrosequences, spanning the V1 and V2 region of 16S rRNA genes, was obtained per sample. Although primer choice in barcoded pyrosequencing did not affect species richness and diversity estimates, detection of Actinobacteria strongly depended on the selected primer. Microbial profiles obtained by pyrosequencing and phylogenetic microarray analysis (HITChip) correlated strongly for fecal and ileal lumen samples but were less concordant for ileostomy effluent. Quantitative PCR was employed to investigate the deviations in profiling between pyrosequencing and HITChip analysis. Since cloning and sequencing of random 16S rRNA genes from ileostomy effluent confirmed the presence of novel intestinal phylotypes detected by pyrosequencing, especially those belonging to the Veillonella group, the divergence between pyrosequencing and the HITChip is likely due to the relatively low number of available 16S rRNA gene sequences of small intestinal origin in the DNA databases that were used for HITChip probe design. Overall, this study demonstrated that equivalent biological conclusions are obtained by high-throughput profiling of microbial communities, independent of technology or primer choice.

Effect of Carrot (Daucus carota) Microstructure on Carotene Bioaccessibility in the Upper Gastrointestinal Tract. 2. In Vivo Digestions

Nutrient bioaccessibility and subsequent absorption will be directly influenced by changes in food structure during gastrointestinal processing. The accompanying paper (Tydeman et al. J. Agric. Food Chem. 2010, 58, doi: 10.1021/jf101034a) reported results on the effect of carrot processing on the release of carotene into lipid phases during in vitro gastric and small intestinal digestions. This paper describes results from in vivo digestion of two of the types of processed carrot used previously, raw grated carrot and cooked carrot mashed to a puree. Ileostomy effluents from human volunteers fed meals containing the carrot material were used to study tissue microstructure and carotene release. Raw carrot shreds and intact cells that had survived the pureeing process were identifiable in ileal effluent. The gross tissue structure in the shreds had not changed following digestion. Carotene-containing particles remained encapsulated in intact cells, but were absent from ruptured cells. Microscopy revealed marked changes to the cell walls including swelling and pectin solubilization, which increased in severity with increasing residence time in the upper gut. These observations were entirely consistent with the in vitro observations. It was concluded that a single intact cell wall is sufficient to reduce carotene bioaccessibility from a cell by acting as a physical barrier, which is not broken down during upper gut digestion.

Model experiments mimicking the human intestinal transit and metabolism of D‐galacturonic acid and amidated pectin

In order to study the human intestinal transit and metabolism of D-galacturonic acid and amidated pectin a number of model experiments were carried out. Both substrates were incubated under aerobic conditions at 37 degrees C using saliva (2 min) and simulated gastric juice (4 h). Under anaerobic conditions the substrates were incubated at 37 degrees C using human ileostomy and colostomy fluids, each obtained from three different donors, for 10 and for 24 h, respectively. D-Galacturonic acid, SCFA (acetic acid, propionic acid, and butyric acid), as well as methanol were analyzed photometrically after carbazole reaction, GC-flame ionization detection (GC-FID), and headspace solid-phase microextraction GC/MS (HS-SPME-GC/MS), respectively. D-Galacturonic acid and amidated pectin were found to be stable during incubations with saliva and simulated gastric juice, whereas both substrates underwent degradation in the course of human ileostomy and colostomy fluid incubations. D-Galacturonic acid was practically completely decomposed within 10 h and SCFA, with acetic acid as the major representative, were formed up to 98% of the incubated substrate in colostomy effluent. The amidated pectin was only degraded in part, revealing stable amounts of 22-35% and 3-17% in ileostomy (after 10 h) and colostomy fluid (after 24 h), respectively. SCFA were generated up to 59% of the applied amidated pectin. In parallel, 19-60% and 52-67% of the available methyl ester groups were cleaved in the course of incubations with ileostomy and colostomy fluids, respectively. The results demonstrate for the first time that D-galacturonic acid and amidated pectin are stable in human saliva and simulated gastric juice. The degradation of both compounds during incubation with ileostomy effluent is highlighted, providing evidence for a considerable metabolic potential of the small intestine.

Fecal Calprotectin Level Measurements in Small Bowel Allograft Monitoring: A Pilot Study

Protocol endoscopy with biopsy is currently the gold standard of small bowel transplantation (SBTx) monitoring, however it is invasive, costly, needs skilled operator, may require anesthesia and may cause complications. We investigated fecal calprotectin level (FCL) as a candidate noninvasive marker for monitoring patients after SBTx. A pilot study was performed to test the use of FCL measurement in following up SBTx patients. Ileostomy effluents were collected at various postoperative days before endoscopy and biopsy. FCLs were measured by enzyme-linked immunosorbent assay and a cut-off level of 100 ng/mg was considered positive. The results were retrospectively evaluated in combination with clinical, endoscopic, and histopathological findings. FCLs are presented as median nanogram per milligram. FCLs were measured in 122 samples that were obtained from 29 patients after SBTx. Only 1 of 69 positive FCL did not accompany abnormal findings. Retrospective evaluation showed that 11 samples from six patients (FCL: 217) coincided with rejection episodes, six samples from three patients (FCL: 125) coincided with viral enteritis, 51 samples from 21 patients (FCL: 207) coincided with nonspecific inflammation, 11 samples from two patients (FCL: 998) coincided with chronic intestinal ulceration, and finally 50 samples from 19 patients (FCL: 43) coincided with normal findings. No significant FCL difference was found between rejection, infection, and inflammation. FCL evolution in individuals showed that FCL can predict rejection days before histopathological diagnosis. FCL is a sensitive test for ongoing organic intestinal allograft pathologies. It might be useful as prescreening marker to avoid unnecessary endoscopies.