Corrigendum to “Bulgarian yogurt relieved symptoms and distress and increased fecal short-chain fatty acids in healthy constipated women: A randomized, blinded crossover controlled trial” [NFS Journal, Volume 22, March 2021, Pages 20–31

Bulgarian yogurt relieved symptoms and distress and increased fecal short-chain fatty acids in healthy constipated women: A randomized, blinded crossover controlled trial

Background: Short chain fatty acids (SCFAs) are the microbially-derived end products of dietary fiber fermentation. The SCFA butyrate is a signaling molecule shown to influence blood pressure (BP) in mouse models. The association of butyrate and other SCFAs on BP in humans is unclear, due in part to predominantly cross-sectional analyses and to differences in which biospecimens (blood vs. fecal) SCFAs were measured. Longitudinal studies including both circulating and fecal SCFAs are lacking. Objective: Investigate the cross-sectional and longitudinal associations of fecal and serum SCFAs with hypertension and changes in BP. Methods: We leveraged existing data from the SPIRIT trial, which randomized 121 adult cancer survivors with overweight/obesity to a behavioral weight loss intervention, metformin, or self-directed weight loss. Of participants with serum and fecal SCFAs measured at baseline (N=111), a subset also had serum (N=93) and fecal (N=89) SCFA measurements 12 months later. We used Poisson regression with robust error variance to estimate associations of serum or fecal SCFAs with hypertension at baseline (adjusting for age, sex, BMI, and fiber intake). In longitudinal analyses, we assessed the percent change in serum or fecal SCFAs from baseline with corresponding 12-month changes in BP (covariates listed in Table ). Results: Participants were aged 60.2±8.9 years, 78.4% female, and 46.9% identified as White. Higher baseline fecal butyrate was inversely associated with prevalent hypertension (standardized PR [95%CI]: 0.71 [0.54, 0.92]). In longitudinal analyses ( Table ), a 10% increase in fecal butyrate from baseline levels was significantly and inversely associated with systolic BP, and a 10% increase in serum butyrate was significantly and inversely associated with systolic and diastolic BPs. Associations of butyrate with BP were not modified by sex or race. Conclusion: Increased serum or fecal butyrate is associated with lowered BP. Butyrate may be a target for SCFA-centered BP-lowering interventions.

Defecation requires relaxation of the internal and external anal sphincters. High anal resting pressure is associated with painful constipation, defecatory disorders, and increased healthcare utilization in constipated patients; the mechanisms are unclear. Perhaps patients with a high anal resting pressure have a less distensible canal, which impedes defecation. In 50 of 64 participants (33 healthy and 17 constipated women), anal pressures and distensibility were measured, respectively, with manometry and balloon distention combined with magnetic resonance imaging; rectal balloon expulsion time (BET) was also studied. The BET (P=.006) was longer, and the mean (SD) rectoanal pressure gradient (-58[40] vs -34[26] mmHg, P=.03) was more negative in constipated than healthy women; anal resting pressure was not different. During anal distention, the balloon expanded rapidly at an opening pressure of 49 (18) mmHg, which was lower (P<.0001) than resting pressure (90 [25] mmHg). The resting pressure was correlated with the opening pressure (r=0.57, P<.0001) and inversely (r=-0.38, P=.007) with maximum volume but not with anal distensibility (volume-pressure slope). In healthy women, the difference (opening-resting pressure) was correlated with anal relaxation during evacuation (r=0.35, P=.04). Anal distensibility and sensory thresholds were not different between constipated and healthy women. Among healthy and constipated women, a greater anal resting pressure is correlated with greater opening pressure and lower maximum volume during distention, and, hence, provides a surrogate marker of anal distensibility. The difference (opening-resting pressure), which reflects anal relaxation during distention, is correlated with anal relaxation during evacuation. Anal resting pressure and distensibility were comparable in healthy and constipated women.

Short chain fatty acids (SCFA) have been associated with improved glycemia and to reductions in inflammatory responses in cell and rodent models. Low grade inflammation has been associated with insulin resistance. We measured fecal and plasma SCFA and inflammatory biomarkers in response to a proprietary oligosaccharide intervention. 22 overweight and obese men and women participated in a three‐way crossover study that included 3 levels of oligosaccharide: 0, 15 and 30 g/d. for 3 weeks. During each treatment, a collection of 5 days' feces were combined and analyzed for SCFA. At the end of each treatment, a meal challenge protocol was employed in which participants consumed half their daily dose of oligosaccharide with breakfast followed by a standard lunch. We measured plasma acetate, propionate and butyrate in 12 blood draws over the course of a 9 hour test day and inflammatory cytokines at 3 time points, 0, 195 and 435 min following the breakfast test meal. All SCFA analyses were conducted by GC‐MS. Inflammatory cytokines were measured on an 8‐plex chemiluminescent MSD plate except for adiponectin and CRP which employed an Integra 400 bio‐analyzer. Breath hydrogen and methane were collected to characterize participants as either methane producers (MP) or non‐producers (NM). Adiponectin was positively associated with fecal SCFA (p&lt;0.05). TNF‐α and IL‐10 were negatively associated with various plasma SCFA AUC segments (p&lt;0.05). Neither CRP nor IL‐6 was associated with either fecal or plasma SCFA. Fecal SCFA concentrations did not correlate well with plasma SCFA AUCs. Fecal propionate and butyrate were lower in MP than NM. Breath methane was negatively associated with SCFA production in MP. Plasma and fecal SCFA may predict some inflammatory responses but differences in the time of collections and the span of collections may reflect different temporal relationships to cytokines.Support or Funding InformationFunding provided by USDA CRIS 2032‐51530‐022‐OOD and the West Coast Metabolomics Center (U24 DK097154)

Introduction: Fecal lactic acid (LA) and short chain fatty acids (SCFA) are produced from fermented unabsorbed carbohydrates by colonic bacterial flora. Recent studies show LA and SCFAs have anti-inflammatory and immunological effects on the gastrointestinal tract. It is known that the differences of intestinal microflora are reflected of the profile of fecal SCFAs. On the other hand, a previous study reported the colonic luminal concentration of butyric acid which is one of the SCFA might be a factor in the etiology of necrotizing enterocolitis (NEC). However, very few studies have been carried out about the beneficial effect of probiotics on fecal SCFA profile especially in premature infants. The aim of this study is to examine the effect of enteral administration of Bifidobacterium breve (B. breve) on fecal LA and SCFA profile in low birth weight infants (LBWI). Methods: Forty eight infants were enrolled in the study, who were divided into three groups depending on their birth weight; 11 term infants (Group I; mean birth weight, 3062g; mean gestational age, 39.5 weeks), 21 LBWI (Group II; mean birth weight, 1812g; mean gestational age, 33.7 weeks) and 16 very low birth weight infants (VLBWI) (Group III; mean birth weight, 1277g; mean gestational age, 31.1 weeks). Fecal samples were collected at three periods; 1–7, 8–14 and 21–35 days of age. LA and SCFA (aceteic acid; AA, propionic acid and butyric acid; BA) concentration were analyzed using high performance liquid chromatography. Additionally both of Group II and Group III were divided into two groups; B. breve administration group (B+; n= 11 in Group II, 11 in Group III) and control group (B− n=10 in Group II, 5 in Group III), respectively. A dose of 1.6×108 cells were administrated to B. breve group twice per day from 1 day of age before feeding. Results: Fecal AA concentrations on 21–35 days were significantly higher than those on 1–7 days in Group I and II. On 21–35 days of age, (1) fecal LA and AA concentrations of Group I were significantly higher than those of Group III (p<0.01, p<0.05, respectively); (2) the fecal AA ratio to total fecal SCFA in B+ infants was significantly higher than that in B− infants in Group II and Group III (p<0.05); (3) the concentration and ratio of fecal BA in B+ infants were significantly lower than those in B− infants in Group III (p<0.01). Conclusion: Our results indicated that enteral administration of B. breve augmented the fecal level of LA and AA in LBWI. Furthermore it reduced the level of BA in VLBWI but not in LBWI. These differences might be associated with the colonization by bifidobacteria in the gastrointestinal tract. Our study suggests that early administration of B. breve to VLBWI prevents them from the development of NEC.

Construction of an ileal reservoir changes the fecal bacterial flora and the fecal composition of bile acids and short-chain fatty acids. We examined the relationships between pouch inflammation (pouchitis) and pouch content, as assessed by analysis of fecal bacteria, bile acids, and short chain fatty acids. Four groups were studied: ileal pouch-anal anastomosis (IPAA) for ulcerative colitis with pouchitis (N = 10), IPAA without pouchitis (N =5), IPAA for familial adenomatous polyposis without pouchitis (N = 5); and Brooke ileostomy for ulcerative colitis, which served as controls (N = 5). Pouchitis was defined as > or = 7 points on an 18-point pouchitis disease activity index. Aerobic and anaerobic bacteria were quantitatively cultured. Total aqueous-phase bile acids were measured by thin-layer chromatography and an enzymatic 3 alpha-OH hydroxysteroid dehydrogenase method. Fecal short chain fatty acids were measured by gas liquid chromatography. All patients with an IPAA had higher ratios of anaerobes/aerobes and concentrations of anaerobic gram-negative rods than did patients with an ileostomy. There were no other differences between patient groups with respect to bacteria, aqueous-phase total bile acids, or fecal short-chain fatty acids. Fecal concentrations of bacteria, bile acids, and short-chain fatty acids were similar in patients with and without pouchitis, indicating that these factors can not be the sole cause of pouchitis.

BackgroundSome probiotics have hypocholesterolemic effects in animal studies, which are mediated, in part, by increases in fecal short chain fatty acids (SCFAs). Clinical trials of probiotics on lipids/lipoproteins are inconsistent.ObjectiveWe examined the effects of Bifidobacterium animalis subsp. lactis BB-12® (BB-12®) (3.16 × 109 CFUs/day) on lipids and lipoproteins and fecal excretion of SCFAs in healthy adults.MethodsIn a randomized, partially blinded, 4-period, crossover study, 30 adults (11 men, 19 women) aged 18–40 years were randomly assigned to: 1) yogurt smoothie with no BB-12® (YS), 2) yogurt smoothie with BB-12® added pre-fermentation (PRE), 3) yogurt smoothie with BB-12® added post-fermentation (POST), 4) BB-12® containing capsule (CAP). We measured serum lipids/lipoproteins, glucose, insulin, C-reactive protein (CRP), and fecal SCFAs at baseline and after each treatment period.ResultsTotal cholesterol (TC), LDL cholesterol (LDL-C), HDL cholesterol (HDL-C), and triglycerides (TGs) did not differ after the PRE, POST, and CAP periods versus the YS or between treatments. Compared to baseline, fecal acetate was significantly increased after the YS (Δ = 211.89 ± 75.87 μg/g, P = 0.007) and PRE (Δ = 204.98 ± 75.70 μg/g, P = 0.009) periods. The percent increase in fecal acetate was significantly greater after the YS versus the POST period (52.2 ± 13.2% vs. 24.5 ± 13.2%, P = 0.023). Fecal total SCFAs, propionate and butyrate did not differ between treatment periods. Fecal total SCFAs were negatively associated with TC (r = -0.22, P = 0.01), LDL-C (r = -0.24, P = 0.004), age (r = -0.33, P < 0.001), and waist circumference (r = -0.25, P = 0.003).ConclusionsBB-12® supplementation did not improve lipids, lipoproteins and total and individual fecal SCFAs. Fecal SCFAs were negatively associated with TC, LDL-C, age, and waist circumference.Trial registrationThis trial was registered at clinicaltrials.gov as NCT01399996.

Age, Dietary Fiber, Breath Methane, and Fecal Short Chain Fatty Acids Are Interrelated in Archaea-Positive Humans1–3

Background: Short-chain fatty acids (SCFAs), mostly found in colon feces, is an important group of gut microbial metabolites from anaerobic fermentation of indigestible carbohydrates. Objectives: To develope an HPLC method to determined SCFAs in rat feces treated with resistant starch. Materials and methods: Sample is the rat feces fed with acetate wheat starch and normal starch; fatty acid hydrazides are derived from SCFAs in feces and measured by HPLC. After validated as guidance of US-FDA, method is applied to identify SCFAs in rat treated two types of starch. Results: the HPLC condition was optimized as follow: Eclipse XDB–C8 (4.6mmx150mm, 5µm) column; mobile phase: methanol, acetonitrile and 0,057 mM acid triflouroacetic (pH 4.5) (0:13:87 – 10:20:70 – 0:13:87, v/v/v) in 40 mins;, examinized wavelength: 396 nm. Method was validated with parameters: system suitability, specificity, linearity, precision, accuracy and stability. The result showed amount ratio of SCFAs in feces of mice group treated with acetate wheat starch containing resistant starch higher than from the diet containing normal starch significantly. Conclusion: This method can be used to investigate SCFAs in the gastrointestinal segments of the living organism. Key words: SCFAs, HPLC, resistant starch, feces

Short-chain fatty acids (SCFAs) produced by gut microbiota are reduced in feces but paradoxically increased in plasma of patients with Parkinson's disease (PD), which may stem from intestinal wall leakage. Gut function should be taken into consideration when conducting microbial-metabolite research. The objective was to investigate synchronous changes of SCFAs in feces and plasma of patients with PD, taking constipation as a confounder to better disentangle the SCFA metabolism exclusively associated with PD. The concentrations of fecal and plasma SCFAs in 33 healthy control subjects and 95 patients with PD were measured using liquid and gas chromatography mass spectrometry, respectively. Patients with PD were divided into patients with PD without constipation (n=35) and patients with PD with constipation (n=60). Gut-blood barrier (GBB) permeability was assessed by plasma/fecal ratio of SCFA concentrations and fecal α1-antitrypsin concentration. Patients with PD displayed decreased concentrations of fecal acetic, propionic, and butyric acid and increased concentrations of plasma acetic and propionic acid. Fecal acetic, isobutyric, and isovaleric acid were lower and plasma acetic and propionic acid were higher in patients with PD with constipation than in patients with PD without constipation. Constipation aggravated GBB permeability in patients with PD. Combined fecal and plasma SCFAs could discriminate patients with PD from healthy control subjects. Fecal SCFAs, except propionic acid, were negatively correlated with disease severity, while plasma acetic, propionic, and valeric acid showed a positive correlation. Our study showed alterations of fecal and plasma SCFAs in patients with PD that were associated with an impaired GBB and might be aggravated by constipation. © 2022 International Parkinson and Movement Disorder Society.

Prebiotics positively affect gut microbiota composition, thus improving gut function. These properties may be useful for the treatment of constipation. This study assessed the tolerance and effectiveness of a prebiotic inulin/partially hydrolyzed guar gum mixture (I-PHGG) for the treatment of constipation in females, as well as its influence on the composition of intestinal microbiota and production of short chain fatty acids. Our study enrolled 60 constipated female health worker volunteers. Participants reported less than 3 bowel movements per week. Volunteers were randomized to treatment with prebiotic or placebo. Treatment consisted of 3 weeks supplementation with 15 g/d IPHGG (fiber group) or maltodextrin (placebo group). Abdominal discomfort, flatulence, stool consistency, and bowel movements were evaluated by a recorded daily questionnaire and a weekly interview. Changes in fecal bacterial population and short chain fatty acids were assessed by real-time PCR and gas chromatography, respectively. There was an increased frequency of weekly bowel movements and patient satisfaction in both the fiber and placebo groups with no significant differences. Total Clostridium sp significantly decreased in the fiber group (p = 0.046) and increased in the placebo group (p = 0.047). There were no changes in fecal short chain fatty acid profile. Consumption of I-PHGG produced clinical results comparable to placebo in constipated females, but had additional protective effects on gut microbiota by decreasing the amount of pathological bacteria of the Clostridium genera.

To determine the effect of olestra on microbial ecology of the gut, faecal short-chain fatty acids (SCFAs) and water content were assessed in 93 subjects in a 36 d parallel, placebo-controlled, double-blind study. Faecal SCFAs and water content were determined at the end of an 8 d low fibre baseline period and 28 d treatment period. The test meal consisted of either a moderate (7 g) or high (24 g) level of fibre and olestra (24 g) or an equivalent amount of conventional fat. Olestra had a significant effect on both faecal water content and SCFAs. The water content decreased and concentrations of some SCFAs increased. The effect of olestra on SCFA concentrations in faeces (mmol/kg) differed with different fibre content of the meal. When the SCFAs were expressed as mmol/1 faecal water, a similar pattern of changes (several SCFAs increased) was observed for both olestra groups. Although all individual SCFA values were within the range of a healthy population, the effects were consistent, and potential reasons for it are discussed. Keywords: olestra, SCFA, faeces, faecal water, microbial ecology.

Short-chain fatty acids (SCFA) in faeces were examined in 18 patients with the irritable bowel syndrome (IBS) during treatment with wheat bran or placebo. In the placebo period, the patients could be classified in accordance with the faecal concentrations of SCFA into one group with low concentrations (mean, 40 mmol/l; range, 19-77 mmol/l; 10 patients) and another with high concentrations (mean, 168 mmol/l; range, 145-187 mmol/l; 8 patients). The concentrations of SCFA differed (P less than 0.001) in both groups from concentrations found in faeces from a reference group of nine normal individuals (mean, 114 mmol/l; range, 93-155 mmol/l). Patients with low levels of SCFA had lower (P less than 0.001) mean stool mass and longer (P less than 0.05) transit times than those with high concentrations of SCFA in faeces. Ingestion of bran, although a precursor of SCFA, did not change faecal concentrations of SCFA. Abdominal pain, distension, and rumbling were not correlated to low or high concentrations of SCFA in faeces, nor did bran improve these symptoms when compared to placebo. The level of SCFA was rather constant intraindividually and independent of the variability of the daily faecal mass. It is concluded that patients with IBS apparently have continuously abnormal concentrations of SCFA in faeces, either high or low, which are unaffected by the treatment with bran and which hypothetically may be of pathophysiologic importance.

Short chain fatty acids (SCFAs) are generated by the degradation and fermentation of complex carbohydrates, (i.e., dietary fiber) by the gut microbiota relevant for microbe–host communication. Here, we present a method for the quantification of SCFAs in fecal samples by liquid chromatography tandem mass spectrometry (LC-MS/MS) upon derivatization to 3-nitrophenylhydrazones (3NPH). The method includes acetate, propionate, butyrate, and isobutyrate with a run time of 4 min. The reproducible (coefficients of variation (CV) below 10%) quantification of SCFAs in human fecal samples was achieved by the application of stable isotope labelled internal standards. The specificity was demonstrated by the introduction of a quantifier and qualifier ions. The method was applied to investigate the pre-analytic stability of SCFAs in human feces. Concentrations of SCFA may change substantially within hours; the degree and kinetics of these changes revealed huge differences between the donors. The fecal SCFA level could be preserved by the addition of organic solvents like isopropanol. An analysis of the colon content of mice either treated with antibiotics or fed with a diet containing a non-degradable and -fermentable fiber source showed decreased SCFA concentrations. In summary, this fast and reproducible method for the quantification of SCFA in fecal samples provides a valuable tool for both basic research and large-scale studies.

BackgroundNon-caloric artificial sweeteners (NCAS) are widely used as a substitute for dietary sugars to control body weight or glycemia. Paradoxically, some interventional studies in humans and rodents have shown unfavorable changes in glucose homeostasis in response to NCAS consumption. The causative mechanisms are largely unknown, but adverse changes in gut microbiota have been proposed to mediate these effects. These findings have raised concerns about NCAS safety and called into question their broad use, but further physiological and dietary considerations must be first addressed before these results are generalized. We also reasoned that, since NCAS are bona fide ligands for sweet taste receptors (STRs) expressed in the intestine, some metabolic effects associated with NCAS use could be attributed to a common mechanism involving the host.ResultsWe conducted a double-blind, placebo-controlled, parallel arm study exploring the effects of pure saccharin compound on gut microbiota and glucose tolerance in healthy men and women. Participants were randomized to placebo, saccharin, lactisole (STR inhibitor), or saccharin with lactisole administered in capsules twice daily to achieve the maximum acceptable daily intake for 2 weeks. In parallel, we performed a 10-week study administering pure saccharin at a high dose in the drinking water of chow-fed mice with genetic ablation of STRs (T1R2-KO) and wild-type (WT) littermate controls. In humans and mice, none of the interventions affected glucose or hormonal responses to an oral glucose tolerance test (OGTT) or glucose absorption in mice. Similarly, pure saccharin supplementation did not alter microbial diversity or composition at any taxonomic level in humans and mice alike. No treatment effects were also noted in readouts of microbial activity such as fecal metabolites or short-chain fatty acids (SCFA). However, compared to WT, T1R2-KO mice were protected from age-dependent increases in fecal SCFA and the development of glucose intolerance.ConclusionsShort-term saccharin consumption at maximum acceptable levels is not sufficient to alter gut microbiota or induce glucose intolerance in apparently healthy humans and mice.Trial registrationTrial registration number NCT03032640, registered on January 26, 2017.3ThJTRkvKM2w8o53nrbAgaVideo abstract