Insight into the mechanism of short chain fatty acid absorption in the ostrich (Struthio camelus) proximal colon.

1. Occurrence of short chain fatty acids (SCFA) in anionic form limits their diffusion across the absorptive membrane. The present study sought to establish the mechanism of SCFA absorption in the ostrich. 2. Epithelial tissues were taken from the sacculated part of the colon and mounted in Ussing chambers in a bathing solution. The tissues were voltage-clamped and allowed to equilibrate to obtain a baseline short circuit current (Isc). 3. Propionate (23 mM) on the mucosal side increased the Isc. The SCFA-induced Isc was completely inhibited by anoxia, ouabain (1 to 2 mM), acetazolamide (0.5 mM) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (0.1 mM) on the mucosal side. 4. These findings indicate that SCFA stimulate hydrogen ion secretion through an electrogenic H(+)-K(+)-ATPase, the source of hydrogen ions being carbonic anhydrase catalysed hydration of CO2. 5. Simultaneous activation of Cl(-)/HCO3(-) exchange prevents intracellular accumulation of bicarbonate ions. This system may provide hydrogen ions for protonation of SCFA anions and subsequent absorption by non-ionic diffusion.

Water absorption in relation to fermentation in the colon of the ostrich (Struthio camelus).

The colon is a major site for fermentation and water absorption in the ostrich. Water absorption along the colon was evaluated and its relationship to osmolality, Na+ concentration, short chain fatty acid (SCFA) concentration and carbohydrate content of digesta analysed. Mean water content decreased from 5.30 +/- 0.99 to 2.51 +/- 0.13 mf/g dry mass in the first 5 m of the colon. Correspondingly, mean carbohydrate content fell from 529.85 +/- 46.61 to 434.99 +/- 29.89 mg/g dry mass. A significant correlation was shown between the decreases in mean carbohydrate and water content along the colon (r2 = 0.997, P < 0.05). Changes in mean osmolality (+/- 10 mOsm/kg) and SCFA concentration (+/- 7 mmol/l) were minimal in comparison to the change in Na+ concentration (-54 mmol/l). These findings reflect a close coupling between SCFA production and absorption on the one hand and water absorption on the other.

Human rectal absorption of short- and medium-chain C2-C10 fatty acids.

BACKGROUND: Current knowledge on the colonorectal absorption of medium-chain fatty acids is limited. The purpose of the present study was to evaluate and compare the human rectal absorption of short- and medium-chain C2-C10 fatty acids in healthy volunteers. METHODS: Dialysis bags containing 20 mmol x l(-1) of the fatty acids acetate, butyrate, hexanoate, octanoate, or decanoate in a phosphate-buffered (pH neutral) isoosmotic electrolyte solution were placed in the rectum for 30 min in 14 healthy volunteers. Absorption rates were calculated for all fatty acids, sodium, potassium, and water. RESULTS: Absorption rates of the fatty acids acetate, butyrate, hexanoate, octanoate or decanoate were the same (1.9 +/- 0.1 = 2.5 +/- 0.2 = 1.7 +/- 0.2 = 1.9 +/- 0.2 = 2.2 +/- 0.1 micromol x cm(-2) x h(-1) (mean +/- standard error of the mean), respectively; P = 0.24). CONCLUSIONS: Medium-chain fatty acids were absorbed in the human rectum at a rate similar to that for short-chain fatty acids. If results can be applied to the human colon, colonic absorption of medium-chain fatty acids could possibly become an important secondary site of absorption in abnormal intestinal conditions such as massive small-intestinal resection or malabsorption syndromes.

Birth and prematurity influence intestinal function in the newborn pig.

Developmental changes in intestinal function occur in the perinatal period of many species. We investigated the hypothesis that gestational age at delivery and the mode of delivery influence intestinal function. Newborn pigs (106-108 or 113-115 days gestation, term = 115 +/- 2 days) were either delivered by caesarean section or born vaginally following induction of parturition with a prostaglandin F2 alpha analogue. The pigs were killed at birth and used for measurements of intestinal ion transport in vitro (using Ussing chambers) or killed at 2 days of age, after being fed porcine colostrum to follow the absorption of intact proteins into plasma. The results indicate that premature birth is associated with increased paracellular permeability to ions. The uptake and net absorption of chloride were higher in the term, vaginally-delivered pigs than in the remaining pigs. Among the newborn pigs, the preterm caesarean-delivered pigs exhibited the lowest chloride secretion in response to the secretagogue, theophylline. The latter pigs also absorbed the lowest amounts of immunoglobulin G and albumin from colostrum. In conclusion, gestational age at delivery and the mode of delivery have significant effects on intestinal transport of ions and intact proteins. However, the observed variation in the magnitude and the direction of responses indicate that (a) prematurity and birth influence the transport of ions and intact proteins through independent regulatory mechanisms and (b) the absorption pathways for ions and intact proteins in the neonatal pig intestine are not closely associated.

Experimental studies of intestinal ion and water transport.

A major advance in transport physiology was H. H. Ussing's development of the voltage-clamp method, and later the Koefoed-Johnsen-Ussing model for Na+ transport. In the same decade, J. C. Skou identified the Na(+)-K(+)-ATPase, which maintains the Na+ and K+ gradients that drive most epithelial transport processes. With this foundation, Danish scientists have pursued the mechanism of ion transport and the resulting solute-linked water flow. Recent contributions have been on isosmotic transport, suggesting solute recycling, and KCl-water cotransport in the basolateral epithelial cell membrane. Efficient small intestinal nutrient absorption is dependent on coupling to the Na+ gradient. Cotransport of Na+ and glucose is quantitatively the most important absorptive mechanism in the small intestine, as illustrated by the success of oral rehydration solutions in diarrhoea. The majority of amino acids are likewise transported by Na+ dependent carriers, but recent experiments have identified a concomitant Cl- dependency for some. Regulation of intestinal secretion, both under normal digestive processes, and in response to enterotoxins, has turned out to be very complex. It involves local and central neuronal regulation through an array of neurotransmitters and local actions of gastrointestinal hormones. Major effectors are the submucosal neurons and the main transmitters serotonin, vasoactive intestinal peptide, acetylcholine, substance P, and neurotensin. Development of antisecretagogues is impeded by the existence of several receptor subtypes and significant species differences. The Na+ and water-conserving properties of the large intestine have been shown to be regulated by adrenocortical hormones, with aldosterone as a potent stimulator of colonic Na+ absorption. A major colonic function is the symbiosis with the anaerobic bacterial population. The fermentation of carbohydrate to short-chain fatty acids, which can be absorbed, supplements small intestinal digestive function.

Stimulation of butyrate absorption in the human rectum in vivo.

BACKGROUND: Models of short-chain fatty acid absorption have focused on the stimulation of sodium absorption, an effect mainly located in the proximal colon of man. With the present efforts to utilize butyrate enemas as a treatment of ulcerative colitis, it seemed important to assess the transport in the rectum. METHODS: Non-equilibrium dialysis of the rectum was applied by placing dialysis bags containing various electrolyte solutions in the rectum of volunteers for 30 min. In this period changes in ion concentrations were linear with time. Net absorption and secretion rates were calculated from the change in fluid composition. RESULTS: Sodium absorption was highest (24 +/- 8 mumol/cm2 h) in the presence of chloride and lowest (16 +/- 2 mumol/cm2 h) in the presence of bicarbonate and butyrate. Butyrate (70 mmol/l) was absorbed at a high rate of 7.1 +/- 2.2 mumol/cm2 h, independent on the presence of chloride, and was accompanied by increased bicarbonate secretion. Butyrate absorption increased to 9.6 +/- 1.8 mumol/cm2 h in sodium-free high-potassium media containing bicarbonate. CONCLUSION: The results show that it is possible to increase butyrate uptake by manipulation of the electrolyte composition in the rectal lumen. Maximal uptake occurred with an electrolyte composition that was similar to the natural rectal content. The information gathered could be useful in designing enemas for trial in ulcerative colitis, provided the findings can be confirmed in these patients.

Butyrate absorption and lactate secretion in ulcerative colitis.

PURPOSE: Fecal electrolytes and organic anion concentrations are altered in ulcerative colitis, presumably reflecting changes in colon epithelial transport. Information of mucosal absorption of butyrate in active ulcerative proctosigmoiditis is not available. METHODS: Dialysis bags containing 70 mmol/liter of butyrate in an isotonic electrolyte solution were placed in the rectum for 30 minutes. Net absorption or secretion rates of butyrate, lactate, and electrolytes were determined in the rectum of 12 patients with active ulcerative colitis (UC) and in 10 patients with quiescent UC and then compared with 10 healthy controls. RESULTS: Net flux rates demonstrated a considerable absorption of butyrate in patients with active inflammation of 7.5 +/- 0.4 mumol/cm2/h and quiescent colitis of 6.6 +/- 0.4 mumol/cm2/h, equal to absorption in healthy controls of 6.3 +/- 0.5 mumol/cm2/h, P = 0.12. Despite normal butyrate absorption, sodium absorption was compromised in active ulcerative colitis (11.5 +/- 1.4 mumol/cm2/h) compared with quiescent (15.4 +/- 1.0 mumol/cm2/h) and controls (18.7 +/- 0.8 mumol/cm2/h) (P = 0.0006). Mucosal secretion of L-lactate was minimal in both healthy controls and quiescent UC but significantly increased in patients with proctosigmoiditis (0.2 +/- 0.1 mumol/cm2/h, 0.2 +/- 0.1 mumol/cm2/h vs. 0.9 +/- 0.2 mumol/cm2/h; P = 0.0001). Appearance of D-lactate was negligible in all three groups. CONCLUSIONS: This study demonstrates that rectal butyrate absorption is normal in UC, and it follows that butyrate supplied in enemas can be expected to be absorbed. The inflamed colonic mucosa secretes L-lactate, and the increased fecal lactate concentrations can be explained by mucosal origin of lactate.

Mucosal acidification and an acid microclimate in the hen colon in vitro.

Experiments were performed on isolated, stripped colonic epithelia of low-salt-adapted hens (Gallus domesticus) in order to characterize acid secretion by this tissue. With symmetric, weak buffer solutions, colonic epithelia acidified both mucosal and serosal sides. Titration measurements of the mucosal acidification rate (pH-stat technique) averaged 1.63 +/- 0.25 microEq.cm-2.h-1. Mucosal acidification was also evident in colons from high-salt-adapted birds and in low-salt-adapted coprodeum, but was completely abolished in the high-salt coprodeum. Mucosal acidification by low-salt-adapted colonic epithelium was unaffected by sodium replacement, mucosal amiloride (10(-3) mol.l-1), and serosal ouabain (5 x 10(-4) mol.l-1), although all three treatments significantly reduced or reversed the short-circuit current. Acetazolamide (10(-3) mol.l-1, serosal) reduced mucosal acidification by 15% and simultaneously increased short-circuit current by a similar amount. Colonic epithelia incubated in glucose-free solutions had significantly lower acidification rates (0.59 +/- 0.13 microEq.cm-2.h-1, P < 0.002 versus controls) and addition of glucose (15 mmol.l-1), but not galactose, partially restored acidification to control levels. Anoxia (N2 gassing) completely inhibited short-circuit current, but reduced acidification by only 30%. A surface microclimate pH, nearly 2 pH units more acidic than the bath pH of 7.1-7.4, was measured in low-salt-adapted colon and coprodeum. The acid microclimate of both tissues was partially attenuated by adaptation to a high-salt diet. Colonic microclimate pH was dependent on the presence of glucose and sensitive to the bath pH.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of propionate on the acid microclimate of hen (Gallus domesticus) colonic mucosa.

1. Short-chain fatty acid absorption in hen colon is protonated across the apical border coupled to an apical electrogenic proton pump. 2. The surface pH of the isolated colonic epithelium was 6.27 +/- 0.05, when incubated in Krebs-phosphate buffer pH 7.0. 3. Propionate 7 and 40 mmol/l in the incubation medium (pH 7.0) increased microclimate pH to 6.47 +/- 0.04 and 6.56 +/- 0.04. Inhibition of metabolic activity by potassium cyanide 1 mmol/l increased surface pH to 6.66 +/- 0.06. 4. The calculated concentration of propionic acid in the microclimate is near-linearly related to the propionate concentration. Thus, the acid microclimate is not responsible for the Michaelis-Menten like kinetics of propionate transport.

Colonic fermentation of ispaghula, wheat bran, glucose, and albumin to short-chain fatty acids and ammonia evaluated in vitro in 50 subjects.

The production of short-chain fatty acids and ammonia was measured in 16.6% fecal homogenates from 50 subjects incubated at 37 degrees C for 6 and 24 hours. All 50 homogenates produced ammonia and short-chain fatty acids of any chain length (C2-C5). Incubation for 24 hours with dietary fiber (ispaghula husk or wheat bran), albumin, or glucose (10 mg/mL) increased the short-chain fatty acid production (43.6 +/- 2.8, 45.4 +/- 2.0, 60.3 +/- 3.2, and 65.8 +/- 3.1 mmol/L, respectively) compared with controls (21.4 +/- 1.3 mmol/L). The degradation of different substrates was associated with the production of different amounts of ammonia and short-chain fatty acids. Ispaghula, wheat bran, albumin, and glucose were fermented to acetate (> 2 mmol/L; 24-hour incubations) in 86%, 96%, 98%, and 98% of the homogenates, to propionate in 80%, 76%, 100%, and 68%, and to butyrate in 32%, 94%, 88%, and 54% of the homogenates, respectively. Isobutyrate, valerate, and isovalerate were produced from albumin in all (100%) of the homogenates, but only in 2 to 4% of the homogenates incubated with ispaghula or glucose. Ammonia was always (100%) produced after the addition of albumin and always (98%) consumed (assimilated) when glucose was fermented. Surgery (sigmoid or right- or left-sided colonic resection) did not change the pattern of ammonia and short-chain fatty acid production from these substrates. This study suggests that the different colonic flora from a large number of subjects share general biochemical characteristics, which metabolize different substrates to specific patterns of ammonia and short-chain fatty acids.

An in vitro study of short-chain fatty acid concentrations, production and absorption in pig (Sus scrofa) colon.

1. Short-chain fatty acid concentration was 180 mmol/l in the proximal colon and decreased to 108 mmol/l in the rectum. 2. Fermentation in chymus from different regions of the colon, showed the pattern of end products to reflect the substrate and not the site of the colon. 3. Isolated mucosa from proximal and distal colon had electroneutral sodium absorption of 4.8 +/- 0.2 and 2.9 +/- 0.8 mueq/cm2 hr in bicarbonate free media, which was abolished in the absence of chloride. 4. Electroneutral sodium absorption was enhanced by short-chain fatty acids in the proximal colon and could be described by Michaelis-Menten kinetics with Km 2.0-11 mmol/l and Jm 1.6-3.6 mueq/cm2 hr. In the distal colon the stimulation was smaller and propionate even inhibited sodium absorption. 5. Butyrate was absorbed in the proximal colon, whereas acetate and propionate, and butyrate in the distal colon had a flux ratio of one. 6. Amiloride (5 mmol/l) inhibited sodium absorption and net butyrate absorption.

The colon in carbohydrate malabsorption: short-chain fatty acids, pH, and osmotic diarrhoea.

Short-chain (C2-C6) fatty acids (SCFA) are the major anions in colonic contents and the result of anaerobic fermentation of mainly saccharides. The effects and regulation of saccharide fermentation were studied in vitro and in vivo. In vitro faecal incubation was used to study the effects of lactose, glucose, and galactose and of pH on SCFA formation. Changing the pH to below 5 or above 11 abolished SCFA formation in the faecal incubates; in the pH 5-9 interval SCFA production was high, with only minor pH dependence. Adding glucose, galactose, or lactose to the incubation system increased SCFA production, but at high saccharide concentrations (100-300 mmol/l) SCFA formation was inhibited by the pH change. In vivo disaccharide malabsorption with increasing doses of lactulose caused a decrease in faecal pH to less than 5, values inhibitory to fermentation, before the appearance of carbohydrate in faeces. In 6 of 12 volunteers diarrhoea occurred suddenly and was caused by malabsorbed non-fermented carbohydrate. The six other volunteers had a gradual increase in faecal output with lactulose dose and developed diarrhoea before the appearance of saccharide in faeces. The intake of lactulose tolerated before diarrhoea ensued varied between individuals, with the majority having diarrhoea of more than 11/day at 160 g lactulose per day. At this dose SCFA absorption was estimated to be in the range 550 to 1150 mmol/day.

Fermentation to short-chain fatty acids and lactate in human faecal batch cultures. Intra- and inter-individual variations versus variations caused by changes in fermented saccharides.

The fermentation to short-chain fatty acids, lactate, and ammonia from several non-starch polysaccharides, glucose, and albumin was investigated in 16.6% faecal homogenates. Increasing concentrations (0-30 mg/ml) of glucose, wheat bran, pectin, ispaghula, cellulose, or albumin incubated for 24 h in homogenates pooled from three individuals increased short-chain fatty acid production linearly. Amounts and ratios of short-chain fatty acids formed were highly dependent on the type of substrate fermented. Fermentable saccharides increased ammonia assimilation, in contrast to the metabolic inert cellulose. Nine faecal homogenates sampled from three individuals at three occasions were incubated for 6 and 24 h. The production of total short-chain fatty acids, acetate, propionate, and butyrate and the accumulation of D- and L-lactate changed considerably in relation to the type of substrate added (cellulose, ispaghula, wheat bran, pectin, gum arabic, and glucose; p less than 10(-4)-10(-7). In contrast, there were no significant (p greater than 0.05) differences in organic acid formation between the nine homogenates, and the intra- and inter-individual variations were of the same magnitude. Variations in fermentation, when measured as organic acid formation, were therefore related to the type of substrate fermented rather than the individual tested.

Localization of sodium absorption and chloride secretion in an intestinal epithelium.

Hen coprodeum absorbs sodium electrogenically and, when stimulated by theophylline, secretes chloride. In this study the vibrating microprobe technique was used to localize the transport of these ions to intestinal villi/folds and crypts. With the isolated, stretched epithelium, controlled by light microscopy and scanning electron microscopy, in open circuit, currents were inward, 40 +/- 7 microA/cm2, 50 microns vertically above villi, and outward, 36 +/- 7 microA/cm2 above crypts. The currents decayed exponentially to near zero at 300 microns with the same length constant. A physical model simulating the observed loci of current sources and sinks predicts potential profiles consistent with our data. Extrapolation of the currents gives a surface potential of 45 microV, negative on villi and positive above crypts. Short circuiting increased villus current to 86 +/- 27 microA/cm2 at 50 microns, and amiloride treatment reduced it to -8 microA/cm2; in both cases crypt currents were abolished. The inward currents are compatible with sodium absorption. Induction of chloride secretion after amiloride treatment, resulted in current circuits similar to those induced by sodium absorption, with villus currents of 23 +/- 7 microA/cm2. This is in accord with chloride secretion at the villi. Quantitative estimates of crypt number (860/cm2) and opening diameter (15 microns), in conjunction with isotopic measurements of active and electrical potential-driven ion fluxes demonstrate, however, that only 4% of the potential-driven co-ion transport occurs through the crypts. This indicates that nearly all chloride secretion comes from the sodium-absorbing villar area. Were the chloride secretion to occur solely from the crypts, the current should have been in the opposite direction and 10,000-fold larger.

Ion transport across isolated pig jejunum.

Sodium, chloride, and glucose transport were studied in stripped pig jejunal epithelium mounted between halfchambers. The epithelium proved to be stable for experimental purposes for up to 2 h. Net sodium absorption was electrogenic and was dependent solely on sodium-glucose cotransport. Both sodium and glucose influx varied with mucosal glucose concentration; when fitted to the Michaelis-Menten equation the two fluxes generated glucose-dependent Michaelis constant values of 1.5 and 9.6 mM, respectively. In the absence of mucosal glucose, the flux ratios for sodium and chloride were of unity, and the unidirectional fluxes were linearly dependent on the ion concentrations. The chloride accompanying sodium in open circuit was driven by the potential generated by the sodium-glucose absorption (-5 mV) and supported an absorption of 1 mumol.cm-2.h-1 NaCl. Substituting 2-hydroxyethanesulfonate for chloride decreased total tissue conductance and reduced the sodium fluxes. This effect cannot be ascribed to sodium-chloride cotransport but reflects the different mobilities of the ion pairs in solution. Chloride secretion can be induced by theophylline.

The degradation of amino acids, proteins, and blood to short-chain fatty acids in colon is prevented by lactulose.

Short-chain (C2-C5) fatty acids account for 60%-70% of the anions in the colon. Acetate (C2) is nontoxic in contrast to C(3)4-C5 fatty acids (propionate, butyrate, isobutyrate, valerate, and isovalerate), which induce coma in animals and may be important in the pathogenesis of hepatic coma in humans. An in-vitro fecal incubation system was used to map out short-chain fatty acid production in the presence of lactulose, amino acids, albumin, or blood. Albumin and blood increased production of all C2-C5 fatty acids. In contrast, lactulose was converted to acetate only and increased fecal acidity. The degradation of amino acids, albumin, and blood to short-chain fatty acids was completely inhibited by 10-25 mM lactulose. This was caused mainly by the acidifying effect of lactulose. pH-independent inhibition of blood and amino acid degradation to short-chain fatty acids required concentrations of lactulose exceeding 50-100 mM. Thus, the effect of lactulose in the treatment of hepatic coma may be related to its rapid fermentation into organic acids at rates exceeding colonic buffering capacity. This probably reduces formation of toxic fatty acids and ammonia from amino acids, polypeptides, and blood in the colon.

Short-chain fatty acids in bowel contents after intestinal surgery.

Short-chain fatty acids are produced in the human colon by bacterial fermentation of dietary fibers and other saccharides escaping absorption in the small bowel. Short-chain fatty acid concentrations were determined together with production rates in 6- and 24-h incubations of intestinal outputs from 56 patients with various types of intestinal resections. Concentrations and 6- and 24-h production rates in feces from 9 healthy persons (controls; median +/- SD) were 98.9 +/- 21.4 mmol/L and 17.2 +/- 5.1 and 9.3 +/- 1.5 mmol/L.h, respectively. Colectomized patients with short bowel syndrome had extremely low concentrations (0.8 mmol/L) compared with controls (p less than 10-5), patients with ileostomy (p = 0.003), and ileal reservoirs (p less than 10-5), and showed low 6- and 24-h production rates (1.5 and 0.9 mmol/L.h, respectively; p less than 10-5 vs. controls). Short-chain fatty acids in ileostomic digesta (11.1 mmol/L) were decreased (p = 0.011) compared with outputs from ileal reservoirs (51.5 mmol/L), although production rates were in the same order of magnitude--all below control values (p less than 0.001). Patients partially colectomized and patients with small bowel bypass or short bowel syndrome with preserved colon had normal fecal concentrations with decreased production rates of short-chain fatty acids vs. controls (p less than 0.01). Only minor changes in ratios between individual acids were found. Reciprocal values of short-chain fatty acid concentrations correlated to volumes of outputs from both small intestine (r = 0.86, p less than 10-6) and colon (r = 0.79, p less than 10-6) when results were cumulated. It is concluded that partial resections of colon and the small bowel do not influence the fecal concentration level of short-chain fatty acids as long as colon is not totally resected.