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A comparative study of nutrient digestibility, kinetics of digestion and passage and rumen fermentation pattern in goats and sheep offered medium quality forages at the maintenance level of feeding.

The in vivo digestibility and the rates of degradation and passage of alfalfa hay (AH) and vetch straw (VS) were compared in three Granadina goats and three Segureña wethers fed at approximately maintenance level, using a change over design. The nylon bag technique was used to estimate the fractional rate of degradation of the feeds in the rumen. The fractional outflow rate of hay and straw particles was determined with chromium as a marker. No significant differences in the digestibility coefficients of nutrients between animal species were found. Nitrogen retention expressed as coefficients of both nitrogen intake and digested nitrogen was lower (P < 0.01) in goats than in sheep when VS was offered. There was no significant difference between goats and sheep in the effective degradability (measured at outflow rates in the range of 0.027 to 0.032 h-1) of dry matter (DM) for either forage. However, the effective degradabilities of the neutral detergent fibre (NDF) and crude protein (CP) of VS were lower (P < 0.01 and P < 0.001, respectively), in goats than in sheep. The degradation rates of DM, NDF and CP were not significantly different (P > 0.05) between goats and sheep. Also, the fractional rate of passage of particles from the rumen was not different (P = 0.056) between species, although there was a tendency for an increased outflow rate in goats. In a second experiment the effects of animal species, type of diet and frequency of feeding on rumen fermentation pattern were studied following a factorial design. Four goats and four wethers were used. They were randomly allocated to three dietary treatments: AH, VS or a 50:50 mixture of AH and VS (AH/VS). Rations were offered in two or three meals daily and samples of rumen content were obtained at 09.00, 11.00, 13.00, 15.00, 17.00 and 19.00 h. No difference in rumen pH was found between goats and sheep. Total volatile fatty acids and ammonia concentrations were higher (P < 0.001) in sheep than in goats. The molar proportions of acetate and isovalerate were higher (P < 0.01 and P < 0.001, respectively) in goats than in sheep, whereas the opposite was true for the molar proportion of n-butyrate. Feeding frequency had a slight effect on the rumen parameters measured. It is concluded that only minor differences in the digestive capacity and rumen activity of sheep and goats can be expected when they are fed on medium quality forages.

Animal Feed↗

Nonbreeding eastern curlews Numenius madagascariensis do not increase the rate of intake or digestive efficiency before long-distance migration because of an apparent digestive constraint.

The possibility of premigratory modulation in gastric digestive performance was investigated in a long-distance migrant, the eastern curlew (Numenius madagascariensis), in eastern Australia. The rate of intake in the curlews was limited by the rate of digestion but not by food availability. It was hypothesized that before migration, eastern curlews would meet the increased energy demand by increasing energy consumption. It was predicted that (1) an increase in the rate of intake and the corresponding rate of gastric throughput would occur or (2) the gastric digestive efficiency would increase between the mid-nonbreeding and premigratory periods. Neither crude intake rate (the rate of intake calculated including inactive pauses; 0.22 g DM [grams dry mass] or 3.09 kJ min(-1)) nor the rate of gastric throughput (0.15 g DM or 2.85 kJ min(-1)) changed over time. Gastric digestive efficiency did not improve between the periods (91%) nor did the estimated overall energy assimilation efficiency (63% and 58%, respectively). It was concluded that the crustacean-dominated diet of the birds is processed at its highest rate and efficiency throughout a season. It appears that without a qualitative shift in diet, no increase in intake rate is possible. Accepting these findings at their face value poses the question of how and over what time period the eastern curlews store the nutrients necessary for the ensuing long, northward nonstop flight.

Animal Migration↗

Degradation products of bran phytate formed during digestion in the human small intestine: effect of extrusion cooking on digestibility.

To investigate the digestion of phytate in the stomach and small intestine in humans, studies were performed in subjects with established ileostomy. A recently developed high performance liquid chromatography method made it possible to analyze phytate and its degradation products in food and digesta. The digestibility of phytate in raw bran and extruded bran was investigated in seven ileostomy patients. Each subject was studied for two 4-d periods while consuming a constant low fiber diet with the addition of either 54 g/d of a bran-gluten-starch mixture or the corresponding extruded product. During passage through the subject's stomach and small intestine 58%, on average, of the phytate in unprocessed bran was hydrolyzed to inositol penta-, tetra- and triphosphates. When bran was subjected to extrusion cooking, 25% of the inositol hexaphosphate was hydrolyzed to penta- and tetraphosphate and the phytase activity ceased. Essentially no phytate digestion occurred when the ileostomy subjects consumed the extruded product. The reduced digestibility might be due to the lost phytase activity or to formation of indigestible phytate complexes during extrusion cooking.

Cooking↗

Recombinant Saccharomyces cerevisiae expressing P450 in artificial digestive systems: a model for biodetoxication in the human digestive environment.

The use of genetically engineered microorganisms such as bacteria or yeasts as live vehicles to carry out bioconversion directly in the digestive environment is an important challenge for the development of innovative biodrugs. A system that mimics the human gastrointestinal tract was combined with a computer simulation to evaluate the survival rate and cinnamate 4-hydroxylase activity of a recombinant model of Saccharomyces cerevisiae expressing the plant P450 73A1. The yeasts showed a high level of resistance to gastric and small intestinal secretions (survival rate after 4 h of digestion, 95.6% +/- 10.1% [n = 4]) but were more sensitive to the colonic conditions (survival rate after 4 h of incubation, 35.9% +/- 2.7% [n = 3]). For the first time, the ability of recombinant S. cerevisiae to carry out a bioconversion reaction has been demonstrated throughout the gastrointestinal tract. In the gastric-small intestinal system, 41.0% +/- 5.8% (n = 3) of the ingested trans-cinnamic acid was converted into p-coumaric acid after 4 h of digestion, as well as 8.9% +/- 1.6% (n = 3) in the stomach, 13.8% +/- 3.3% (n = 3) in the duodenum, 11.8% +/- 3.4% (n = 3) in the jejunum, and 6.5% +/- 1.0% (n = 3) in the ileum. In the large intestinal system, cinnamate 4-hydroxylase activity was detected but was too weak to be quantified. These results suggest that S. cerevisiae may afford a useful host for the development of biodrugs and may provide an innovative system for the prevention or treatment of diseases that escape classical drug action. In particular, yeasts may provide a suitable vector for biodetoxication in the digestive environment.

Computer Simulation↗

Effects of source and level of dietary neutral detergent fiber on feed intake, ruminal fermentation, ruminal digestion in situ, and total tract digestion in beef cattle fed pelleted concentrates with or without supplemental roughage.

The effects of source and level of dietary NDF on intake, ruminal digestion in situ, ruminal fermentation, and total tract digestion were evaluated in Hereford steers using a replicated 5 x 5 Latin square design. Diets contained 62 to 64% TDN and included 1) 80% control concentrate (contained pelleted ground grains) and 20% timothy hay (traditional diet), 2) 80% control concentrate and 20% alfalfa cubes, 3) 90% control concentrate and 10% cubes, 4) a completely pelleted diet using corn cobs as the primary NDF source, and 5) 80% textured (rolled instead of ground grains) concentrate and 20% hay. Dry matter intake differed (P less than .05) between the traditional and cube diets due to limited acceptance of alfalfa cubes. Increased (P less than .05) ruminal osmolality, total VFA, and NH3 N and lower (P less than .01) ruminal pH in steers fed corn cob and cube diets relative to steers fed the traditional diet were due to preferential consumption of concentrate over supplemental roughage and the resultant rapid fermentation of concentrates. Potentially degradable DM in the traditional diet exceeded (P less than .06) all other diets, resulting in the increased (P less than .10) extent of DM disappearance despite a slower (P less than .05) rate of DM disappearance. Rate of NDF disappearance and all in situ starch disappearance parameters were similar between the traditional, corn cob, and cube diets. All ruminal digestion parameters involving NDF disappearance were similar between hay diets and between cube diets, whereas rate and extent of starch disappearance differed (P less than .05) between hay diets. Although formulation of diets with different sources of dietary NDF did not affect total tract digestion of nutrients, nutrient availability and ruminal fermentation were altered due to dietary differences in sources of dietary NDF and preferential selection of feedstuffs by steers.

Ammonia↗

Effects of saturation and esterification of fat sources on site and extent of digestion in steers: digestion of fatty acids, triglycerides, and energy.

Five steers (mean BW 526 kg) fitted with ruminal, duodenal, and ileal cannulas were used in a 5 x 6 Youden square design with 14-d periods. Diets contained chopped alfalfa hay, corn silage, and concentrate (25:35:40, DM basis). Treatments were 1) control (no added fat); 2) tallow (T), iodine value (IV) = 51.5; 3) partially hydrogenated tallow (PHT), IV = 30.7; 4) hydrogenated tallow (HT), IV = 6.9; 5) blend (1: 1) of HT and hydrogenated free fatty acids (HTHFA), IV = 9.0; and 6) hydrogenated free fatty acids (HFA), IV = 11.2. Fats replaced cornstarch in the control diet to supply 5% added fatty acids. Intake was restricted to 90% of ad libitum; DMI was similar among diets (mean 9 kg/d). Total fatty acid intake averaged 170, 500, 506, 525, 489, and 491 g/d for treatments 1 to 6, respectively. Flows of total C16, total C18, and total fatty acids to the duodenum were increased by supplemental fat; flows of total C18 and total fatty acids were greater than their intake for all treatments. Flow of total fatty acids associated with ruminal bacteria accounted for 50 and 17% of the total duodenal fatty acid flow for the control and fat-supplemented diets, respectively. Digestibility of total fatty acids entering the small intestine (74, 71, 62, 39, 53, and 63% for treatments 1 to 6, respectively) was greater for the control diet than for fat-supplemented diets and decreased as either saturation (T < PHT < HT) or esterification (HFA < HTHFA < HT) increased. Digestibilities of fatty acids in the total tract followed similar patterns. Ruminal lipolysis of dietary triglycerides decreased linearly as the degree of saturation of fat sources increased. Small intestinal disappearance of triglycerides (89, 75, 51, 44, 64, and 73% of duodenal flow for treatments 1 to 6, respectively) decreased linearly as either saturation or esterification increased. Flows and digestion of gross energy followed patterns similar to those for fatty acids and triglycerides. Resistance to ruminal and small intestinal lipolysis is a major factor contributing to the poor digestibility of highly saturated triglycerides.

Animal Feed↗

[Regulatory properties of digestive enzymes and the biology of multiple substrate digestive processes].

Characteristics of functional organization and autoregulation of natural polysubstrate digestion in various animals are largely defined by regulatory properties of the enzymes ensuring membrane digestion. This has been demonstrated for both the animals which are at different evolutional levels and those specialized for different types of food. The effective organization of the digestive and transport processes in time and space is provided by an interaction of food substances at the stage of membrane digestion.

Alkaline Phosphatase↗

Anaerobic co-digestion of olive mill wastewater with olive mill solid waste in a tubular digester at mesophilic temperature.

Anaerobic co-digestion is a well established process for treating many types of organic wastes, both solid and liquid. In this study we have investigated, on a laboratory scale, the anaerobic co-digestion of olive mill wastewater (OMW) with olive mill solid waste (OMSW) using semi-continuous, feeding, tubular digesters operated at mesophilic temperatures. Each digester was fed with an influent, composed of OMW and OMSW, at an organic loading rate (OLR) varying between 0.67 and 6.67 g COD/l/d. The hydraulic retention times (HRT) were 12, 24 and 36 days. The TCOD concentrations of OMW used as the main substrate were 24, 56 and 80 g COD/l; the amount of the dry OMSW used as a co-substrate was fixed to approximately 56 g/l of OMW. The results indicated that the best methane production was about 0.95 l/l/day obtained at an OLR = 4.67 g COD/l/d, corresponding to influent TCOD = 56 g COD/l at an HRT = 12d. In contrast, the maximum TCOD removal efficiency (89%) was achieved at an OLR = 0.67 g COD/l/d, corresponding to influent TCOD = 24 g COD/l at an HRT = 36 d. Moreover, the inhibition of biogas production was observed at the highest OLR studied.

Anaerobiosis↗

Effect of digestive health care services integration on resource use and outcomes in patients with digestive disorders.

BACKGROUND & AIMS: Academic medical centers are under pressure to contain resources. The University of Virginia Digestive Health Service Center (DHSC) is a fully integrated inpatient service combining gastroenterology, hepatology, surgery, pathology, radiology, and nutritional services. The aim of this study was to evaluate whether integration of digestive health services can contain resource use among gastrointestinal (GI) inpatient admissions. METHODS: This is a cohort study of 2934 inpatient cases admitted to a referral academic medical center between January 1, 1998, and June 30, 2000, with a primary diagnosis of an esophageal disorder, appendicitis, abdominal hernia, intestinal obstruction, diverticulitis, biliary tract disease, liver disease, a pancreatic disorder, or GI hemorrhage. Cases were grouped by admitting service (DHSC vs. other nonintegrated inpatient services). Total cost, length of stay, and 30-day readmission rate were adjusted for differences in baseline patient characteristics and compared. RESULTS: DHSC admission was associated with significant cost savings (P = 0.0363) and reduced length of stay (P < 0.0001). Cost savings were attributable to patients admitted for liver disease (P = 0.0077), GI hemorrhage (P = 0.0031), and diverticulitis (P = 0.0497); reductions in length of stay were seen for patients with liver disease (P = 0.0314), GI hemorrhage (P = 0.0212), diverticulitis (P = 0.0017), esophageal disorders (P = 0.0006), and abdominal hernia (P = 0.0458). DHSC patients with pancreatic disorders had increased total cost (P = 0.0247), but no difference in length of stay (P = 0.7504) and a lower 30-day readmission rate (P = 0.0478). CONCLUSIONS: Integration of digestive health services reduces resource use for patients with digestive diseases and may improve outcomes for patients with pancreatic disorders. A multidisciplinary service center may represent a more efficient model for health care delivery for these patients.

Academic Medical Centers↗

Presence of sorbin in human digestive tract and endocrine digestive tumours.

BACKGROUND: Sorbin, a 153 amino acid peptide isolated from porcine intestine, was localised by immunohistochemistry in endocrine cells of the intestinal mucosa and pancreas and in the enteric nervous system in the pig. AIMS: To identify sorbin cells in normal human digestive tissues and to explore the expression of sorbin in 37 digestive endocrine tumours: 14 intestinal carcinoid tumours and 23 endocrine pancreatic tumours including six insulinomas. METHODS: Two polyclonal antibodies against the C-terminal and the N-terminal sequences of porcine sorbin raised in rabbit were used to evaluate sorbin expression by immunohistochemistry. RESULTS: In the human digestive tract, sorbin, characterised by both C-terminal and N-terminal immunoreactivity, was found in enterochromaffin cells of the gastric and intestinal epithelium from the pyloric junction to the descending colon. C-Terminal sorbin immunoreactivity alone was found in plexii from the enteric nervous system and in some insulin-containing cells of normal pancreas. C-Terminal and N-terminal antibodies disclosed sorbin in five of 14 intestinal carcinoid tumours; C-terminal antibody alone disclosed a C-terminal sorbin peptide in two of six insulinomas and three of 17 endocrine pancreatic tumours. The presence of sorbin was not associated with a specific clinical syndrome. CONCLUSIONS: Sorbin is present in the digestive tract in several forms. It is expressed in some intestinal and pancreatic endocrine tumours.

Aged↗

Sau3A in situ digestion of human chromosome 3 pericentrometric heterochromatin. I. Differential digestion of alpha-satellite and satellite 1 DNA sequences.

In situ digestion with the restriction endonuclease (RE) Sau3A (Sau3A REISD) uncovers a polymorphism for the pericentromeric heterochromatin of human chromosome 3, which can be positively stained (3+) or not (3-), and has proven useful to differentiate donor and recipient cells after sex-matched bone marrow transplantation and to analyze the so-called hemopoietic chimerism. The aim of the present investigation was to obtain insight into the molecular basis of such polymorphism to optimize its use for chimerism quantification using methodological approaches other than REISD. To this end, fluorescence in situ hybridization (FISH) assays using probes for the satellite DNA sequences that mainly constitute chromosome 3 pericentromeric heterochromatin (alpha-satellite and satellite 1 DNA) were performed on control and Sau3A-digested chromosomes. The results obtained suggest that chromosome 3 alpha-satellite DNA is digested in all individuals studied, irrespective of the karyotype obtained by Sau3A REISD (3++, 3+-, 3--), and thus it does not seem to be involved in the polymorphism uncovered by Sau3A on this chromosome. Satellite 1 DNA is not digested in any case, and shows a polymorphism for its domain size, which correlates with the polymorphism uncovered by Sau3A in such a way that 3+ chromosomes show a large domain (3L) and 3- chromosomes show a small domain (3S). It seems, therefore, that the cause of the polymorphism uncovered by Sau3A on the pericentromeric region of chromosome 3 is a difference in the size of the satellite 1 DNA domain. Small satellite 1 DNA domains fall under the resolution level of REISD technique and are identified as 3-.

Centromere↗

Up-to-date modification of the anaerobic sludge digestion process introducing a separate sludge digestion mode.

Sewage treatment plants in Japan are subjected to advanced treatment to remove nutrients and hence control eutrophication problems in lakes and bays. This paper discusses the advantages and disadvantages of the separate digestion treatment mode for sludge generated from advanced wastewater treatment. In the separate digestion only primary sludge is digested and the excess activated sludge is directly dewatered. Separate digestion can reduce the return load of nutrients to approximately one third, and has major potential for the beneficial use of sludge.

Bioreactors↗

Anaerobic co-digestion of sewage sludge and food waste using temperature-phased anaerobic digestion process.

This study was performed to overcome the low efficiency of anaerobic digestion of sewage sludge and food waste by combining temperature-phased digestion, sequencing batch operation, and co-digestion technology. It was demonstrated that the temperature-phased anaerobic sequencing batch reactor (TPASBR) system for the co-digestion of sewage sludge and food waste resulted in enhanced volatile solids (VS) reduction and methane production rate. At the organic loading rate (OLR) of 2.7 g VS/l/d, the TPASBR system showed the higher VS reduction (61.3%), CH4 yield (0.28 l/g VS(added)) and CH4 production rate (0.41 l CH4/l/d) than those (0.29 l CH4/l/d) of the mesophilic two-stage ASBR (MTSASBR). In the specific methanogenic activity (SMA) tests on thermophilic biomass of the TPASBR system, the average SMA of acetate (93 ml CH4/gVSS/d) was much higher than those of propionate (46 ml CH4/g VSS/d) and butyrate (76 ml CH4/g VSS/d). Also, higher specific hydrolytic activity (SHA, 217 mg COD/g VSS/d) of the biomass supported fast hydrolysis under thermophilic conditions. The track study revealed that the most active period of the 24 h cycle was between 6 and 12 h. The enhanced performance of the TPASBR system could be attributed to longer solids retention time, fast hydrolysis, higher CH4 conversion rate, and balanced nutrient condition of co-substrate. It was verified that this combination could be a promising and practical alternative for the simultaneous recycling of two types of organic fraction of municipal solid waste (OFMSW) with high stability.

Anaerobiosis↗

Effect of intestinal microflora on digestible energy and fiber digestion in chickens fed a high-fiber diet.

The effect of intestinal microflora on digestible energy (DE) value and fiber digestion was studied in single-comb White Leghorn chickens fed a low-fiber diet (experiment 1) or a high-fiber diet with low or adequate metabolizable energy (ME) value (experiment 2). Fecal energy excretion was calculated from the difference between total energy excretion in urinary and fecal droppings and urinary energy excretion, which was estimated from the energy values for individual urinary nitrogenous compounds extracted with Li2CO3. When the birds were fed the low-fiber diet, no differences in growth, DE, or ME were observed between germ-free and conventional environments. Of birds fed the high-fiber diet, growth of those in the conventional environment was similar to that of the birds in the germ-free environment at the adequate ME value, whereas birds in the conventional environment grew faster than the birds in the germ-free environment at the low ME value. Changes in observed dietary ME values of the high-fiber diets, being higher in birds in the conventional environment than in birds in the germ-free environment (experiment 2), were almost entirely accounted for by those in dietary DE values, most of which was contributed by crude fiber digestion. It was concluded, therefore, that by means of fiber digestion, the intestinal microflora may benefit the host bird by supplying extra energy, which would result in growth promotion, particularly when the bird is deficient in energy.

Animals↗

Comparison of microwave digestion with conventional wet ashing and dry ashing digestion for analysis of lead, cadmium, chromium, copper, and zinc in shellfish by flame atomic absorption spectroscopy.

A closed vessel microwave digestion procedure was developed for shellfish samples. This procedure was compared with wet and dry ash procedures for levels of lead, cadmium, chromium, copper, and zinc. Results obtained for microwave and conventional wet ash digestion were comparable. The dry ashing procedure produced results consistently lower than either of the other methods. Recoveries ranged from 80-92% for microwave and wet ashing procedures and 54-72% for the dry ashing procedure. Accuracy was also determined by analyzing lobster hepatopancreas marine reference material. Values for Pb, Cd, and Cr fell within the range specified for the reference material for all 3 digestion procedures; however, values were lower for Cu and Zn. Results of this study show that microwave digestion is comparable to wet ashing.

Animals↗

Studies on digestive physiology and feed digestibilities in captive Indian rhinoceros (Rhinoceros unicornis).

We performed intake, digestibility and ingesta passage studies in 11 Indian rhinoceroses (Rhinoceros unicornis) from four zoological institutions, using total faecal collection for the quantification of faecal output. The regularly fed zoo ration of roughage and concentrates (ration RC) and a roughage-only ration (ration R) were used; the roughage source differed between the facilities and comprised grass hay, grass silage, straw and lucerne hay. Dry matter intake ranged between 0.8 and 1.3% of body weight on ration RC and 0.5-1.2% on ration R. Digestibility coefficients achieved were similar to those reported for horses on diets of comparable composition. Endogenous losses as determined by linear regression analysis were within the range reported for horses. Measurements of faecal volatile fatty acids, faecal lactate and faecal pH also showed similarity to similar measurements in horses. The mean retention times of fluids (Co-EDTA) and particles (Cr-mordanted fibre <2 mm) in the whole gastrointestinal tract averaged 42 and 61 h, respectively, and were the longest ever recorded in a monogastric ungulate with this marker system. The results suggest that the horse is a useful model animal for designing diets for Indian rhinoceroses. Why digestive parameters are similar between these species in spite of enormous differences in body weight and retention times remains to be answered.

Animal Feed↗

Effect of combinations of organic acids in weaned pig diets on microbial species of digestive tract contents and their response on digestibility.

The effect of combinations of organic acids on digestive microbial population and total tract digestibility of piglets was studied. Thirty 19-21 days weaned pigs (5.4 +/- 0.23 kg, six pigs per treatment) were given a mixed feed with 10 ml water/kg (CTL) or 200 mEq/kg of formic acid (FOR), 1:1 formic:fumaric (FOFU), 1:1 formic:lactic (FOLA) or 2:1 formic:lactic (2FOLA). After 6-8 days, animals were slaughtered. In gastric contents, pH was higher (p = 0.01) with FOFU and lactobacilli tended (p = 0.08) to be lower with FOFU than CTL and FOLA, but coliform counts did not differ (p = 0.14). Acetate proportion was lower (p = 0.03), and propionate (p = 0.05) and butyrate (p = 0.01) higher, with FOFU than CTL, FOLA and 2FOLA. Intestinal coliform counts were higher (p = 0.03) with CTL than FOLA and 2FOLA, but there were no differences on lactobacilli. In intestinal contents, acetate tended (p = 0.06) to be lower with FOR than FOLA and 2FOLA, and butyrate was higher (p = 0.001) with FOR. Although not significantly, dry matter digestibility was 0.03-0.05 lower with CTL than with the other treatments. Combinations of organic acids in piglet diets modify gastric and intestinal microflora, the mixtures of formic:lactic appearing as the most interesting.

Animals↗

Susceptibility of Kintoki bean lectin to digestive enzymes in vitro and its behavior in the digestive organs of mouse in vivo.

The enzymatic digestion of a Kintoki bean lectin in vitro resulted in neither the extensive hydrolysis nor complete inactivation of the lectin. The majority of [3H]lectin administered to mice by stomach-intubation was found in the digestive tract at levels of 88.7%, 99.4%, 99.5% and 78.6%, after 0.5, 2, 5 and 24h of intubation, respectively. Twenty to forty percent of the administered radioactivity was found combined with the mucosa of the small intestine. After 24 h of intubation, part of the radioactivity was found in the feces. The recovery test of hemagglutinating activity and the molecular weight determination of the lectin administered to and recovered from mice revealed that more than 70% of the radioactivity detected in the digestive tract represented intact lectin. Therefore, it was concluded that most of the Kintoki bean lectin survived its passage through the gastrointestinal tract. The lectin-binding sites in the intestinal mucosa were traced with fluorescence microscopy using the lectin conjugated with fluorescein isothiocyanate. The fluorescent sites were found located at the top and upper sites of the villi, but not at the crypt.

Animals↗