The profiling of amino acids in crop milk and plasma and mRNA abundance of amino acid transporters and enzymes related to amino acid synthesis in the crop tissue of male and female pigeons during incubation and chick-rearing periods.

The present study was carried out to investigate the changes in amino acid (AA) contents of crop milk and plasma and mRNA abundance of AA transporters and AA synthesis-related enzymes in the crop tissue of male and female pigeons during incubation and chick-rearing periods. Forty-two pairs of adult White King pigeons with 2 fertile eggs per pair were randomly divided into 7 groups by different breeding stages. The AA content of crop milk decreased from day 1 (R1) to day 25 (R25) of chick rearing (P < 0.05). In both male and female adult pigeons, the contents of Thr, Leu, Val, His, Asp, and Pro in plasma increased to maximum levels on R25. Parental sex effect and interaction between stage and sex were observed in the AA contents of pigeon plasma (P < 0.05). For AA transporters, the mRNA abundances of SNAT2, ASCT1, LAT1, and y+LAT2 in the male crops reached the highest value on day 17 of incubation (I17), and the peak mRNA levels of PAT-1, xCT, b0,+AT, and CAT1 were found on R7 (P < 0.05). In females, the abundances of ASCT1, B0AT1, asc-1, and CAT1 mRNA peaked on R1, whereas the maximum levels of LAT1, PAT-1, b0,+AT, and y+LAT2 were observed on R7. For enzymes involved in AA synthesis, the highest gene expressions of glutamate dehydrogenase 1, acetolactate synthase in both parent pigeons, and L-threonine 3-dehydrogenase in female pigeon crops were attained on I17. The expressions of ornithine-δ-aminotransferase, glutamic-oxal(o)acetic transaminase 1, glutamic-oxal(o)acetic transaminase 2, asparagine synthetase, serine hydroxymethyltransferase 2, and glutamic-pyruvic transaminase 2 in both sexes and argininosuccinate lyase and L-threonine 3-dehydrogenase in males were the highest on R1. In conclusion, AA used for pigeon crop milk formation may originate from plasma and intracellular synthesis. The genes involved in AA transport and synthesis varied significantly with sexual effects, indicating that other factors should be considered in future explorations of the mechanism of protein formation in crop milk.

Leucine promotes the growth of squabs by increasing crop milk protein synthesis through the TOR signaling pathway in the domestic pigeon (Columba livia).

Leucine (Leu) plays a critical regulatory role in protein synthesis, however, the effects and molecular mechanisms of Leu on crop milk protein in the domestic pigeons (Columba livia) are still unknown. Therefore, the study aimed to investigate the effects of dietary Leu supplementation on crop milk protein synthesis and the growth performance of squabs and the possible underlying mechanism. A total of 240 pairs of breeding pigeons (1102.3 ± 9.5 g/pair) were randomly assigned to 1 of 5 treatments, including a positive control (PC) diet that had adequate crude protein (crude protein, CP = 18%; Leu = 1.30%), a negative control (NC) diet that was low in CP (CP = 16%, Leu = 1.30%), and NC diets supplemented with Leu at 0.15%, 0.45%, or 1.05%. Compared with the NC diet, 0.15 to 0.45% Leu supplementation decreased BW loss and increased relative crop weight, crop thickness, and protein levels in the crop tissue and milk of breeding pigeons. However, dietary supplementation with 1.05% Leu inhibited ADFI in breeding pigeons. Dietary supplementation with 0.15 to 0.45% Leu decreased the mortality rate and increased the BW, eviscerated yield, and breast muscle yield of young squabs. The protein expression levels of the target of rapamycin (TOR), ribosomal protein S6 kinase 1 (S6K1), ribosomal protein S6 kinase (S6), eukaryotic initiation factor 4E binding protein 1 (4EBP1), and eukaryotic translation initiation factor 4E (eIF4E) were upregulated in the crop tissue of breeding pigeons in PC, 0.15% and 0.45% Leu-supplemented groups. Collectively, these results indicated that 0.15 to 0.45% Leu supplementation could decrease BW loss, increase milk protein synthesis in the crop of breeding pigeons, and enhance the survival rate and growth performance of young squabs through the TOR signaling pathway.

Physiological responses to central and peripheral injection of polyinosinic-polycytidylic acid in chicks.

1. The purpose of the present study was to determine if intracerebroventricular (ICV) and intraperitoneal (IP) injection of polyinosinic-polycytidylic acid (poly I:C), a viral mimetic that binds to toll-like receptor-3 (TLR3), affects food intake, voluntary activity, cloacal temperature, plasma corticosterone (CORT) and glucose concentrations, and crop emptying rate in chicks (Gallus gallus). 2. Both ICV and IP injection of poly I:C significantly decreased food intake. 3. IP but not ICV injection of poly I:C significantly suppressed voluntary activity, whereas ICV injection decreased time spent sitting. Both ICV and IP injection of poly I:C significantly increased plasma CORT and glucose concentration. Neither ICV nor IP injection of poly I:C significantly affected cloacal temperature. 4. In addition, ICV injection of poly I:C significantly reduced crop emptying rate, whereas IP injection had no effect. 5. These results suggested that central TLR3 is related to anorexia, stress response and retardation of crop emptying while peripheral TLR3 is related to anorexia, change in behaviour and stress responses during viral infection in chicks.

Compound 48/80 reduces the crop-emptying rate, likely through a histamine-associated pathway in chicks.

Infectious conditions are associated with reduced food passage through the digestive tract in both mammals and chicks; however, the precise mechanism mediating this response in chicks remains unclear. The purpose of the present study was to determine if mast cells, a blood cell type which plays an important role in the immune system, might affect food passage through the digestive tract in chicks. Specifically, we performed intraperitoneal (IP) injections of compound 48/80, an inducer of mast cell degranulation, and measured crop emptying. The IP injection of compound 48/80 significantly reduced the crop-emptying rate, but it did not affect the proventriculus to small intestine transit rate or the number of defecations. We also found that IP-injected histamine, which is secreted by mast cells, also reduced the crop-emptying rate. In addition, IP injection of 2-pyridylethylamine (histamine H1 receptor agonist), but not dimaprit, (R)-(-)-α-methylhistamine, and VUF8430 (histamine H2, H3, and H4 receptor agonists, respectively), reduced the crop-emptying rate, implying that histamine reduces the crop emptying rate via the histamine H1 receptor. Finally, we found that IP injection of compound 48/80 reduced mRNA expression of histidine decarboxylase, a rate-limiting enzyme for histamine synthesis, in the esophagus and proventriculus at 1 h and the proventriculus and duodenum at 3 h after the injection. In sum, the present study suggests that the degranulation of mast cells causes a reduction in the crop-emptying rate, possibly via the histamine pathway in chicks.

Influence of the incorporation mode of sugar beet pulp in the finishing diet on the digestive tract and performances of geese reared for foie gras production.

The aim of this work was to study the effects of incorporating sugar beet pulp (SBP) into the diet of geese in two feeding systems (complete pelleted feed or loose-mix feeding system) on crop development and performance. A total of 480 1-d-old male geese were divided into three groups whose diet differed from d 56 to 90: a complete pelleted diet containing 50% corn (control diet: AMEn 11.5 MJ/kg; CP 161 g/kg), and no SBP; a complete pelleted diet containing 50% corn and 10% SBP (SBPcp diet: AMEn: 11.5 MJ/kg; CP: 161 g/kg); and a mix in the same feeder (SBPlm diet) of 500 g/kg of protein-rich pellets containing 20% SBP (SBPprp: AMEn: 9.0 MJ/kg; CP: 250 g/kg) and 500 g/kg of whole corn (WC: AMEn: 14.0 MJ/kg; CP: 72 g/kg). Body traits, including crop volume, were measured at d 91. From d 91 to 106, 88 birds/group were overfed with a mixture containing mainly corn and water before slaughter to measure fatty liver performance. Feed intake from d 56 to 90 was higher (+10%; P = 0.004) in the SBPcp group than the other two, but at d 90, the body weight (BW) of the birds was higher (+7%; P = 0.002) in the SBPlm group than the other two. At d 91, the volume of the crop was greater in the SBPcp group (80.4 mL/kg of BW, P < 0.001) than in the control group (60.3 mL/kg of BW), the SBPlm group being intermediate (64.1 mL/kg of BW). Feed intake (13,321 g), weight gain (2,733 g), and feed-to-gain ratio (4.9) during the overfeeding period, as well as fatty liver weight (963 g) and commercial grading, were similar (P > 0.05) between the three groups. In conclusion, the use of sugar beet pulp in the diet of finishing geese helps the adaptation of the digestive tract to the overfeeding period, even in a loose-mix feeding system based on whole corn.

Differential expression of fatty acid transporters and fatty acid synthesis-related genes in crop tissues of male and female pigeons (Columba livia domestica) during incubation and chick rearing.

1. The growth performance of squabs reared solely by male or female parent pigeons was measured, and the changes of lipid content of crop milk and the expression profiles of genes potentially involved in lipid accumulation by crop tissues of parent pigeons were evaluated during incubation and chick rearing. 2. Squabs increased in body weight during 25 d of rearing, whereas both male and female pigeons lost weight after finishing rearing chicks, and the weight loss of male pigeons was significantly greater than that of female parent pigeons. Lipid content of crop milk from both parent pigeons gradually decreased to the crude fat level in the formulated diet after 10 d (R10) of chick rearing. 3. The gene expression of fatty acid translocase (FAT/CD36), fatty acid-binding protein 5 (EFABP) and acyl-CoA-binding protein (ACBP) in male pigeon crop tissue were the greatest at 17 d (I17) of incubation. In female pigeons, FAT/CD36 expression was the highest at I14, and both EFABP and ACBP expression peaked at I14 and R7. The expression of acetyl-CoA carboxylase and fatty acid synthase in male pigeons reached the maximum level at R1, while they peaked at I14 and I17, respectively in female pigeons. The gene expression of peroxisome proliferators-activated receptor-gamma (PPARγ) was the greatest at I17 in the male, while it was at I14 in the female. However, no regular changing pattern was found in PPARα gene expression in male pigeons. 4. These results indicated that male and female pigeons may make different contributions in rearing squabs. The gene expression study suggested that fatty acids used in lipid biosynthesis of crop milk probably originated from both exogenous supply and de novo synthesis. The sex of the parent pigeon affected the lipid content of crop milk and the expression profiles of genes involved in fatty acid transportation and lipogenesis.

Aviisotricha hoazini n. gen., n. sp., the Morphology and Molecular Phylogeny of an Anaerobic Ciliate from the Crop of the Hoatzin (Opisthocomus hoazin), the Cow Among the Birds.

The hoatzin is the only known avian species that has evolved a foregut fermentation system similar to that of ruminant animals. Due to the closeness of the bird's fermentation chamber, the crop, to the bird's beak it exudes an unpleasant odour; therefore, the hoatzin is called the "cow among the birds". In addition to Eubacteria and Archaea, responsible for digestion of the vegetation they consume, the bird's crop contains a holotrich ciliate, described here for the first time in detail. Cytological staining of this isotrichid-like ciliate with the Chatton-Lwoff and Protargol staining procedures, as well as SEM and TEM, justified the establishment of the new genus Aviisotricha n. gen. with its new type species Aviisotricha hoazini n. gen., n. sp.. Phylogenetic analyses of a portion of the small subunit rRNA gene supported the taxonomic placement of this new genus and species in the family Isotrichidae. Aviisotricha is compared with Balantidium, Dasytricha and Isotricha with special reference to their dorsal brushes, which show similarity to the paralabial organelle of the Entodiniomorphida. The possible phylogenetic origin of Aviisotricha is discussed and a taxonomic revision of the family Isotrichidae is given.

Comparison of brain urocortin-3 and corticotrophin-releasing factor for physiological responses in chicks.

Corticotrophin-releasing hormone (CRH) plays an important role in response to stress, and exerts a physiological effect via its receptor, CRH receptor type-1 (CRH-R1) and CRH receptor type-2 (CRH-R2) with high affinity to CRH-R1 in mammals. Urocortin-3 (UCN-3), a CRH family peptide, is an endogenous ligand for CRH-R2 in mammals. The physiological roles of UCN-3 and CRH-R2 have been investigated in mammals, although their roles still need to be clarified in chicks (Gallus gallus). Few studies have been performed comparing the physiological responses of CRH and UCN-3 in chicks. Therefore the present study was conducted to investigate the effect of intracerebroventricular (ICV) injection of UCN-3 on food intake, rectal temperature, crop-emptying rate and behaviors in chicks, and to compare these physiological responses with the effects resulting from CRH injection. The ICV injection of 20 and 80 pmol UCN-3 decreased food intake, increased rectal temperature and decreased crop-emptying rate and the results were similar to those achieved with CRH. The injection of both UCN-3 and CRH increased spontaneous activity but the behavioral patterns were different: CRH increased the number of vocalizations while UCN-3 increased the number of jumps, wing-flaps and scratching behaviors. These results suggest that UCN-3 regulates food intake, body temperature, and gastric emptying via the CRH-R2 in the brain of chicks, and these effects were similar to those induced by CRH.

Peripheral "chicken" obestatin administration does not affect feed intake and gut muscle contractility of meat-type and layer-type chicks (Gallus gallus domesticus).

Obestatin has recently been discovered in the rat stomach. As for ghrelin, the 23-amino acid obestatin is also derived from post-translational processing of the prepro-ghrelin gene but seems to have opposite effects on feed intake. In avian species, ghrelin is mainly present in the proventriculus and decreases feed intake, as opposed to its orexigenic properties in mammals. An obestatin-like sequence was also found in the avian ghrelin precursor protein but the potential involvement of this peptide in appetite regulation of chickens is unclear. We therefore investigated the effects of a single peripheral administration of this predicted "chicken" obestatin peptide on voluntary feed intake of 7- to 9-day-old meat-type and layer-type chicks. "Chicken" obestatin was injected intraperitoneally or intravenously at a dose of 1 nmol or 10 nmol/100 g body weight and feed intake was measured up to 4 h post injection. None of these treatments did reveal any effect of the putative "chicken" obestatin on appetite of either meat-type of layer-type chicks. Furthermore, "chicken" obestatin also failed to affect the in vitro contractility of muscle strips from crop and proventriculus. In conclusion, in the given experimental settings, the putative "chicken" obestatin has indistinctive physiological effects on feed intake and in vitro muscle contractility of gut segments, and hence its functional properties in ingestive behavior of avian species remain obscure.

Fossil evidence of avian crops from the Early Cretaceous of China.

The crop is characteristic of seed-eating birds today, yet little is known about its early history despite remarkable discoveries of many Mesozoic seed-eating birds in the past decade. Here we report the discovery of some early fossil evidence for the presence of a crop in birds. Two Early Cretaceous birds, the basal ornithurine Hongshanornis and a basal avian Sapeornis, demonstrate that an essentially modern avian digestive system formed early in avian evolution. The discovery of a crop in two phylogenetically remote lineages of Early Cretaceous birds and its absence in most intervening forms indicates that it was independently acquired as a specialized seed-eating adaptation. Finally, the reduction or loss of teeth in the forms showing seed-filled crops suggests that granivory was possibly one of the factors that resulted in the reduction of teeth in early birds.

Nutrient utilization and functionality of the anterior digestive tract caused by intermittent feeding and inclusion of whole wheat in diets for broiler chickens.

Two experiments were carried out to study the effect of intermittent feeding, diet structure, and their interaction on the performance, feeding pattern, and digestive adaptation of broiler chickens. In experiment 1, 40 Cobb broiler chickens were fed, either ad libitum or intermittently, a commercial starter diet diluted with 150 g/kg of ground or whole wheat in a 2 × 2 factorial arrangement. Ad libitum feeding consisted of continuous access to feed in a room with 18 h of light and 6 h of complete darkness, whereas birds on intermittent feeding had restricted access to feed from 7 d of age, with 4 one-hour feeding bouts/d and one 2-h feeding bout/d from d 14. No interaction was found between feed structure and feeding regimen for any of the measurements. Although BW gain was not significantly different among any of the treatments, birds given whole wheat or fed intermittently had significantly better feed efficiency than those given ground wheat and fed ad libitum. Inclusion of whole wheat instead of ground wheat also markedly increased (P < 0.001) the AME(n) value and fecal starch digestibility. In experiment 2, 60 Ross broiler chickens were trained to meal feeding on a wheat-based diet containing a commercial phytase, and 5 birds were killed every 15 min after having access to feed, to collect crop contents. Phytate was gradually degraded in the crop, with a 50% reduction in inositol 6-phosphate after a 100-min retention time. In conclusion, these results indicate that broiler chickens have a remarkable ability to maintain growth rate during intermittent feeding because the crop is used as an intermediate storage organ, and that improvements in feed efficiency may occur, among others through increased efficacy of exogenous enzymes. Even though stimulation of the gizzard through a coarse feed structure improves feed efficiency, it does not appear to affect the ability of the bird to handle intermittent feeding.

Dispersal of viable row-crop seeds of commercial agriculture by farmland birds: implication for genetically modified crops.

To address some concerns about the expansion of genetically engineered pharmaceutical and industrial crops to outdoor plantings and potential impacts on the human food supply, we determined whether commercial agriculture seeds of maize or corn Zea mays L., barley Hordeum vulgare L., safflower Carthamus tinctorius L. and rice Oryza sativa L. are digested or pass viably through the digestive tract, or are transported externally, by captive mallard ducks Anas platyrhynchos L., ring-necked pheasants Phasianus colchicus L., red-winged blackbirds Agelaius phoeniceus (L.) and rock pigeons Columba livia Gmelin (with the exception of whole maize seeds which were too large to feed to the blackbirds). These crop seeds, whether free-fed or force-fed, did not pass through the digestive tract of these bird species. The birds nonetheless did retain viable seeds in the esophagus/crop and gizzard for several hours. For example, after foraging for 6 h, mallards had retained an average of 228 +/- 112 barley seeds and pheasants 192 +/- 78 in the esophagus/crop, and their germination rates were 93 and 50%, respectively. Birds externally transported seeds away from the feeding location, but in only four instances were seeds found attached to their muddy feet or legs and in no case to feathers. Risk of such crop seeds germinating, establishing and reproducing off site after transport by a bird (externally or internally) or movement of a carcass by a predator, will depend greatly on the crop and bird species, location, environmental conditions (including soil characteristics), timing, and seed condition.

Central, but not peripheral, glucagon-like peptide-1 inhibits crop emptying in chicks.

We investigated the effect of central and peripheral glucagon-like peptide-1 (GLP-1) on crop emptying in growing chicks. Intracerebroventricular injection of two concentrations of GLP-1 (15 and 60 pmol) similarly suppressed crop emptying, compared with control chicks. The delay in crop emptying induced by GLP-1 (15 pmol) was partly attenuated by co-administration with exendin (5-39) (600 pmol), a GLP-1 receptor antagonist, although exendin (5-39) alone did not affect crop emptying. On the other hand, intraperitoneal administration of several doses of GLP-1 (120, 300 and 3000 pmol) did not alter crop emptying. The present study revealed that central, but not peripheral, GLP-1 inhibits crop emptying in chicks.

Release and endogenous actions of the gastrin/cholecystokinin (CCK) family in the chicken.

The regulation of cholecystokinin (CCK) and gastrin release in the chicken and their endogenous actions are summarized. Both dietary protein and amino acids stimulated CCK releases. Among dietary fat sources, medium-chain triacylglycerol (MCT) was a potent stimulator of CCK release compared with long-chain triacylglycerol (LCT). However, it is difficult to explain that endogenous CCK released by those stimulators has an important role in the avian gastrointestinal physiology. Luminal acids may be an important regulator in pancreatic enzyme and fluid secretion. Gastrin (a regulator of luminal acid secretion) release was stimulated by food components, strongly by MCT, but not by LCT, and weakly by some amino acids, and was inhibited by luminal acids. Luminal acids controlled food passage from the crop. In conclusion, gastrointestinal physiology may be directly regulated by luminal acid rather than by the gastrin/CCK family in the chicken.

Effect of ketone bodies on crop emptying in the chicken.

1. The effect of ketone bodies on crop emptying was studied in chickens in 2 experiments. In the first, the effect of beta-hydroxybutyrate, acetoacetate or acetone on relative crop content was measured. The effects of dietary medium and long chain triacylglycerols upon serum beta-hydroxybutyrate were investigated in the second. 2. beta-Hydroxybutyrate, but not acetoacetate and acetone, delayed crop emptying in a dose dependent fashion. Serum beta-hydroxybutyrate concentration was high in chicks given medium chain triacylglycerol, when compared with long chain triacylglycerol. 3. The results suggest that delayed crop emptying induced by medium chain triacylglycerol could partly be explained by an enhanced concentration of serum beta-hydroxybutyrate, which is the result of the rapid oxidation of medium chain fatty acids.

The influence of environmental temperature on in vivo limestone solubilization, feed passage rate, and gastrointestinal pH in laying hens.

Several studies were conducted to determine whether suppression of in vivo limestone solubilization was partially responsible for the reduction of shell quality under conditions of high environmental temperatures. In Experiment 1, excreta from hyperthermic and thermoneutral hens fed three levels of Ca (3.5, 4.3, and 5.2%) at two cycling environmental temperatures (averaging 28.3 and 26.1 C) were analyzed for percentage in vivo limestone solubilization. Hens in Experiment 2 received diets containing 3.9% Ca at 32.8 and 18.3 C. Rate of feed passage and gastrointestinal pH were also measured in Experiment 2. Experiment 3 evaluated the influence of temperature (22.2 and 30.0 C) and feed consumption on percentage limestone solubilization when Ca intake was held constant. In vivo limestone solubilization was influenced by Ca level in Experiment 1, but not by temperature. Hyperthermic hens solubilized a higher percentage of limestone than thermoneutral hens in Experiment 2, but it was concluded that this was due to a difference in Ca consumption and not due to temperature directly. When Ca intake was held constant in Experiment 3, there was no difference in limestone solubilization at the temperatures tested. Rate of feed passage was 16.6% slower in the hyperthermic hens. Crop, proventriculus, and upper small intestine pH were similar at each temperature, but gizzard pH was significantly lower in the hyperthermic hens. It was concluded that the high environmental temperatures used in the present studies did not suppress in vivo limestone solubilization.

Nocturnal digestion, cloacal excretion, and digestion-related thermogenesis in pigeons (Columba livia).

Two indicators of nocturnal digestive activity were identified in pigeons. Experiment 1 showed that a sizable amount of food empties from the crop while pigeons are inactive during the night. Experiment 2 showed that the number and volume of cloacal droppings during the night were directly related to the volume of food consumed during the day. The temporal pattern of cloacal droppings in the night was systematically related to features of the nocturnal body temperature (Tb) curves, suggesting that excretory activity is thermogenic in its own right and/or that it is a marker for a thermogenic process in the upper digestive tract. Questions about the relationship between digestion-related thermogenesis and shivering thermogenesis during the night in birds are highlighted by these findings.

Effects of partially hydrolysed guar gum on feeding behaviour and crop emptying rate in chicks.

1. The effects of partially hydrolysed guar gum (PHGG) or intact guar gum (GG) on feeding behaviour and crop emptying rate in growing chicks were investigated. 2. Several combinations of dietary PHGG and GG at 50 g per kg diets were prepared for a feed intake experiment. Birds (1 7-d-old) were given diets for 3 h after 16 h fasting, and food consumption was measured at 1 h intervals. The food intake rapidly decreased as the dietary GG content increased even at 1 h after feeding. 3. The rate of food passage from the crop was also investigated with birds (20-d-old after 16 h fasting. Birds were tube-fed diets having several ratios of dietary PHGG and GG. After 1 h of feeding, the diet remaining in the crop was measured after drying. The crop emptying rate decreased linearly as dietary PHGG concentration decreased. 4. The present study suggests that partial hydrolysis of dietary GG improve both feeding behaviour and food passage from the crop in growing chicks.

Intestine fullness influences feeding behaviour and crop filling in the domestic turkey.

Image-intensification radiology showed that artificial introduction of food slurry into the intestines of 6-12 wk-old turkey hens significantly increased the proportion of boli entering the crop during an evening meal, hence decreasing the proportion of boli travelling directly to the gizzard. Since bolus movement is directed by esophageal motility, esophageal motility may be partially mediated by stretch or chemoreceptors in the distal duodenum. Increased crop filling during afternoon meals lends support to the widely held belief that the crop increases gut storage capacity and helps "tide birds over" the nightly fast. Artificial filling of the distal duodenum paradoxically increased food intake in birds observed by radiology while eating a single meal (morning and evening), and in birds observed by eye over a 3 h period in the morning. This phenomenon is similar to that previously reported for rabbits and domestic chicken. Conversely, duodenal filing reduced food intake over a full day (11 h), more than compensating for the introduced food. Rapid filling of the small intestine (within 25 min. of the start of the meal) suggests a modification of the function of the domestic turkey duodenum to serve as a "mixing chamber," possibly enhancing digestive efficiency. The ways in which this modification affects digestion and absorption in the duodenum warrant investigation.