Diet and the evolution of digestion and renal function in phyllostomid bats

Bat species in the monophyletic family Phyllostomidae feed on blood, insects, small vertebrates, nectar, fruit and complex omnivorous mixtures. We used nitrogen stable isotope ratios to characterize bat diets and adopted a phylogenetically informed approach to investigate the physiological changes that accompany evolutionary diet changes in phyllostomids. We found that nitrogen stable isotopes separated plant-eating from animal-eating species. The blood of the latter was enriched in (15)N. A recent phylogenetic hypothesis suggests that with the possible exception of carnivory, which may have evolved twice, all diets evolved only once from insectivory. The shift from insectivory to nectarivory and frugivory was accompanied by increased intestinal sucrase and maltase activity, decreased trehalase activity, and reduced relative medullary thickness of kidneys. The shift from insectivory to sanguinivory and carnivory resulted in reduced trehalase activity. Vampire bats are the only known vertebrates that do not exhibit intestinal maltase activity. We argue that these physiological changes are adaptive responses to evolutionary diet shifts.     

The fetal and postnatal development of small intestinal disaccharidases in the rabbit

The development of small intestinal enzymes (lactase, acid- and hetero beta-galactosidases, cellobiase, maltase, trehalase, and sucrase) was studied from 18 days after conception until birth in 24 rabbit fetuses, and during the postnatal period in 15 newborn, juvenile, and adult rabbits. Lactase, acid- and hetero beta-galactosidases, cellobiase, and trehalase activities increased significantly during the fetal stage, while changes in sucrase and maltase activities were not substantial. In the postnatal period, lactase and cellobiase activities decreased significantly whereas maltase, sucrase, and trehalase activities increased significantly to reach adult values by 30 days of age. The acid- and hetero beta-galactosidases remained unchanged.     

A quantitative study of some digestive enzymes in the rabbitfish, Siganus canaliculatus and the sea bass, Lates calcarifer

Digestive enzyme distribution and activity in the digestive tracts of the rabbitfish, Siganus canaliculatus and the sea bass, Lates calcarifer were studied. Quantitative determinations of digestive enzymes in the guts of both fishes showed that they were capable of digesting carbohydrates and proteins in their diet. The carbohydrases, amylase, laminarinase, maltase, sucrase and trehalase were detected in the rabbitfish; their activities being mainly in the stomach, intestine and pyloriccaeca. Amylase, maltase, trehalase and chitinase activities were recorded in the gut of the sea bass, primarily in the intestine and the pyloriccaeca. Their activities were significantly lower than those in the rabbitfish. Proteases (pepsin, chymotrypsin, elastase, leucine aminopeptidase and trypsin) were found in both the rabbitfish and the sea bass. Pepsin activity however, was higher in the sea bass; while trypsin and chymotrypsin activities were higher in the rabbitfish. The activities of the various digestive enzymes in both fishes are discussed in relation to their feeding habits.     

Circadian rhythms in digestive enzymes in the small intestine of rats. I. Patterns of the rhythms in various regions of the small intestine.

The activities of the digestive enzymes, maltase [EC 3.2.1.20], sucrase [EC 3.2.1.26], trehalase [EC 3.2.1.28], Leucine aminopeptidase [EC 3.4.11.1], and alkaline phosphatase [EC 3.1.3.1] were measured in various regions of the small intestine of rats. The activities of all these enzymes were much higher in the jejunum than in the ileum, and in the distal regions of the ileum no sucrase, trehalase or alkaline phosphatase activity was detected. In the jejunum, the activities of all the enzymes tested exhibited clear circadian variations with the highest activity at 0000-0400 h and the lowest at 1200 h when the rats were fed ad libitum. In the ileum, maltase and sucrase also exhibited circadian variations, but the amplitude of the rhythm was smaller than that in the jejenum. Trehalase and alkaline phosphatase did not show any circadian variation in the ileum. Leucine aminopeptidase showed a circadian variation in the ileum with the same amplitude as in the jejunum. The phase of the circadian variations shifted about half a day when the rats were fed in the daytime, but the amplitude of the rhythm did not change.     

Origin of intestinal disaccharidases of Ascaris suum

Disaccharidases from the gut of Ascaris suum were investigated to determine whether they were synthesized by the worm or whether they were host enzymes adsorbed to the worms' intestinal cells. Alpha-d-glucoside glucohydrolase (maltase) (EC 3.2.1.20), Beta-d-fructofuranoside fructohydrolase (invertase) (EC 3.2.1.26) and 1-glucohydrolase (trehalase) (EC 3.2.1.28) from Ascaris were studied in both a membrane (brush border)-bound and solubilized form with regard to temperature stability and pH optima. Data collected were compared to similar data on hog intestinal enzymes. Worm maltase and trehalase were relatively heat labile, whereas the hog enzymes were more stable to heat inactivation. Worm invertase was heat stable in comparison to the hog enzyme. The pH optima for Ascaris maltase and invertase were different from those of hog disaccharidases, whereas the pH optimum for trehalase from both parasite and host were similar. Tissue homogenates of second-stage larvae contained measurable maltase, but not sucrase, or trehalase activity. Results suggested that Ascaris intestinal disaccharidases represent three distinct enzymes of parasite rather than host origin.     

Food composition and digestive enzymes in the gut of pond-cultured Clarias isheriensis (Sydenham 1980), (Siluriformes: Clariidae)

Food composition, diestive enzyme distribution and activity in the gut of pond-cultured Clarias isheriensis were studied It had an omnivorous diet but fed mainly on plancon (particularly Cyanophyceae) and detritus. Qualitative determinations of digestive enzymes in the gut showed that it is capable of digesting carbohydrates, proteins and lipids in its diet. Carbohydrases (amylase, cellulase, maltase, salicinase, sucrase and trehalase) were detected and their activities restricted to the stomach, duodenum and ileum. The incidence of cellulase activity could be responsible for the capacity of this fish species to digest large quantities of Cyanophyceae present in the pond. Lower activity of proteases (chymotrypsin, pepsin and trypsin) and lipases were recorded. Enzyme activity was not recorded in either oesophagus or rectum. The relative distribution and activity of the various digestive enzymes were possibly induced by the nutritional requirements of this catfish species.     

Activity of renal hydrolases in pre- and postnatal development of mice

The fetal and postnatal activity patterns of different hydrolytic enzymes (alkaline phosphatase, gamma-glutamyltransferase, trehalase, maltase, glucoamylase, lactase, and sucrase) have been examined in mouse renal homogenates. Alkaline phosphatase and gamma-glutamyltransferase activities presented approximately similar changes. They increased from 18 days of gestation up to 30 days after birth. These activities showed marked increases during the 3rd and 4th postnatal weeks. A similar important rise was observed for trehalase activity at the end of the suckling period. Maltase activity increased gradually after birth. Traces of lactase, sucrase, and glucoamylase activities were detected at each developmental stage.     

Are levels of digestive enzyme activity related to the natural diet in passerine birds?

Digestive capabilities, such as the rates nutrient hydrolysis and absorption, may affect energy intake and ultimately feeding behavior. In birds, a high diversity in gut biochemical capabilities seems to support the existence of a correlation between the morphology and physiology of the intestinal tract and chemical features of the natural diet. However, studies correlating the activity of digestive enzymes and the feeding habits at an evolutionary scale are scarce. We investigated the effect of dietary habits on the digestive physiological characteristics of eight species of passerine birds from Central Chile. The Order Passeriformes is a speciose group with a broad dietary spectrum that includes omnivorous, granivorous and insectivorous species. We measured the activity of three enzymes: maltase, sucrase and aminopeptidase-N. Using an autocorrelation analysis to remove the phylogenetic effect, we found that dietary habits had no effect on enzymatic activity. However, we found that granivorous and omnivorous species had higher levels of disaccharidase activities and insectivores had the lowest. The major difference in enzymatic activity found at the inter-specific level, compared to the reported lower magnitude of enzyme modulation owing to dietary acclimation, suggests that these differences to some extent have a genetic basis. However, the lack of a clear association between diet categories and gut physiology suggested us that dietary categorizations do not always reflect the chemical composition of the ingested food.     

Intestinal brush border membrane-bound disaccharidases of the American alligator, Alligator mississippiensis

1. The disaccharidase activities of the small intestines of American alligators (Alligator mississippiensis) were studied in epithelial scrapes and brush-border membrane preparations. 2. Maltase, isomaltase and trehalase activities were found. Activities of these enzymes were higher in the proximal small intestine and decreased distally. 3. Disaccharidase activities were enriched 12-15 times in brush-border membrane preparations, compared with mucosa/enterocyte crude homogenates and were co-enriched with the brush-border membrane marker alkaline phosphatase. 3. The pH optima were: maltase 6.5; isomaltase 5.6; and trehalase 5.8. The Q10 of maltase, the most active enzyme, was equal to 1.82. 4. In reptiles, as in mammals, disaccharidase activities may be correlated with feeding habits. The co-occurrence of sucrase and isomaltase may not be a common feature of vertebrates.     

Enzymes of the human intestinal brush border membrane. Identification after gel electrophoretic separation

The position of a number of human intestine brush border membrane enzyme activities in polyacrylamide gels after electrophoresis has been determined. These activities are, in order from the origin, maltase/glucoamylase, lactase/phlorizin hydrolase, maltase/sucrase/isomaltase, enteropeptidase, trehalase and γ-glutamyltransferase. Leucylnaphthylamide hydrolyzing activity was inactivated by sodium dodecylsulfate and its position was not determined. The positions of the activities have been correlated with the positions of protein bands previously determined. One such band situated between enteropeptidase and alkaline phosphatase has not been identified.     

Enzymes of the human intestinal brush border membrane. Identification after gel electrophoretic separation.

The postition of a number of human intestine brush border membrane enzyme activities in polyacrylamide gels after electrophoresis has been determined. These activities are, in order from the origin, maltase/glucoamylase, lactase/phlorizin hydrolase, maltase/sucrase/isomaltase, enteropeptidase, trehalase and gamma-glutamyl-transferase. Leucylnaphthylamide hydrolyzing activity was inactivated by sodium dodecylsulfate and its position was not determined. The positions of the activities have been correlated with the positions of protein bands previously determined. One such band situated between enteropeptidase and alkaline phosphatase has not been identified.     

The level and distribution of disaccharidases in the camel (Camelus dromedarius) intestine

1. The disaccharidases, cellobiase, isomaltase, lactase, maltase, sucrase and trehalase were investigated for presence in the camel (Camelus dromedarius) intestine and pancreas. All, except sucrase, were present. 2. Their levels of activities were measured at different positions of the small and large intestines and the location of maximum level of activity for each enzymes along the intestinal tract was established. 3. High levels of activities were determined in the contents of the intestinal lumen and, therefore, it is absorbed into the cells of the epithelial villi and hydrolyzed there. 4. The possibility of carbohydrate digestion in camel intestine is discussed.     

Carbohydrase activities in the bovine digestive tract

1. The carbohydrase activities of homogenates of mucosa from the abomasum, small intestine, caecum and colon, and of the pancreas of cattle were studied. 2. The disaccharidase activities were located mainly in the small intestine and showed a non-uniform pattern of distribution along the small intestine; trehalase activity was highest in the proximal part, lactase and cellobiase activities were highest in the proximal and middle parts and maltase activity was highest in the distal part. 3. The intestinal lactase and cellobiase activities were highest in the young calf and decreased with age, whereas the intestinal maltase and trehalase activities, which were very low compared with the lactase activity, did not change with age. 4. No intestinal sucrase or palatinase activity was detected in the calf or in the adult cow. 5. Homogenates of intestinal mucosa also exhibited amylase and dextranase activity. 6. Homogenates of the pancreas possessed a strong amylase activity and a weak maltase activity. The maltase activity did not change with age, whereas the amylase activity increased with age. 7. No marked differences were observed between the carbohydrase activities of calves fed solely on milk and those of calves given a concentrate-hay diet from 6 weeks of age.     

Insulin accelerates the development of intestinal brush border hydrolytic activities of suckling mice

The influence of insulin on the postnatal development of intestinal brush border membrane disaccharidase (sucrase, maltase, trehalase, lactase) and peptidase (leucylnaphthylamidase, γ-glutamyltranspeptidase) activities has been studied in mice. At 8 days of age, the animals received either 5, 10, or 12.5 mU insulin/g body wt/day during 3 days. A premature appearance of sucrase activity was noted, the level of sucrase activity being dependent of the amount of insulin injected. Maltase and lactase activities were both increased while trehalase activity was affected only by the highest dose of insulin. The behavior of the two peptidases was quite different as γ-glutamyltranspeptidase was prematurely increased and leucylnaphthylamidase was unaffected by insulin. The hormonal effect is exerted along the entire small intestine. The time course of the responses of the disaccharidases in relationship to cellular migration along the crypt-villus axis has also been studied. By 24 hr after administration of a single injection of 12.5 mU insulin/g body wt, sucrase activity was already present and an increased maltase and trehalase activities were observed. During the subsequent 72 hr no further increase of enzymatic activity was noted even though the epithelial cells are moving up on the villi at a faster rate than in controls, thus indicating that there is no relationship between the enzymatic responses and the cellular migration. The present data show that a premature increase of the circulating level of insulin influences the development of intestinal mucosa in suckling mouse.     

Effect of tannic acid on brush border disaccharidases in mammalian intestine

Tannic acid is a glucoside (penta-m-digallolyl-glucose), which exhibits a wide variety of physiological functions. Around neutral pH, 0.4 mM tannic acid produced 84% inhibition of rat brush border sucrase activity, but 35-40% enzyme inhibition was observed in the rabbit intestine at 0.08 mM concentration. In the mice, 74-77% enzyme inhibition was observed at 0.05 mM concentration of tannic acid. The observed inhibition was reversible in rat intestine. Tannic acid (0.2 mM) also inhibited lactase (18% in adult and 71% in suckling animals), maltase (76%) and trehalase (88%) activities in rat intestine. pH versus activity curves showed that 0.2 mM tannic acid inhibited enzyme activity in rat by 91% at pH 5.5 which was reduced to 14% at pH 8.5 compared to the respective controls. In the rabbit 18-60% enzyme inhibition was noticed below pH 7.0, however at pH 8.5, it was of the order of 38%. Kinetic analysis revealed that tannic acid is a competitive inhibitor of rat brush border sucrase at pH 6.8. Effect of tannic acid together with various -SH group reacting reagents revealed that the enzyme inhibition is additive in nature, suggesting the distinct nature of binding sites on the enzyme for these compounds. The results suggest that tannic acid is a potent inhibitor of intestinal brush border disaccharidases, and could modulate the intestinal functions.     

Test of the adaptive modulation hypothesis in rodents: dietary flexibility and enzyme plasticity.

The phenotypic response of digestive enzymes was assessed in two species of rodents with different foods habits. Species were Phyllotis darwini (omnivorous) and Octodon degus (herbivorous). The activity of sucrase, maltase and aminopeptidase-N were determined in vitro in animals feeding two contrasting diets. No effect of dietary chemistry on sucrase and maltase activities was observed. Nevertheless, aminopeptidase-N showed a reversible response to diet in P. darwini but not in O. degus. Through Principal Component Analysis we separated the specific and non-specific modulation of the enzymes. The analysis showed that aminopeptidase-N activity is up-regulated by dietary protein in P. darwini. Differences in the phenotypic response of this species apparently reflect the historic levels of specific substrates of the natural diets for this enzyme, linking dietary flexibility and digestive plasticity in an evolutionary context.     

Intestinal disaccharidases in five species of phyllostomoid bats.

1. Intestinal disaccharidases were studied in nectarivorous (Leptonycteris curasoae and Glossophaga soricina), frugivorous (Artibeus jamaicensis and Sturnira lilium), and insectivorous (Pteronotus personatus) adult bats. 2. Adult bats lacked measurable lactase activity. With the exception of trehalase activity, which was present only in P. personatus, nectar- and fruit-eating bats exhibited higher disaccharidase activities standardized by intestinal nominal area than insect-eating P. personatus. 3. Maltase and sucrase activities were significantly linearly correlated. 4. Apparent affinity of sucrase varied almost 5-fold among species. This variation may reflect unstirred layer effects resulting from sucrase being a membrane bound enzyme rather than differences in the "true" affinity of sucrase in solution. 5. Passerine birds showed higher maltase activity per unit of sucrase activity than bats and hummingbirds. Maximal sucrase and maltase activities standardized per intestinal nominal area are 1.5-2 times higher in hummingbirds than in nectar-feeding bats.     

Maltase-glucoamylase and trehalase in the rabbit small intestine and kidney brush border membranes during postnatal development, the effects of hydrocortisone

Kidney and intestinal brush border membranes were isolated from 14-day-old rabbits and papa?n solubilized maltase-glucoamylase was purified to almost homogeneity from both membranes. Maltase-glucoamylase from kidney and intestine have the same molecular weight (669,000 daltons by AcA 22 gel filtration) and the same Km (4 mM, for maltose). Tris (Ki = 12.5 mM, for maltose) is a non-competitive inhibitor for both enzymes. In intestine, maltase and glucoamylase have low activity during the first two postnatal weeks and then undergo a sharp increase during the next 2 weeks. In contrast, for trehalase, adult levels are reached about 6 days after birth. Hydrocortisone injection to 10 days rabbits causes precocious increases in the specific activities of trehalase (3.6 x), maltase (5.2 x) and glucoamylase (7.4 x). Conversely, kidney maltase, glucoamylase and trehalase activities rise gradually from birth, reaching adult levels by the end of the third week. Administration of hydrocortisone to suckling rabbit does not affect either trehalase or maltase and glucoamylase in kidney brush border membrane.     

Effect of glucose and insulin on small intestinal brush border enzymes in fasted rats

Fasting reduced small intestinal length. It also decreased mucosal weight, DNA and protein content, and concentrations of enterokinase, maltase, and sucrase in both duodenal and jejunal segments. In contrast, the concentrations of lactase and leucine aminopeptidase were not affected. Concomitantly, serum insulin levels dropped to one-fifth of the control levels while serum glucose concentrations showed a lesser degree of reduction. Glucose supplementation alone raised the serum insulin level, prevented the decrease in DNA content, and showed a protective effect on mucosal protein, mucosal weight, mucosal thickness, and villus height. Glucose also protected the sucrase and maltase concentrations; more significantly for maltase in the jejunal segment. Insulin alone, although it increased the serum insulin level to that found with glucose supplementation alone, had no protective effect on the loss in protein, DNA, and most enzymes except for maltase concentration in the jejunal segment. Addition of insulin to glucose did not modify the glucose effect on the contents of DNA, protein, and concentrations of sucrase and maltase. These results suggest that the glucose effect on the mucosa is not mediated by insulin. In addition, the retention of both maltase and sucrase activities through only glucose supplementation suggests the loss of maltase and sucrase in fasting is due to nutrient rather than specific substrate restriction.