Intestinal Digestive Enzyme Activities Research Articles

Effects of practical dietary protein to lipid levels on growth, digestive enzyme activities and body composition of juvenile rice field eel (Monopterus albus)

A 10-week feeding experiment was conducted to evaluate the effects of dietary protein to lipid ratios (P/L) on growth, intestinal digestive enzyme activities and body composition in juvenile rice field eel (Monopterus albus) (initial mean body weight of 65.76 ± 1.07 g, mean ± SEM). Nine test diets were formulated in a 3 × 3 factorial design to contain three protein levels (350, 400 and 450 g kg−1) for each of three lipid levels (40, 80 and 120 g kg−1), respectively. Each diet was randomly assigned to triplicate groups of 60 fish per net cage (1.5 × 2.0 × 1.5 m). Results showed the survival was above 96 % and was not affected by dietary treatments. Fish fed the diet with 450 g kg−1 protein and 40 g kg−1 lipid showed the best weight gain (WG) (103.95 %) and feed conversion ratio (1.60) (P < 0.05). WG, protein efficiency ratio and energy retention increased with the increasing in lipid at 350 g kg−1 protein level (P < 0.05). However, WG showed a little decline with increasing dietary lipid when fish fed the diets with 400 and 450 g kg−1 protein level, but no significant difference was observed (P < 0.05). Hepatosomatic index, visceralsomatic index and intestinal lipase activity increased with the increasing of dietary lipid level irrespective protein level. Intestinal trypsin activity increased with the increasing of dietary lipid level when fish fed the diets with 350 g kg−1 protein, but showed converse trend when fish fed the diets with 400 and 450 g kg−1 protein. Serum triglyceride, body lipid and energy were positively correlated with the dietary lipid. Results of the present study showed that the dietary protein/lipid ratio of 450/40 g kg−1 is considered optimum for rice field eel under culture conditions, and the increase in dietary lipid level has no efficient protein-sparing effect when fish fed the diets with 400 and 450 g kg−1 protein level.

Effect of dietary supplementation with an ethanolic extract of propolis on broiler intestinal morphology and digestive enzyme activity

The present study aimed to evaluate the effect of different levels of an ethanolic extract of propolis (EEP) on broiler performance, carcass characteristics, weight of gastrointestinal organs, intestinal morphometry and digestive enzyme activity. 1020 male broiler chicks were assigned in a completely randomised experimental design to six treatments (EEP supplement levels of 0, 1000, 2000, 3000, 4000 and 5000 ppm) and five replications, and 34 birds per experimental unit. The experimental diets were administered from 1 to 21 days of age, and the birds were subsequently provided a ration based on corn and soybean meal. EEP supplementation from 1 to 7 days negatively affected (p < 0.05) the weight gain and feed intake. The proventriculus weight at 7 days exhibited a quadratic response (p < 0.05), which predicted a lower weight at a dose of 2865 ppm of the EEP. For the duodenum at 21 days of age, the response pattern (p < 0.05) predicted that birds that were fed 2943 and 3047 ppm of the EEP would exhibit an improved crypt depth and villus-to-crypt ratio respectively. The villus height, crypt depth and villus-to-crypt ratio in the jejunum and the ileum were not affected (p > 0.05). With increased EEP doses, the duodenal sucrase activity linearly decreased at 7 days of age and linearly increased in the jejunum at 21 days of age (p < 0.05), while pancreatic enzyme activity was unaffected (p > 0.05). Although the carcass and cut yields did not improve, the percentage of abdominal fat decreased (p < 0.05). The supplementation of the broiler pre-starter diet with 1000-5000 ppm of the EEP impaired performance at this stage, most likely due to the decreased sucrase activity. However, the EEP supplementation from 3000 ppm improved intestinal morphophysiology at 21 days of age and did not affect the performance or carcass yield at 42 days of age.

Application of autochthonous Bacillus bioencapsulated in copepod to grouper Epinephelus coioides larvae

Probiotic administration to fish larvae via live feeds such as rotifers and Artemia has been reported extensively. However, no information is available about the application of probiotics via copepods, one of the most appropriate live feeds for many marine fish larvae. This study evaluated the application of autochthonous probiotic Bacillus clausii DE5 and Bacillus pumilus SE5 bioencapsulated in copepods to orange-spotted grouper Epinephelus coioides larvae. The larvae were fed with control copepod (Pseudodiaptomus annandalei) or probiotic-enriched copepods from day 40 to day 68 after hatching. Increased total length and body weight were observed in larvae fed with the two probiotics for 14days. The improved growth may be related to the enhanced activities of intestinal digestive enzymes, such as alkaline phosphatase and acid phosphatase. Improved survival was also observed in probiotic fed larvae and may be associated with changes in immune defense, as evidenced by increased lysozyme and SOD activities. On the other hand, several potentially pathogenic bacteria such as Aliivibrio fischeri, Vibrio scophthalmi and Vibrio sp. were suppressed and a desirable gut microbiota was induced by probiotic B. clausii DE5 and B. pumilus SE5, which may contribute to the improvement of health and the development of gastrointestinal (GI) tract in larval grouper E. coioides. Therefore, our results suggest that copepod (P. annandalei) is suitable to act as vector of probiotics in marine fish larvae.

Effects of Neutral Phytase Supplementation on Biochemical Parameters in Grass Carp, Ctenopharyngodon idellus, and Gibel Carp, Carassius auratus gibelio, Fed Different Levels of Monocalcium Phosphate

AbstractTwo 8‐wk studies were conducted to evaluate the effects of neutral phytase supplementation on hemato‐biochemical status, liver biochemical parameter, and intestinal digestive enzyme activity of grass carp, Ctenopharyngodon idellus, and gibel carp, Carassius auratus gibelio, fed with different levels of monocalcium phosphate (MCP). The control diet was prepared with 2% MCP but without phytase (P2.0). The other three experimental diets were prepared with the addition of 1.5, 1.0, and 0.5% MCP, respectively, when supplemented with 500 U/kg neutral phytase in each diet and designated as PP1.5, PP1.0, and PP0.5, respectively. The results indicated that the serum alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate transaminase (AST) activities, as well as the albumin (ALB) content were increased in grass carp (P &lt; 0.05) and gibel carp (P &gt; 0.05) fed with phytase‐supplemented diets. Meanwhile, the serum cholesterol, high‐density lipoprotein, and total protein contents of the two species of fish were increased in comparison to the control. In addition, dietary phytase inclusion did not significantly affect hepatic ALP, ALT, and AST activities in the two species of carp fed with different levels of MCP. Amylase activity increased in foregut and hindgut of both species when fed with the phytase‐supplemented diets while lipase activity was reduced in the foregut and hindgut in both fish. This study suggests that neutral phytase supplementation increases serum ALP, ALT, and AST activities but does not notably affect these enzyme activities in the liver of the two species of carp when fed different levels of MCP. On the other hand, amylase activity increased while lipase activity was reduced in the intestine of the species of carp fed with phytase‐supplemented diets.

The in vivo infusion of hydrogen peroxide induces oxidative stress and differentially affects the activities of small intestinal carbohydrate digestive enzymes in the neonatal pig1

Chronic fatigue syndrome (CFS) is characterized by persistent and relapsing fatigue that involves oxidative stress in its pathogenesis. We tested the hypothesis that a decrease in key carbohydrate-digesting enzyme activity in the gut is one of the major biological mechanisms of developing CFS in liquid formula-fed neonatal pigs with in vivo infusion of H(2)O(2). Piglets at 7 to 10 d of age were fitted with an intraperitoneal catheter, allowed a 3-d post surgical recovery, and infused with either H(2)O(2) at 5 mmol/kg BW (PER; n = 8) or the same volume of saline (CON; n = 8) in six 20-ml doses daily for a period of 10 d. During this period, animal behavior was monitored, blood samples collected, and jejunal enzyme activity kinetic experiments for lactase, sucrase, maltase, and maltase-glucoamylase were conducted. Plasma concentration of reduced glutathione remained similar (P > 0.05) to the pre-infusion level over the study duration in the CON group whereas this was 65% lower (P < 0.05) than the pre-infusion level in the PER group. Piglets experiencing oxidative stress had an overall lower (P < 0.05) physical mobility and the maximal jejunal specific activities [μmol/(mg protein · min)] for lactase (PER, 6.54 ± 0.68 vs. CON, 12.65 ± 0.69) and maltase (PER, 57.39 ± 1.02 vs. CON, 75.60 ± 1.04), respectively. However, differences were not observed (P > 0.05) in the maximal specific activities [μmol/(mg protein · min)] of sucrase (PER, 10.50 ± 1.37 vs. CON, 12.40 ± 1.55) and maltase-glucoamylase (PER, 0.71 ± 0.08 vs. CON, 0.70 ± 0.07) between the 2 groups. In conclusion, infusion of a suitable dose of H(2)O(2) induced CFS in the neonatal pigs. Oxidative stress in vivo differentially affected the maximal activities of important small intestinal carbohydrate-digesting enzymes in neonatal pigs fed a dairy milk-based liquid formula.

Effects of Replacement of Fish Meal by Soy Protein Isolate on the Growth, Digestive Enzyme Activity and Serum Biochemical Parameters for Juvenile Amur Sturgeon (Acipenser schrenckii).

An 8-wk experiment was conducted to evaluate the effect of replacing fish meal (FM) with soy protein isolate (SPI) on the growth, digestive enzyme activity and serum biochemical parameters of juvenile Amur sturgeon (Acipenser schrenckii). SPI was used to replace 0, 25, 50, 62.5, 75, 87.5, 100% of dietary FM and 100% replacement supplemented crystalline amino acid. Healthy sturgeon with an average initial weight of 26.38±0.24 g were randomly assigned to 24 aquaria (8 treatments with triplicates each) at an initial stocking density of 11 fish per aquarium and cultured for 8 wks. The results showed that 75.00% or more substitution resulted in a poor weight gain rate, feed conversion ratio and survival rate compared to that of fish fed the control diet (p<0.05), whereas no significant differences were observed between diets of 25.00% to 62.50% substitution. Protease, lipase and amylase activity in foregut, mid-gut and hindgut were significantly (p<0.05) decreased by diets where SPI replacement levels were 62.50% or more. Levels of serum total protein (TP) and globulin decreased significantly from 21.03, 10.34 to 14.05, 5.63 g/L with the increasing dietary SPI (p<0.05), but alkaline phosphatase activity significantly increased (p<0.05). In addition, supplemental crystalline amino acid in the FM absence diet did not improve growth performance, intestine digestive enzyme activities and serum biochemical parameters. In conclusion, the results from this study showed adverse effects of inclusion of SPI in diets on growth performance, feed utilization and serum biochemical parameters in juvenile Amur sturgeon. Based on WGR and replacement ratio presented in this report, a 57.64% replacement level was recommended.

Partial replacement of monocalcium phosphate with neutral phytase in diets for grass carp, Ctenopharyngodon idellus

Summary A 9-week experiment was designed to study the effects of partial replacement of monocalcium phosphate (MCP) with neutral phytase on growth, body compositions, serum biochemical statuses and intestinal digestive enzyme activities of grass carp, Ctenopharyngodon idellus. The control diet (designated as P2.0) was prepared with 2.0% MCP but without phytase. The three other diets (designated as PP1.5, PP1.0 and PP0.5, respectively) were supplemented with 1.5, 1.0 and 0.5% MCP, respectively, along with 500 FTU of neutral phytase kg−1 diet in each. After a 9-week feeding trial, fish (initial body weight: 43.44 ± 2.37 g) fed with PP1.5 and PP1.0 had no significant change in weight gain (WG), specific growth rate (SGR), protein efficiency rate (PER) or feed conversion ratio (FCR) compared with the control (P > 0.05) whereas fish fed with PP0.5 showed significantly lower growth performance in the above parameters. The crude lipid content in whole body or muscle of the fish fed with PP1.5 was significantly lower than the control while significantly higher in fish fed with PP0.5 (P < 0.05), whereas no obvious change was observed in the fish fed with PP1.0. For serum indices, higher serum alkaline phosphatase (Alkp), phosphorus (P) and calcium (Ca) contents were observed in fish fed with phytase-supplemented diets in comparison with the control. In addition, dietary phytase supplementation increased amylase activity and decreased lipase activity in both foregut and hindgut. The present study suggests that dietary MCP can be reduced when neutral phytase is added to the grass carp diet, and that the maximum MCP reduction level can be up to 1% when neutral phytase is supplemented at 500 FTU kg−1 diet.

Impaired Small-Bowel Barrier Integrity in the Presence of Lumenal Pancreatic Digestive Enzymes Leads to Circulatory Shock

In bowel ischemia, impaired mucosal integrity may allow intestinal pancreatic enzyme products to become systemic and precipitate irreversible shock and death. This can be attenuated by pancreatic enzyme inhibition in the small-bowel lumen. It is unresolved, however, whether ischemically mediated mucosal disruption is the key event allowing pancreatic enzyme products systemic access and whether intestinal digestive enzyme activity in concert with increased mucosal permeability leads to shock in the absence of ischemia. To test this possibility, the small intestinal lumen of nonischemic rats was perfused for 2 h with either digestive enzymes, a mucin disruption strategy (i.e., mucolytics) designed to increase mucosal permeability, or both, and animals were observed for shock. Digestive enzymes perfused included trypsin, chymotrypsin, elastase, amylase, and lipase. Control (n = 6) and experimental animals perfused with pancreatic enzymes only (n = 6) or single enzymes (n = 3 for each of the five enzyme groups) maintained stable hemodynamics. After mucin disruption using a combination of enteral N-acetylcysteine, atropine, and increased flow rates, rats (n = 6) developed mild hypotension (P < 0.001 compared with groups perfused with pancreatic enzymes only after 90 min) and increased intestinal permeability to intralumenally perfused fluorescein isothiocyanate-dextran 20 kd (P < 0.05) compared with control and enzyme-only groups, but there were no deaths. All animals perfused with both digestive enzymes and subjected to mucin disruption (n = 6) developed hypotension and increased intestinal permeability (P < 0.001 after 90 min). Pancreatic enzymes were measured in the intestinal wall of both groups subjected to mucin disruption, but not in the enzyme-only or control groups. Depletion of plasma protease inhibitors was found only in animals perfused with pancreatic enzymes plus mucin disruption, implicating increased permeability and intralumenal pancreatic enzyme egress in this group. These experiments demonstrate that increased bowel permeability via mucin disruption in the presence of pancreatic enzymes can induce shock and increase systemic protease activation in the absence of ischemia, implicating bowel mucin disruption as a key event in early ischemia. Digestive enzymes and their products, if allowed to penetrate the gut wall, may trigger multiorgan failure and death.

Posthatch development of intestinal morphology and digestive enzyme activities in domestic pigeons (Columba livia)

The gastrointestinal tract plays an important role in studies of avian ontogenies because of its key function in nutrient intake. The pattern of intestinal development during postnatal growth in domestic pigeons (Columba livia), as an altricial bird, would be different from the pattern observed in precocial poultry. In this study, the intestinal morphology, mucosal enzyme activities, and pancreatic enzyme activities were determined from hatch to 14 d of age in pigeons. Rapid morphological development was observed immediately after hatching. Villus area and crypt depth increased in parallel in the duodenum and jejunum and more slowly in the ileum. Mucosal enzyme activities per gram of duodenum, jejunum, and ileum varied significantly with segment and age. There were also intestinal segment × age interactions, in which enzyme activity increased most dramatically in the jejunum as compared with the duodenum and ileum for sucrase and maltase activity from day of hatch to 8 d, and for alkaline phosphatase activity through 14 d. However, aminopeptidase-N activity increased most dramatically from day of hatch to 3 d in the ileum as compared with the duodenum and jejunum. Changes in carbohydrate-digesting enzymes activity in duodenum, jejunum, ileum, and pancreas are more pronounced than for protein-digesting enzymes activity from 3 d after hatching. Intestinal total enzyme activities exhibited a steady increase, which was highly correlated with BW. Pancreatic enzymes indicated patterns somewhat similar to those for intestinal enzymes. It was concluded that development of the absorptive area and the capacity for hydrolysis in the duodenum, jejunum, and ileum was rapid immediately after hatching, although rates of development were different in the 3 segments. Young pigeons differ from poultry, in whom intestinal and pancreatic digestive enzyme activities per gram of tissue are generally constant or decline during postnatal development. Also, the data seem consistent with the idea that intestinal hydrolysis may be a determining step in digestion and hence growth.

Effect of feeding on the function and structure of the digestive system in juvenile southern catfish (Silurus meridionalis Chen)

Postprandial physiological and morphological responses to feeding were examined in juvenile southern catfish (Silurus meridionalis Chen) that had consumed a loach (Misgurnus anguillicaudatus Cantor) meal equivalent to 6% of the body mass of the catfish. The gastric evacuation rate (GER) peaked at 4h postfeeding, averaging 0.36g food weighth(-1), at which time 14% of the ingested meal had passed into the intestine. Less than 10% of the ingested meal remained in the stomach at 24h postfeeding. Pepsin activity peaked at 8h postfeeding, reaching a level approximately twofold higher than the prefeeding level. Pancreatic trypsin activity peaked at 16h postfeeding, reaching a level 4.5-fold higher than the prefeeding level. Peaks in lipase activity in both the proximal and middle intestinal segments occurred at 16h, reaching 2.8- and 2.4-fold higher levels than the prefeeding level, respectively, while the activity in the distal intestine segment reached a level 2.9-fold higher than the prefeeding level at 24h postfeeding. With respect to amylase activity, only the middle intestinal segment exhibited a change, first an increase and then a decrease, after feeding. Feeding also triggered an approximately 200% increase in the metabolic rate and resulted in 44.6kJkg(-1) being expended on specific dynamic action, equivalent to 16.1% of the meal's energy. In terms of organ size, the wet mass of the liver increased by 11% at 24h postfeeding, whereas the wet mass of the pancreas did not change. Except for a decrease in the thickness of the submucosa in the middle intestinal segment, the thickness of the intestinal fold, mucosa, submucosa, muscularis and serosa of each intestinal segment did not change significantly with feeding. These results suggest that the continuum of physiological responses observed with respect to metabolic increases, GER, regulation of pancreatic and intestinal digestive enzyme activities and liver wet mass to feeding corresponds to the changes in the demand on the digestive system in S. meridionalis. Moreover, species maintained stable gastrointestinal tract morphology during the short interval of repeated feeding.

Effects of Wei Chang An pill on enzyme activity and levels of vasoactive peptide and substance P in the small intestine of rats with compound diarrhea

ObjectiveTo investigate the regulatory effects of Wei Chang An Pill (WCAP) on enzyme activity and gastrointestinal hormones in the small intestine of rats with compound diarrhea. MethodsForty Wistar rats were randomly divided into a control, diarrhea model, and WCAP high, medium, and low dose groups. The control group was not treated, and the model group was administered intragastric distilled water. The WCAP groups were given WCAP suspension, 80, 60 or 40 mg · kg−1 · d−1, for 4 days. Stool properties were observed. After the experiment, thymus and spleen indices were measured, and the activities of lactate dehydrogenase (LDH), malate dehydrogenase (MDH), and disaccharidase (lactase) in the small intestinal mucous membrane, and levels of substance P (SP) and vasoactive peptide (VIP) in the colon were determined. ResultsCompared with the control group, thymus and spleen indices were significantly decreased, LDH, MDH, and disaccharidase activity in the small intestine was decreased, and SP and VIP levels in the colon were significantly increased in the diarrhea model group. Compared with the model group, thymus and spleen indices were significantly increased, and LDH, MDH, and disaccharidase activity in the small intestine and SP and VIP levels in the colon were significantly decreased in the WCAP medium dose group. ConclusionThe diarrhea model rats exhibited pathological changes including atrophy of the thymus and spleen, decreased enzyme activity in the small intestine, and gastrointestinal hormone disturbance. WCAP can increase the activity of intestinal digestive enzymes and regulate gastrointestinal hormones, thereby relieving diarrhea.

Effects of xylooligosaccharide on growth performance, activities of digestive enzymes, and intestinal microflora of juvenile Pelodiscus sinensis

The effects of xylooligosaccharide (XOS) on growth performance, activities of digestive enzymes, and intestinal microflora of Chinese soft-shelled turtle Pelodiscus sinensis were investigated by adding different concentrations of XOS in the diet of juvenile P. sinensis. The turtles with an average initial bodyweight of 12.26±0.32 g were randomly divided into five groups, which were fed with diets containing 0 mg/kg (control), 50 mg/kg (Trial I), 100 mg/kg (Trial II), 200 mg/kg (Trial III), and 500 mg/kg (Trial IV) XOS product (95%), respectively. The feeding trial lasted for 30 days. Indices described above were determined at the end of the experiment. The results showed that the diet supplemented with XOS could enhance the growth of P. sinensis, and the growth rate of Trial III was the highest. Feed conversion ratio (FCR) of the trials was lower than that of the control. All the trials had higher activities of intestinal digestive enzymes than the control. The amylase activity of Trial II was the highest and was significantly different from that of the control, while the protease activity of the trials was not significantly but slightly higher than that of the control. The numbers of total aerobic bacteria and Enterobacteria of all the trials were significantly lower than those of the control. The number of Bifidobacteria of Trial II ranked the largest, significantly higher than that of the control, whereas the number of Bifidobacteria of all the other trials was higher but not more significant than that of the control. The results demonstrated that XOS supplementation in the diet of P. sinensis could enhance its growth and decrease FCR. Furthermore, it could enhance activities of intestinal protease and amylase and optimize its intestinal microflora. The recommended supplemental concentration of XOS in the diet of Juvenile P. sinensis should be 100–200 mg/kg.

Evaluations of the nutritional value of Jatropha curcas protein isolate in common carp (Cyprinus carpio L.)

Jatropha curcas seeds are rich in oil and protein. The oil is used for biodiesel production. Jatropha seed cake (JSC) obtained after oil extraction is rich in protein; however, it is toxic (phorbol esters content 1.3 mg/g) and consists of 50-60% shells, which are indigestible. The principle of isoelectric precipitation was used to obtain Jatropha protein isolate (JPI) from JSC and it was detoxified (DJPI). Carp (n = 45, 20.3 ± 0.13 g) were randomly distributed into five groups with three replicates and for 12-week fed iso-nitrogenous diets (crude protein 38%): Control [fishmeal (FM)-based protein]; J(50) and J(75) (50% and 75% of FM protein replaced by DJPI); S(50) and S(75) (50% and 75% of FM protein replaced by soy protein isolate). Growth performance and nutrient utilisation parameters were highest in S(75) group and not significantly different to those in J(50) and S(50) groups but were significantly higher than those for all other groups. Similar trend was observed for protein and energy digestibilities of experimental diets, whereas opposite trend was observed for the feed to gain ratio. Activities of intestinal digestive enzymes did not different significantly between the five groups. In conclusion, DJPI is a good quality protein source for carp.

Changes in Small Intestinal Morphology and Digestive Enzyme Activity with Oral Administration of Copper-Loaded Chitosan Nanoparticles in Rats

The experiment was conducted to evaluate the effect of copper-loaded chitosan nanoparticles on the small intestinal morphology and activities of digestive enzyme and mucosal disaccharase in rats. Forty male Sprague-Dawley rats, with average body weight of 82 g, were randomly allotted to five groups (n = 8). All rats were received a basal diet (control) or the same basal diet added with 80 mg/kg BW CuSO(4), 80 mg/kg BW chitosan (CS-I), 80 mg/kg BW copper-loaded chitosan nanoparticles (CSN-I), 160 mg/kg BW copper-loaded chitosan nanoparticles (CSN-II), respectively. The experiment lasted 21 days. The results showed that the villus heights of the small intestinal mucosa in groups CSN-I and CSN-II were higher than those of the control, group CuSO(4) or CS-I. The crypt depth of duodenum and ileum mucosa in group CSN-I or CSN-II was depressed. Compared with the control, there were no significant effects of CuSO(4) or CS-I on the villus height and crypt depth of small intestinal mucosa. Supplementation with CSN improved the activities of trypsin, amylase and lipase in the small intestinal contents and maltase, sucrase and lactase of duodenum, jejunum, and ileum mucosa while there were no significant effects of CuSO(4) on the digestive enzyme activities of the small content compared with the control. The results indicated that intestinal morphology, activities of digestive enzyme in digesta and mucosal disaccharase were beneficially changed by treatment of copper-loaded chitosan nanoparticles.

High Affinity Small Protein Inhibitors of Human Chymotrypsin C (CTRC) Selected by Phage Display Reveal Unusual Preference for P4′ Acidic Residues

Human chymotrypsin C (CTRC) is a pancreatic protease that participates in the regulation of intestinal digestive enzyme activity. Other chymotrypsins and elastases are inactive on the regulatory sites cleaved by CTRC, suggesting that CTRC recognizes unique sequence patterns. To characterize the molecular determinants underlying CTRC specificity, we selected high affinity substrate-like small protein inhibitors against CTRC from a phage library displaying variants of SGPI-2, a natural chymotrypsin inhibitor from Schistocerca gregaria. On the basis of the sequence pattern selected, we designed eight inhibitor variants in which amino acid residues in the reactive loop at P1 (Met or Leu), P2' (Leu or Asp), and P4' (Glu, Asp, or Ala) were varied. Binding experiments with CTRC revealed that (i) inhibitors with Leu at P1 bind 10-fold stronger than those with P1 Met; (ii) Asp at P2' (versus Leu) decreases affinity but increases selectivity, and (iii) Glu or Asp at P4' (versus Ala) increase affinity 10-fold. The highest affinity SGPI-2 variant (K(D) 20 pm) bound to CTRC 575-fold tighter than the parent molecule. The most selective inhibitor variant exhibited a K(D) of 110 pm and a selectivity ranging from 225- to 112,664-fold against other human chymotrypsins and elastases. Homology modeling and mutagenesis identified a cluster of basic amino acid residues (Lys(51), Arg(56), and Arg(80)) on the surface of human CTRC that interact with the P4' acidic residue of the inhibitor. The acidic preference of CTRC at P4' is unique among pancreatic proteases and might contribute to the high specificity of CTRC-mediated digestive enzyme regulation.

Effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity in growing rat

In this study, a growing rat model of zinc deficiency was established to investigate the effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity as well as to provide a scientific basis for zinc supplementation therapy in patients with diarrhea. Three-week-old weaned Sprague-Dawley male rats (n = 30) were randomly divided into 3 groups with 10 in each: rats in the control group (ZA) were fed with a normal diet containing 30 µg/g zinc; rats in the zinc deficient group (ZD) were fed with a zinc-deficient diet containing 0.4 µg/g zinc (refer to AIN-76 formula); and rats in the paired fed group (PF) were fed with a normal diet, but the food intake was limited to intake of rats in ZD group in the previous day. All rats were provided with deionized water for drinking. Their body weight was measured and the food intake during the previous day was recorded early in the morning of the following day. Symptoms of zinc deficiency, such as anorexia, diarrhea, dermatitis, and growth retardation, were observed. Two weeks later, the rats were sacrificed and serum zinc concentration was measured. Jejunal mucosa was taken for biopsy and was stained with hematoxylin and eosin (HE). The height ratio of the jejunal mucosal villi and crypts was measured. In addition, the activity of lactase in the jejunal mucosal brush border, γ-glutamyl peptidase (GGT), and aminopeptidase N (APN) were measured. The average weight of the rats in the ZA, ZD, and PF groups at the beginning of the experiment was (67.4 ± 5.3) g, (64.7 ± 4.8) g, and (66.5 ± 4.1) g, respectively, and the average daily food intake was (11.2 ± 1.0) g, (11.6 ± 1.6) g, and (11.2 ± 1.4) g, respectively. The intergroup differences were not significant. On the 7(th) day of experiment, no significant differences in average food intake were observed between the ZD group and the ZA and PF groups, but the average body weight in the ZD group was significantly lower than that in the ZA and PF groups (P < 0.01). At the end of the experiment (2 weeks), the average weight in the ZD group (112.0 ± 11.5) g was significantly lower than that in the ZA (164.0 ± 15.9) g and PF groups (137.5 ± 16.2) g. The average food intake in the ZD group (13.4 ± 5.1) g was significantly lower than that in the ZA group (18.2 ± 2.4) g (P < 0.01). Serum zinc level in the ZD group (733 ± 231) µg/L was significantly lower than that in the ZA (1553 ± 159) µg/L and PF groups (1457 ± 216) µg/L (P < 0.01). The height ratio of jejunal mucosa villus and crypt in the ZA, ZD, and PF groups was 2.98 ± 0.5, 2.77 ± 0.5, and 2.81 ± 0.7, respectively, and lactase activity was (26.1 ± 15.0) U/mg, (27.4 ± 12.8) U/mg, and (40.8 ± 18.5) U/mg, respectively, without significant intergroup differences. The GGT activity in the jejunal mucosa in the ZD group (12.7 ± 6.5) U/g was significantly lower than that in the ZA (19.1 ± 10.4) U/g and PF groups (18.5 ± 7.7) U/g, but the difference was not significant. The activity of APN in the jejunal mucosa in the ZD group (25.5 ± 7.5) U/g was significantly lower than that in the ZA (48.7 ± 16.8) U/g and PF groups (43.9 ± 14.5) U/g (P < 0.01). Zinc deficiency can cause loss of appetite, weight loss, and decreased activity of peptidase in the jejunal mucosal brush border. Zinc deficiency has little effect on the height ratio of the villus and crypt and lactase activity, thereby indicating that zinc deficiency may first affect protein digestion and absorption.

Use of probiotics Bacillus coagulans, Rhodopseudomonas palustris and Lactobacillus acidophilus as growth promoters in grass carp (Ctenopharyngodon idella) fingerlings

The present research evaluated the effects of three probiotics on the growth performance and intestinal digestive enzyme activity in fingerlings of grass carp, Ctenopharyngodon idella. Three treatments (T-1, T-2 and T-3) were fed with diets containing different viable bacteria with a final concentration 106 CFU g−1 feed (Bacillus coagulans, Rhodopseudomonas palustris and Lactobacillus acidophilus, respectively) and the control was fed with basal diet without probiotics. All the diets supplemented with probiotics resulted in better (P 0.05) in protease and cellulase activities compared with the control. The protease activity was higher in the foreintestine than in the hindintestine (P < 0.05), and the reverse was observed in cellulase activity. As for amylase activity, there was no difference between foreintestine and hindintestine. In conclusion, the three selected probiotics increased the growth performance of grass carp fingerlings. Furthermore, different digestive enzyme activity was observed in different intestine segment.

Intrauterine Growth Restriction Delays Feeding-Induced Gut Adaptation in Term Newborn Pigs

Background: Neonates born after intrauterine growth restriction (IUGR) show higher mortality and morbidity and greater feeding problems postnatally. Objectives: We tested the hypothesis that IUGR affects the intestinal structure, function, microbiology and proinflammatory cytokine expression during the immediate neonatal period. Methods: We firstly compared organ weights and intestinal digestive enzyme activities between control and IUGR newborn piglets delivered by cesarean section at full term or prematurely (91% gestation). Next, we compared intestinal structure, function and microbiota in spontaneously delivered control and IUGR term piglets during the period 0–5 days of age, when intestinal adaptation is normally very rapid. Results: At the time of birth, organ weights and intestinal enzyme activities were not notably affected by IUGR, neither for preterm nor term pigs, except that IUGR was associated with a relatively long and thin intestine. Between birth and 5 days of age, the normal developmental pattern in the ileum and colon appeared to be delayed in term IUGR piglets, as indicated by lower ileal density (weight per unit length) and villous area (–20 to –30%), higher expression of the peptide transporter PEPT1 (+150% on day 2) and enhanced bacterial adhesion and translocation during days 2–5 (all p < 0.05). Further, expression of the proinflammatory cytokine IL-6 was modified in the intestine of term IUGR piglets at birth without any change in IL-1β. Conclusion: IUGR is associated with a longer and thinner intestine at birth, and during the immediate postnatal period, intestinal adaptation and bacterial colonization are altered in IUGR piglets born at full term.

Stable isotopes document mainland–island divergence in resource use without concomitant physiological changes in the lizard Liolaemus pictus

Shifts in feeding ecology are believed to promote island-mainland divergence. The lizard Liolaemus pictus has several different subspecies on Chilean islands and mainland. These subspecies inhabit contrastingly different habitats both in different islands and mainland, which suggests the potential for habitat related dietary variation. We investigated the dietary habits of L. pictus by both stomach content analyses and by nitrogen stable isotope analyses (delta(15)N), which we used as a proxy variable for trophic level. We also compared the morphology of the digestive tract and the activity of intestinal digestive enzymes of mainland and island lizards. We hypothesized differences in diet and trophic level among populations and that these differences would predict the expression of the morphological and biochemical features of the digestive tract. More specifically, we predicted shorter intestines and higher levels of peptidases in more insectivorous than in more frugivorous/herbivorous lizards. The diet of L. pictus was characterized by the consumption of a wide diversity of food types, including fruit and insects, in all populations. Stable isotopes revealed higher trophic level, and hence probably higher protein intake, in mainland than in island populations, but contrary to our prediction, they had shorter intestines and higher relative activity of intestinal peptidases than mainland lizards. Furthermore, the proportion of fruit items in the stomach content was higher in the population that exhibited the lowest tropic level. These results suggest that morphological and physiological differences among populations of L. pictus are not correlated with feeding ecology, suggesting that the lizard's first responses to the selective pressure represented by a diet shift are behavioral.

Effects of Diets Supplemented with Recombinant Epidermal Growth Factor and Glutamine on Gastrointestinal Tract Development of Early-weaned Piglets

This study attempted to determine effects of recombinant porcine epidermal growth factor (pEGF) and glutamine (Gln) supplement on the growth performance and intestinal development of piglets weaned at 14 days of age. A total of ninety-six piglets were allotted to one of four dietary treatments which comprised inclusion of 1.0 mg pEGF supematant/kg diet or 0.5% Gln both alone and in combination. Each treatment consisted of four replicates with six pigs per pen for a 28 days experimental period. Two pigs per replicate were sacrificed and gastrointestinal tract samples were collected on day 14. Data showed that dietary treatment failed to promote growth performance. On day 14, diets supplemented with pEGF elevated pancreatic chymotrypsin, jejunal alkaline phosphatase, sucrase, lactase and maltase activities (p<0.05), but failed to alter the small intestinal villus morphology, DNA, or protein content of gastrointestinal mucosa. Diets supplemented with Gln increased pancreatic chymotrypsin activity, tended to enhance the protein contents of gastric (p = 0.08) and jejunal mucosa (p = 0.09) but did not influence the serum IgA level or the enzyme activity in the gastrointestinal tract. On day 28, the diets supplemented with Gln increasedt (p<0.05) serum IgA and the proliferation of peripheral blood mononuclear cells by PHA stimulation. However, a combination of pEGF and Gln did not have a synergistic effect on these biomarkers in early-weaned piglets. The results demonstrate that diets supplemented with recombinant pEGF supernatant indeed improve intestinal digestive enzyme activity and diets supplemented with Gln increases the immune response in early-weaned piglets.