Influence of roundup herbicide on the activities of peptidases in the intestines of various fish species

It has been revealed that, as a rule, Roundup herbicide inhibits activities of peptidases in the intestine mucosa and chyme in various fish species, both benthivorous (common carp Cyprinus carpio, silver bream Blicca bjoerkna, roach Rutilus rutilus, and crucian carp Carassius carassius) and piscivorous (perch Perca fluviatilis, pike Esox lucius, and sander Sander lucioperca). The tolerance of peptidases to the herbicide is species-specific. The magnitude and vector of the effects of Roundup depend on its concentration and localization of an enzyme. In the mucosa, the enzymes of common carp and pike are most tolerant, while the enzymes of common carp, silver bream, and pike are most tolerant in the chyme.     

Effect of pH on the activity of proteinases in intestinal mucosa,chyme, and microbiota of fish from the Cuciurgan reservoir

Proteolytic activities of the intestinal mucosa, chyme, and enteral microbiota have been studied in a wide range of pH values in five fish species from the Cuciurgan reservoir (Moldova). Differences in pH dependence of the intestinal proteinase activity of fish are determined by their feeding type. The maximum activity of proteinases is found in the pumpkinseed Lepomis gibbosus. The minimum activity of proteinases has been demonstrated by the zander Zander lucioperca. The pH optimum of the mucosa and chyme in all fish species (except for the European perch Perca fluviatilis) is 10. The pH optimum of the intestinal microflora varies from 6.0 (in the common carp Cyprinus carpio) to 10 (in the crucian carp Carassius carassius), whereas that in the perch from the Cuciurgan and Rybinsk reservoirs is 7. The majority of fish species, mostly Zander lucioperca and Lepomis gibbosus, are characterized by high proteniase activity of the microbiota, in the pH ranging from 6 to 9. It is assumed that proteinases in the enteral microbiota of fish are able to make up for the relatively low activity of those synthesized by their digestive system in the range of low pH values.     

The effect of temperature on chyme proteinase activities in perch Perca fluviatilis L. and roach Rutilus rutilus (L.)

The effect of temperature on the chyme proteinase activities of the perch Perca fluviatilis L. and the roach Rutilus rutilus (L.) was investigated. The two fish species showed different characters of temperature dependence and different values of activation energy for caseinolytic and hemoglobinolytic chyme proteinases. The perch was characterized by a higher level of relative proteinase activities and lower activation energy for protein hydrolysis at a temperature of 0–10°C than the roach.     

Effects of temperature,pH, and heavy metals (copper,zinc) upon proteinase activities in digestive tract mucosa of typical and facultative piscivorous fish

Species-specific differences and effects of natural (temperature, pH) and anthropogenic (zinc, copper) factors upon activities of casein-and hemoglobinlytic proteinases have been studied in the gut mucosa in pike (Esox lucius), zander (Stizostedion lucioperca), burbot (Lota lota) and perch (Perca fluviatilis). It is revealed that the intestine mucosa proteinase activities differ considerably from those ones in the fish stomach mucosa and that studied factors affect these activities to different extents. The stomach mucosa proteinases are most tolerant to the effects of studied factors, which is especially true for pike. The least tolerant are the intestine caseinlytic proteinases (except for the enzymes in perch). The effects of temperature, pH, and heavy metals depend on fish species, factors’ combinations (mono-factor, bi-factor, or multi-factor action) and on factor variability.     

Effect of temperature on proteinase activities in intestinal chyme and mucosa of fish of different ecological groups

Effect of temperature on activities of proteinases in intestinal chyme and mucosa was studied in three fish species (pike-perch, zope, roach) belonging to different ecological groups by their nutrition type. There was revealed a significant difference of dependence of enzyme activities in chyme on temperature in the benthophage, roach (a higher level of relative activity in the range of lower temperatures and a wider zone of temperature optimum) as well as of values of apparent energy of activation of the protein hydrolysis process as compared with that in planktoand ichtyophages, zope and pike-perch, which indicates a significant effect of the enteral microbiota proteinases and of nutrition objects on characteristics of hydrolases functioning in fish intestine.     

Effect of temperature on proteinase activities in the intestinal chyme and the mucosa of various ecological fish groups

Effect of temperature on activities of proteinases in intestinal chyme and mucosa was studied in three fish species (pike-perch, zope, roach) belonging to different ecological groups by their type of feeding. There was revealed a significant difference of dependence of enzyme activities in chyme on temperature in the benthophage roach (a higher level of relative activity in the zone of lower temperatures and a larger zone of temperature optimum) as well as of values of apparent energy of activation of the protein hydrolysis process as compared with that in plankto- and ichthyophages--zope and pike-perch, which indicates a significant effect of the enteral microbiota proteinases and of nutrition objects on characteristics of hydrolases functioning in fish intestine.     

Effect of pH on the proteinase activities in the intestine mucosa,chyme, and enteral microbiota in the piscivorous fish differing in their ecological traits

The effect of pH on the activity of proteinases of intestine mucosa, chyme, and enteral microbiota was studied in three piscivorous fish species of the Rybinsk Reservoir differing in their ecological traits: pike Esox lucius, burbot Lota lota, and zander Sander lucioperca. In all fish species, studied pH of ten is optimal for the functioning of proteinases of intestine mucosa; in chyme proteinases, optimal pH values vary from six to ten. Optimal pH for functioning of proteinases of enteral microbiota is seven for zander and pike; pH of nine, for burbot. High activity of chyme and microbiota proteinases within the diapason of pH values of six to nine is a characteristic of burbot. Relative activity of proteinases of intestine mucosa in all fish species is not more than 15% of the maximal activity; that of chyme and enteral microbiota is lower than 40% in zander, close to 50% in burbot, and 80 and 50%, respectively, in pike. It is suggested that diversity of the patterns of pH-dependence of enteral microbiota proteinases relates to the specificity of microbiota in various ecological zones of the reservoir (littoral, sublittoral, and bathyal).     

Influence of biogenic metals (Cu,Zn) on the activity of carbohydrases in juvenile fish in vitro

It was revealed that intestine and whole-body amylolytic activity (AA) in juvenile fish serving as potential feeding items for piscivores significantly decreases in the presence of Cu and Zn ions within a wide range (0.1–25 mg/l) of concentrations in vitro. In most of the studied fish species, Cu and Zn decrease the activities of carbohydrases in the intestine mucosa stronger than in the whole-body. On the other hand, in piscivore-facultative benthivore perch, which have the least AA, the inhibiting effect of Cu and Zn ions is 1.5 to 2 times higher in the whole body than in the intestine. The study data suggest that Cu and Zn ions at concentrations found in organisms that serve as fish food may not only reduce the rate of initial stages of carbohydrates hydrolysis in the juvenile fish intestine, but also considerably decrease the potential contribution of the food objects to the digestion processes in typical and facultative piscivores.     

The influence of testosterone on activity of glycosidases and proteinases in the intestine of the sterlet (Acipenser ruthenus)

Observation of the influence of testosterone (0.7 mg/kg) on the activity of glycosidases and proteinases, which function in the chyme and the mucosa of the sterlet intestine (Acipenser ruthenus), revealed a decrease in enzyme activity of both chains as against that one typical for intact individuals. The activity of the investigated enzymes changes in both the experimental and control groups of fish. Nevertheless, they have unlike dynamics of this activity. In a number of cases, we indicated a significant increase in enzyme activity of both chains in comparison to the control group. We also observed an increase in activity of chyme glycosidases, which was significant on the 21st day as compared to intact fish.     

Transport of fat through jejunal absorptive epithelium in vitro

The mucosa of excised pieces of jejunum of fasting rats was exposed for 10 min to fresh chyme obtained from other rats which had been digesting either buttered bread or bread alone. The mucosa was then processed for electron microscopy of lipid material. After exposure to fatty chyme, fat globules were scattered throughout the cytoplasm of absorptive epithelial cells, and concentrated in both supranuclear and infranuclear components of the Golgi apparatus. They were also present within lymphatics of the villus. After exposure to both types of chyme, the basal parts of the cells were much elongated through collection of a finely granular deposit of non-lipid material. This collection was probably due to failure of clearance of absorbed material from the cells in the absence of blood flow through the villus.     

Multiple regulation of peptide YY secretion in the digestive tract

In the last two decades, multiple aspects of the peptide YY (PYY) secretion have been investigated. Besides fat and fatty acids, many luminal nutrients in the distal intestine appear to induce PYY release. Some studies have shown that bile acid, but not nutrients, plays a crucial role in the regulation of PYY secretion. Moreover, chyme in the proximal intestine also regulates the peptide release by indirect action through humoral and neuronal factors. Gastrin, cholecystokinin, and the vagus nerve are major candidates for mediators of these indirect actions. Several growth factors have been shown to regulate PYY synthesis in mucosa of the distal intestine. This review is aimed at presenting an overview of these recent studies on PYY secretion in the distal intestine.     

An in vitro analysis of intestinal ammonia handling in fasted and fed freshwater rainbow trout (Oncorhynchus mykiss)

Ammonia transport and metabolism were investigated in the intestinal tract of freshwater rainbow trout which had been either fasted for 7 days, or fasted then fed a satiating meal of commercial trout pellets. In vivo, total ammonia concentrations (T _amm) in the chyme were approximately 1 mmol L^?1 across the entire intestine at 24 h after the meal. Highest chyme pH and P _NH3 values occurred in the posterior intestine. In vitro gut sac experiments examined ammonia handling with mucosal (Jm_amm) and serosal (Js_amm) fluxes under conditions of fasting and feeding, with either background (control ≤0.013 mmol L^?1) or high luminal ammonia concentrations (HLA = 1 mmol L^?1), the latter mimicking those seen in chyme in vivo. Feeding status (fasted or fed) appeared to influence ammonia handling by each individual section. The anterior intestine exhibited the greatest Jm_amm and Js_amm values under fasted control conditions, but these differences tended to disappear under typical post-feeding conditions when total endogenous ammonia production (Jt_amm = Js_amm ? Jm_amm, signs considered) was greatly elevated in all intestinal sections. Under fasted conditions, glutamate dehydrogenase (GDH) and glutaminase (GLN) activities were equal across all sections, but the ammonia-trapping enzyme glutamine synthetase (GS) exhibited highest activity in the posterior intestine, in contradiction to previous literature. Feeding clearly stimulated the total rate of endogenous ammonia production (Jt_amm), even in the absence of a high luminal ammonia load. This was accompanied by an increase in GDH activity of the anterior intestine, which was also the site of the largest Jt_amm. In all sections, during HLA exposure, either alone or in combination with feeding, there were much larger increases in endogenous Jt_amm, most of which was effluxed to the serosal solution. This is interpreted as a response to avoid potential cytotoxicity due to overburdened detoxification mechanisms in the face of elevated mucosal ammonia. Thus T _amm of the intestinal tissue remained relatively constant regardless of feeding status and exposure to HLA. Ammonia production by the gut may explain up to 18 % of whole-body ammonia excretion in vivo under fasting conditions, and 47 % after feeding, of which more than half originates from endogenous production rather than from absorption from the lumen.     

Effect of dexamethasone on activity of glycosidases and proreinases of intestine of sterlet <Emphasis Type="Italic">Acipenser ruthenus</Emphasis>

Effect of dexamethasone (2.0 mg/kg) on activity of enzymes hydrolyzing proteinaceous and carbohydrate food components in the intestine of the starlet Acipenser ruthenus is studied. Dexamethasone modifies the activity of proteinases more than of glycosidases. As a rule, the hormone significantly decreases the level of proteolytic and general amylolytic activity of the intestinal mucosa and chyme in comparison with intact fish on the first day into the experiment and increases it on the 7th or 14th day. The dynamics of activity was different in enzymes of different chains (glycosidase and proteinase) and preparations (the mucosa and chyme).     

Activity dynamics of proteinases and glycosidases of fish chyme with exposure in fresh and brackish water

Activity of proteinases of the content of intestines (chyme) of the benthos-eater crucian carp Carassius carassius fed different diets during prolonged exposure to water is studied. In the process of exposure of the chyme to water, the activity of proteinases decreases. Activity of glycosidases may increase, maximally during the first three days of exposure. This phenomenon suggests the important role of enzymes of the enteral microflora in processes of destruction of protein and carbohydrate components of the suspension and especially of organic detritus.     

Influence of various concentrations of phenol and its derivatives on the activity of fish intestinal peptidases

The effects of phenol and its derivatives (4-chlorophenol, 4-nirophenol and 2,4-dinitrophenol) on the activity of peptidases in five species of fish differing in feeding modes have been studied. The power of the effects depends on the fish species and localization of the enzyme (mucosa or chyme). As a rule, in the in vitro conditions the phenols in concentrations of 0.06?0.5 μM/L considerably decreases activities of the intestinal peptidases in bream; silver bream; and, especially, in pike. In some cases, phenol and its derivatives induce an inconsiderable increase in peptidase activities. In zander and perch, in fact, no changes in the enzymatic activity under impact of phenols are observed. It is suggested that the effect of phenol and its derivatives depends to a high extent on the species specificity of peptidases: in the fish of fam. Percidae, the enzymes are relatively tolerant; in fam. Cyprinidae and Esocidae, they are sensitive to the studied toxicants.     

Leucine aminopeptidase activity in the digestive tract of perch, Perca fluviatilis L.

The distribution of the enzyme leucine aminopeptidase (LAP, EC 3.4.1.1) in the digestive tract of perch, Perca fluviatilis L., was investigated histochemically. The enzyme was present in the mucosa of the entire intestine and was absent in the oesophagus, stomach and pyloric sphincter. In the intestine, the enzyme was localized in the supranuclear cytoplasm of the columnar cells and was strongest in the brush border area. Enzyme activity was also present in the lumen of the intestine. The activity of the enzyme in the intestine decreased slightly towards the rectum. In perch, the final digestion of peptides and aminopeptides is both extracellular and intracellular and takes place along the entire length of the intestine.     

Impact of polychlorinated biphenyls on the activity of intestinal proteinases and carbohydrases in juvenile roach Rutilus rutilus (L.)

The chronic effects that polychlorinated biphenyls (PCBs) have on the activities of proteinases and carbohydrates in intestinal mucosa and chyme in juvenile roach Rutilus rutilus (L.) have been studied for the first time. Upon consuming food with PCB 50.8 ng/g wet weight for 218 days, the exposition of fingerlings in aquariums with dirt bottoms (contents PCB 425.6 ng/g dry weight) reduces the proteolytic activity of mucosa to a greater degree than amylolytic activity; the activity of sucrase changes in different directions. In 2-year-old fish receiving PCB only with food (50.8 ng/g wet weight bottom), proteolytic and amylolytic activities of the mucosa are reduced 18 and 35% in comparison with the control (52 days); the sucrose activity is reduced 13% (52 days) and 22% (169 days). In most cases the relation of amylolytic activity to proteolytic activity in fish of the experimental groups is less than in the control. The enzyme activities in chyme change in different directions, depending on the terms and conditions of the experiment.     

Effect of temperature on proteinase activities of enteral microbiota and intestinal mucosa of fish of different ecological group

Effect of temperature on proteinases activities of enteral microbiota and of intestinal mucosa was studied in five fish species (roach Rutilus rutilus, crucian carp Carassius carassius, common perch Perca fluviatilis, pike-perch Zander lucioperca, and pike Esox lucius) belonging by the nutrition type to different ecological groups. Essential differences of temperature characteristics of proteinases of intestinal mucosa and of enteral microbiota are revealed in fish belonging by the nutrition type to different ecologic groups. The character of the t0-function of proteinases of intestinal mucosa and enteral microbiota by casein and hemoglobin as a rule is different. The highest values of relative proteinases activities for casein in the zone of low temperatures (38 and 45.3 % of the maximal activity) are found at study of proteinases of enteral microbiota in common perch and crucian carp. The latter indicates a significant adaptability of the enteral microbiota proteinases of common perch and crucial carp to functioning at low temperatures.     

Non-specific carboxylic esterase activity in the digestive tract of the perch, Perca fluviatilis L.

The distribution of non-specific carboxylic esterases (Ec 3.1.1) in the digestive tract of perch, Perca fluviatilis L., was investigated histochemically using 1-naphthyl acetate as the substrate. Strong enzymatic activity was present in the gastric glands and surface cells of the stomach, intestinal mucosa of the pyloric caeca, upper and middle intestine, pancreas (exocrine cells) and liver. The enzymatic activity in the lower intestine and rectum was weak. The activity was not demonstrated in the oesophagus or pyloric sphincter. In the intestine, the activity was localized in the columnar cells especially in the supranuclear cytoplasm. The enzymatic activity demonstrated in the digestive tract of perch using 1 -naphthyl acetate represents combined esterolytic and lipoproteolytic activity.     

Influence of diurnal rhythms of feeding during intestine total amylolytic activity and activity of alkaline phosphatase in juvenile fish

The total amylolytic activity and activity of alkaline phosphatase in the intestine mucosa of larvae and fries of roach, blue bream, and perch change both during the process of individual development and during the day. Maximal intensities of juvenile feeding was observed primarily in the morning and evening hours. The pattern of diurnal alkaline phosphatase activity correlates to a greater extent to intensity of juvenile feeding, in comparison to the pattern of total amylolytic activity. In planktivorous blue bream, such regularity is more pronounced than in benthivorous roach. The total amylolytic activity in fries of roach, blue bream and perch correlates to fish type of feeding.