Specific Activity Of Trypsin Research Articles

Stage-Specific Ontogeny of Digestive Enzymes in the Cultured Common Dentex (Dentex dentex) Larvae

Stage-specific ontogeny of main digestive enzymes in common dentex, Dentex dentex, were assayed during early life development according to notochord flexion studied from hatching to 45 days after hatching (DAH). The notochord was straight in yolk-sac larvae of D. dentex and preflexion stage was observed between 0 and 18 DAH as 2.75 and 6.27 mm at total length (TL). Then, flexion of the notochord was estimated between 19 and 30 DAH as 6.27 and 10.9 mm at TL. Important alterations in specific activities of all digestive enzymes measured during period of this study were mostly related with metamorphosis and co-feeding. Trypsin and chymotrypsin specific activities were firstly detected on day 3 and were related with mouth opening, whereas they slightly increased until 25 DAH respectively. Pepsin was firstly detected on day 28 and related with stomach formation and a sharp increase was observed until 30 DAH. Both amylase and lipase were measured on day 2 and 4, respectively and they also sharply increased during the first week of life. Additionally, except pepsin, most of the digestive enzyme activities were firstly secreted in the preflexion stage and also gastric secretion started and functional development digestive system were occurred in in flexion and postflexion stages, respectively.

Development of Enzymes andIn VitroDigestibility during Metamorphosis and Molting of Blue Swimming Crab (Portunus pelagicus)

The work focuses on development of digestive enzymes (amylase, total protease, trypsin, and chymotrypsin) and activity ratio of trypsin to chymotrypsin (T/C ratio) for digestive efficiency and growth, in blue swimming crab (Portunus pelagicus) during metamorphosis and molting. Specific activities of all enzyme parameters studied were associated with growth during metamorphosis, while only those of trypsin and T/C ratio were associated during molting cycle where trypsin and chymotrypsin specific activities associated with consumption rate with especially high levels during late intermolt and early premolt stages. About 50% increased weight gain was observed with at least double increased T/C ratio at the end of molting period, compared to the stages prior to molting. Growth of carapace would be more significant after finishing molting. Carapace width gain and T/C ratio were highest at the first crab stage. Studies ofin vitroprotein digestibility of different feed raw materials indicated thatArtemia, Rotifer, andMoinaare the best for larval stages. Otherwise, the use of shrimp feed andArtemiaflake could be the alternatives. Incorporating of cassava meal into the feed formula for early adult stage (juvenile) could be an advantage. The proteins from animals are more beneficial for adult crab culture than the proteins from plants and bacteria. The digestible quality of dietary protein is very important during larval stages, while the protein level of diet is more important during adult stages with fully developed digestive enzymes.

Partial characterization of digestive proteases in tropical gar Atractosteus tropicus juveniles

Tropical gar (Atractosteus tropicus) is an economically and socially important freshwater species from Southeastern Mexico, with a high aquaculture potential. With this in mind, the purpose of this study was to characterize the digestive proteases of tropical gar juveniles through biochemical and electrophoretic analyses. Twenty specimens with an average weight of 73.6±12.7g were used to obtain stomach and intestinal tissue from which multienzymatic extracts were prepared. The general activities of the acid and alkaline proteases were evaluated, as well as the specific activities of trypsin, chymotrypsin, leucine aminopeptidase and carboxypeptidase A. The effect of the pH and temperature on the proteases was also analyzed, together with the composition of the multienzymatic extracts using protease inhibitors and electrophoretic tests. Results showed that A. tropicus have a functional stomach in which protein hydrolysis starts with pepsin and which contains endo- and exopeptidases (trypsin, chymotrypsin, leucine aminopeptidase and carboxypeptidase A) and proteases that are resistant to high temperatures (45 and 55°C for alkaline and acid proteases, respectively) and pH values. Using zymogram technique, we found two acid protease isoforms (0.35 and 0.71rf) and five alkaline protease isoforms (83.7, 43.7, 27.5, 24.0 and 19.4kDa), which decrease or disappear with the different inhibitors. Thus, this species is considered to be a carnivore capable of adapting to its environment by consuming different types of proteins from preys and also could adapt rapidly to consume a compound diet with different animal protein sources.

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Evaluation of red monascal rice supplementation on growth, digestive function and oocyte maturation were investigated in Siamese fighting fish (Betta splendens). Completely randomized design with different dietary levels of red monascal rice (0.00, 0.25, 0.50, 1.00 and 2.00%) was conducted for six weeks. The growth of fish fed a control diet was not statistically different (P > 0.05) from a diet containing 0.25% of red monascal rice. However, significantly lower values (P < 0.001) were observed in fish fed more than 0.50% red monascal rice, in a dose-dependent manner. Muscle RNA concentrations were higher in fish fed control diet than in fish fed red monascal rice, while protein concentration and RNA/protein ratio were similar. Body composition and fatty acid profiles (saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, total omega-3 and total omega-6) were unchanged among dietary treatments. Digestive functions were perturbed by decreasing the specific activities of amylase, total protease, trypsin and chymotrypsin (P < 0.001), and increasing the specific activity of lipase (P < 0.005). Reproductive consummation of oocytes was significantly different in fish fed red monascal rice, due to increasing RNA concentration and RNA/protein ratio (P < 0.003) and decreasing specific activities of trypsin- and chymotrypsin-like enzymes (P < 0.03). These findings suggest the toxicological effects of red monascal rice by interfering with growth, digestive function and oocyte maturation in Siamese fighting fish. Key words: Digestive enzyme, fatty acid, growth, muscle, oocyte, red monascal rice, Siamese fighting fish.

Effects of Aflatoxins on Performance and Exocrine Pancreas of Broiler Chickens

The aim of this research was to evaluate, on a weekly basis, the effects of aflatoxins on the activity of digestive enzymes (alpha-amylase, lipase, and trypsin) in the pancreas as well as on the performance and histology of pancreas in broiler chickens over the course of 42 days. One thousand and eighty 1-day-old male Cobb broilers were divided into four treatments with 18 replicates and 15 birds per replicate (i.e., 270 broilers per treatment). Treatments were established according to the amount of aflatoxins added to the diet, as follows: T1 = 0 mg of aflatoxins per kilogram of feed (mg/kg); T2 = 0.7 mg/kg; T3 = 1.7 mg/kg; and T4 = 2.8 mg/kg. Pancreas sample collection was performed from one bird out of each replicate at 7, 14, 21, 28, 35, and 42 days of experiment, which yielded a total of 18 samples per treatment on each collection. Each sample was homogenized in distilled water, frozen in liquid nitrogen, lyophilized, and stored at -20 C until analysis. Performance parameters (body weight, feed consumption, and feed conversion rate) were measured at 21, 35, and 42 days of experiment. At the end of the experiment (42 days), six birds from each treatment were randomly chosen for histologic evaluation of the pancreas. The presence of aflatoxins in the diet induced a negative effect on all performance parameters. The pancreatic activity of lipase and alpha-amylase were significantly increased in treatments T3 and T4, while the specific activity of trypsin was only affected during treatment T4. In addition, several histologic changes were observed in the pancreas of birds receiving aflatoxin-contaminated feed. Aflatoxins present in the feed determined an increase in the activity of pancreatic enzymes in broilers, affecting the digestibility of the diet, thereby leading to losses in performance and productivity.

Alpha 1-antitrypsin activity is markedly decreased in Wegener’s granulomatosis

Alpha 1-antitrypsin (A1AT) is the most abundant proteinase inhibitor in plasma and the main inhibitor of Proteinase 3, the target antigen of antineutrophil cytoplasmic antibodies (ANCAs) that predominant in Wegeners' granulomatosis. Α1AT deficiency correlated with ANCA-associated vasculitis. This study explores the trypsin inhibitory capacity (TIC), specific activity, and phenotypic deficiency of Α1AT in Wegener's granulomatosis. Twenty-seven WG patients were studied. ANCA was tested by IIF and ELISA. Serum a1-anti-trypsin levels were quantified in WG patients and healthy controls by immunoturbidimetric assay. Serum TIC was assessed by the enzymatic colorimetric assay. Phenotypes of A1AT were detected by Isoelectric Focusing. A1AT concentration was equivalent in patients and controls; however, serum TIC (P = 0.001) and specific activity of A1AT (P = 0.001) were dramatically lower in WG patients. Five patients had deficient phenotypes of A1AT: MZ (n = 3), MS (n = 1) and SS (n = 1). This was correlated with an increase in the prevalence of deficient phenotypes of A1AT in WG (P = 0.01). Trypsin inhibitory capacity and specific activity of A1AT were decreased in WG patients and may be involve in disease pathogenesis and can worsen the clinical manifestations. This A1AT deficiency probably resulted from oxidative inactivation and/or enzymatic degradation of A1AT. This could result in localized deficiency of A1AT in vessel wall interfaces and lead to severe disease.

Effects of tannic acid on trypsin and leucine aminopeptidase activities in gypsy moth larval midgut

The effects of allelochemical stress on genetic variations in the specific activities of gypsy moth digestive enzymes (trypsin and leucine aminopeptidase) and relative midgut mass (indirect measure of food consumption), as well as variability in their plasticity, were investigated in fifth instar gypsy moths originating from two populations with different trophic adaptations (oak and locust-tree forests). Thirty-two full-sib families from the Quercus population and twenty-six full-sib families from the Robinia population were reared on an artificial diet with or without supplementation with tannic acid. Between population differences were observed as higher average specific activity of trypsin and relative midgut mass in larvae from the Robinia population. Significant broad-sense heritabilities were observed for the specific activity of trypsin in the control state, and for specific activity of leucine aminopeptidase in a stressful environment. Significantly lower heritability for relative midgut mass was recorded in larvae from the Robinia population reared under stressful conditions. Significant variability of trypsin plasticity in larvae from both populations and significant variability of leucine aminopeptidase plasticity in larvae from the Robinia population point to the potential for the evolution of enzyme adaptive plastic responses to the presence of stressor. Non-significant across-environment genetic correlations do not represent a constraint for the evolution of enzyme plasticity.

Effects ofQuillajasaponins on growth, feed efficiency, digestive enzyme activities and metabolism of common carp (Cyprinus carpioL)

Common carp were fed diets containing various levels of Quillaja saponins (QS) (0, 150, 300 and 450 mg kg−1 dry diet) in a completely computerized respirometric system for 4 weeks. Fish fed diets containing QS exhibited significantly higher ABW and specific growth rate than did those fed the control diet; those fed diets containing QS 150 grew fastest but were not significantly different from those fed diets with QS 300 and QS 00450. All the utilization efficiency indices, namely food conversion efficiency (FCE), protein productive value and PG were increased by QS supplementation. There were no significant differences in the average routine metabolic rate between treatments, indicating that dietary QS at the levels tested were not toxic to the carp. Increases in amylase and trypsin specific activities were observed at QS 300 and QS 450. Enzymes of carbohydrate metabolism such as G6PDH, 6-phosphogluconate dehydrogenase and pyruvate kinase were not significantly affected by dietary QS. Activities of the aerobic enzyme Cox and to a limited extent that of the anaerobic enzyme lactate dehydrogenase were significantly increased by dietary QS but the net effect was a shift towards aerobic metabolism, indicating absence of stress and favouring the anabolic processes. Thus, Quillaja saponin was beneficial as a feed supplement in the common carp.

Ecological Effects on Food Utilization, Trypsin Isozymes, and Performance Qualities of Growth and Maturation in Northeast Arctic Cod (Gadus morhua L.)

The work demonstrates, for the first time, natural biological changes in fish during maturing processes using a unique combination of different biochemical techniques. Northeast Arctic cod caught by demersal trawls were studied in three Barents Sea areas during February-March 2008. Older larger cod from area A (between North Kanin Bank and Eastern Basin) had higher food utilization efficiency (with females showing highest white muscle protein synthesis capacity) than those from areas B (Kanin Bank) and C (Central Bank). Populations in areas A and C living in separate environments showed parallel regressions of pyloric caecal slope T/C ratio, with different elevations of trypsin specific activities and trypsin isozymes expressions. Approximately 30 trypsin isozymes were observed, with 13 isozymes of possibly exogenous trypsin isozyme fragments from prey items found at higher percentage of cod in areas A and B with higher food varieties. Larger maturing cod required higher energy from carbohydrate (probably phytoplankton), as amylase specific activity correlated with body weight. White muscle RNA levels were varied among females from different areas, but not among males. RNA and RNA/protein ratio levels were higher in oocytes than white muscle, and these levels decreased in higher developed oocytes. Oocyte trypsin-like specific activity in areas B A > C) illustrated that females from area B (youngest with lowest oocyte protein turnover) had highest maturation rate, and would reach their spawning area(s) before those from areas A and C. Younger females (probably also males) living closer to coastal area with higher temperature and food varieties would reach sexual maturity faster than older cod and those living far from coastal area.

Adaptive differences in digestive enzyme activity in the crab Neohelice granulata in relation to sex and habitat

Neohelice granulata (Dana, 1851) is a semi-terrestrial burrowing crab that inhabits estuaries and saltmarshes feeding on grasses or sediment depending on the microhabitat they occupy (vegetated saltmarsh or bare mudflat). Specific cellulolytic, amylolytic, and proteolytic enzyme activities were analyzed in midgut gland homogenates of males and females from each microhabitat fed in the laboratory with Spartina densiflora leaves and sediment, respectively, in order to detect sex, food and microhabitat related differences. The presence of β-1,4-glucosidase, endo-β-1,4-glucanase, α amylase, trypsin and chymotrypsin were confirmed. Specific cellulolytic activity was higher in crabs fed on leaves than in those fed on sediment or in no fed controls and variable differences between sexes were observed. Specific amylase activity of crabs fed on leaves was the lowest recorded. Trypsin and chymotrypsin specific activities were higher in saltmarsh crabs fed on leaves than in mudflat crabs fed on sediment. Different mechanisms of enzyme regulation to explain the observed differences among groups were suggested. Additionally, differences between sexes suggest different metabolic needs related to gonad maturation. It is concluded that N. granulata has the ability to adapt digestive enzyme production to support its physiological and metabolic needs based on the different food sources available at each microhabitat.

Effect of starvation and refeeding on digestive enzyme activities in juvenile roach, Rutilus rutilus caspicus

We evaluated the effects of starvation and refeeding on digestive enzyme activities in juvenile roach, Rutilus rutilus caspicus. Fish were divided into four feeding groups (mean mass 1.68 ± 0.12 g). The control group was fed to satiation twice a day throughout the experiment with formulated diet (SFK). The other three groups were deprived of feed for 1(S1), 2(S2), and 3(S3) weeks, respectively, and then fed to satiation during the refeeding period. The results showed that trypsin specific activity was not affected significantly either by starvation or refeeding, in all experimental groups. Chymotrypsin specific activity did not change significantly in S1 fish during the experimental period. In S2 and S3 fish no significant changes were observed during the starvation period. Upon refeeding, the activity increased in S2 fish, while it decreased in S3 fish. Amylase specific activity decreased significantly during the starvation period in all experimental groups. Upon refeeding, the activity increased. Alkaline phosphatase specific activity did not change significantly during the experiment period in S3 fish, while it showed significant changes during the starvation and refeeding period in the S1 and S2 fish. Starvation also had a significant effect on the structure of the intestine.

Dietary reduction of fish meal, supplementation of vitamin C and fish oil may promote ovary maturation by improving digestibility and immunity inProcambarus clarkii(Crustacea: Decapoda)

Fish meal, vitamin C and fish oil are known to play essential roles in reproduction and immunity in animals. To investigate how the diet affect reproduction under the condition of reducing costs, we designed an experimental diet that decreased the ratio of fish meal with soybean meal from 1:1 to 1:1.5, and supplemented with 0.06% vitamin C and 3.8% fish oil for the crayfish, Procambarus clarkii. After a 100-day feeding experiment, female crayfish fed the test diet had significantly greater gonadosomatic index (GSI) compared with those fed the basal diet (P < 0.05). At the middle stage of ovarian development, the test diet gave significantly higher trypsin-specific activity (340.94 ± 57.32 U mg−1 protein) in hepatopancreas of the crayfish than the basal diet (89.48 ± 10.01 U mg−1 protein) (P < 0.05). However, hepatosomatic index (HSI) was remarkably decreased for those females fed test diet (P < 0.05). In addition, the experimental diet resulted in markedly lower superoxide dismutase (SOD) activity and malonaldehyde (MDA) levels, especially in the ovaries (P < 0.01). These results suggest that dietary reduction of fish meal, supplementation of appropriate amounts of vitamin C and fish oil may promote ovary development probably by increasing digestibility, as well as by promoting transferring of nutrients from hepatopancreas to ovary and raising immunity of P. clarkii.

Effect of Three Probiotics Administered Through Live Feed on Digestive Enzyme Activity in California Halibut, Paralichthys californicus, Larvae

The objective of this study was to evaluate the effect of probiotics as digestive system maturation promoters in larvae of the California halibut, Paralichthys californicus, during ontogeny. Three commercial probiotics: Levucell®, Bactocell®, and live yeast, were supplied at a concentration of 50 mg/L, using enriched rotifers and Artemia nauplii as vectors. Total and specific activities of trypsin, leucine‐aminopeptidase (LAP), lipase, chymotrypsin, and pepsin were assessed throughout the development. Growth in terms of length was not significantly different between treatments. Total pepsin and chymotrypsin activity were significantly higher at the end of the experiment, 46‐d posthatch (dph) in larvae supplied with Bactocell. However, no significant effect of the probiotic was detected for trypsin, lipase, and LAP. On the basis of the results, California halibut larvae seemed to have adequate alkaline proteolytic activity early in development, whereas acid proteolytic activity was not detected until the complete formation of a functional stomach. The feeding protocol and probiotics evaluated were unable to improve the morphological development of digestive system or the digestive enzyme activity early on. Nevertheless, the increase in activity of pepsin and chymotrypsin observed at 46 dph suggests a potential use of these probiotics once the metamorphosis is completed, and thus warrants further evaluation.

Digestive Enzymes in Larvae and Juveniles of Farmed Sharpsnout Seabream (Diplodus puntazzo) (Cetti, 1777)

The ontogenetic development and pattern of main digestive enzyme activities (proteases, carbohydrases and lipases) were investigated in farmed sharpsnout seabream Diplodus puntazzo larvae and juveniles, during the passage from the larval stage to adult (from 21 to 277 days after hatching - DAH). Larvae showed exponential growth, thus confirming that Rotifers and Artemia nauplii are suitable preys for early feeding of D. puntazzo larvae. Pepsin activity was low in the larvae, but it increased afterwards in relation with stomach development. In juveniles, levels of specific activity of trypsin, chymotrypsin and elastase were lower compared to that recorded in larvae. A decrease in the activity of the alkaline proteases could be related to an increase in the functionality of stomach. Qualitative and quantitative assay of glycosidases (� -amylase, cellulase and maltase) showed high glycosidase activity, indicating that D. puntazzo is capable of digesting carbohydrates in its diet. Levels of lipase activity in juveniles were higher than those found in larvae. Our results didn't show an inductive character of lipases, since in juveniles the activity levels were almost constant even if the fat content of compound diet varied (from 7.0 % to 20 %). The rapid changes in the digestive activities of D. puntazzo larvae supported the existence of well established induction and regulation digestive mechanisms. Present results also confirm that D. puntazzo larvae have complete digestive enzymatic equipment that increased during larval ontogeny and enable larvae to digest and absorb nutrients contained in live preys once exogenous feeding occurred. The enzymatic pattern is correlated with omnivorous feeding habits of the species. Moreover, the patterns of activity of the main digestive enzymes of D. puntazzo larvae and juveniles indicated the early functional development of their digestive system and suggested this species as a good candidate for Mediterranean aquaculture using the compound diet.

The optimal feeding frequency of large yellow croaker (Pseudosciaena crocea, Richardson) larvae

A 30-day feeding experiment was conducted to determine the effects of feeding frequency on the growth, survival and specific activity of digestive enzymes in large yellow croaker larvae (13days after hatch) with an initial weight of 4.08±0.1mg. Feeding frequency of 2, 4, 8 and 12 times daily as four treatments were evaluated against a control without feeding in triplicate tanks. The fish were fed to satiation each time. The results showed that treatment of 8 meals per day (meals d−1) had significantly higher survival (P<0.05) compared with other treatments. Specific growth rate (SGR) in 8 meals d−1 treatment was significantly higher than those in 2 meals d−1 or 4 meals d−1 treatments (P<0.05), but was not significantly different from that in 12 meals d−1 treatment (P>0.05). No significant differences in coefficient variations (CV%, P>0.05) of fish larvae body length were observed among the treatments, but there was a significant negative dependency between CV and feeding frequency (CV=−0.1476×feeding frequency+17.55, R2=0.9352). Both crude protein and crude lipid in fish larvae significantly increased with increasing feeding frequency. There were no significant differences in fatty acid composition of fish larvae. The specific activity of trypsin, both in pancreatic and intestinal segments, first significantly increased (from 2 meals d−1 to 8 meals d−1) and then kept at a constant level (from 8 meals d−1 to 12 meals d−1). There were no significant differences in specific activities of amylase among dietary treatments. Results of this study suggest that the optimal feeding frequency was 8 meals d−1 in the culture of large yellow croaker larvae.

Neuroligin Trafficking Deficiencies Arising from Mutations in the α/β-Hydrolase Fold Protein Family

Despite great functional diversity, characterization of the alpha/beta-hydrolase fold proteins that encompass a superfamily of hydrolases, heterophilic adhesion proteins, and chaperone domains reveals a common structural motif. By incorporating the R451C mutation found in neuroligin (NLGN) and associated with autism and the thyroglobulin G2320R (G221R in NLGN) mutation responsible for congenital hypothyroidism into NLGN3, we show that mutations in the alpha/beta-hydrolase fold domain influence folding and biosynthetic processing of neuroligin3 as determined by in vitro susceptibility to proteases, glycosylation processing, turnover, and processing rates. We also show altered interactions of the mutant proteins with chaperones in the endoplasmic reticulum and arrest of transport along the secretory pathway with diversion to the proteasome. Time-controlled expression of a fluorescently tagged neuroligin in hippocampal neurons shows that these mutations compromise neuronal trafficking of the protein, with the R451C mutation reducing and the G221R mutation virtually abolishing the export of NLGN3 from the soma to the dendritic spines. Although the R451C mutation causes a local folding defect, the G221R mutation appears responsible for more global misfolding of the protein, reflecting their sequence positions in the structure of the protein. Our results suggest that disease-related mutations in the alpha/beta-hydrolase fold domain share common trafficking deficiencies yet lead to discrete congenital disorders of differing severity in the endocrine and nervous systems.

Organogenesis of exocrine pancreas in sharpsnout sea bream (Diplodus puntazzo) larvae: characterization of trypsin expression

The ontogeny and differentiation stages of digestive systems related with trypsin expression in larvae of sharpsnout sea bream, Diplodus puntazzo, were investigated from hatching to 40 DAH (days after hatching), and total lengths and weights of larvae were determined. Histologic and enzymatic techniques were used to explain the functional development of the pancreas including trypsin activity. The pancreas was identified as a compact structure located in the region slightly posterior to the liver. At 3 DAH, first anus and then mouth opened. Incipient pancreas secretion polyhedral cells could be first observed as zymogen granules. During larval metamorphosis, the pancreas became diffuse, spreading throughout the mesentery in proximity to the stomach, the anterior intestine and the pyloric caeca. The specific activity of trypsin (42.54±6.8 mU/mg protein(-1)) was found as early as after hatching at larvae size of 2.87±0.34 mm at 0 DAH. Activity further increased until 10 DAH, especially after exogenous feeding. The highest trypsin activity was detected at 25 DAH as 119.26±11.6 mU/mg protein(-1). It is concluded that exocrine pancreas organogenesis is the main critical step in the development of digestive system that results in zymogen granules accumulation and increased trypsin activity.

Exocrine Pancreas Development and Trypsin Expression in Cultured European Sea Bass (Dicentrarchus labrax) Larvae

The ontogenesis and formation stages of exocrine pancreas in European sea bass (D. labrax) larvae were investigated from hatching to 40 days after hatching (DAH). Histological and enzymatical techniques were used to explain the functional development of the pancreas in D. labrax with the expression of trypsinogen activity. The incipient pancreas appeared as a lamination of the dorsal wall of the digestive tract. It was observed that the primary visible indication of exocrine cell differentiation was polarization. The first zymogen granules and pancreas with exocrine polyhedral cells appeared on 6 DAH and became abundant as a compact structure located dorsal and slightly posterior to the liver. At the same time, firstly, anus and then mouth were opened, and total lengths of larvae were determined as 3.47±0.26 mm. Until larval metamorphosis, the pancreas became diffuse, spreading throughout the mesentery enclosure, the stomach, the upper intestine and the pyloric caeca. On the other hand, zymogen granules were more numerous and larger, and a greater quantity of material was carried by the ducts, indicating an increased cellular activity. The specific activity of trypsin was determined as early as after hatching (42.54±6.8 mU/mg protein-1) at 4.28±0.2 mm total length of larvae and increased immediately during the following days especially after exogenous feeding. The highest tryptic activity was detected on 30 DAH as 122.45±11.76 mU/mg protein-1. It is concluded that exocrine pancreas organogenesis is the main critical step of the zymogen granules and trypsin activity is present as early as after hatching and continuously increasing with larval period of D. labrax.

Molecular characterization of trypsinogens and development of trypsinogen gene expression and tryptic activities in grass carp (Ctenopharyngodon idellus) and topmouth culter (Culter alburnus)

This study examined the gene structures and expression of trypsinogens, as well as the trypsin activities of the grass carp Ctenopharyngodon idellus (herbivorous) and the topmouth culter Culter alburnus (carnivorous), which are commercially important freshwater species of the family Cyprinidae in China. Isolated full-length trypsinogen cDNA clones were 869 bp and 857 bp. The deduced amino acid sequences were 242aa and 247aa long, both containing the highly conserved residues essential for serine protease catalytic and conformational maintenance. The results from isoelectric and phylogenetic analyses suggest that grass carp trypsinogen is grouped with teleost trypsinogen group I, while topmouth culter trypsinogen is grouped with group II. The expression pattern of trypsinogen mRNA was similar between these two species, appearing 2 days post-hatching (dph) and reaching peaks at 11 and 23 dph. The trypsin-specific activities in both species were detected 2 dph and reached the major peaks at 8 dph, however the minor peaks were observed at 20 dph in the grass carp and 17 dph in the topmouth culter. The trypsin-specific activity was significantly higher in the grass carp than in the topmouth culter, which may be attributed to the nature of their different nutritional habits.

Nutritional condition of anchovy Engraulis encrasicolus larvae in connection with mesozooplankton feeding catabolism in the southern Bay of Biscay, NE Atlantic

The study examines the spatial variations over the French continental shelf of the southern Bay of Biscay (North-East Atlantic) in the nutritional condition of anchovy ( Engraulis encrasicolus L.) larvae which was estimated through their DNA/C index. The same cruise yielded quantitative estimates of two metabolic descriptors of the functioning of mesozooplanktonic communities, the enzymes pyruvate kinase (PK) and trypsin, for the first time evaluated simultaneously. The aim was to appraise quality and quantity of matter assimilated by the mesozooplankton and to specify whether the main components of prey ingested by copepods are principally carbohydrates, through PK activity, or protein, under the action of the digestive endopeptidase trypsin. The larval DNA/C index was significantly correlated with PK specific activity but not with trypsin specific activity in the mesozooplankton. This is consistent with previous data on concomitant variations, although on quite different scales, in mesozooplankton PK activity and the larval DNA/C index.