Some proteolytic properties of hatching liquid in the sea trout,Salmo trutta m. trutta L.

Proteolytic activity of sea trout hatching liquid was examined towards casein and azocazein as a function of pH and temperature. The optimum pH for caseinolytic and azocaseinolytic activities were 9.4, and 9.0, respectively. At alkaline pH the enzyme was activated by low concentrations of Zn²⁺ ions (10⁻⁵ M). Maximum proteolytic activity of the hatching liquid was observed at 25°C. Temperatures exceeding 30°C caused a rapid reduction in enzyme activity. Proteolytic activity observed at 10°C was approximately 50% of that observed at 25°C. In general, a pseudo-Arrhenius plot indicated a Q₁₀ of 1.6 between 6 and 25°C.     

Low temperature acclimation decreases rates of protein synthesis in rainbow trout (Oncorhynchus mykiss) heart

Protein synthesis was assessed in rainbow trout (Oncorhynchus mykiss) hearts perfused with medium containing ³H phenylalanine. Isolated hearts from fish acclimated to 5° and 15°C were used as the model system, and were perfused at variable test temperatures and pH. Protein synthesis expressed as nmol PHE mg protein^–1 h^–1 was two fold higher in hearts from fish acclimated to 15°C and tested at 15°C and extracellular pH 7.6 than in hearts from fish acclimated to 5°C and tested at 5°C and extracellular pH 8.0. The prime determinant of the decreased rate of protein synthesis was thermal history. Fish acclimated to 5°C had lower levels of RNA mg protein^–1 than fish held at 15°C. There was a direct linear relationship between the rate of protein synthesis in nmol PHE mg protein^–1 h^–1 and RNA content. RNA activity (nmol PHE g RNA^–1 h^–1 remained constant regardless of thermal history or perfusion condition. Elevated pH resulted in only a marginal decrease in protein synthesis. Test temperature had no effect on in vitro rates of protein synthesis.     

Changes in Na+,K+-ATPase activity and gill chloride cell morphology in the grouper Epinephelus coioides larvae and juveniles in response to salinity and temperature

The activity of the enzyme Na⁺,K⁺-ATPase and morphological changes of gill chloride cells in grouper, Epinephelus coioides larvae and juveniles were determined 6–48 h after abrupt transfer from ambient rearing conditions (30–32 ppt, 26.5–30 °C) to different salinity (8, 18, 32, 40 ppt) and temperature (25, 30 °C) combinations. Na⁺,K⁺-ATPase activity in day 20 larvae did not change at salinities 8–32 ppt. Activity decreased significantly (P <0.01) after exposure to 40 ppt at 25–30 °C, which was accompanied by an increase (P <0.05) in density and fractional area of chloride cells. Enzyme activity in 40 ppt did not reach a stable level and larvae failed to recover from an osmotic imbalance that produced a low survival at 25 °C and death of all larvae at 30 °C. Enzyme activity and chloride cell morphology in day 40 groupers did not change in 8–40 ppt at 25 °C and 8–32 ppt at 30 °C. A significant decrease and a subsequent increase in Na⁺,K⁺-ATPase activity in 40 ppt at 30 °C was associated with the increase in chloride cell density resulting in an increased fractional area but a decreased cell size. Enzyme activity and chloride cells of day 60 grouper were unaffected by abrupt transfer to test salinities and temperatures. These results demonstrate that grouper larvae and juveniles are efficient osmoregulators over a wide range of salinities. Salinity adaptation showed an ontogenetic shift as the larvae grew and reached the juvenile stage. This development of tolerance limits may reflect their response to actual conditions existing in the natural environment.     

First multi-generation culture of the tropical cuttlefish Sepia pharaonis Ehrenberg, 1831

Sepiapharaonis, the pharaoh cuttlefish was cultured through multiplegenerations in the laboratory (5 consecutive generations) using closed,recirculating water filtration systems. The eggs of the original parentalgeneration (G_P) were spawned by a wild caught Gulf of Thailandfemale in alocal fisheries laboratory, then packed and shipped air cargo to Texas wherehatching occurred. The culture temperature ranged 25°–28°C, except for one generation that was chilled intentionallyto21 °C and then warmed to 25 °C after 9.6months. Spawning occurred as early as day 161. Spawning output was high in allgenerations except the group that was cultured at 21 °C. Eggfertility was low in captivity (< 20%), but hatchling survival was high(>70%). The average egg incubation time was 13.6 d at 25–28°C. The largest spawn resulted in 600 viable hatchlings andthesmallest resulted in 11 hatchlings. The cuttlefish ate a wide variety ofestuarine crustaceans and fishes as well as frozen shrimp. There were noapparent disease problems since survival from hatching to maturity was over70%.The average life span for cuttlefish cultured at 25–28°Cwas 8.9 months and 12.3 months at 21 °C. Size at hatching wasmeasured for fourth generation (G₄) hatchlings; the mean weight athatching was 0.103 g and the mean mantle length was 6.4mm. The largest cuttlefish cultured was a male 300 mmML and 3,045 g; the oldest cuttlefish lived 340 d.This cuttlefish species presents an excellent choice for commercial mariculturebecause of its rapid growth, short life span, tolerance to crowding andhandling, resistance to disease and feeding habits.     

Influence of pH and temperature on force development and shortening velocity in skinned muscle fibres from fish

Three species of fish were studied: Atlantic cod (Gadus morhua), sculpin (Myoxocephalus scorpius) (from the North Sea, temperature 2 to 12°C) andNotothenia neglecta (from Antarctica, temperature –2 to +2°C). Single fast muscle fibres were isolated from anterior myotomes and skinned with detergent in order to directly determine the effects of pH and temperature on force production and shortening velocity.In all species maximum force production (Po) was independent of pH over the range 7.3–8.0. Decreasing the pH from 7.3 to 6.6 reduced maximum force by 28% in fibres fromG. morhua andN. neglecta but had no effect on fibres fromM. scorpius. The depression in maximum force with acidosis was accompanied by a proportional decrease in stiffness and an increase in the rate of force recovery after stretch.Unloaded contraction velocity of cod fibres (Vmax) showed a pH optimum at around pH 7.6 decreasing by 31% at pH 6.6. Vmax of fibres from the other species was independent of pH over the range 6.6–8.0.The effects of pH on Po and Vmax were similar at 0 and 10°C. Thus for maximally activated fibres both force and contraction velocity are independent of temperature induced changes in pH. In some species acidosis depresses contractility and is likely to be a contributory factor to muscle fatigue.     

Some properties of the intestinal proteases of the rabbitfish,Siganus canaliculatus (Park)

Some properties of the intestinal proteases of the rabbitfish were examined. At 25°C, both trypsin and chymotrypsin showed pH optima of 8.0. Leucine aminopeptidase, however, displayed maximum activity in the pH range, 7.0–9.0. Leucine aminopeptidase had the highest optimum temperature (60°C), and chymotrypsin, the lowest (30°C). The optimum temperature of trypsin was 55°C. The activation energy, E_a, was found to be 8.24 for trypsin and 8.50 kcal mol^–1 for chymotrypsin. The E_a for leucine aminopeptidase was 6.29 kcal mol^–1 above 40°C and 1.73 kcal mol^–1 below 40°C. Substrate concentration-velocity plots showed that all three enzymes followed Michaelis-Menten kinetics; the K_m and V_max were estimated for the three enzymes. The effects of various protease inhibitors on enzyme activity were also examined and confirmed the protease classes to which each enzyme belonged. The three proteases examined have similar properties to proteases in other fishes.     

Rates of oxygen consumption and ammonia excretion of juvenile Eurasian perch Perca fluviatilis L.

The impact of feeding, fish size (body weight from 18.5 to 56.5 g) and water temperature (20 and 23 °C) on oxygen consumption (OC, mg O₂ kg^–1 h^–1) and ammonia excretion (AE, mg TAN kg^–1 h^–1) was studied in Eurasian perch held in recirculation systems. OC for both fed and feed-deprived (3 days) fish was higher at 23 °C (278.5 and 150.1 mg O₂ kg^–1 h^–1) than at 20 °C (249.3 and 135.0 mg O₂ kg^–1 h^–1; P < 0.01). AEs for both fed and feed-deprived fish were also significantly higher at 23 °C than at 20 °C (P < 0.001). Water temperature and fish size had a significant impact on the oxygen:feed ratio (OFR, kg O₂ kg^–1 feed fed day^–1) and ammonia:feed ratio (AFR, kg TAN kg^–1 feed fed day^–1; P < 0.001). Their average values at temperatures of 20 and 23 °C were 0.17 and 0.19 kg O₂ kg^–1 feed fed day^–1 and 0.009 and 0.011 kg TAN kg^–1 feed fed day^–1, respectively.     

Development of wolffish eggs at different temperature regimes

Embryonic development of common wolffish (Anarhichas lupus L.) was studied at constant temperatures 5.0, 7.0, 9.0, 11.0, 13.0 and 15.0°C. Duration of development from egg activation to several morphological stages including 50% hatching was determined. At 5.0–11.0°C, the survival rate of eggs to hatching ranged from 51 to 88% with a tendency to increase at 5.0 and 7.0°C. Morphological anomalies, bacterial contamination and large mortalities were observed in eggs incubated at 13.0 and 15.0°C. The period of hatching lasted from 10 to 50 d in different egg groups. Embryo length and yolk sac volume at identical morphological stages of development showed only slight relation to temperature. At lower temperatures newly hatched larvae were longer and at more advanced stages of ontogeny. Normal numbers of fin rays in larvae (mean values 74 for dorsal fin and 46 for anal fin) were observed at 5.0 and 7.0°C and in most larvae at 9.0°C. At 11.0 and 13.0°C, many rays were absent, with mean values for dorsal fin 60 and 39 respectively and for anal fin 28 and 4 respectively. The approximate upper limit for normal development of fin rays appeared to be 9.0 °C.     

Characterization of the microsomal epoxide hydrolase of hepatic microsomes of the common dab,Limanda limanda

Epoxide hydrolase of microsomal membranes of the common dab (Limanda limanda) has been characterized using p-nitrostyrene oxide as substrate. Under the conditions of assay used, the turnover number with this substrate was higher than found for the more frequently used styrene oxide and steady state kinetics were observed. The enzyme had a K_M of 0.12 mM and optima for pH and temperature between pH 8–10.2 and 50–60°C respectively. Enzyme activity was unaffected by low concentrations of ionic and non-ionic detergents but was inhibited by higher concentrations of Lubrol and Brij. The enzyme protein did not react with monospecific antibodies to rat or human microsomal epoxide hydrolase during Western blotting. Large inter-individual variation in enzyme activity was found but the enzyme does not appear to be expressed in a gender-specific way. Fish were administered a wide range of hydrocarbons which are known to alter the expression of cytochrome P450 1A but these had no effect other than benzothiophene which caused a small increase in enzyme activity.Abbreviations mEH microsomal epoxide hydrolase - pNSG p-nitrostyrene glycol - pNSO p-nitrostyrene oxide     

Kurokura Solution as Immobilizing Medium for Spermatozoa of Tench (Tinca tinca L.)

This study tested KUROKURA solution (Kurokura et al., 1984, Aquaculture 37, 267–274) and its modifications (by increasing NaCl content to 160, 180 and 200 mM) on immobilizing properties for sampling and short-term preservation of potential motility of tench spermatozoa. The immobilizing solution is used because, when collected, the sperm of most samples is contaminated by urine, causing spermatozoa to be of poor quality, with low motilities and velocities (almost 0), thus resulting in a worsened fertilization and hatching rate. Sperm was sampled with a syringe containing an immobilizing solution (IS), allowing an IS:sperm ratio of 2:1, under aerobic conditions at 0–4°C. This sperm solution was stored for 10 h and untreated sperm was collected prior to fertilization as a control. Spermatozoa quality was evaluated for the cell motility and velocity parameters and also for fertilization ability and hatching rate. Results obtained for tench sperm motility, velocity, fertilization and hatching rate showed that only sperm collected in the various immobilizing solutions can be successfully used for artificial insemination and preservation after 10 h at 0–4°C. The best immobilizing solution was found to be KUROKURA 180 (180 mM NaCl, 2.68 mM KCl, 1.36 mM CaCl₂· 2H₂O and 2.38 mM NaHCO₃), giving a fertility and hatching rate of 41%, with no change in rates after 10 h storage of sperm. Control sperm without immobilizing solution showed a fertilization and hatching rate of only 6–7%.     

Actions of epinephrine on the contractility of fast and slow skeletal muscle fibres in teleosts

Fast and slow muscle fibers were isolated from the myotomes of atlantic cod (Gadus morhua L.) and sculpin (Myoxocephalus scorpius L.). Epinephrine was found to have no effect on twitch or sub-tetanic contractions in fast muscle fibres. Isoprenaline (10^–6M) had no effect on the contractility of slow muscle fibres. In contrast, epinephrine elicited a dose-dependent decrease in the half-time for twitch relaxation (t_1/2r), and in most cases a decrease in twitch amplitude. The maximum decrease in t_1/2r was around 5–20% of control values (at 10^–6M epinephrine), with a half maximal response at about 30 nmol l^–1. Responses to epinephrine were unaffected by propranolol and reversed by phentolamine, consistent with the stimulation of -adrenoreceptors. 10^–6M epinephrine produced a rise in cAMP levels from 1.8 to 3.1 pmol mg dry wt^–1 in cod slow fibres. However, the cellular mechanism underlying the action of epinephrine is unclear since forskolin, a potent activator of adenylate cyclase activity, where it has been investigated, was found to increase not decrease twitch duration and amplitude. The responses of fast and slow fibres to epinephrine and its antagonists were similar in summer (13°C) and winter acclimatized (5–6°C) sculpin.It is suggested that epinephrine may act to modulate the active state of slow muscle fibres at high cruising speeds and thereby increase swimming performance.     

Metabolic biochemistry of cardiac muscle in three tuna species (bigeye, Thunnus obesus ; yellowfin, T. albacares ; and skipjack, Katsuwonus pelamis ) with divergent ambient temperature and oxygen tolerances

Bigeye tuna (Thunnus obesus) have much greater vertical mobility than yellowfin (T. albacares) and skipjack (Katsuwonus pelamis) tunas, due to an apparent greater tolerance of the changes in ambient temperature and oxygen occurring with depth. In an attempt to identify physiological processes (e.g., effects of temperature on cardiac function) responsible for these behavioral differences, we examined enzyme activities (at 12 °C, 17 °C, and 25 °C) of cardiac muscle in all three species. Contrary to our expectations, we found few differences and no clear explanatory patterns in maximum enzyme activities (V_max) or enzyme activity ratios. For example, citrate synthase (CS) activity was the same in bigeye and skipjack tunas, but 40% lower in yellowfin tuna, whereas carnitine palmotoyltransferase (CPT) activity in skipjack tuna was approximately double that in the other two species. The ratio of CPT to pyruvate kinase (PK) activity, a measure of the tissues preference for fatty acids as metabolic substrates, was the same in bigeye and yellowfin tunas, but elevated skipjack tuna. The ratios of lactate dehydrogenase (LDH) to CS activity and of PK to CS activity (anaerobic–aerobic enzyme activity ratios – taken as measures of the tissues ability to tolerate hypoxia) were both elevated in yellowfin tuna cardiac tissue relative to the other two species. We also found no differences in temperature sensitivity (Q₁₀ values) when comparing cardiac enzyme activities across species, nor effects of temperature on the substrate affinity (K_m) of LDH. In sum, our results do not suggest any clear metabolic difference in the cardiac muscle that would explain the apparent greater tolerance of bigeye tuna to acute hypoxia and ambient temperature changes or their substantially greater vertical mobility.     

Drinking activity of the newly hatched larvae of codGadus morhua L.

The drinking rate of cod larvae 1–7 days post hatching was measured from the uptake of³H-labelled dextran (MW = 70000) admixed in the incubation seawater (34 ppt 5°C). The drinking rate increased gradually from 0.15% to 0.59% of the larval body weight on day 1 and day 7 respectively. This increase in drinking rate correlated with an observed decrease in the volume of the yolk sac and its water store. Autoradiographs showed the labelled dextran to be confined to the intestine. Electron micrographs showed an open mouth communicating with the oesophagus and the intestine in cod larvae at the time of hatching. Chloride cells were present on the opercular folds but not on the vestigial, developing gills. The data indicate that the water acquisition mechanism of larval cod is similar to that of adult marine fish.     

Temperature Tolerance and Metabolic Depression of a Convict Child Under the Influence of Enhanced Ultraviolet-A (320–400 nm) Irradiation

The effect of an enhanced (5%) sub-lethal UVA irradiation on the total metabolism of a sub- tropical convict cichlid (Cichlasoma nigrofasciatum) is described. The specific oxygen consumption (vo₂) of related but differently treated populations was determined from 23 °C to 35 °C, to detect their tolerance limits toward environmental temperature. The experiments cover two generations (G1 and G2) of inbred derived populations. The fish were measured at different stages of development covering the period before and after maturation. The fish were grown at and adapted to 27–29 °C. The results showed that the population which was grown under an enhanced UVA irradiation since hatching (G2-UVA) had developed a general metabolic depression (MD) at all ranges of experimental temperature. The MD was especially expressed after the fish reached maturation. No MD was shown for the non-irradiated control populations (G2-control and G1-control) nor for the mature fish of G1-UVA. The G1-UVA population was grown under an enhanced UVA irradiation from 4 months of age. The vo₂ increased with increasing temperature, reaching a maximum at 31–33 °C for the G1- and G2-control populations. For the G1-UVA, a small peak of vo₂ was obtained at 29 °C reaching its maximum level at 33 °C. For the G2-UVA population, a maximum vo₂ was reached at 33 °C for the young and mature fish and at 29 °C during the maturation period. It can be concluded that the accumulation of UVA irradiation from early life was responsible for the MD in this fish. A wider temperature tolerance observed in the G2-UVA population might possibly be due to a reduction of total metabolic activity. The mechanisms underlying the MD observed in this fish are not yet known, but natural selection and adaptation processes might be involved.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Effect of water hardness on egg hatchability and larval viability of Clarias gariepinus

Studies were conducted to determine the effect of water hardness onClarias gariepinus egg hatchability and larval viability.The fertilized eggs were incubated at 28 °C and with waterhardnesses ranging from 10–700 mg/l CaCO₃. Themean hatching rate fluctuated between 42.31% at hardness of 10mg/land 64.66% at 2000 mg/l. Abnormalities in the larvae were observedbeyond 200 mg/l and increased with increase in water hardness. Thehighest larval survival of 71.05% was recorded at 60 mg/l waterhardness. Based on statistics performed with analysis of variance (ANOVA) andfurther compared with Duncan's multiple range test (p = 0.05), the resultsimplythat very soft water (0–10 mg/l) and very hard water (300mg/l and above) are not suitable for Clariasegg incubation and larval rearing. A water hardness of 30–60mg/l CaCO₃ is recommended for optimal normal hatching,high viability and maximum larval development of Clariasgariepinus.     

Surface disinfection of Atlantic halibut and turbot eggs with glutaraldehyde: evaluation of concentrations and contact times

To evaluate the effects of glutaraldehyde treatment at different disinfection temperatures, Atlantic halibut, Hippoglossus hippoglossus (L.), and turbot, Scophthalmus maximus (L.), eggs, incubated at 5 and 12°C, respectively, were disinfected with 400–1200 mg glutaraldehyde l-1 at three different contact times (2.5, 5 and 10 min). Egg batches of both poor and good quality were tested for halibut. Positive effects were more pronounced in poor than in good-quality batches at hatching. Egg disinfection had a highly positive effect on the viability of yolk-sac larvae in both types of batches. A level between 400 and 800 mg l-1 at a contact time of 5–10 min was optimal for halibut: at lower levels, the bactericidal effect was reduced, and at higher levels, there were indications of toxic effects. Halibut eggs disinfected with optimal doses of glutaraldehyde had furthermore a reduced hatching time and more synchronous hatching as compared with untreated eggs. Turbot was more sensitive to higher doses than halibut, and the best larval performance was obtained for 400–800 mg glutaraldehyde l-1 at a contact time of 2.5 min. A further evaluation should, however, be performed before recommendations are given for species incubated at temperatures higher than 5°C.     

Temperature shifts induce adaptive changes in the physical state of carp (Cyprinus carpio L.) erythrocyte plasma membranes in vitro

Blood, freshly collected from warm- and cold-acclimated carp, Cyprinus carpio L., was cooled to 5°C for 4h or warmed to 25°C for 4h, respectively, and the fluorescence anisotropy of washed red blood cells was recorded using the fluorescent dye 3-(p-(6-phenyl-1,3,5-hexatrienyl) phenyl propionic acid [DPH-PA] (which is restricted to the outer leaflet of the plasma membrane) before and after the temperature shift. Despite individual variation, the plasma membrane of cold-exposed erythrocytes became more fluid while that of warm-exposed cells became more rigid following the temperature shift. This response was rapid and reversible. Cold-exposed cells from warm-acclimated fish became more fluid within 40–60 minutes and reverted to their original fluidity within the same time on warming, at a rate of 1°C/min; erythrocytes, from cold-adapted carp displayed an opposite change in fluidity over a similar time period. Cells from warm-acclimated, temperature down-shifted carp hyperfluidized their plasma membranes in the cold, whereas cells from cold-acclimated fish up-shifted in temperature showed no similar effect. These cells showed a complete adjustment of membrane physical state to the temperature. Total phospholipids obtained from warm-acclimated temperature down-shifted cells became more rigid than they were, when assayed at the acclimation temperature. In contrast, phospholipids obtained from cold-acclimated cells became more rigid when exposed to increasing temperatures. No significant changes occurred to the polar head groups, or to the fatty acid composition of the total phospholipids. It was concluded that the lipids play only a secondary role in the control of the physical state of plasma membrane in carp erythrocytes, and that some non-lipid components of these structures might be involved in these regulatory processes.     

Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.)

Proteolytic activity in the different parts of the digestive tract of the turbot (Scophthalmus maximus L.) were studied in this work. One pure protease was isolated from turbot stomach and its behavior was studied. Results showed the optimum pH for proteases in the different parts of the digestive tract of the turbot were pH 2.0 for the stomach, pH 8.0 for the pylorus cecum, pH 8.0 for the foregut, pH 8.5 for the midgut, and pH 8.0 for the hindgut. The activity of proteases in the different parts of the digestive tract were in the sequence pylorus cecum protease > stomach protease > foregut protease > midgut protease > hindgut protease. The stomach protease was purified by ammonium sulfate precipitation and column chromatography on DEAE-Sepharose F.F. and Sephadex G-100. The purified enzyme gave a single band in SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Its molecular weight was found to be approximately 42,000 Da. The enzyme is stable at pH 1.0–9.0 and at temperatures below 40°C. Its activity was maximum at pH 2.0 and 40°C. When reaction time was prolonged the optimum temperature of the enzyme tended to decline. The enzyme was activated by Mn²⁺ and Cu²⁺ and inactivated by Fe³⁺. It was fully inhibited by pepstatin and partially inhibited by PMSF, TPCK, PCMB, and NBS. These results imply the enzyme is a pepsin.     

Abnormalities in the juvenile stage of sea bass (Dicentrarchus labrax L.) reared at different temperatures: types, prevalence and effect on growth

Osteological malformations are always considered an important problem in intensive aquaculture. This work studies the effect of rearing temperature on malformations in sea bass, Dicentrarchus labrax. To this end, two batches of the species were subjected to the following incubation/cultivation temperatures: 15 °C/natural and 19/19 °C, from fertilization until 190 days after hatching. The different malformations were studied in 1643 juveniles at 190 days and classified by categories. The Chi-square statistic was calculated to verify the relationship between the presence of anomalies and the application of different temperatures. The percentage of anomalies observed in individuals reared at a high temperature (19/19 °C) was 66.44%. In both temperature systems, opercular malformations were those which caused a greater delay in growth. The results found indicated that temperature played a very important role in the development of deformations, which may be of interest from the viewpoint of aquaculture.