Lysozyme,chitinase and exo-N-acetyl-β-D-glucosaminidase (NAGase) in lymphomyeloid tissue of marine fishes

Lymphomyeloid (haemopoietic) tissues produce or store blood cells — among these leucocytes rich in lysosomal enzymes. The thymus, unlike the other lymphomyeloid tissues found in fish, produces exclusively lymphocytes. The carbohydrate-hydrolysing enzymes lysozyme (EC 3.2.1.17), chitinase (EC 3.2.1.14) and exo-N-acetyl--D-glucosaminidase (NAGase; EC 3.2.1.30) were assayed in various lymphomyeloid tissues of the cartilaginous fish Chimaera monstrosa, Squalus acanthias, Etmopterus spinax and Raja radiata, and in the thymus of the marine teleost Lophius piscatorius. Lysozyme activity was high in the cranial lymphomyeloid tissue of C. monstrosa; in Leydig's organ (oesophageal lymphomyeloid tissue) and the spleen from E. spinax; and in Leydig's organ, the epigonal organ and the spleen from R. radiata. Little or no lysozyme activity was found in Leydig's organ and the epigonal organ of S. acanthias, or in the thymus of C. monstrosa, R. radiata and L. piscatorius. The pH optima for lysozyme activities lay between 4.8 and 5.4 when assayed photometrically. Chitinase was most active at pH 1 in Leydig's organ from R. radiata, and at pH 2.7 in the epigonal organ from S. acanthias. The chitinolytic activity in Leydig's organ of E. spinax may be due to lysozyme. The optimum for NAGase activity in Leydig's organ from R. radiata lay at pH 4.0, that from S. acanthias and E. spinax at pH 4.5. The role of the enzymes in the defense against microorganisms and parasites is discussed.     

Chitinolytic enzymes in the digestive system of marine fishes

Chitinase, exo-N-acetyl--D-glycosaminidase (NAGase) and lysozyme activities were assayed in the digestive tract of 6 species of marine fishes: Myxine glutinosa (cyclostome), Chimaera monstrosa (holocephalan), Squalus acanthias, Etmopterus spinax, Raja radiata (elasmobranchs) and Coryphaenoides rupestris (teleost). Strong chitinase activity was found in the gastric mucosa of the elasmobranchs (S. acanthias, E. spinax and R. radiata) and the teleost (Coryphaenoides rupestris). A remarkably high chitinase activity occurred in the pancreas of the stomachless holocephalan fish Chimaera monstrosa. NAGase activity was strong in the digestive tract of all species. It could be concluded that marine fishes with diets consisting largely of chitinous invertebrates may display high chitinase and NAGase activities in their digestive system; however, only low chitinase activity was found in the intestine of the cyclostome Myxine glutinosa. Coryphaenoides rupestris gastric mucosa chitinase had one optimum activity at pH 1.25, whereas S. acanthias chitinase had two optima, at pH 1.6 and 3.6. The NAGase pH-activity curves from S. acanthias and R. radiata gastric mucosa displayed similar optima, at pH 4.5 and 4.25 respectively. Chimaera monstrosa pancreatic chitinase had a very strong optimum around pH 8 to 10, and one less strong at pH 3. These enzyme activities could not be separated by gel filtration or isoelectric focusing. The pI (isoelectric point) was approximately 4.9 for both enzymes. The molecular weight of the C. monstrosa pancreatic chitinase was estimated to be approximately 43 000. Lysozyme activity was absent or extremely weak in the material studied.     

Role of bacteria with regard to chitin degradation in the digestive tract of the cod Gadus morhua

Numbers of aerobic, heterotrophic and chitinolytic bacteria in stomach, pyloric caeca and intestine of eight specimens of freshly caught cod (Gadus morhua L.) were determined by the spread plate method (medium: sea water agar, including 1% chitin). In crude enzyme extracts, chitinase activity was assayed according to the end product measurement. Results revealed no correlation between the number of chitinoclasts and the level of chitinase activity in any part of the digestive tract of cod. In another series of studies, antibiotics (Ampicillin/Sisomicin, or Chloramphenicol) were applied via tank water and feed. In most cases, antibiotic treatment resulted in the complete elimination of bacteria from the digestive tract. In contrast, up to 1.0×10⁹ chitinoclasts per g wet wt were determined in control fish. Chitinase did not reflect the absence or presence of bacteria: no significant difference in enzyme activities of cod treated with Ampicillin/Sisomicin and non-treated fish could be demonstrated. A decrease of chitinase activities of cod exposed to Chloramphenicol is interpreted as a toxic effect of this antibiotic on the fish. It is concluded that chitinase activities measured in the digestive tract of G. morhua primarily originate from fish tissues.Results presented in this paper are part of the author's Ph.D. dissertation. Authorization for pre-publication was given by the Graduation Committee of the Biology Department, University of Hamburg     

Effects of environmental hypercapnia on hemato-immunological parameters,carbonic anhydrase,and Na+, K+-ATPase enzyme activities in rainbow trout (Oncorhynchus mykiss) tissues

In this study, the effects of environmental hypercapnia on hemato-immunological parameters and the activities of respiratory enzymes such as carbonic anhydrase (CA) and Na⁺, K⁺-ATPase were investigated in rainbow trout (Oncorhynchus mykiss) tissues (gill, liver and kidney). Batches of 12 fish were exposed to 4.5 mg L^?1 (control) and 14 mg L^?1 CO₂. No mortalities occurred during the 14 days of the experimental period. Red blood cell (RBC), hemoglobin (Hb), and hematocrit (Ht) levels, and innate immune parameters such as nitro blue tetrazolium (NBT), lysozyme, and myeloperoxidase activities, and the melano-macrophage frequency were negatively affected by elevated CO₂ levels. Patterns of change in CA activity differed among the gill, liver, and kidney. Compared with the activities of CA in the control group, the CA enzyme was significantly stimulated at day 7 in the gill tissue, whereas it was stimulated at day 14 of the experiment in the liver tissue of fish exposed to 14 mg L^?1 CO₂ (P < 0.05). In contrast to the pattern of CA enzyme activities, the Na⁺, K⁺-ATPase enzymes were stimulated significantly in the liver after day 7 but inhibited in the kidney and gill (P < 0.05). These results suggest that a subchronic exposure to hypercapnia of rainbow trout tissues may lead to adaptive changes in the respiratory enzymes and negatively affects hemato-immunological parameters.     

Bacterial chitinase in the stomachs of marine fishes from Yaquina Bay,Oregon, USA

The occurrence of chitinase in the stomach contents of Enophrys bison and Platichthys stellatus was investigated. The highest chitinase activity and the greatest percentage of chitinoclastic bacteria in relation to the total bacterial flora were associated with fish whose stomach content was composed primarily of chitinous animals. Stomach contents lacking visible chitin possessed low or no detectable levels of chitinase activity and few chitinoclastic bacteria. Juvenile E. bison treated with chloramphenicol to remove their indigenous bacterial flora had no detectable level of chitinase in their stomach contents while non-treated juveniles showed inducible chitinase activity, indicating the bacterial origin for the chitinase.Published as Technical Paper No. 4435, Oregon Agricultural Experiment Station. This research was supported by NIH training grant 5TO1 GM-00704 and NSF grant DES73-06611 AO2.     

Incidence and estimation of chitinase activity associated with marine fish and other estuarine samples

A procedure for the determination of chitinase activity was adapted for the seawater environment. Preliminary data indicate that the controlled bacterial environment within the digestive tracts of marine fishes and possibly other marine animals plays a significant role in the decomposition and recycling of chitin. It is estimated in the stomachs of a single population of Enophrys bison (buffalo sculpin) of 1×10⁵ fish that ca. 16 metric tons of chitin could be decomposed annually.Published as Technical Paper No. 4434, Oregon Agricultural Experiment Station. This research was supported by NIH training grant 5TO1 GM-00704 and NSF grant DES73-06611 AO2.     

Fluids and jellies of the acusticolateralis system in relation to body fluids in Coryphaenoides rupestris and other fishes

The marine deep-water fish Coryphaenoides rupestris has a lymph-like fluid in its large cranial lateral-line canals. A similar canal endolymph is also found in a few other fishes possessing closed lateral-line canals. We have analysed, chemically, the canal endolymph from 3 species of teleosts. In C. rupestris, the ionic composition resembles that of blood plasma, but the content of potassium is slightly lower. In Lota lota, the canal endolymph has a high potassium concentration, as does the inner-ear endolymph. In Glyptocephalus cynoglossus, the ionic composition of the canal endolymph resembles sea water. In all species, the concentrations of glucose and protein are much lower than in the blood plasma. Analyses were further made on endolymph and perilymph of the inner ear, on jelly-like material from the acusticolateralis system, and on body fluids from a small number of fish species, even elasmobranchs and a holocephalan. The water phase of the cupulae of the lateral-line system of C. rupestris had an ionic composition similar to that of the endolymph surrounding the cupulae. High potassium and low values for glucose and protein were found in the inner-ear endolymph of all species investigated. No carbonic anhydrase activity was found in the lateral-line system of C. rupestris, whereas a low activity of this enzyme was found in the inner ear of Gadus morrhua. The investigation shows that the composition of the fluids in contact with the mechano-receptors of the lateral-line system differs between different species of fishes. A few observations were made on the structural organization of the cranial lateral-line system in C. rupestris.     

Cloning and expression of high choriolytic enzyme,a component of the hatching enzyme system,during embryonic development of the marine ornamental fish<Emphasis Type="Italic"> Chrysiptera parasema</Emphasis>

In the present study, a cDNA for the hatching enzyme of a marine tropical fish, Chysiptera parasema, was cloned. This is the first demonstration of hatching enzyme cDNA from a marine tropical fish. The amino acid (aa) sequence deduced from the cDNA consisted of an 18-aa signal sequence, a 53-aa propeptide sequence and a 196-aa mature enzyme portion, having a consensus active site sequence for astacin family proteases. Phylogenetic analysis showed that the C. parasema enzyme was included in the clade of HCEs (high choriolytic enzymes), one of the hatching enzymes of freshwater fishes such as medaka (Oryzias latipes), masu salmon (Oncorhynchus masou) and zebrafish (Danio rerio), but not in the group of LCEs (low choriolytic enzymes), another type of hatching enzymes identified in the medaka. The developmental expression patterns of the C. parasema HCE gene were highly similar to that of the medaka HCE gene. The results suggested that the hatching enzyme system is highly conserved between these marine and freshwater fish species.Communicated by R. Cattaneo-Vietti, Genova     

Concentration of ascorbic acid and innate immune effectors in <Emphasis Type="Italic">Engraulis ringens</Emphasis> and <Emphasis Type="Italic">Strangomera bentincki</Emphasis> during their main spawning period (2007–2008) in the Humboldt current system off Chile

In this study, we determined variations in ascorbic acid (AA) concentrations in the gonads and liver, and of the innate immune response (lysozyme activity and antiprotease) in the plasma of Engraulis ringens (anchoveta) and Strangomera bentincki (common sardine) during their reproductive seasons of 2007 and 2008 in the central area of the Humboldt Current. During the main spawning season in 2007, colder environmental conditions, higher phytoplankton biomasses, low dinoflagellate abundance and the dominance of large copepods in winter-early spring and of small copepods in late spring contributed to explain higher AA concentrations in anchoveta tissues. During the warmer year of 2008, lower phytoplankton biomasses, a notable increase of dinoflagellate abundance and the dominance of small-size copepods in winter and of larger copepods in spring occurred along with a drastic drop in AA concentrations in anchoveta tissues. These results contrasted with those in common sardine, a species in which AA did not vary as much as in anchoveta. An inverse trend between AA concentration in the liver and the gonadosomatic index suggested the AA utilization during gonadic maturation. Innate immune parameters were determined for the first time in small pelagic fish and constitute a new baseline data to evaluate their natural response to environmental changes. Changes in the distribution and abundance of edible taxa containing AA and edible for fish along with variations in AA in fish tissues during their reproductive season could become good indicators of the overall fish physiological condition resulting from variations in the trophic web structure.     

A preliminary approach to epidermal antimicrobial defense in the Delphinidae

In three delphinid species (Pacific spotted dolphin, Stenella attenuata; common dolphin, Delphinus delphis; harbor porpoise, Phocoena phocoena) the enzyme lysozyme and the peptide group -defensins are demonstrated for the first time in cetaceans as products of the thick integument. Lysozyme was found between the lamellae of the stratum corneum, in the cells of the stratum spinosum, free cells of the dermis, and in endothelial cells of dermal blood vessels. -defensins (especially type 3) were clearly located in cells of the upper stratum spinosum, and concentrated between the upper five or six layers of the stratum corneum. The occurrence and localization of such substances that may serve as a non-specific defense against bacteria, fungi, algae, and ectoparasites, was known until now only for haired marine mammals with typical skin glands, like in pinnipeds.Communicated by O. Kinne, Oldendorf/Luhe     

Distribution of enzymes of glycolysis and gluconeogenesis in fish tissues

Gluconeogenesis in fishes has been demonstrated in whole animals and liver preparations. However, at present, the relative physiological importance of possible substrates such as lactate, pyruvate and amino-acids or the precise sites of gluconeogenesis are unclear. In mammals, gluconeogenesis takes place in the liver and kidney, and the same could occur in fishes although it has been proposed that fish red muscle is also capable of reconverting lactate (derived from white muscle) to glucose. In this present study, the activities of 3 key glycolytic (hexokinase, phosphofructokinase and pyruvate kinase) and 2 key gluconeogenic (fructose diphosphatase and phosphoenolpyruvate carboxykinase) enzymes were investigated in tissues of the rainbow trout Salmo gairdneri, the cod Gadus morhua, and the plaice Pleuronectes platessa in order to elucidate the relative glycolytic/gluconeogenic capacities of the individual fish tissues. The glycolytic enzymes were found in all tissues, the relative potential being skeletal muscle>heart, brain >kidney, gills>liver. The gluconeogenic enzymes were not present in all tissues, and were mainly concentrated in the liver and kidney. Hence the results indicate that the liver, and to a lesser degree, the kidney are the major sites of gluconeogenesis in fishes, and that the process is unlikely to occur in skeletal muscle.     

Sulfide tolerance and detoxification in shallow-water marine fishes

Hydrogen sulfide is a potent inhibitor of aerobic respiration. Sulfide is produced in sediments, and many species of fish live in association with the bottom. Tolerance tests, enzyme assays, and chromatography of sulfur compounds in thirteen species of shallow-water marine fishes (collected in San Diego, California, USA in 1987–1988) indicate adaptations to sulfide that vary with habitat and lifestyle. Tidal-marsh inhabitants, like Gillichthys mirabilis and Fundulus parvipinnis, have higher tolerance to sulfide (96 h LC₅₀ at 525 to 700 M) relative to outer-bay and open-coast inhabitants (surviving <12 h at much lower concentrations). The cytochrome c oxidase of all species shows high activity and susceptibility to sulfide poisoning, with 50% inhibition at 30 to 500 nM in various tissues. The two marsh species are able to survive at sulfide concentrations already inhibitory to their cytochrome c oxidase and fatal to other species. All species detoxify sulfide by oxidizing it to thiosulfate. All have sulfide-oxidizing activity in the blood, spleen, kidney, liver and gills, which correlates significantly with heme content. Thiosulfate appears in the tissues of sulfide-exposed fish and builds up to high concentrations (up to 2 mM) with stronger and longer exposure. Unexposed fish contain little or no thiosulfate. Sulfide is barely detectable in the tissues, even in high-sulfide exposure tests. We suggest that fish blood, in having high sulfide-oxidizing activity and no cytochrome c oxidase, can act as a short-term first line of defense against sulfide, and thus minimize the amount that reaches the vital organs. The results of this study indicate that sulfide is a significant environmental factor influencing the ecological distribution of marine fishes.     

Biotransformation rates of xenobiotic compounds in relation to enzymes activities: A critical review

Many biotransformation reactions and many activities of enzymes involved in the biotransformation of xenobiotics in fish have been described. Therefore, the prediction of biotransformation of (other) xenobiotics through literature data seems to be possible. However, very few data allow correlation of in vitro enzyme activities with in vivo biotransformation rates. This is mainly due to a lack of quantitative data. Suggestions are made for future research, which may help to relate enzyme activities and biotransformation in fish.     

Biochemical indices of aerobic and anaerobic capacity in muscle tissues of California elasmobranch fishes differing in typical activity level

Biochemical indices of aerobic and anaerobic metabolic capacity were measured in white myotomal muscle of eight marine elasmobranch fish species representing a broad range of locomotor performance, and in red myotomal muscle and heart of three of those species. The objectives were to determine if metabolic capacities vary with typical fish activity level, to compare the endothermic mako shark with ectothermic pelagic sharks, and to compare elasmobranchs with teleosts in order to test the hypothesis that elasmobranchs have lower aerobic capacities, metabolic rates, and swimming speeds. In white myotomal muscle, activities of the enzymes citrate synthase (an index of aerobic capacity), pyruvate kinase, and lactate dehydrogenase (LDH, an index of anaerobic capacity) covaried with typical activity level, and the ability to tolerate intracellular acidification (nonbicarbonate buffering capacity) corresponded with LDH activity. Enzyme activities in red muscle and heart did not show a consistent pattern with respect to fish activity. In comparison with ectothermic sharks, the mako shark had greater aerobic and anaerobic capacities in white muscle, but no significant differences were found in red muscle or heart. This pattern has also been found in teleosts. Thus, endothermic fishes elevate the temperature of red muscle, a tissue specialized for high aerobic performance, whereas white muscle biochemical characteristics are adjusted to support high rates of contraction both aerobically and anaerobically. Muscle enzymic activities of elasmobranchs and teleosts with comparable locomotor habits are similar, thus refuting the hypothesis that elasmobranchs are sluggish, with lower metabolic capacities than teleosts.     

Blood sugar metabolism during moulting in the isopod crustacean Ligia exotica

Blood sugar metabolism of the isopod Ligia exotica Roux was studied in relation to chitin synthesis during the moult cycle. The blood sugars were found to occur free, as well as bound with blood proteins, during all moult stages, as in Emerita asiatica. The variety of free blood sugars is smaller in L. exotica; it comprises only glucose and glucose-6-phosphate. The protein-bound blood sugars are glucose and glucosamine. The blood-volume values do not differ markedly during the moult cycle. Estimations of blood sugars showed a marked rise of sugar levels in the blood during the premoult phase, and a subsequent fall during the freshmoult phase, when chitin is actively synthesised. The chitin values also showed a definite correlation with the blood-sugar values. The increased resorption of chitin during the premoult stage, and its liberation into the blood stream as glucosamine, is indicative of its reutilisation in chitin synthesis as in E. asiatica and the genus Cecropia. As more immediate precursors of chitin, such as uridine diphosphate and uridine diphophate acetylglucosamine compounds, were absent in the blood, a thorough biochemical study of the epidermis is suggested to locate the intermediate precursors of chitin which are possibly derived from blood sugars.     

Mercuric chloride induced proteotoxicity and structural destabilization in marine fish (Therapon jarbua)

The reaction byproducts derived from lipid peroxidation (LPO), as well as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) enzyme activities were measured in tissues of marine fish (Therapon jarbua) exposed to mercuric chloride (HgCl) in water dispersions of 0.125 or 0.25 ppm. LPO was significantly increased in various tissues relative to control values after 96-h exposure. SOD and GPx activities significantly decreased after exposure to first two doses but significantly elevated CAT in Dose II kidney and liver tissues. The elevated levels of LPO, decreased activities of SOD and GPx, and increased CAT activities in all tissues examined in T. jarbua are an index of oxidative stress in fish. Structural analysis by scanning electron microscopy studies revealed the structural deformation in HgCl₂-exposed animals. These observations suggest that HgCl acts as a mediator in free radical generation. The increase in CAT and decrease in SOD and GPx activities in these tissues may be an adaptive response.     

Two distinct forms of the chitin-degrading enzyme N-acetyl-β-d-glucosaminidase in the Antarctic krill: specialists in digestion and moult

In the Antarctic krill Euphausia superba two forms of the chitinolytic enzyme N-acetyl-β-d-glucosaminidase (NAGase, EC 3.2.1.52) have been described, previously identified as NAGase B and NAGase C. Here, we demonstrate the organ-specific distribution and physiological relevance of both forms using a polyclonal antibody preparation which allows them to be distinguished immunologically. While NAGase B was localized in the integument and displayed a pattern of activity related to the moult cycle, the activity of NAGase C was independent of the moult cycle and was predominantly found in the gastrointestinal tract. Accordingly, NAGase B played a significant role in chitin degradation during the krill's moult, whereas NAGase C participated in the digestion of chitin-containing dietary components. Chromatographic elution profiles of isolated organs confirmed the immunological results by displaying characteristic organ-specific patterns in NAGase activity. The molecular characteristics of the moulting form, NAGase B, may further indicate a vesicular transport of moulting enzymes from the epidermis into the ecdysial space. Based on our results we develop a hypothesis explaining the concurrent processes of simultaneous chitin degradation and chitin synthesis occurring during moult. Received: 30 December 1998 / Accepted: 23 April 1999     

Acute toxicity of waterborne manganese to Rutilus caspicus (Yakovlev, 1870) – gill histopathology,immune indices,oxidative condition,and saltwater resistance

The acute toxicity of manganese (Mn) in Caspian roach Rutilus caspicus was determined. The 96 h-LC₅₀ was calculated to be 300 (240–360) mg L^?1. Then, fish were exposed for 96 h to 300 (96 h-LC₅₀), 150 (50% of 96 h-LC₅₀), 60 (20% of 96 h-LC₅₀), and 0 (control group) mg L^?1 Mn. Exposure to all Mn concentrations led to fish gill damages, including epithelial lifting, hyperplasia, lamellar fusion, aneurism, and lamellar curling. Also, the fish exposed to all Mn concentrations showed decreased plasma alternative complement and lysozyme activity. Whole body superoxide dismutase, catalase, and malondialdehyde showed significant increases in the Mn-exposed fish which also exhibited lower tolerance to saltwater than the control group.     

氨氮胁迫下罗非鱼（GIFT Oreochromis niloticus）机体免疫力的变化及其对海豚链球菌易感性的影响

研究了不同作用时间（0、24、48、96和120 h）下不同质量浓度氨氮（对照组、1、3、5和10 mg·L-1）对罗非鱼（GIFT Oreochromis niloticus）血清SOD、AKP、溶菌酶和补体C3活性的影响,以及不同质量浓度氨氮（对照组、1、3、5和10 mg·L-1）胁迫下罗非鱼对海豚链球菌的易感性。结果显示：在1.0 mg·L-1氨氮质量浓度下,罗非鱼血清SOD活性在24 h受到诱导;在3.0、5.0和10.0 mg·L-1氨氮质量浓度下,罗非鱼血清SOD活性均受到抑制,最大抑制率分别达18.85%、23.47%和38.46%。在1.0 mg·L-1氨氮质量浓度下,罗非鱼血清AKP活性在0-96 h呈先降低后恢复的趋势;5.0和10.0 mg·L-1实验组的血清AKP活性在24-120 h则一直受到抑制。1.0 mg·L-1实验组的罗非鱼血清溶菌酶活性在24-120 h一直无显著变化,3.0、5.0和10.0 mg·L-1实验组的血清溶菌酶活性随着氨氮质量浓度的升高而降低。1.0 mg·L-1实验组的罗非鱼血清补体C3活性在各个采样时间下均大于对照组,而5.0和10.0 mg·L-1实验组补体C3活性始终受到抑制。罗非鱼对海豚链球菌的易感性实验研究显示,置于3.0、5.0和10.0 mg·L-1的罗非鱼的累积死亡率分别为43.33%、46.67%和63.33%,显著高于对照组和1.0 mg·L-1实验组,表明氨氮质量浓度超过3.0 mg·L-1能够增加罗非鱼对海豚链球菌的易感性。本实验认为,超过3 mg·L-1的氨氮质量浓度胁迫能够显著降低罗非鱼的免疫力,增加对海豚链球菌的易感性。因此在罗非鱼健康养殖中,调控并降低氨氮质量浓度非常重要。     

Effects of long-term acclimation to environmental hypercapnia on extracellular acid–base status and metabolic capacity in Mediterranean fish <Emphasis Type="Italic">Sparus aurata</Emphasis>

In the context of future scenarios of anthropogenic CO₂ accumulation in marine surface waters, the present study addresses the effects of long-term hypercapnia on a Mediterranean fish, Sparus aurata. By equilibration with elevated CO₂ levels seawater pH was lowered to a value of 7.3, close to the maximum pH drop expected in marine surface waters from atmospheric CO₂ accumulation. Intra- and extracellular acid–base parameters as well as changes in enzyme profiles were studied in red and white muscles and the heart under both normocapnia and hypercapnia. The activities of pyruvate kinase (PK), lactate dehydrogenase (L-LDH), citrate synthase (CS), malate dehydrogenase and and 3-hydroxyacyl CoA dehydrogenase (HOAD) reflect the pathways and capacity of oxidative processes in metabolism. Long-term hypercapnia caused a transient reduction in blood plasma pH (pH_e) as well as in intracellular pH (pH_i). Compensation of the acidosis occurred through increased plasma and cellular bicarbonate levels. Changes in enzymatic activities, especially the increase in the activity of L-LDH, paralleled by a drop in CS activity in white and red muscles reflect a shift from aerobic to anaerobic pathways of substrate oxidation during long-term acclimation under hypercapnia. The present results suggest that moderate environmental hypercapnia changes the metabolic profile in tissues of S. aurata. Consequences for slow processes like growth and reproduction potential as well as potential harm at population, species and ecosystem levels require further investigation.