Temperature-dependent pharmacokinetics and tissue distribution of oxolinic acid in sea bass, Dicentrarchus labrax L., after a single intravascular injection

The pharmacokinetics and tissue distribution of oxolinic acid following an intravascular administration (15 mg kg^?1 fish) were determined in sea bass, Dicentrarchus labrax L. (110 g), at 13 °C and 22 °C water temperature. The kinetic profile of the drug was found to be temperature dependent, with increased temperature having a greater effect on distribution after equilibrium and the elimination phase than on the distribution process. The distibution half‐life of oxolinic acid was 1.15 and 2.76 h at 22 °C and 13 °C respectively, whereas the elimination half‐life of the drug was 55 h at 22 °C and 315 h at 13 °C. The values of the apparent volume of distribution (1.44 L kg^?1 at 22 °C and 3.31 L kg^?1 at 13 °C) and the volume of distribution at steady state (5.2 and 14.7 L kg^?1 at the high and low temperature respectively) were considerably different between the two tested temperatures. The total body clearance of the antibiotic was found to be low (1.47 L kg^?1 day^?1 at 22 °C and 0.80 L kg^?1 day^?1 at 13 °C). Lower rates of elimination were found for the liver compared with muscle, the difference increasing with increasing temperature, while elimination rates from the serum were higher than those of other tissues, especially at the high temperature.     

Pharmacokinetics and bioavailability of flumequine and oxolinic acid after various routes of administration to Atlantic salmon in seawater

The present preclinical study was performed to investigate the pharmacokinetics of flumequine in Atlantic salmon (Salmo salar L.) in seawater after administration of different doses and dosage formulations. Flumequine was administered intravenously (dose 4.9 mg/kg fish) and orally from the drug delivery system Aqualets as Apoquin 5 g/kg (dose 25 mg/kg) and 10 g/kg (dose 50 mg/kg), respectively. Experiments were carried out with oxolinic acid administered in the same way for the purpose of comparing the two compounds. The seawater temperature was 5±0.2°C in all experiments.

The pharmacokinetic calculations showed that the distribution half-life for flumequine was and for oxolinic acid . The drugs were absorbed rapidly, and flumequine reached a plasma concentration of C_max = 2.26 μg/ml after a single oral dose of 25 mg/kg, whereas oxolinic acid reached C_max = 0.99 μg/ml. The apparent bioavailability of flumequine was found to be 40–45%, whereas the apparent bioavailability of oxolinic acid varied from 25% at a dose of 50 mg to 40% at a dose of 25 mg/kg body weight of fish. The distribution profile of flumequine in the various compartment of fish appeared to be different from that of oxolinic acid. After a single oral dose (25 mg/kg) the areas under the concentration-time curves showed that flumequine was 2.3 times more concentrated in plasma and 2.6 times more concentrated in liver compared to oxolinic acid. In muscle the difference was less pronounced, flumequine being 1.4 times more concentrated than oxolinic acid.     

Pharmacokinetics and tissue residues of vancomycin after oral administration in crucian carp

The pharmacokinetics and tissue residues of vancomycin in crucian carp (Carassius auratus) were studied after oral administration of a single dosage of 1 mg/kg body weight at water temperature of 18°C. The drug concentration‐time data were fitted using a two‐compartment open model with first‐order absorption. The vancomycin concentrations in plasma and tissues including muscle, liver and kidney were analyzed by means of liquid chromatography‐tandem mass spectrometry. As a result, the plasma absorption rate constant of vancomycin was 3.75/hr and the time point of maximum concentration in plasma was 3.0 hr respectively. The maximum concentration of vancomycin in plasma was 216.58 μg/L. The distribution half‐life and the elimination half‐life in plasma were 6.69 hr and 190 hr, as well as the absorption half‐life of plasma was 0.19/hr. The area under the plasma concentration‐time curve of plasma was 15.12 mg hr/L. The apparent volume of distribution of plasma was estimated to be 4.45 L/kg and the total body clearance was computed as 0.061 L hr^?1 kg^?1. In addition, the maximum tissue concentrations of vancomycin were in the order of liver > kidney > muscle. To guarantee the safety of tissue samples for consumption, it was suggested that the withdrawal time should not be less than 30 days at 18°C. All these results may provide guidance for future vancomycin treatment in aquaculture.     

A single-dose pharmacokinetic study of oxolinic acid and vetoquinol, an oxolinic acid ester, in Atlantic halibut, Hippoglossus hippoglossus L., held in sea water at 9 °C

The pharmacokinetic properties of the antibacterial agent oxolinic acid were studied after intravenous, intraperitoneal and oral administration to 1.5–3.0 kg Atlantic halibut, Hippoglossus hippoglossus L., held in sea water at 9 °C. Following intravenous injection, the plasma drug concentration-time profile showed two distinct phases. The terminal elimination half-life was estimated to be 52 h, whereas total body clearance (Cl_T) was determined to be 0.044 L kg^–1 h^–1. The volume of distribution at steady state, V_d(ss), was calculated to be 3.0 L kg^–1, indicating good tissue penetration of oxolinic acid in Atlantic halibut. The peak plasma concentration (C_max) and the time to peak plasma concentration (T_max) were estimated to be 1.2 and 2.7 μg mL^–1, and 21.5 and 80 h, respectively, following oral administration of medicated feed or intraperitoneal injection. The corresponding bioavailabilities were calculated to be 15% and 92%, respectively. Oral administration of vetoquinol, the carbitol ester of oxolinic acid, increased the bioavailability of oxolinic acid to 64% and the total bioavailability (oxolinic acid + vetoquinol) to 82%, whereas C_max and T_max values of 6.7 μg mL^–1 and 14.5 h, respectively, for oxolinic acid, and 1.0 μg mL^–1 and 6.3 h, respectively, for vetoquinol were obtained. Based on a minimum inhibitory concentration (MIC) of 0.0625 μg mL^–1 for susceptible strains, a single intraperitoneal injection of 25 mg kg^–1 of oxolinic acid maintains plasma levels in excess of 0.25 μg mL^–1, corresponding to four times the MIC value, for ≈12 days. The corresponding values for a single oral dose of 25 mg kg^–1 of oxolinic acid and vetoquinol were 5 and 10 days, respectively. For resistant strains with a MIC of 1 μg mL^–1, a single oral dose of vetoquinol (25 mg kg^–1) maintained plasma levels in excess of 4 μg mL^–1 for 34 h.     

Prevention of thaw-rigor during frozen storage of bigeye tuna Thunnus obesus and meat quality evaluation

Thaw-rigor is often found in frozen meat of bigeye tuna Thunnus obesus. Excessive amounts of drip loss and stiffness greatly lower the commercial value of tuna meat. In order to prevent thaw-rigor in meat stored at −60°C post-capture, we adapted a temperature shift technique that stores the meat at −7°C for 1 day or −10°C for 7 days before thawing. Biochemical changes in muscle of bigeye tuna before and after the temperature shift to −7 or −10°C were characterized. Contents of ATP, NAD⁺, glycogen, and creatine phosphate decreased after the temperature shift. NAD⁺ levels decreased faster than ATP levels and were highly correlated with the rigor index. Thaw-rigor occurred in muscle containing NAD⁺ at 1 μmol/g and ATP at 7 μmol/g. On the other hand, the meat color of tuna during frozen storage changed to brown depending on the storage temperature and reflected the rate of metmyoglobin (met-Mb) formation. Met-Mb formation increase was dependent on the decrease in NADH levels during the frozen storage. A temperature shift technique with storage at −7°C for 1 day or −10°C for 7 days before thawing prevented thaw-rigor and met-Mb formation.     

Pharmacokinetics of oxolinic acid in black tiger shrimp, Penaeus monodon Fabricius, and the effect of cooking on residues

This study examined the pharmacokinetics and bioavailability of oxolinic acid (OA) in black tiger shrimp Penaeus monodon Fabricius, in brackish water (salinity 10 g L^?1) at 28–29°C, after intra‐sinus (10 mg kg^?1) and oral (50 mg kg^?1) administration and also investigated the net changes of OA residues in the shrimp after cooking (boiling, baking and frying). The haemolymph concentrations of OA after intra‐sinus dosing were best described by a two‐compartment open model. The distribution and elimination half‐lives were 0.84 and 17.7 h respectively. The apparent volume of distribution at a steady state and the total body clearance were estimated to be 2061 mL kg^?1 and 90.1 mL kg^?1 h^?1 respectively. The bioavailability of OA after an oral administration was 7.9%. The peak haemolymph concentration, the time to peak haemolymph concentration and the elimination half‐life after oral administration were 4.20 μg mL^?1, 4 h and 19.8 h respectively. Oxolinic acid muscle and shell levels increased to a maximum (muscle 1.76 μg g^?1 and shell 8.17 μg g^?1) at 4 h post administration and then decreased with the elimination half‐life value of 20.2 and 21.9 h respectively. Residual OA in muscle and shell was reduced by 20–30% by each cooking procedure examined.     

An increase in the concentration of hepatic iron during the metamorphosis of the lamprey Geotria australis is accompanied by increased superoxide dismutase activity

The concentration of total non-haem iron and its ferritin iron component, and the activity of the enzyme superoxide dismutase (SOD), were measured in the livers of ammocoetes, metamorphosing animals (stages 1–7) and recently metamorphosed downstream migrants of the lamprey Geotria australis. Total non-haem iron in the liver rose significantly from 0.15–0.55 μg.mg wet weight⁻¹ in ammocoetes and metamorphosing stages 1–3 to 2.2–2.9 μg.mg⁻¹ in stages 5–7 and to 8.8 μg.mg⁻¹ in downstream migrants. The comparable values for ferritin iron were 0.06–0.26, 1.4–2.0, and 5.3 μg.mg⁻¹. Superoxide dismutase activity fell sharply from 0.39 μg.mg⁻¹ in large ammocoetes to between 0.07 μg.mg⁻¹ in stage 1 and 0.15 μg.mg⁻¹ in stage 6, before rising significantly to 0.26 μg.mg⁻¹ in stage 7 and 0.35 μg.mg⁻¹ in downstream migrants. The sharp fall in SOD activity at the beginning of metamorphosis is assumed to be related to the marked decline in plasma iron which occurs at the onset of this non-trophic phase in the life cycle. It is proposed that the subsequent increase in SOD activity in the liver of G. australis with increasing iron represents a mechanism aimed at reducing the potentially toxic effects of iron accumulation. This view is consistent with the significant and positive correlation found between both total non-haem and ferritin iron and SOD activity in the liver of non-trophic animals.     

Seasonal variations of trimethylamine oxide and urea in the blood of a cold-adapted marine teleost,the rainbow smelt

Trimethylamine oxide (TMAO), which has previously not been known to occur in significant amounts in the blood of marine teleosts, rose to concentrations of approximately 50 mM in the blood of winter-acclimatized rainbow smelt, Osmerus mordax. Urea also increased in the blood of cold-acclimatized smelt, and, with TMAO, contributed significantly to the winter freezing point depression. TMAO and urea also varied seasonally in muscle and liver tissues. TMAO and urea appeared to be reabsorbed from the urine. Losses of TMAO and urea from the head region of the fish, where most of the losses appeared to occur, were approximately 9 μmol and 8 μmol 100 g⁻¹ h⁻¹, respectively. Despite the effluxes, TMAO and urea levels in both the blood and muscle either increased or were maintained in starved, cold-acclimated fish, suggesting that they were synthesized in response to cold temperature. TMAO was also found in the blood of some other cold-hardy teleosts.     

Protein-sparing effect of dietary lipid in practical diets for blunt snout bream (Megalobrama amblycephala) fingerlings: effects on digestive and metabolic responses

This study aimed to evaluate the protein-sparing effect of dietary lipid on digestive and metabolic responses of fingerling Megalobrama amblycephala. Fish were fed nine practical diets with three protein levels (270, 310 and 350 g kg⁻¹) and three lipid levels (40, 70 and 100 g kg⁻¹) for 8 weeks. Weight gain was significantly affected only by dietary lipid levels with the highest found in fish fed 70 g kg⁻¹ lipid. Relative feed intake and whole-body protein content showed little difference among all the treatments. Activities of intestine lipase and amylase increased significantly as dietary lipid levels increased, whereas little difference was observed in protease activities. Liver lipid content was significantly affected only by protein levels with the lowest found in fish fed 310 g kg⁻¹ protein. Liver aspartate aminotransferase (GOT) activities increased significantly with decreasing lipid levels, whereas the highest GOT activity was obtained in fish fed 310 g kg⁻¹ protein in terms of dietary protein levels. Activities of liver lipoprotein lipase, total lipase and plasma cholesterol concentration of fish fed 350 g kg⁻¹ protein were significantly lower than that of the other groups, whereas the same was true for plasma 3, 5, 3′-triiodothyronine level of fish fed 270 g kg⁻¹ protein. The results indicated that an increase of dietary lipid content from 40 to 70 g kg⁻¹ can enhance the growth and digestive enzyme activities of this species and reduce the proportion of dietary protein catabolized for energy without inducing hepatic steatosis; meanwhile, decreasing protein level from 350 to 310 g kg⁻¹ leads to the increase of lipase activities both in intestine and liver coupled with the reduced liver lipid content.     

Stress responses and disease resistance in salmonid fish: Effects of chronic elevation of plasma cortisol

Basal levels of plasma cortisol in unstressed salmonid fish are normally in the range 0–5 ng ml⁻¹. An acute stress such as handling or 1 h confinement caused a temporary elevation of the plasma cortisol levels of both brown trout,Salmo trutta L., and rainbow trout,Salmo gairdneri Richardson, in the range 40–200 ng ml⁻¹ with a return to basal levels within 24–48 h. The extent of the cortisol elevation in response to an acute stress was dependent upon both the species and strain of trout. Chronic stresses, such as prolonged confinement or crowding, resulted in an elevation of plasma cortisol levels to approximately 10 ng ml⁻¹. Under these circumstances, blood cortisol levels remained elevated for periods of up to 4 weeks before acclimation finally occurred. It is shown, by means of intraperitoneal implantation of cortisol, that chronic elevation of plasma cortisol levels in the brown trout results in a dose-dependent increase in mortality due to common bacterial and fungal diseases. This effect is apparent at plasma cortisol levels as low as 10 ng ml⁻¹, levels below those often reported as being representative of ‘unstressed’ fish. These findings are discussed in relation to the known immunosuppressive effects of corticosteroids in teleost fish.     

Evolution of drug resistance and minimum inhibitory concentration to enrofloxacin in <Emphasis Type="Italic">Tenacibaculum maritimum</Emphasis> strains isolated in fish farms

The in vitro susceptibility of 63 isolates of Tenacibaculum maritimum from four fish farms to eight chemotherapeutic agents used for the treatment of bacterial diseases in fish were assessed. The results indicated that all strains were resistant to oxolinic acid and susceptible to amoxicillin, nitrofurantoin, florfenicol, oxytetracycline and trimethoprim-sulphamethoxazole. However, some isolates presented resistance to enrofloxacin and flumequine, ranging from 10 to 30%, and from 25 to 60%, respectively, depending on the farm sampled. These data were used in an attempt to predict whether the resistance to enrofloxacin was static or evolved during the time of sampling from 2003 to 2004. A relationship between the use of enrofloxacin and levels of resistance was detected in the studied farm, increasing significantly from no resistant isolates in 2003 to 44.8% resistant strains in 2004, the year in which this drug was commonly employed. This result was accompanied by a marked decline of about 29.2% of the inhibition zone sizes for the T. maritimum strains in comparison to the initial values (average 21.5 mm). Minimum inhibitory concentration (MIC) of enrofloxacin for 100 T. maritimum strains was determined by the microdilution method. Twenty isolates were resistant to enrofloxacin (> 256 μg ml⁻¹), while the remaining strains showed a bimodal distribution, which ranged from 0.5 to 32 μg ml⁻¹. Our interpretation of the enrofloxacin MIC data suggests that the breakpoint for T. maritimum should be 4 μg ml⁻¹. However, similar studies in other laboratories are necessary to validate this breakpoint value.     

Plasma levels of insulin and liver glycogen contents in one- and two-year old Atlantic salmon (Salmo salar L.) during the period of parr-smolt transformation

Plasma levels of insulin were measured by specific radioimmunoassay in 1-year and 2-year old Atlantic salmon (Salmo salar) parr during the period of parr-smolt transformation. The two-year old fish were of two different categories; silvering pre-smolts and previously mature male parr. If insulin plays an important role in parr-smolt transformation and/or subsequent osmoregulatory changes it was expected that the pre-smolts would show a different insulin profile compared to the mature male parr and one-year old parr, both of which show impaired hypoosmoregulatory ability compared to smolts. Measurements were taken during two separate years. Between January and April both categories of two-year old fish had generally higher plasma levels of insulin compared to the non-smolting one-year old parr. In the pre-smolts insulin levels ranged from 4.0 to 7.9 ng ml⁻¹, and from 7.8 to 16.7 ng ml⁻¹ in 1990 and 1992 respectively, while in the previously mature males the same respective values were from 4.3 to 10.0 ng ml⁻¹, and from 6.6 to 24.1 ng ml⁻¹. In the two-year old fish, whether pre-smolts or mature males, plasma insulin levels peaked between 1–2 months before final smoltification, after which insulin titers declined sharply. In 1990, the 1-year old parr showed a dual peak in plasma insulin. Insulin first peaked in February (7.8 ng ml⁻¹), and then again in April–May (7.7 ng ml⁻¹), while in 1992 the 1-year old parr showed a number of smaller transient peaks (5–7 ng ml⁻¹) between March–May, followed by sharp elevation of insulin levels in June. Liver glycogen contents were at their highest (3.5–5.0 g 100 g⁻¹ I liver wet weight) in March in both 1-year and 2-year old fish. Glycogen levels were low during the later stages of parr-smolt transformation, before rising again in June in both the 1-year old and precociously mature parr, but not in the smolts.     

A single-dose pharmacokinetic study of oxolinic acid and vetoquinol,an oxolinic acid ester,in cod,Gadus morhua L., held in sea water at 8 degrees C and in vitro antibacterial activity of oxolinic acid against Vibrio anguillarum strains isolated from diseased cod

The pharmacokinetic properties of the antibacterial agent oxolinic acid and vetoquinol, the carbitol ester of oxolinic acid, were studied after intravenous (i.v.) and oral (p.o.) administration to 100-150 g cod, Gadus morhua L., held in sea water at 8 degrees C. Following i.v. injection, the plasma drug concentration-time profile showed two distinct phases. The distribution half-life (t1/2alpha) was estimated at 1.3 h, the elimination half-life (t1/2beta) as 84 h and the total body clearance (Cl(T)) as 0.047 L kg(-1) h(-1). The volume of distribution at steady state, Vd(ss) was calculated to be 5.5 L kg(-1), indicating good tissue penetration of oxolinic acid in cod. Following p.o. administration of oxolinic acid or vetoquinol, the peak plasma concentrations (C(max)) of oxolinic acid and the time to peak plasma concentrations (T(max) were estimated to be 1.2 and 2.5 microg mL(-1) and 24 and 12 h, respectively. The bioavailabilities of oxolinic acid following p.o. administration of oxolinic acid and vetoquinol were calculated to be 55 and 72%, respectively. The in vitro minimum inhibitory concentration (MIC) values of oxolinic acid against three strains of Vibrio anguillarum isolated from diseased cod were 0.016 microg mL(-1) (HI-610), 0.250 microg mL(-1) (HI-618) and 0.250 microg mL(-1) (HI-A21). Based on a MIC value of 0.016 microg mmL(-1) a single p.o. administration of 25 mg kg(-1) of oxolinic acid maintains plasma levels in excess of 0.064 microg mL(-1), corresponding to four times the MIC-value, for approximately 12 days. The analogous value for a single p.o. dose of 25 mg kg(-1) of oxolinic acid administered as vetoquinol was 13 days.     

The influence of feeding rate on growth, feed efficiency and body composition of juvenile grass carp (Ctenopharyngodon idella)

An 8 weeks growth study was conducted to estimate the optimal feeding rate for juvenile grass carp (3.08±0.03 g, mean ± SD). Fish were fed with a casein purified diet (360 g protein, 56 g lipid and 3000 kcal total energy/kg dry diet) at six feeding rates: 1.0, 1.5, 2.0, 2.5, 3.0, 3.5% body weight per day (BW d⁻¹). Each feeding rate was randomly assigned to three tanks of fish with 30 fish per tank (50W × 50H × 100L, cm). Fish were maintained in recirculating systems at a water temperature of 24.97±2.23 °C and were fed four times per day. After 2 weeks, fish fed on 3.5% BW d⁻¹ could not finish the diet and this treatment was cut-off. Analysis of variance showed that growth performance was significantly (p<0.05) affected by different feeding rates. The nutrient compositions of whole body, muscle and liver were also significantly different among treatments. The body weight gain (WG), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), apparent digestibility coefficiency (ADC), retention of protein (PR), mesenteric fat index, body moisture and protein content were significantly (p<0.05) affected by feeding rate. The WG, SGR and digestion rate were highest at 2% BW d⁻¹, although the FE and PER decreased with increasing feeding rate. Broken line analysis on specific growth rate indicated that the optimum feeding rate of juvenile grass carp is 1.97% body weight day⁻¹.     

Protein and energy utilization and the requirements for maintenance in juvenile mulloway (<Emphasis Type="Italic">Argyrosomus japonicus</Emphasis>)

This study describes the digestible protein (DP) and digestible energy (DE) utilization in juvenile mulloway, and determined the requirements for maintenance. This was achieved by feeding triplicate groups of fish weighing 40 or 129 g held at two temperatures (20 or 26°C), on a commercial diet (21.4 g DP mJ DE⁻¹) at four different ration levels ranging from 0.25% of its initial body weight to apparent satiation over 8 weeks. Weight gain and protein and energy retention increased linearly with increasing feed intake. However, energy retention efficiency (ERE) and protein retention efficiency (PRE) responses were curvilinear with optimal values, depending on fish size, approaching or occurring at satiated feeding levels. Maximum predicted PRE was affected by body size, but not temperature; PRE values were 0.50 and 0.50 for small mulloway, and 0.41 and 0.43 for large mulloway, at 20 and 26°C respectively. ERE demonstrated a similar response, with values of 0.42 and 0.43 for small, and 0.32 and 0.34 for large mulloway at 20 and 26°C respectively. Utilization efficiencies for growth based on linear regression for DP (0.58) and DE (0.60) were independent of fish size and temperature. The partial utilization efficiencies of DE for protein (k _p) and lipid (k _l) deposition estimated using a factorial multiple regression approach were 0.49 and 0.75 respectively. Maintenance requirements estimated using linear regression were independent of temperature for DP (0.47 g DP kg^−0.7 day⁻¹) while maintenance requirements for DE increased with increasing temperature (44.2–49.6 kJ DE kg^−0.8 day⁻¹). Relative feed intake was greatest for small mulloway fed to satiation at 26°C and this corresponded to a greater increase in growth. Large mulloway fed to satiation ate significantly more at 26°C, but did not perform better than the corresponding satiated group held at 20°C. Mulloway should be fed to satiation to maximize growth potential if diets contain 21.4 g DP mJ DE⁻¹.     

The content of total mercury and methylmercury in common carp from selected Czech ponds

The aim of the present study was to investigate concentrations of total mercury (THg) and methylmercury (MeHG) in the common carp from Czech ponds of Rožmberk, Spolsky, Nezmar, and Velky Bědny. Seven common carps (Cyprinus caprio) from each of the ponds were caught. Muscle tissue, the liver, and the soft and hard roe were used for the tests. Total mercury and methylmercury were found in all the muscle tissue samples examined. Detection limits for total mercury and methylmercury determination methods were 0.001 mg kg^–1 and 0.013 mg kg⁻¹ respectively. Methylmercury levels in the liver and gonads were below the method’s limits of detection. THg and MeHg concentrations in muscle tissues were 0.018–0.063 mg kg^–1 w.w. and 0.019–0.063 mg kg^–1 w.w. respectively. MeHg made up 90–100% of THg in muscle tissues. Total mercury and methylmercury concentrations were significantly (P < 0.05) higher in fish from the Spolsky pond than from the Nezmar pond. MeHg/THg ratios were significantly (P < 0.05) higher in fish from the Rožmberk, Spolsky and Nezmar ponds compared with fish from the Velky Bědny pond.     

Effect of ethanolic extract of propolis on growth performance and plasma biochemical parameters of rainbow trout (Oncorhynchus mykiss)

This study was conducted to evaluate the effect of ethanolic extract of propolis (EEP) on growth performance and plasma biochemical parameters of rainbow trout (Oncorhynchus mykiss). Graded levels of EEP [0 (control), 1, 2, and 4 g kg⁻¹ diet] were fed to trout juveniles (mean weight 7.73 ± 0.17 g) for 10 weeks. Dietary EEP supplementation regardless of inclusion level significantly improved the specific growth rate of fish. Similarly, supplemental EEP generally improved the feed efficiency ratio and protein efficiency ratio, but no significant differences were observed between the 1 g kg⁻¹ EEP group and the control group. In addition, dietary EEP supplementation generally increased the plasma superoxide dismutase, lysozyme, total antioxidant capacity, glutathione peroxidase, and catalase activities, but decreased the plasma malondialdehyde level. The plasma triglycerides level was significantly lower in the 1 or 4 g kg⁻¹ EEP group as compared with the control group. Dietary EEP supplementation generally decreased the plasma aspartate aminotransferase and alanine aminotransferase activities, but increased the hepatic aspartate aminotransferase and alanine aminotransferase activities. These results indicate the potential to use the EEP as a growth promoter, hepatoprotective agent, and immunostimulant for rainbow trout.     

The role of glycogen phosphorylase in the regulation of glycogenolysis by insulin and glucagon in isolated eel (Anguilla rostrata) hepatocytes

The effects of porcine, scombroid, and salmon insulins, and bovine and anglerfish glucagons on glycogen depletion and glycogen phosphorylase (GPase) activities were examined in freshly isolated American eel (Anguilla rostrata) hepatocytes. Eel liver GPase in crude homogenates was activated (increase in % GPase a) by phosphorylating conditions and was rapidly inactivated (less than 1 h) when a phosphatase inhibitor (fluoride) was absent. Caffeine inhibits, and AMP activates, the b form of GPase consistent with their effects on rat liver GPase. Both mammalian and fish glucagons increased glucose production in eel hepatocytes, but had more ambiguous effects on glycogen levels and GPase activities. The magnitude of bovine glucagon effects were dependent on the initial glycogen content of the cells; only at glycogen concentrations less than approximately 70 μmoles.g⁻¹ did glucagon significantly increase % GPase a. Anglerfish glucagon significantly increased cyclic AMP (cAMP) concentrations by 90% at 10⁻⁷ M, but had no effects at 10⁻⁹ M and 10⁻⁸ M. Scombroid and salmon insulins maintained hepatocyte glycogen concentrations and decreased glucose production, with these effects more pronounced at low (10⁻⁹ to 10⁻⁸ M) rather than high (10⁻⁷ M) hormone concentrations. Porcine and salmon insulins decreased total GPase and % GPase a activities, and salmon insulin decreased CAMP levels, but only at 10⁻⁸ M (by 44%). Glycogen is, therefore, depleted by glucagon and maintained by insulin in freshly isolated American eel hepatocytes, and these changes are accomplished, at least in part, by changes in the activities of GPase. Changes in cAMP do not explain all of the observed hormone effects.     

Pharmacokinetics of ivermectin in sea bream, Sparus aurata using a direct competitive ELISA

The pharmacokinetics of ivermectin in the serum of cultured sea bream, Sparus aurata, after a single intraperitoneal injection of 100 μg kg⁻¹ body weight was studied by the use of direct competitive ELISA. Pharmacokinetic analysis of the serum concentrations versus time points obtained was performed using non-comparmental analysis and a compartmental pharmacokinetic model approach. In the latter case a two-compartment open model with a lag time gave the best fitting. The maximum peak serum concentration was 308.4 ng ml⁻¹ at 2 h post treatment. The AUC of ivermectin was 10700 ng h ml⁻¹ and the elimination half-life 15.37 h, indicating a rapid uptake, high bioavailability and fast elimination of the drug by sea bream. This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of ionic composition on acid-base regulation in rainbow trout (Oncorhynchus mykiss) exposed to environmental hypercapnia

The influence of ambient calcium, bicarbonate and chloride levels on acid-base regulation was investigated in rainbow trout acclimated and exposed to hypercapnia in five different water types. In soft water (low [Ca⁺⁺] and [HCO₃ ⁻]), compensation of the respiratory acidosis was slow and incomplete within 72h. High ambient [HCO₃ ⁻] clearly improved extracellular HCO₃ ⁻ accumulation, and pH recovery was accomplished within 24h. This may result from stimulation of branchial HCO₃ ⁻ (influx)/Cl⁻ (outflux) exchange. Elevation of ambient [Cl⁻] had a small, positive effect on pH compensation. High ambient [Ca⁺⁺] improved the degree of pH compensation. Plasma [HCO₃ ⁻] and [Cl⁻] showed an inverse 1:1 relationship in all acclimation groups, revealing an ubiquitous chloride-mediated acid-base regulation. Ventilation activity was increased by hypercapnia and only returned to control values in hard water (high [HCO₃ ⁻]and [Ca⁺⁺]). During progressive hypercapnia (up to 3% CO₂), hard water acclimated fish obtained significantly higher plasma [HC0₃ ⁻] (51.2 mM) than fish acclimated to low [Ca⁺⁺]/high [HCO₃ ⁻] (44.7 mM). This suggests an additive effect of ambient Ca⁺⁺ on plasma HCO₃ ⁻ accumulation. At levels of CO₂ above 1%, some mortality was induced in low [Ca⁺⁺]/high [HCO₃ ⁻] water. Dying fish could be distinguished from surviving fish by an excessive Cl⁻loss and increasing extracellular anion gap.