Effect of feeding on the tissue free amino acid concentrations in rainbow trout (Oncorhynchus mykiss Walbaum)

This study investigates whether tissue free amino acid (FAA) pools in rainbow trout,Oncorhynchus mykiss (Walbaum), are altered following feeding and the relationships between the amount of food consumed and the FAA pool size. Trout were starved for 7 days to provide baseline data and then refed on day 8. Individual food intake was measured by radiography and the consumption of amino acids (AA) calculated from dietary protein consumption. Total FAA concentrations in the stomach, liver and white muscle were little changed at various times after the meal and this pattern was repeated for the majority of individual FAA. Overall, the most notable change was a reduction in essential FAA concentrations (principally in valine, leucine and isoleucine) in the white muscle following feeding. However, in the caeca total FAA, total essential FAA and a number of individual FAA were significantly elevated at 4, 9 and 15h following feeding. There were few significant correlations between dietary amino acid consumption and total tissue FAA and essential FAA concentration in the stomach, caecum and white muscle; correlations were stronger in the liver. In order to explain the relative constancy of total FAA concentrations in the tissues following food intake (which represents over 100% of the total FAA pool) a model is presented that quantifies the AA flux through the free pools and considers the role of protein turnover in regulating FAA pool size.     

Supplementation of potassium, magnesium and sodium chloride in practical diets for the Pacific white shrimp, Litopenaeus vannamei, reared in low salinity waters

The culture of Litopenaeus vannamei in inland low salinity waters is currently being practiced in various countries around the world. These environments are often deficient in key ions essential for normal physiological function, including potassium (K⁺) and magnesium (Mg²⁺). Farmers have sometimes been able to counteract ionic deficiencies in the water profile by adding mineral salts containing sources of K⁺ and Mg²⁺. The purpose of this study was to explore the possibility of correcting deficiencies of K⁺ and Mg²⁺ in the water profile with dietary supplementation of these minerals. Two separate 7‐week experiments were conducted in 4.0 g⁻¹ artificial low salinity water to evaluate the effects of mineral supplements (K⁺, Mg²⁺ and NaCl) to diets of L. vannamei reared in low salinity waters. In trial 1 seven diets were formulated (10 g NaCl kg⁻¹, 20 g NaCl kg⁻¹, 150 mg kg⁻¹ Mg²⁺, 300 mg kg⁻¹ Mg²⁺, 5 g K⁺ kg⁻¹, 10 g K⁺ kg⁻¹, and a basal diet to serve as a control). Minerals were added in the form of purified potassium chloride (KCl), magnesium chloride (MgCl₂·6H₂O) and NaCl. Trial 2 evaluated the use of a coating agent for the Mg²⁺ and NaCl treatments, while a K⁺ amino acid complex was utilized in the K⁺ treatments to reduce mineral leaching. Trial 2 was performed using similar treatment levels as trial 1. Shrimp survival and growth were assessed in both experiments. Results from trial 1 indicated no significant differences in survival, growth or percent weight gain. Results from trial 2 revealed no significant differences in survival and growth in the NaCl and Mg²⁺ treatments. However, significant differences in growth (P < 0.05) were observed when using the 10 g K⁺ kg⁻¹ treatment, suggesting that dietary supplementation of a K⁺ amino acid complex may help improve growth of the species in low salinity waters.     

Effects of steroid hormones on in vitro oocyte maturation in white sturgeon (Acipenser transmontanus)

The in vitro effects of several steroids on the maturation of intact white sturgeon (Acipenser transmontanus) ovarian follicles were investigated. At the highest concentration (1024 ng ml^–1 for the C21 steroids and 1139 ng ml^–1 for the C19 steroids), all of the C21 steroids tested, progesterone (P4), 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,(20,21-trihydroxy-4-pregnen-3-one 20-S), 11-deoxycortisol (S) and cortisol (F), as well as testosterone (T) induced germinal vesicle breakdown (GVBD) at 14 and 22 h. At 6 h, only P4 and 17,20-P induced maturation at the highest concentration (1024 ng ml^–1). At 14 and 22 h, 11-deoxycortisol was the most potent steroid inducer of GVBD followed by P4, 17OHP, 17,20-P, and 20-S. The steroid 11-hydroxytestosterone (11OHT) was completely ineffective at all concentrations and exposure times. The C21 steroids induced oocyte maturation at concentrations ranging from 4 to 1024 ng ml^–1, whereas T induced GVBD at 225 to 1139 ng ml^–1. Calculation of the mean effective concentration that induced 50% GVBD (EC50) from the 22 h incubations revealed the following order of potencies: S > P4 > 17OHP > 17,20-P > 20-S >> F > T. These bioassay results, together with previous findings on the endogenous production of steroids by ovarian follicles from gonadotropin-primed females, indicate that more than one steroid has a biological role in the resumption of meiosis in sturgeon oocytes and provides empirical evidence for P4, 17OHP, S, 20-S, and 17,20-P as maturation-inducing steroids in white sturgeon.     

Dietary protein level and natural food management in the culture of blue (Litopenaeus stylirostris) and white shrimp (Litopenaeus vannamei) in microcosms

The effect of dietary protein level and natural food management on the production parameters of blue and white shrimp, as well as on water quality, was evaluated in a microcosms system (plastic pools simulating aquaculture ponds). Two experimental trials were carried out in the facilities of DICTUS, University of Sonora, Northwest México. Treatment with low protein diet (LP) consisted of a low protein input (diet with 250 g kg^?1 crude protein) through the culture period; treatment with high protein diet (HP) consisted of a high protein input (diet with 400 g kg^?1 crude protein) through the trial, and finally treatment VP consisted of an adjustment of protein input (diets with 250, 350 or 400 g kg^?1 crude protein), depending on the abundance of biota (zooplankton and benthos) in the system. Each species responded differently to the treatments. For blue shrimp, low protein input resulted in the lowest final body weight (12.9 ± 0.6 g) and biomass (696.0 g pool^?1). Survival and feed conversion ratio were similar in the three treatments. For white shrimp, the best growth, biomass and food conversion ratio were obtained in the low protein input treatment. Water quality parameters such as nitrate, ammonia and organic matter during the two trials, were better for LP and VP treatments. White shrimp seems to have lower protein requirements than blue shrimp. For the blue shrimp culture, adjusting protein input according to natural food abundance (zooplankton and benthos) in the system, seems to be advantageous because of the possibility of getting a production similar to that obtained with a high protein input through the farming period, but at lower feed cost, and with a lower environmental impact. It is concluded that a high protein input through the whole farming period is not the best feeding strategy for any of the two species.     

Characterization and application of monoclonal antibodies against white spot syndrome virus

Three hybridoma clones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with white spot syndrome virus (WSSV) isolated and purified from Penaeus monodon (Fabricius), collected from north-eastern Taiwan. By sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE), the protein profile of this isolate contained four major proteins with sizes of approximately 35 (VP35), 28 (VP28), 24 (VP24), and 19 kDa (VP19). Western blot analysis revealed that two MAbs (1D7 and 6E1) recognized epitopes on VP28 and one MAb (3E8) recognized an epitope on VP19. The MAb 6E1 isotyped to the IgG₁ class was used in both an indirect immunofluorescence assay (IFA) and in an immunochemical staining protocol for successful identification and localization of WSSV in infected shrimp tissues. Antigenic similarity of isolates from Indonesia and Malaysia to the Taiwan isolate was illustrated by IFA with MAb 6E1. A MAb (2F6) which bound specifically to two shrimp proteins, 75 and 72 kDa, and reacted to the healthy and non-target tissues of WSSV in infected shrimp, such as hepatopancreas, is also described here and shows the necessity for specific identification of antibodies.     

Digestive protease activities and free amino acids in white muscle as indicators for feed conversion efficiency and growth rate in Atlantic salmon (Salmo salar L.)

The aim of the present experiment was to screen several biochemical indices in fish and their interrelations in order to select variables for future studies of growth rate and feed conversion. Several parameters [trypsin activity, chymotrypsin activity, free amino acids (FAA) in plasma and white muscle, and RNA and RNA/protein ratio in the white muscle] were measured together with specific growth rate (SGR), feed intake and feed conversion efficiency (FCE) in four groups of diploid or triploid Atlantic salmon (Salmo salar L.) reared under different light regimes. SGR was measured on individually tagged fish, whereas feed intake and feed conversion was estimated on tank basis. A principal component analysis (PCA) explained 80.6% of the variance in the data, using all measured parameters, regardless of ploidy and light regime. Muscle free hydroxyproline showed the highest correlation, alone explaining 55% of SGR variability. The SGR also significantly correlated with trypsin activity (r=0.34), the activity ratio of trypsin to chymotrypsin (T/C) (r=0.39), plasma essential FAA (EAA) (r=0.39), plasma total FAA (TFAA) (r=0.37), the ratio of essential to non-essential FAA (EAA/NEAA) in the white muscle (r=–0.45), muscle RNA (r=–0.45) and RNA/protein ratio (r=–0.41). Tank FCE correlated positively (r=0.97) with SGR, T/C ratio and muscle free hydroxyproline, and negatively (r=–0.90) with muscle EAA/NEAA. The groups reared under continuous light (LL) regime showed significantly higher SGR than simulated natural photoperiod (SNP) groups, and with an apparently higher FCE. A higher growth rate was associated with either a higher consumption rate and/or a higher feed utilization. A negative correlation between muscle RNA concentration and SGR may indicate that increased growth rate under LL regime was not caused by an increased protein deposition rate.     

Does the aerobic capacity of fish muscle change with growth rates?

To ascertain whether growth rate modifies the oxidative capacity of fish white muscle, we examined the effects of individual growth rate on the activities of four mitochondrial enzymes in white muscle of the fast growing Atlantic cod,Gadus morhua. Growth rates were individually monitored in cod held at three acclimation temperatures during experiments repeated in four seasons. The size dependence of citrate synthase (CS), cytochrome C oxidase (CCO) and β-hydroxyacyl CoA dehydrogenase (HOAD) activities was established using wild cod ranging from 115 to 17,350 g. Given their negative allometry, CS and CCO activities in the experimental cod were corrected to those expected for a 1.2 kg animal. HOAD activities did not change with size. The specific activities of CCO and CS were positively correlated with growth rate. However, for both enzymes, season explained more of the variability than growth rate or temperature. Season was the only factor to significantly affect the activity of HOAD, while temperature and season interacted to determine glutamate dehydrogenase activity. CS activity was positively correlated with the initial condition of the cod, which differed among the seasons. The other enzymes did not show this relationship. The independent changes of these enzymes suggest that mitochondria undergo qualitative modifications with changes in growth rate, season and size. Although growth rate and the activities of CCO and CS are positively correlated, the activity of the mitochondrial enzymes is more affected by size, physical condition and season.