Role of molecular species catabolism in the temperature-induced restructuring of phosphatidylcholines in liver microsomes of thermally-acclimated rainbow trout (Oncorhynchus mykiss)

Most previous studies of the temperature-induced restructuring of phospholipid molecular species composition have examined steps in the biosynthesis of phospholipids to explain the accumulation of unsaturated fatty acids in membranes of cold-acclimated poikilotherms. In contrast, the present study explores the role of phospholipases in this restructuring process by determining the rates of degradation of specific molecular species of phosphatidylcholine, using enzymes (microsomes) freshly isolated from the liver of rainbow trout. (Oncorhynchus mykiss) acclimated to either 5° or 20°C. The substrate preparation employed to assay phospholipase activity possessed a range of molecular species, all radiolabeled with 1-¹⁴C-palmitic acid at thesn-1 position, similar to that present in native trout liver microsomes. After defined periods of incubation (120 and 240 min at 5°C; 60 and 120 min at 20°C), phospholipids were extracted from the reaction mixture and the distribution of radioactivity among the molecular species of phosphatidylcholine was determined by HPLC/liquid scintillation counting. In general, molecular species catabolism was not significantly influenced by either assay or acclimation temperature. Only in 20°C-acclimated fish did a reduction in assay temperature (from to 20 to 5°C) result in significantly increased proportions of radioactivity being recovered in one polyunsaturated fatty acid-containing species (16:0/22:6-PC). It is concluded: 1) that phospholipase specificity, assayed under conditions approximating thosein situ, is not significantly influenced by temperature; and 2), that the increased proportions of unsaturated fatty acid-containing molecular species of phosphatidylcholine observed at low temperatures must reflect the specificity of biosynthetic rather than degradative processes.     

虹鳟肝癌一例

本文报道了对虹鳟肝脏肿瘤进行的组织学检验情况,认定为恶性肿瘤。病因为饲料中含黄曲霉毒素及鱼体老龄化。     

Dynamics of plasma free amino acids in rainbow trout (Oncorhynchus mykiss under variety of dietary conditions

Plasma amino acid profiles in the systemic circulation of rainbow trout acclimated to diets containing different protein sources, with and without supplementation with free amino acids, were monitored for up to 120 hours after consumption of the diets. Plasma concentrations of lysine, methionine, and isoleucine increased more rapidly after feeding a diet supplemented with these amino acids in free form and, depending upon the increase in plasma concentration, remained at concentrations above those in fish fed the basal diet for at least 24h after feeding. Dietary supplementation with isoleucine increased plasma concentrations of leucine and valine as well as of isoleucine. Maximum plasma concentrations for most amino acids were attained between 12 and 24h postprandial. Dietary inclusion of gelatin caused more rapid elevations in plasma glycine and serine than did free glycine. Feeding at three hour intervals resulted in stable plasma concentrations of individual amino acids in contrast to the fluctuations occurring when fish were fed once daily. Frequent feeding favoured a higher proportion of protein to lipid in body composition.     

Distribution and induction of cytochrome P450 1A1 in the rainbow trout brain

Cytochrome P450 (CYP) 1A1 participates in the activation as well as detoxification of environmental pollutants such as aromatic hydrocarbons. This CYP form is also efficiently induced by aromatic hydrocarbons. The presence of CYP 1A1 in the brain might thus be of physiological and toxicological importance. In the present investigation on rainbow trout, the distribution of 7-ethoxyresorufin-O-deethylase (EROD) activity, a cytochrome CYP 1A1 catalyzed reaction, was measured in whole tissue homogenates from brain parts. In control fish, a relatively high activity was found in the rainbow trout olfactory bulb compared to the other brain parts. Although an EROD induction (3 to 7-fold) by β-naphthoflavone (BNF) was recorded in all brain parts from the rainbow trout, the highest induced activity was measured in the olfactory bulbs. To ascertain the distribution of EROD activity in cells, whole brain tissue was subfractionated by differential centrifugation. The fractionation scheme separated mitochondria (P2 fraction) and microsomes (P3 fraction) as determined by marker enzymes and electron microscopy. In control rainbow trout, a low EROD activity could be measured in the P2 fraction. BNF induced the EROD activity in both P2 and P3 fractions. Western blotting showed the induction by BNF of a protein band in the P2 and P3 fractions with a molecular mass around 58,000 when highly specific anti-cod CYP 1A1 antibodies were used. ELISA measurements confirmed the induction of CYP 1A1 protein in the rainbow trout brain subcellular fractions.     

Variable versus constant temperature acclimation regimes: Effects on hemoglobin isomorph profile in goldfish,Carassius auratus

Goldfish,Carassius auratus, were acclimated for 2 to 3.5 weeks to three temperature regimes: [1] temporally-constant (10, 20 and 30°C), [2] diurnally-cycling (20 ± 10°C) and [3] randomly-fluctuating (± 2°C at approximately 2h intervals between extremes of 10 and 30°C). No significant differences in hematocrit were evident. Hemoglobin levels in fish at constant 30°C and under randomly fluctuating temperature were significantly elevated. Of the three hemoglobin isomorphs observed, the two minor components (G₁, G₃) tended to decrease in relative abundance with increase in constant temperature, but increased under varying temperature regimes. The converse was true of the principal hemoglobin, G₂. Extent of isomorph variation was correlated with extent of temperature variability. These observations confirm that temperature variability significantly effects thermoacclimatory response. The functional significance of changes in isomorph abundances during the acclimatory process is considered.     

Quantification of presumptive Na+/H+ antiporters of the erythrocytes of trout and eel

The presumptive Na⁺/H⁺ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[³H]isobutyl)-amiloride (³H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC₅₀ = 20.1 ± 1.1 mol l^–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific ³H-MIA binding (IC₅₀ = 257 ± 8.2 nmol l^–1, N = 3) than amiloride, which possessed a proton extrusion IC₅₀ of 26.1 ± 1.6 mol l^–1 (N = 6) and a binding IC₅₀ of 891 ± 113 nmol l^–1 (N = 3). The specific Na⁺ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific ³H-MIA binding. Trout erythrocytes suspended in Na⁺-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable ³H-MIA binding sites per cell (B_max, presumptive Na⁺/H⁺ antiporters) with an apparent dissociation constant (K_D) of 244 ± 29 nmol l^–1 (N = 6). Acute hypoxia (PO₂ = 1.2 kPa; 30 min) did not affect the K_D, yet resulted in a 65% increase in the number of presumptive Na⁺/H⁺ antiporters. Normoxic eel erythrocytes, similarly suspended in Na⁺-free saline, possessed only 17,133 ± 3,716 presumptive Na⁺/H⁺ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar K_D (246 ± 41 nmol l^–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na⁺/H⁺ exchange sites.     

Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

From a comparison of 125 bacterial isolates recovered from the digestive tract of rainbow trout, Oncorhynchus mykiss, and carp, Cyprinus sp., a culture was obtained which was effective at preventing clinical disease caused by Lactococcus garvieae and Streptococcus iniae when used as a feed additive. The culture, Aeromonas sobria GC2, was incorporated into the feed and fed to rainbow trout (average weight = 20 g) for 14 days at a dose equivalent to 5 x 10(7) cells g(-1) of feed. Whereas the untreated controls experienced losses of 75-100% when challenged intraperitoneally with L. garvieae and S. iniae, the probiotic-treated groups remained healthy with total mortalities of only 0-6%. Formalized and sonicated preparations of GC2 and cell-free supernatant fared less well. The mode of action reflected stimulation of innate immunity, namely an increased number of leucocytes and enhanced phagocytic and respiratory burst activity.