Effect of sub-lethal nitrite on selected haematological parameters in fingerling Catla catla (Hamilton)

A sub‐lethal nitrite toxicity trial was conducted using static conditions for a period of 96 h with fingerlings of Catla catla (21.4±3.6 g). Fingerlings were exposed to five concentrations of nitrite, that is, 1, 2, 4, 8 and 10.4 mg L^?1 and a nitrite‐free control to study changes in haematological parameters. Nitrite caused an increase in immature erythrocyte population (7–24%) in lower concentrations (0–4 mg L^?1) at 6 h while they were absent in higher concentrations. The total erythrocyte count was reduced at 6 h followed by an increase at 12 h with further reduction up to 96 h in all concentrations of nitrite. The 96‐h exposure resulted in 21.2–31.8% reduction in erythrocyte population in 1–10.4 mg L^?1 nitrite. The haemoglobin content decreased progressively with increasing nitrite concentrations as well as exposure periods. Total leukocyte count decreased initially at 6 h in all treatments followed by an increase after 12 h, signifying development of a protective response of the body to nitrite stress. Blood glucose decreased initially up to 24 h followed by an increase through 96 h. Serum protein level decreased continuously with increasing exposure period. The study revealed that exposure to nitrite caused changes in almost all the haematological parameters in the fingerlings depending on the concentration as well as exposure period. Nitrite being one of the important inorganic nutrients often recorded at higher levels in intensively cultured ponds, the present study highlights its adverse impact on fish and stressed the need for the management of this nutrient in culture ponds.     

Stable isotope variations in otoliths of Pacific halibut (Hippoglossus stenolepis) and indications of the possible 1990 regime shift

Stable oxygen and carbon isotope ratios (δ¹⁸O and δ¹³C) from archived otoliths of Pacific halibut, Hippoglossus stenolepis, were measured to examine the most recent regime shifts in British Columbia and the Gulf of Alaska. δ¹⁸O values of these otoliths ranged from −1.5 to +2.8‰ VPDB, and were consistent with the life stages and migration behavior of halibut. δ¹³C varied from −3.3 to +0.9‰ VPDB, but did not show a transition from the juvenile to the adult stage as does δ¹⁸O. Evaluation of δ¹⁸O and δ¹³C values of mature halibut (ages 8–12) indicated that the 1977 regime shift might have a warming impact on the northeast Pacific fish stocks. In contrast, a possible regime shift around 1990 with a bottom seawater temperature decrease of about 2 °C might have occurred in both the areas. The connection between stable isotope variations in otoliths and the climate regime shifts is thus potentially useful in studying the population dynamics of Pacific halibut, and decadal scale (e.g., the last 20–30 years) ocean environmental changes.     

Dietary inclusion of crystalline D- and L-methionine: effects on growth, feed and protein utilization, and digestibility in small and large Atlantic salmon (Salmon salar L.)

Use of crystalline methionine to optimize amino acid profiles in feeds for Atlantic salmon (Salmo salar L.) was studied in three experiments. A basic experimental diet with 25% of dietary protein from marine origin and the remaining 75% from soya protein concentrate was used either with or without supplementation of L ‐methionine, D ‐methionine or DL ‐methionine. A fishmeal‐based diet was used as control. In experiment 1, growth performance of 100 g salmon in seawater was improved by supplementation with the different methionine products. Growth performance was, however, inferior to that of fish fed the control. Feed conversion ratios (FCR) exhibited similar trends to specific growth rate (SGR) with methionine supplementation, but again not as the control. Protein retention, measured as productive protein value (PPV), was significantly better in the control and D ‐methionine groups as compared with either L ‐methionine or the nonsupplemented group. These trends were confirmed in experiment 2, using fish of the same size, where an absorption and digestibility study of D ‐ and L ‐methionine clearly indicated that D ‐methionine was absorbed as well as L ‐methionine. Results indicated that racemization of D ‐ into L ‐methionine occurs in liver and kidney in Atlantic salmon. Ornithine decarboxylase (ODC) activity in liver was significantly affected by dietary treatment and there was a good correlation with growth and feed utilization data. In experiment 3, a growth study with large salmon (1 kg) was performed using increasing supplementation of DL ‐methionine to the basic diet. Results indicated a methionine requirement for maximum growth of 9.2 g kg^–1 feed dry matter (DM) kg^–1 growth. This is slightly higher than that reported in the literature. The present study clearly shows that optimizing the amino acid profile by inclusion of a single crystalline amino acid (crystalline methionine) does not give the same protein utilization as that of a diet in which the amino acids are mainly protein bound. D ‐methionine was found to result in better protein utilization than L ‐methionine when measured as PPV.     

Fast growth, protein turnover and costs of protein metabolism in yolk-sac larvae of the African catfish (Clarias gariepinus)

In fish larvae the costs of rapid growth may be accommodated by a decrease in the rate of protein turnover or by a reduction in the costs of protein synthesis. Protein growth, synthesis and degradation were measured in yolk-sac larvae of Clarias gariepinus and the costs of protein synthesis and protein growth were estimated. Growth rates were over 100% protein weight day^-1. Protein synthesis retention efficiency (retained protein per unit of synthesis) was estimated to be 69.6%, a value comparable to that of larger fish. The larvae used 43% of their oxygen consumption for protein synthesis. Nevertheless, protein synthesis costs were close to theoretical minima. Therefore, the high growth rates of catfish yolk-sac larvae seem to be possible through minimisation of the costs of protein synthesis. These low costs are associated with high rates of protein synthesis (138%protein weight day^-1), and elevated RNA concentrations (107 µg RNA mg^-1protein), which together suggest very high RNA efficiencies (12.9 g protein synthesized g^-1RNA day^-1).     

鲤鱼血清蛋白,血红蛋白,LDH和EST同工酶的电泳分析

采用聚丙烯酰胺凝胶电泳技术,分析了鲤鱼性成熟期雌雄个体的血清蛋白、血清脂蛋白和血红蛋白,结果是雌雄个体血清蛋白电泳图谱有差异,雌性(14条)比雄性(13条)多1条蛋白带;血清脂蛋白和血红蛋白无性别差异。同时,测定了鲤鱼血清、肝胰脏、心脏、背部肌肉和眼晶体的乳酸脱氢酶(LDH)同工酶和酯酶(EST)同工酶,LDH具有组织特异性,除在肝胰脏分离出9条酶带外,在血清中也分离出8条酶带,表明有LDH—c基因的作用,EST同工酶表现出明显的多态性,表明在鲤鱼各组织中有多个基因座位起作用。     

In vitro synthesis of 20α-reduced and of 11- and 21-oxygenated steroids and their sulfates by testes of the goldfish (Carassius auratus): Testicular synthesis of corticosteroids

Testes from spermiating goldfish were incubated with [³H]17-hydroxyprogesterone. The major products in the unconjugated fraction were identified as androstenedione, androstenetrione, 11-β-hydroxyandrostenedione, 11-ketotestosterone, 17,20α-dihydroxy-4-pregnen-3-one (17,20αP) and 11-deoxycortisol. Testosterone was present predominantly in the glucuronide fraction, but yields were low (1–3%). The major components of the sulfate fraction were 17,20αP and 11-deoxycortisol. The identification of cortisone in low yield (< 2.5010) in both the free and sulfate fractions is the first report of corticosteroid biosynthesis by a teleost testis. The high yields of 17,20αP and 11-deoxycortisol and their sulphates suggests that their possible role in spermiation of the goldfish should be further investigated.     

Preparation and comparison of biological properties of recombinant carp (Cyprinus carpio) growth hormone and its Cys-123 to Ala mutant

Carp growth hormone (cGH) cDNA, in which Cys-123 was mutated to Ala, was prepared, transferred to the expression vector, expressed in Escherichia coli and the mutant was purified to homogeneity. The mutation only slightly improved yield of the monomeric fraction, indicating that Cys-123 is not involved in improper refolding. As compared to cGH, the mutant (cGH-C123A) exhibited lower binding affinity toward homologous liver receptors and lower bioactivity in a 3T3-F442A preadipocyte bioassay despite the fact that both hormones exhibited almost identical cross-reactivity with anti-cGH antibodies. These results, along with those of a structural comparison to hGH, suggest that Cys-123 is located in the hydrophobic core of the hormone, and is most likely affecting the conformation of the binding site. Dimeric forms of the hormone and its mutant were less active than their respective monomers. Homologous binding experiments using a carp liver microsomal fraction revealed a single receptor population with Kd = 0.77 nM and B_max = 241 fmol/mg microsomal protein.

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Résumé Un ADN complémentaire (cDNA) de l'hormone de croissance de carpe (cGH), dans lequel l'acide aminé Cys-123 a été muté en Ala, a été préparé, inséré dans un vecteur d'expression, et exprimé dans Escherichia coli. Le mutant a ensuite été purifié jusqu'à homogénéité. La mutation améliore seulement faiblement la production de la fraction monomérique, indiquant que le Cys-123 n'est pas impliqué dans un repliement erroné. Comparé à la cGH, sa forme mutée (cGH-C123A) montre une plus faible affinité de liaison vis à vis de récepteurs hépatiques homologues, et une plus faible activité biologique dans un test réalisé sur des préadipocytes 3T3-F442A; cela en dépit du fait que les deux hormones présentent des réactions croisées presque identiques avec un anticorps anti-cGH polyclonale. Ces résultats, associés à une comparaison à la structure de l'hGH, suggèrent que le Cys-123 est localisé dans la partie hydrophobique de l'hormone, et affecte, le plus vraisemblablement, la conformation du site de liaison. Les formes dimériques de l'hormone et de sa forme mutée sont moins actives que leurs monomères respectifs. Les études de liaison homologue, réalisées avec des fractions microsomales de foie, révèlent une population unique de récepteurs de Kd = 0,77 nM et de B_max = 241 fmol/mg de proteine microsomale.

     

Studies on the green liver in cultured red sea bream fed low level and non-fish meal diets: Relationship between hepatic taurine and biliverdin levels

SUMMARY: The aim of this study was to investigate the cause of green liver in red sea bream fed substitute protein diets. Red sea bream Pagrus major was given either of the following diets for 28 weeks: (1) control diet (50% fish meal), (2) low level fish meal diet (15% fish meal), and (3) non-fish meal diet (0% fish meal). The green liver was observed in all groups tested, but the incidence was much higher in the experimental diet groups. The feeding of substitute protein diets reduced plasma triglyceride and cholesterol levels. However, there was no significant difference in plasma hepatic enzyme activities and plasma bile salts concentration among the treatments. Fish fed the substitute protein diets showed low hepatic taurine levels with an appearance of a biliverdin in the liver. Moreover, the proportion of ditaurobilirubin to total biliary bile pigments was significantly lower in fish fed the substitute protein diets. These data indicate that feeding of substitute protein diets did not induce any cholestatic hepatobiliary obstructions and that the low hepatic taurine level was one of the probable factors responsible for the occurrence of green liver in red sea bream fed substitute protein diets.     

Control of ion transport by the sperm duct epithelium of brook trout (Salvelinus fontinalis)

The sperm duct epithelium of brook trout (Salvelinus fontinalis), mountedin vitro in Ussing-style epithelial chambers actively absorbs Na⁺ (measured as the short-circuit current, I_sc) and secretes K⁺ (measured using⁸⁶Rb⁺ as tracer). Dibutyryl-cyclic-adenosine monophosphate (db-cAMP) and 3-isobutyl-1-methylxanthine (IMX) produce a rapid, sustained stimulation of both ion transport processes, but the hormone connected to the response is unknown. Purified sockeye salmon CON A2 gonadotropin (GtH) produces a dose-dependent, rapid and sustained rise in Na⁺ uptake and K⁺ secretion. The time course, electrophysiological and transport characteristics are similar to those evoked by IMX. Carbohydrate-poor (chum salmon CON A1) GtH is ineffective. Pretreatment of fish with 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-P) significantly increases milt volume but is without effect on resting or stimulated (IMX + db-cAMP) levels of sperm duct ion transport. This is the first indication of a direct, rapid action of GtH on ion transport by the vertebrate blood-testis barrier. The results suggest direct involvement of GtH in control of later stages of sperm maturation.