A proposed nucleotide sequence for the 5S ribosomal ribonucleic acid of rainbow trout (Salmo gairdneri).

Rainbow trout cell cultures have been exposed to 32P-labelled inorganic phosphate and the labelled RNA has been isolated. The 5S ribosomal ribonucleic acid (5S rRNA) was purified by polyacrylamide gel electrophoresis, then digested with RNase T1 or pancreatic RNase. The products of complete digestion were separated and their sequences determined. These analyses have allowed a sequence to be proposed which differs in eight positions from that of mammalian 5S rRNAs.     

Bioconversion pathway of astaxanthin into retinol2 in mature rainbow trout (Salmo Gairdneri Rich.)

• 1.1. Carotenoid and retinoid forms were analysed by HPLC in various tissues of mature rainbow trout fed for 11 months (7–9°C) with astaxanthin (50 and 100 mg/kg diet) or canthaxanthin (100 mg/kg diet).
• 2.2. Decreasing concentrations of canthaxanthin, echinenone and β-carotene, but no retinol₁, were found in the liver and skin of canthaxanthin-fed fish.
• 3.3. Higher retinol₂ concentrations were found in ovaries and testes of astaxanthin-fed fish compared to canthaxanthin and control groups.
• 4.4. A new metabolic pathway for direct conversion of astaxanthin into retinol₂ in gonads is proposed.

     

A radio-gas chromatographic method for the determination of 11-oxotestosterone in fish plasma: its use in confirming estimations by radioimmunoassay

A radio-gas chromatographic method has been devised for the estimation of 11-oxotestosterone (17beta-hydroxyandrost-4-ene-3, 11-dione) in fish plasma samples which provides an independent means of validating the radioimmunoassay described earlier. Estimates of the concentration of 11-oxotestosterone in a sample of male rainbow trout plasma by radio-gas chromatography using peak height and peak weight measurements were 6.9 microgram/100 ml and 7.4 microgram/100 ml respectively, in good agreement with that of 7.1 microgram/100 ml determined by radioimmunoassay.     

Hepatectomy in rainbow trout (Salmo gairdneri). A preliminary study

1. Partial hepatectomy (25% of liver mass removed) has been performed on rainbow trout using a procedure allowing a high level of survival. 2. The liver regeneration was complete from the point of view of liver weight, at the 30th post-operative day. 3. During the first weeks after surgery, the NH-excretion rate in both hepatectomized and shamoperated trout was higher than in intact controls. Likewise, the food intake was lower in the operated animals. 4. Both facts can be attribute to the stress situation caused by the surgical manipulations rather than the removal of the liver.     

Validation of radioimmunoassay systems for the measurement of 11-keto- and 11 beta-hydroxytestosterone in teleost blood

Highly specific antisera for 11-keto- and 11 beta-hydroxytestosterone have been raised in sheep. Assay systems for the simultaneous measurement of 11-ketotestosterone, 11 beta-hydroxytestosterone, testosterone, progesterone and estradiol-17 beta were validated for Ictalurus nebulosus plasma and Carassius auratus serum. In males of both species 11-ketotestosterone and testosterone were the major steroids detected. In females, testosterone and estradiol-17 beta were the predominant steroids measured. Data from samples taken at different stages of the annual cycle suggest that seasonal fluctuations in gonadal steroid secretion occur in I. nebulosus and C. auratus.     

Salmon gonadotropin (sGTH) immunoreactivity and 11-oxotestosterone secretion of mature rainbow trout (Salmo gairdneri) testes in vitro: an alternative to radio-receptor assay for sGTH-binding studies

Summary After three intraperitoneal injections of salmon gonadotropin (sGTH) or bovine serum albumin (BSA), testicular tissue of fully mature rainbow trout was prepared for in vitro incubation with or without sGTH. The secretion of 11-oxotestosterone was measured and the tissue was fixed for light-microscopical localization of sGTH immunoreactivity (ir). Tissue from sGTH-treated males showed an increased basal secretion and sGTH-stimulated androgen secretion but the stimulated versus control ratio was higher without sGTH treatment in vivo. When the tissue had had contact with exogenous sGTH, sGTH-ir showed similar distribution patterns regardless of the sGTH treatment regimes. The extralobular compartment showed a staining of interstitial cells and capillary walls. The staining in the intralobular compartment was less clear. Many Sertoli-cells carried a faint nuclear label, whereas intralobular germ cells appeared to be unlabeled.     

Patterns of transgene inheritance in rainbow trout (Oncorhynchus mykiss).

There have been very few studies of the inheritance of introduced genes (transgenes) in fish. We have followed the inheritance of the mammalian fusion gene MTrGH from founder generation transgenics (originating from eggs microinjected with the MTrGH DNA) to offspring in crosses with control fish. Initial screening of the founder generation transgenics was by analysing DNA from blood samples. Only three out of six fish which carried the novel gene in blood DNA transmitted it to their offspring, despite the presence of the gene in DNA extracted from the sperm of all four male fish in this group. The frequency of transgenics in the progeny groups from the three fish which transmitted the gene varied widely: in one of these groups more than one type of MTrGH restriction pattern was found. These results suggest widespread mosaicism in founder generation transgenics.     

Tooth replacement in the upper jaw of the rainbow trout (Salmo gairdneri).

The present paper describes the pattern of tooth replacement in the upper jaw of two size ranges of rainbow trout (standard lengths 12-15 cm and 20-23 cm) as determined from wax impressions of the dentitions taken twice weekly on anaesthetised fish. There was considerable variation in the nature of replacement waves (lines constructed on dental charts linking eruption times of alternate teeth) between fish for both the premaxillary/maxillary and vomerine/palatine rows. The wave patterns obtained showed the same consistency of form for any one animal which is to be expected when consecutive generations show constant or proportionately changing life spans. Data relating to the life span of the teeth is given. The results are compared with those derived for the lower jaw and the significance of wave form discussed.     

A putative beta2-adrenoceptor from the rainbow trout (Oncorhynuchus mykiss). Molecular characterization and pharmacology.

Extensive molecular characterization of mammalian beta-adrenoceptors has revealed complex modes of regulation and interaction. Relatively little attention, however, has focused on adrenoceptors from early branching vertebrates such as fish. Using an RT-PCR approach we have cloned a rainbow trout beta2-adrenoceptor gene that codes for a 409-amino-acid protein with the same seven transmembrane domain structure as its mammalian counterparts. This rainbow trout beta2-adrenoceptor shares a high degree of amino-acid sequence conservation with other vertebrate beta2-adrenoceptors. The conclusion that this sequence is a rainbow trout beta2-adrenoceptor is further supported by phylogenetic analysis of vertebrate beta-adrenoceptor sequences and competitive pharmacological binding data. RNase protection assays demonstrate that the rainbow trout beta2-adrenoceptor gene is highly expressed in the liver and red and white muscle, with lower levels of expression in the gills, heart, kidney and spleen of the rainbow trout. The lack of regulatory phosphorylation sites within the G-protein-binding domain of the rainbow trout beta2-adrenoceptor sequence suggests that the in vivo control of trout beta2-adrenoceptor signaling differs substantially from that of mammals.     

A radioimmunoassay for corticosterone and its application to the measurement of stress in poultry.

A radioimmunoassay for corticosterone was developed using an antibody to corticosterone-21-hemisuccinate:bovine serum albumin. The assay possessed good specificity, sensitivity and reproducibility and required minimal sample preparation. Tests of adrenal function showed that stimulation of the adrenal with exogenous ACTH and with dexamethasone caused an increase and decrease, respectively, in plasma concentrations of corticosterone. Exposure to cold environmental temperatures caused an increase in plasma corticosterone. Handling and the removal of blood samples by venepuncture had no effect upon the concentration of corticosterone. It was concluded that this assay would accurately measure the response to stresses which affect the pituitary-adrenal axis.     

A new method for the simultaneous determination of androstenedione, testosterone, 11-oxotestosterone and 11 beta-OH-testosterone in fish plasma using combined techniques of Celite chromatography and radioimmunoassay

A new method for the simultaneous determination of androstenedione, testosterone, 11-oxotestosterone and 11 beta-hydroxy-testosterone in teleost plasma has been developed. Steroids extracted from the plasma were first separated by Celite chromatography using ethanediol as the stationary phase and different concentrations of ethyl acetate in iso-octane as the eluting solvents. Androstenedione, testosterone, 11-oxotestosterone and 11 beta-OH-testosterone were eluted successively with 0, 10, 15 and 40% of ethyl acetate in iso-octane. The eluted steroid fractions were then quantatively determined by radioimmunoassay. Data on the accuracy, precision, and sensitivity were presented showing that this new assay system was precise and reproducible. As an illustration of its application, this method was used in the present study to determine the plasma androgen levels in Monopterus albus and in methyl-testosterone treated Tilapia mossambicus.     

Circannual rhythms of gonadal maturation in female rainbow trout (Oncorhynchus mykiss).

Individually identified, 2-year-old female rainbow trout were maintained for up to 51 months on a constant schedule of 6 hr light and 18 hr darkness (LD 6:18), constant temperature (8.5-9.0 degrees C), and constant feeding rate. The fish exhibited free-running circannual rhythms of gonadal maturation and ovulation, which were self-sustaining for up to three cycles. The periodicity of the rhythm showed variation between fish and in successive cycles for the same fish, ranging from approximately 11 to 15 months.     

Oxidative stress generated by dietary Zn-deficiency: studies in rainbow trout (Oncorhynchus mykiss).

To date, there is scarce information on the metabolic and biochemical repercussions of Zn-deficiency in fish. In this work, the effect of dietary Zn-deficiency on the diet utilization and the metabolism of activated oxygen species in rainbow trout (Oncorhynchus mykiss) has been studied. Fish were randomly separated in different lots according to their Zn-starvation and diet intake. In crude extracts of liver, gut and muscle, total and isoenzymatic superoxide dismutase and catalase activities were analysed. Lipid peroxidation was also determined in the same tissues. Western blotting was performed using antibodies against manganese- and copperzinc-containing superoxide dismutase. Lots fed on the Zn-deficient diet and with low intake showed significantly lower weight gain and feed-conversion efficiency indexes than control trouts. However, these parameters returned to control values when trouts were recovered by feeding them a control diet ad libitum. In control trouts, three independent copperzinc superoxide dismutase isozymes were detected in liver, whereas only one isozyme was present in the other lots. However, by Western blotting analysis the presence of a manganese superoxide dismutase was found in liver from all lots except in control trouts. Catalase activity and lipid peroxidation values were mainly detected in liver and gut, respectively, and both parameters increased in all lots with respect to the control group. Our results thus suggest that in rainbow trout an oxidative stress appears to occur as a consequence of Zn-deficiency.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Some of the blood constituents of male, female and immature rainbow trout of the Shasta variety, maintained in known environmental and dietetic conditions, were examined and the results statistically treated. The parameters for male and female trout were similar, only the erythrocyte count exhibiting a significant difference. Large but expected differences were evident between the parameters for mature and immature fish.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Fifty rainbow trout of the Kamloops strain were examined for 12 haematological parameters: erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, leukocyte count, differential leukocyte count, plasma total protein and plasma glucose concentration. The fish had been held under known environmental and dietetic conditions, and at the time of sampling were 14 months old. The majority of results for erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, total protein and differential leukocyte count fell within narrow ranges. The total leukocyte counts and glucose levels were more widely spread. The results are discussed and compared with those already published for Idaho and Shasta strains. It is impossible to say whether the differences that were observed between Kamloops and these other varieties were due to strain alone, since other variables were present. Some problems associated with establishing normal ranges for these parameters in fish are discussed.     

A literature review of the blood chemistry of rainbow trout, Salmo gairdneri Rich.

The literature on the blood chemistry of rainbow trout, Salmo gairdneri Rich., is reviewed from the aspects of experimental methods, electrophoresis, normal values, environmental and endogenous factors including toxic substances and diseases.