Temperature-dependent index of mitotic interval τ0 in greenling Hexagrammos otakii

ABSTRACT:   The aim of this study was to establish effective procedures for chromosome manipulation in greenling Hexagrammos otakii Jordan et Starks, which has enormous aquacultural potential. To accomplish this, temperature-dependent measurements of the mitotic intervals (τ₀) were carried out. The τ₀ in this fish was determined by averaging the duration of the first and third embryonic divisions at temperatures ranging 5–25°C. At higher temperatures, eggs developed faster and underwent more identical development. For greenling, τ₀ were 341.1 ± 3.60 min at 5°C, 275.5 ± 4.53 min at 10°C, 189.7 ± 6.93 min at 15°C, 99.2 ± 8.27 min at 20°C and 34.2 ± 8.74 min at 25°C. There were strong, negative correlations between the τ₀ and water temperatures at all temperatures studied ( Y  = −79.3 X  + 425.3, R ² = 0.9968, where Y is the mitotic interval and X is the temperature).     

Induced meiotic gynogenesis in shovelnose sturgeon

Gynogenesis was induced in three shovelnose sturgeon (Scaphirhynchus pIatorynchus) by heat shock after egg activation with UV-treated paddlefish (Polyodon spathula) milt. Ultraviolet dosage (J m–2) for the pooled milt samples was calculated using the following linear regression equation: Dosage = 2405.27 – 352.80X 19.78X2 (X = percent transmittance of milt). Activated eggs were incubated at 18 °C until shocking at 35 °C. Shock duration was applied at 0.050 intervals from 0.15 to 0.40 0 (8.25 to 22.00 min post-fertilization; 0 at 18 °C = 55 min). The highest yield of gynogenotes (16%) was observed at 0.25 0 for female 3, 10 % at 0.30 0 for female 2 and 12% at 0.35 0 for female 1. The percentage of viable gynogenotes responded quadratically to the tau index (s/0) when shock treatments were applied. The higher yields of viable diploid sturgeon gynogenotes were achieved when eggs were heat shocked at embryological ages ranging from 0.25 to 0.35 0 (approximately 14 to 19 min post-activation at 18 °C). No viable hybrids were produced in the control fertilization of sturgeon eggs with intact paddlefish sperm which verified the gynogenetic origin of the offspring produced. © Rapid Science Ltd. 1998     

Kinetics of Nitrogen Excretion by Cultured Flounder Paralichthys olivaceus

This study was undertaken to quantify the rate of nitrogen excretion by the cultured Japanese flounder as a function of body weight and water temperature. Measurements of the did rate of nitrogen excretion for the cultured flounder were made at mean water temperature of about 16, 19, and 23 C. The excretion rate of nilmgen was estimated by adding the nitrogen excreted in the forms of ammonia and urea nitrogens. The diurnal pattern in the excretion rate of nitrogen was governed mainly by the feeding time. The proportions of excreted ammonia to total nitrogen were 77.9% at 16 C, 84.5% at 19 C, and 83.2% at 23 C. The average was 81.9%. The relationship between the body weight and excretion rate of nitrogen ( E_N , mg N/fish per h) is expressed as E_N =α,.W', where α, = nitrogen excretion rate at temperature 0 . The parameter α, is dependent on water temperature, such that α, =α₂₀ Q₀₁^(0–200) where α₂₀= nitrogen excretion rate at 20 C, and Q₀₁, = temperature coefficient. The values of the kinetics parameters for the cultured flounder were found to be γ = 0.57, α₂₀= 0.060, and Q₀₁= 1.12.     

Induction of interspecific allo-tetraploids of Megalobrama amblycephala♀×Megalobrama terminalis♂ by heat shock

Interspecific allo-tetraploidy was induced by a combination of hybridization of Megalobrama amblycephala ♀× Megalobrama terminalis ♂ and thermal shock. A thermal shock at 40 °C for 2 min applied to eggs 33 or 26 min (τ₀: 1.40) after fertilization resulted in a significantly ( P <0.01) higher yield of allo-tetraploids than hybrid crosses without heat shock. The yield of allo-tetraploids in embryos from mid-gastrula to hatching stages was 15.7–24.4% in different thermal shock treatment groups. The allo-tetraploidy rate in 330-day-old juveniles was 9.3% and 6.7% in different year treatment groups. After 2 years of rearing, allo-tetraploidy males attained sexual maturity with milk-like milt at the age of 2+, while allo-tetraploidy females at the age of 2+, and even at 3+, did not show any signs of sexual maturation exhibiting obviously delayed maturity. Only a few allo-tetraploidy females (three individuals) showed maturation characters at the age of 4+ and these females were induced to spawn.     

A pharmacokinetic study of flumequine in sea bass, Dicentrarchus labrax (L.), after a single intravascular injection

The pharmacokinetic properties of flumequine following a single intravascular injection (10 mg kg^–1 fish) were studied in sea bass, Dicentrarchus labrax (L.), 120 g held at 18 °C. The absorption half life ( t _1/2α) and the elimination half life ( t _1/2β) of the drug were calculated to be 1.05 and 10.71 h, respectively. Tissue penetration of flumequine seemed to be moderate because both the apparent volume of distribution of the drug at steady-state ( V _d(ss)) and the extensive apparent volume of the central compartment ( V _c) were found to be small (1.51 and 0.626 L kg^–1). The mean residence time ( MRT ) was short (09.73 h) and the total clearance (CL_T) of the drug was rapid (0.156 L kg^–1 h^–1).     

In vitro hydroxylation of vitamin D3 and 25-hydroxy vitamin D3 in tissues of Atlantic salmon Salmo salar, Atlantic mackerel Scomber scombrus, Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua

The in vitro metabolism of ¹⁴CD₃ and ³H25OHD₃ was investigated in different tissues from Atlantic salmon Salmo salar , Atlantic mackerel Scomber scombrus , Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua . The tissues analysed were liver, kidney, head kidney, gills, spleen and intestine. The metabolites were extracted in methanol–chloroform and separated by normal-phase high-pressure liquid chromatography (HPLC) followed by scintillation counting. Identification of the metabolites was by comigration with standards on normal and reversed-phase HPLC systems and by protein-binding assays. All tissues from all species analysed produced hydroxylated derivatives identified as 25OHD₃, 24,25(OH)₂D₃ and 1,25(OH)₂D₃. In addition, some unidentified derivatives were recorded, one probably being 25,26(OH)₂D₃. Organs producing great amounts of one metabolite also produced considerable amounts of the other possible derivatives, suggesting a lower degree of specificity in fish organs than in human organs. The predominating metabolite was 24,25(OH)₂D₃ in all organs from salmon and mackerel during incubation with ¹⁴CD₃ and within most organs from all species during ³H25OHD₃ incubation. The latter observation probably results from the need for decreasing rather than increasing the calcium absorption in these species, which live at least some periods of life in a marine environment.     

A comparison of gastrically and surgically implanted telemetry transmitters in shovelnose sturgeon

Abstract  Transmitter retention and the effects gastrically and surgically implanted telemetry transmitters had on condition and mortality of shovelnose sturgeon, Scaphirhynchus platorhynchus (Rafinesque), >450-mm fork length were examined. Four, 35-day trials were conducted with approximately equal numbers of control fish, fish receiving dummy transmitters gastrically and fish receiving dummy transmitters surgically. Four sturgeon expelled gastrically implanted transmitters during the observation period. Two mortalities occurred among fish receiving gastrically implanted transmitters during the study period and one mortality was recorded for control fish. No mortalities were recorded for fish receiving surgically implanted transmitters. Condition change (Δ K _n ) was significantly different between fish receiving surgical implantation (mean Δ K _n  ± SE; −5.17 ± 0.94), fish receiving gastric implantation (−2.98 ± 0.72) and control fish (−0.65 ± 1.09). Surgical transmitter implantation is recommended for telemetry studies on shovelnose sturgeon because of increased transmitter expulsion and occurrence of mortality in fish receiving gastric implantation.     

Induced diploid gynogenesis and polyploidy in crucian carp, Carassius auratus gibelio (Bloch), x common carp, Cyprinus carpio L., hybrids.

Abstract. Results of a 16 year (1977–1992) study of hybrids obtained from crossing two cyprinid fish, crucian carp, Carassius auratus gibelio (Bloch) (bisexual form), and common carp, Cyprinus carpio L., are presented. The ability to produce diploid eggs in F₁ females (resulting from chromosome endoreduplication in early oogenesis) was responsible for the high yield of spontaneous diploids in gynogenetic progenies of F₁ females and for triploidy in hybrids from back-crosses of F₁ females with males of the parent species. Different types of hybrids, including diploid and triploid gynogenetic, tetraploid (amphidiploid) and androgenetic progenies, were obtained. Diploid and some triploid hybrids (females only) were reproduced inter se by induced gynogenesis in several subsequent gynogenetic generations. Amphidiploid hybrids were obtained by crossing triploid F₈ females with diploid common carp males or by crossing diploid gynogenetic females with diploid inverted hybrid males producing diploid sperm. Results of cytogenetic investigations and fish culture properties in hybrids are presented. The scientific and practical significance of the data obtained are discussed.     

Early survival of marble trout Salmo marmoratus: evidence for density dependence?

Abstract –  The role of endogenous and exogenous factors in regulating population dynamics of freshwater salmonids is still a matter of debate. The aim of the present work was to assess the relative importance of density-dependent and -independent factors in determining the survival of marble trout ( Salmo marmoratus ) yearlings in two populations living in Slovenian streams (Zakojska and Gorska). The investigation was performed by combining a classical life table analysis with Monte Carlo simulation. Size-dependent fecundity was estimated by stripping wild adults in the fish farm. A significant positive relationship was found between length of marble trout females and the number of eggs produced. Egg density was the major determinant of survival from eggs to age 1+ ( σ ₀) in both streams. Residuals of the relationship between σ ₀ and egg density were positively correlated with rainfall only in Zakojska, probably because, within a certain range, more intense rainfalls increases stream flow and, consequently, suitable habitat for trout. Our study shows how density-dependent and environmental factors can interact to determine the survival of marble trout during the first year of life.     

Nephrocalcinosis in rainbow trout Salmo gairdneri Richardson; the effect of exposure to elevated CO2 concentrations

Abstract. Rainbow trout exposed to environmental free C0₂ concentrations ranging from 12–60 mg/1 for a minimum of 275 days developed nephrocalcinosis. Gross, histopathological and chemical examinations demonstrated that prevalence and severity of nephrocalcinosis increased with increasing CO₂ concentrations. The condition was characterized by calcification and granuloma formation in kidney and stomach. Diet, but not water, PO₄ -P concentration appeared to modify the prevalence and severity at each CO₂ concentration tested.     

Seasonal changes in respiration rates of copepodite stage V Calanus finmarchicus (Gunnerus)

Copepodite stage V Calanus finmarchicus were collected at locations on the continental shelf north of Scotland, in the Faroe–Shetland Channel and west of Ireland on six occasions covering winter, spring and summer from October 1993 to June 1995. Oxygen consumption by the overwintering and active spring/summer population of animals was determined at temperatures close to in situ temperatures. Laboratory measurements of oxygen consumption were also made at standardized temperatures (0°C, 5°C, 7°C and 12.5°C) to determine the sensitivity of animals to temperature change in the different seasons. Rates of oxygen consumption were very low (7–30 μmol O₂ gC⁻¹ h⁻¹) at in situ temperatures during the winter and early spring and significantly higher (105–219 μmol O₂ gC⁻¹ h⁻¹) for the active surface population in May and June. Animals collected from the overwintering population showed no significant response to changes in temperature. Due to the low respiration rates, the calculated rate of decrease in carbon content in diapausing copepodite stage CV was very low (approximately 0.250 μgC day⁻¹). The respiration rates were used to construct a model to estimate survival of the animals with an initial carbon content equivalent to that expected of animals in October. The results showed that in order to survive during winter and have enough energy for moulting and migration to the surface in the spring, these animals have to live at temperatures close to 0°C and be in a diapause state.     

Age,growth and age at sexual maturity of fan ray <Emphasis Type="Italic">Platyrhina sinensis</Emphasis> (Batoidea: Platyrhinidae) in Ariake Bay,Japan

Abstract:   Age, growth and sexual maturity of the fan ray Platyrhina sinensis in Ariake Bay, Japan were determined from specimens collected from May 2002 to September 2006. Age determination was conducted by vertebral centrum analysis using soft X-radiography. Annual band pair deposition was determined by marginal increment and edge analyses. The von Bertalanffy growth model best described the overall pattern of growth for both males and females (males L _∝  = 455.2, k  = 0.56, t ₀ = −1.09; females L _∝  = 555.8, k  = 0.28, t ₀ = −1.77; L _∝ is the theoretical asymptotic total length in mm, k is the growth rate coefficient and t ₀ is the theoretical time at zero length). Parameter estimates suggest that females attain a larger asymptotic total length and grow more slowly than males. The observed maximum ages were 5 years for males and 12 years for females. Age at 50% sexual maturity was 2.1 years for males and 2.9 years for females. The results indicate that this species is relatively fast-growing, short-lived and early maturing compared with many batoid species.     

Haematological responses in the European eel, Anguilla anguilla L., to sublethal infestation by Anguillicola crassus in a thermal effluent of the Swedish Baltic

Abstract. Haematological variations were studied in a European eel population, Anguilla anguilla L., naturally infested by the sanguivorous nematode Anguillicola crassus. This occurred off the Swedish Baltic coast in an area receiving heated cooling water from a nuclear power station. The erythrocyte count, haematocrit, leucocrit, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentrations, the various cell types in peripheral blood, serum iron concentration, the amount of total serum protein, and scrum protein fractions α ₁ , β₁ , α ₂, β ₂ and γ were all measured. Each variable was correlated to: (1) the intensity of infestation, i.e. number of parasites per host; and (2) a parasitization index, i.e. the weight of parasites per somatic weight of the host. Most variables showed no or only minor reactions to the infestation. However, a marked increase in the γ -fraction of scrum proteins, reduced lymphocyte numbers and increased granulocyte numbers are indicative of a humoral and cellular immune response.     

Comparison of horse mackerel and tilapia surimi gel based on rheological and <Superscript>1</Superscript>H NMR relaxation properties

ABSTRACT:   Horse mackerel and tilapia surimi were subjected to six different heat and pressure treatments in order to compare gelation characteristics of easy- and difficult-setting gels, that is, temperature dependence, by observing rheological properties and microscopic molecular mobility. The stress–relaxation and proton spin–spin relaxation time (¹H T ₂) of water were measured for all treated gels. Horse mackerel gel demonstrated higher elasticity, large distribution of the stress–relaxation process, and smaller water ¹H T ₂ than tilapia in both heat and pressure treatments. The water ¹H T ₂ was steeply increased in the pressure treatment at around 294 MPa for both fishes. In contrast, the ¹H T ₂ rarely changed in the heat treatment in spite of the considerable change in rheological properties. From the experimental results, it is considered that the gelation of horse mackerel (easy setting) surimi is induced by highly unfolding and re-aggregation of protein, which contributes to the formation of a strong network structure compared with tilapia in both heat and pressure treatments, and that pressure treatments hardly improve the gel strength of tilapia (difficult setting) surimi. The water ¹H T ₂ measurement was used effectively in order to study gelation characteristics of easy- and difficult-setting fish through observing its molecular dynamics.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Studies on cryopreservation of eggs from rainbow trout (Salmo gairdneri) and coho salmon (Oncorhynchus kisutch)

The effect of pre-freezing treatments as well as freezing of inseminated, not water-activated eggs from rainbow trout Salmo gairdneri, and coho salmon, Oncorhynchus kisutch, was investigated in relation to survival and further development.Effects above freezing temperatures included: the temperature at insemination, viability of inseminated and unactivated eggs after storage, suitability of an incubation medium and the tolerance of eggs to various levels of the cryoprotectant dimethylsulfoxide (DMSO). Freezing experiments included: investigating the action of DMSO (0, 1, 2 mole) and the tolerance of coho eggs to temperatures between ?4.6 to ?30°C. Insemination temperatures between 0.5°C and 9.8°C (coho eggs) as well as incubation in an artifical medium (1-0°C) for 80 min (rainbow trout eggs) and 170 min (coho eggs) did not influence subsequent fertility. Storage of inseminated and unactivated rainbow trout eggs for 135 min and beyond reduced egg fertility. DMSO at 2 and 4 mole was detrimental to coho eggs (1-0°C). One mole DMSO had no (coho) or reduced (rainbow trout) influence on egg fertility when it was added gradually.In the presence of 1 mole DMSO most eggs remained unfrozen (67–89%) when kept for 10 min in frozen artificial medium (?4.6%) and 27–32% subsequently reached the eyed stage (control = 100). Further cooling (0.3°C/min) to ?10°C was still tolerated (62% unfrozen, 22% eyed eggs) but not to ?20°C (6% unfrozen, no development) and ?30°C (no survival). Use of 2 mole DMSO did not improve the results.     

Effects of estradiol-17β and 17α-methyltestosterone on gonadal sex differentiation in the F2 hybrid sturgeon, the bester

The effects of giving oral estradiol-17β (E₂) and 17α-methyltestosterone (MT) on gonadal sex differentiation in the F₂ hybrid sturgeon, the bester ( Huso huso female ×  Acipenser ruthenus male), are investigated. Giving E₂ at 10 μg/g diet to fish from 14 months until 31 months of age induced incomplete feminization and resulted in approximately 40% abnormal ovary development in which oocytes were observed without ovarian lamellar structures and gonadal shape was similar to normal testis. Giving MT at 25 μg/g diet for the same duration failed to induce masculinization, and resulted in approximately 30% undeveloped gonads even at 30–37 months of age. In contrast, E₂ and MT at only 1 μg/g diet given from 3 to 18 months of age was sufficient to induce feminization and masculinization, respectively. In these fish, feminization and masculinization were observed at 9 months, when most putative ovaries and testes were histologically distinguishable by the shape of the gonadal surface. These results indicate that sex reversal can be induced in these fish by hormone treatment that is started at 3 months age, before morphological differentiation occurs on the stroma of the gonads.     

Exotoxin-induced consumptive coagulopathy in Atlantic salmon, Salmo salar L.: inhibitory effects of exogenous antithrombin and α2-macroglobulin on Aeromonas salmonicida serine protease

Abstract. Consumptive coagulopathy was induced within 4 h in Atlantic salmon, Salmo salar L., by injecting purified serine protease from Aeromonas salmonicida into the dorsal aorta. Pretreatment with a bolus intravascular injection of (human) antithrombin (AT) or (bovine) α₂-macroglobulin (α₂M) just prior to injection of the protease alleviated the in vivo pro-coagulant effects of the enzyme, but could not hinder the development of consumptive coagulopathy. In fish receiving only saline as pretreatment, the coagulopathy was evident even after 28h, but the fish were not overtly sick. The addition of the exogenous inhibitors increased the fish's natural protection against the bacterial exotoxin, suggesting that both AT and α₂M are of importance for the outcome of the pathologic process. Results further indicate that while AT in vivo was mainly directed against generated thrombin and activated coagulation factor X (FX_a), α₂M inhibited the protease directly.     

Plasma protein levels as potential marker traits for resistance to furunculosis

Abstract. The feasibility of including individual records on correlated traits in a family selection programme which aims to increase resistance to furunculosis in Atlantic salmon was studied; markers were selected because of their potential role in the pathogenesis of the disease. Fibrinogen and α₂-antiplasmin both show genetic variation; both are correlated with survival after challenge with Aeromonas salmonicida , the correlation being 0·44 and 0·37 ( P < 0·05), respectively, and it is possible to measure both on a large scale at low costs. Contrary to α₂-antiplasmin, fibrinogen was negatively correlated with survival due to furunculosis within the 10 most resistant families and within the 10 most susceptible families in contrast to an overall positive correlation. This inconsistency could be attributable to the presence of different allelic phases in different families, and of major linked loci influencing survival and fibrinogen levels. Thus, only α₂-antiplasmin fulfils the requirements for a marker trait for resistance to the disease suitable for individual selection at the population level, whereas the use of fibrinogen would be restricted to within family selection. The full statistical model explained 51% of the variation in resistance to furunculosis, and α₂-antiplasmin contributed 15% to this variation when considered as a separate entity. Thus, the additional gain from including individual records on α₂-antiplasmin in a family selection programme could be significant.     

Influence of incubation temperature on body movements of Atlantic cod (Gadus morhua L.) embryos and on size at hatch

Body movements of cod (Gadus morhua L.) embryos reared from fertilization to hatch at 5.4°C were observed at various stages of development and at six experimental temperatures ranging from 0–10°C. Frequency of cod embryo body movements increased from zero at 42 degree‐days post fertilization to maximal at 73–82 degree‐days (1 or 2 days prior to hatch). Embryos were most active at 2°C (mean of 5.5 movements per 10 min), with activity declining to less than 1/10 min at 8–10°C. Lengths of hatched cod larvae reared at a series of constant temperatures (from 4–10°C) from fertilization to hatch were greater at lower incubation temperatures. Incubation temperatures of 2–4°C were found to be optimal for incubation of cod eggs.