Progress in Controlled Maturation and Spawning of Summer Flounder Paralichthys dentatus Broodstock

Abstract— Wild-caught, adult summer flounder Paralichthys dentatus (avg. wt. = 740 g; range = 264–1,540 g; N = 60), collected in northeastern US coastal waters during October 1994, were transported to Vero Beach, Florida in March 1995 and held in 2.6-m³ indoor tanks through November 1995 under two artificial photothermal regimes: (1) natural regime, simulating natural habitat conditions; and (2) accelerated thermal regime, with seasonal temperature changes advanced by one month. A third group of fish was held in outdoor tanks under ambient photothermal conditions. Under all photothermal conditions, onset of vitellogenesis was associated with declining daylength and temperature, beginning in the accelerated group, then progressing to the natural and the ambient groups. From 20 September to 28 November 1995, 23 vitellogenic stage females from the accelerated and natural regimes were implanted with a cholesterol-cellulose pellet containing LHRH-a (100 μg/kg body wt). Females with initial mean oocyte diameters ranging from 258–456 μm spawned voluntarily 2.5–5.5 d postimplantation, while no maturational response was obtained from females with mean diameters ranging from 165–231 μm. Two females were spawned twice during the study period by LHRH-a pellet implantation. Infrequent, natural spawning without hormone intervention was also obtained. Females released from 22.7–396.9 × 10³ eggs on the first day of spawning, with fertilization and hatching rates of 0–93.470 and 0–81.1%, respectively.     

Effect of Oxygen Saturation in Water on Reproductive Performances of Pacu Piaractus brachypomus

Broodstock pacu Piaractus brachypomus as well as their eggs during their embryonic development were exposed to either normoxia (5.5–7.5 mg O₂/L) or hypoxia (2.0–4.5 mg O²/L) conditions. The plasma concentrations of 11-ketotestosterone in males and estradiol-17β in females, as well as that of their precursor testosterone (T) were significantly ( P < 0.01) higher in fish maintained under normoxic conditions than in fish exposed to hypoxia. After ovulation and spermiation induced by hormonal treatments, the plasma concentrations of T and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) significantly ( P 0.05) increased in both sexes under both normoxic and hypoxic conditions. The plasma levels of T and 17,20βP achieved under normoxic conditions were higher than the ones recorded under hypoxia, except for those of 17,20βP in males. Males responded positively to the hormonal treatments, and the concentration of spermatozoa was 10.5 ± 0.8 10⁹/mL under both oxygen conditions. Hypoxia resulted in significantly lower survival of embryos (17.3 ± 28%) in comparison to normoxic conditions (68.5 ± 25%). Moreover, larval deformities were found when exposed to hypoxia (91.6 ± 6%). During embryonic development of this species 4 mg/L of oxygen is tolerated at 26–27 C without negative impact. We conclude that despite the highly adaptable nature of adult pacu to environmental hypoxia, oxygen concentrations below 4 mg/L severely impacted survival of embryos.     

Changes of plasma steroid concentrations associated with spontaneous or induced ovulation in yellow perch Perca flavescens

This study examined the changes in plasma steroids during natural (Experiment 1) and induced (Experiment 2) final maturation in yellow perch Perca flavescens. In experiment 1, ovulating yellow perch were stripped of eggs and blood samples collected to determine the concentrations of testosterone (T), estradiol-17β (E2), and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP). Eggs from individual females were weighed and fertilized. Fertilization rate was determined at the embryo eyed stage. In experiment 2, females were randomly assigned to one of the following treatment groups: (1) saline (0.7% NaCl), (2) des-Gly¹⁰[D-Ala⁶] LHRH-ethylamide (100 μg LHRHa/kg), and (3) LHRHa plus 17,20βP (100 μg LHRHa/kg + 2 mg 17,20βP/kg). Fish were injected intraperitoneally with two doses at a two-day interval. Blood was collected prior to injections and at the time of ovulation/spawning and concentrations of T, E2, and 17,20βP (free and conjugated) were determined. In experiment 1, low concentrations of 17,20βP were recorded at spawning. In experiment 2, all surviving fish injected with LHRHa (5 of 5) released their eggs spontaneously during the week following injections. None of the surviving control fish (0 of 5) ovulated during this period, whereas only 1 of 3 surviving fish injected with LHRHa + 17,20βP released eggs. In the control group, concentrations of E2 and 17,20βP did not show significant differences over the experimental period, whereas plasma T concentrations increased significantly. In fish injected with LHRHa, the concentrations of T and 17,20βP increased significantly after the first injection but then declined at ovulation/spawning. It also appears that 17,20βP was conjugated to its sulfated form. Mortality reached 62.5% in the group injected with LHRHa + 17,20βP indicating that this treatment was severe. Thus, LHRHa alone appears highly effective in inducing ovulation in yellow perch. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of exogenous hormones on ovulation and gonadal steroid plasma levels in starry flounder, <Emphasis Type="Italic">Platichthys stellatus</Emphasis>

The present study was designed to examine the potential for inducing ovulation in starry flounder (Platichthys stellatus) using gonadotropin-releasing hormone analog (GnRHa) and human chorionic gonadotropin (hCG) to assess whether starry flounder are differentially responsive to GnRHa and hCG. Female starry flounder were injected or implanted with different doses of hCG or GnRHa pellets to examine their ovulation-inducing potential and effects on plasma levels of testosterone (T), 17β-estradiol (E2), and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP). Blood samples were collected for up to 10 or 25 days post-injection or post-implantation in two separate experiments designed to mimic the early and middle stages of spawning, respectively. Fish treated with the GnRHa pellets (100 µg) showed a significant increase in the total number of stripped eggs relative to the controls. GnRHa administration had no effect on the floating rate or fertilization rate of ovulated eggs in the both experiments, whereas hCG injection affected both of these rates. Plasma T levels were not significantly different between the exogenous hormone-treated and control fish. In contrast, the plasma E2 level was elevated in those fish treated with GnRHa, regardless of injection or implantation, and was accompanied by increased numbers of stripped eggs in both experiments. Treatment with GnRHa resulted in higher 17,20βP levels compared to the controls, and there was a positive relationship between elevated plasma 17,20βP and an increase in ovulated eggs in response to GnRHa treatment. The implantation of starry flounder with GnRHa-containing pellets was effective at inducing sustained ovulation compared to hCG treatment.     

The enrichment and retention of ascorbic acid in rotifers fed microalgal diets

The enrichment and retention of ascorbic acid (AA) was investigated in rotifers Brachionus plicatilis fed on microalgae ( Nannochloropsis oculata and Isochrysis sp. (T.ISO)) and baker's yeast Saccharomyces cerevisiae . The concentrations of AA of the rotifer diets used in the study differed significantly: 4200 μg g⁻¹ of dry weight in Isochrysis sp. (T.ISO), 2600 μg g⁻¹ in N. oculata and only 77 μg g⁻¹ in S. cerevisiae . Rotifers contained 620 μg AA g⁻¹ prior to the experimental feeding. When subsequently fed for 3 h on microalgae at a ration of 0.13 mg dry microalgae per 10⁶ rotifers rapidly and efficiently increased their content of AA: Isochrysis sp.-fed rotifers contained 1600 μg AA g⁻¹ and N. oculata -fed rotifers contained 1100 μg AA g⁻¹. Concentrations were boosted by a further feeding of a second ration of algae at three times the initial feeding ration; 21 h later, Isochrysis sp.-fed rotifers contained 2500 μg AA g⁻¹ and N. oculata -fed rotifers contained 1700 μg AA g⁻¹. This represented a 180% and 310% increase in the pre-feeding vitamin concentrations in Isochrysis sp. and N. oculata -fed rotifers, respectively. There were no significant changes in AA concentration in rotifers fed a similar ration of yeast throughout the feeding period (520-620 μg AA g⁻¹). Rotifers retained AA during a subsequent 24 h non-feeding period, with no significant changes in the concentrations in any of the rotifer groups. The production of rotifers rich in AA may be particularly valuable for the culture of fish larvae that have a high requirement for the vitamin.     

Effect of antibiotic treatment on the growth and survival of juvenile northern Chilean scallop, Argopecten purpuratus Lamarck (1819), and associated microflora in experimental cultures

Juvenile northern Chilean scallops of 937±55 μm shell height were exposed to five different concentrations of chloramphenicol (CHL) (5, 10, 25, 50 and 100 μg mL⁻¹), plus a control without antibiotics. To determine the effect of CHL on the accompanying microflora, the number of colony-forming units (CFU) that grew on TGE culture medium was counted in the seawater of containers with juveniles, and in containers with microalgae used as food. Both were exposed to the same concentrations of CHL. The growth rates of juveniles treated with CHL and the control without antibiotic showed highly significant differences ( P =0.0001). The growth rate was inversely proportional to the CHL concentration. The control sample presented the highest growth rate (84.4±14.3 μm day⁻¹), followed by the sample treated with 5 μg mL⁻¹ (64.2±14.3 μm day⁻¹). The survival in the control and in the treated samples with 5–50 μg mL⁻¹ was rather high, with a mean value of 95%. Only the sample treated with 100 μg mL⁻¹ had a low survival (36.7%). The CFU count was larger in the containers with juveniles, when compared with the ones with food. The CFU count tended to decrease with increasing CHL concentration in the juveniles.     

Dose-dependent influences of dietary β-1,3-glucan on innate immunity and disease resistance of hybrid striped bass Morone chrysops×Morone saxatilis

To investigate potential use of dietary β-1,3-glucan for health management of hybrid striped bass, juvenile fish were fed diets supplemented with yeast glucan (MacroGuard^®) at 0.05%, 0.1% or 0.2% of diet for 4 weeks, followed by immune response assays and a bath challenge with Streptococcus iniae . Dietary glucan significantly ( P <0.05) enhanced neutrophil oxidative radical production, and fish fed 0.1% glucan had a significant ( P <0.05) reduction in mortality (10%) after bacterial challenge compared with fish fed the control diet (46.7%). However, accumulative mortality of fish fed 0.2% glucan was not significantly different from that of fish fed the control diet. To further elucidate this observation, macrophages from sub-adult hybrid striped bass were isolated and cultured in L-15 medium with 10% foetal calf serum and penicillin/streptomycin supplemented with 0.2, 0.5, 1, 2, 5, 20 and 100 μg soluble glucan (MacroGuard^®) mL⁻¹ for 24 and 48 h. Intracellular superoxide anion production was significantly ( P <0.001) increased by 0.5 μg glucan mL⁻¹, but significantly ( P <0.001) suppressed by doses >5 μg glucan mL⁻¹. It is concluded that dietary yeast glucan has potential for use in diet formulations of hybrid striped bass to limit the adverse effects of S. iniae , but dosage should be an important consideration in administration.     

The efficacy of emamectin benzoate as an oral treatment of sea lice, Lepeophtheirus salmonis (KrÒyer), infestations in Atlantic salmon, Salmo salar L.

The efficacy of emamectin benzoate as an oral treatment of sea lice, Lepeophtheirus salmonis (KrÒyer), infestations in Atlantic salmon, Salmo salar L., was evaluated in a dose titration study and two dose confirmation studies. Replicated groups of salmon with induced infestations of sea lice were given emamectin benzoate on pelleted feed at doses of 0, 25, 50 and 100 μg kg⁻¹ biomass day⁻¹ for seven consecutive days. Sea lice were counted at 7, 14 and 21 days from the start of treatment, and comparisons made with control fish given the same diet without emamectin benzoate. Total numbers of sea lice were significantly reduced at all doses in all three studies when compared to control fish. There was no significant difference between doses of 50 and 100 μg kg⁻¹, but the 50 μg kg⁻¹ dose resulted in significantly fewer lice than the 25 μg kg⁻¹ dose. Emamectin benzoate was highly effective in reducing numbers of preadult and adult lice and prevented the maturation of chalimus to motile stages. The optimum therapeutic dose was selected as 50 μg kg⁻¹ day⁻¹ for seven days. Treatment reduced the incidence of epidermal damage by sea lice and, in one study, survival of treated fish was 48% higher than in control groups. No fish mortalities or adverse effects were attributed to treatment with emamectin benzoate at any of the doses tested.     

Overripening of ovulated eggs in goldfish,Carassius auratus: II. Possible involvement of postovulatory follicles and steroids

Changes in steroid hormone levels in the serum and ovarian fluid during overripening were studied in goldfish. Ovulated eggs retained in the ovarian cavity became overripe at around 12h after ovulation based on loss of fertilizability, with advanced degeneration by 24h. Blood and ovarian fluid were taken at 0, 3, 6, 12, 18 and 24h after ovulation. Both serum and ovarian fluid progesterone (P) showed a highly significant decline by 18h with a further decline by 24h; P levels were higher in the ovarian fluid. Serum 17,20-dihydroxy-4-pregnen-3-one(17,20-P) levels showed a progressive and more rapid decline, decreasing significantly by 12h with further decreases by 18h and 24h. Serum testosterone (T) levels increased significantly at 3h and remained high till 18h, thereafter they declined to the 0h level. No significant changes in estradiol-17 (E₂) levels were observed in the serum, except for a significant difference between 6 and 24h. There were no significant changes in ovarian fluid E₂, T or 17,20-P levels.The postovulatory follicles (POFs) showed degenerative changes which corresponded to the decline in P and 17,20-P. The hypothesis that overripening may be associated with declining levels of P or 17,20-P was tested in vivo by immersing just ovulated females (0h) in P or 17,20-P solutions for 12h, and in vitro by immersing just-ovulated eggs (0h) in ovarian fluid with an anti-serum against P (Anti-P). In the former, P at 0.05 ppm significantly enhanced the fertilization rate of ovulated eggs stripped from the females at 12h while 17,20-P did not produce a significant effect at the tested concentrations (0.025 and 0.05 ppm) on the fertilization rate. In the latter, anti-P significantly lowered the fertilization rate of 0h ovulated eggs after 6h incubation. The evidence suggests a role of P in the maintenance of viability of ovulated eggs.     

Multigenerational effects of 17β-estradiol and nonylphenol on euryhaline cladoceran <Emphasis Type="Italic">Diaphanosoma celebensis</Emphasis>

ABSTRACT:     The effects of parental exposure to 17β-estradiol (E2) and 4-nonylphenol (4-NP) on life history parameters of three successive generations of the euryhaline cladoceran Diaphanosoma celebensis were assessed under laboratory conditions. Less than 24-h-old neonates (P) were exposed to five sublethal concentrations of E2 (0.1, 1, 10, 100 and 1000 μg/L) and 4-NP (0.01, 0.1, 1, 10 and 50 μg/L) at 25°C. Age at first reproduction, reproductive period, fecundity and lifespan were investigated. Successive generations (F₁–F₃) were monitored further in the absence of toxicants. Results showed that cladocerans exposed to 10–1000 μg/L E2 produced more neonates at a younger age compared to the control. The same effects were also observed in the F₁ and F₂ but ceased in F₃. Cladocerans exposed to 1 μg/L 4-NP also produced significantly more neonates compared to the control, but this effect was not found in successive generations. The results suggest that vertebrate hormone can modify the reproduction of D. celebensis , and the effects are multigenerational.     

Effects of Rhizopus extract administration on somatic growth and sexual maturation in lacustrine sockeye salmon Oncorhynchus nerka

ABSTRACT: The filamentous fungi Rhizopus , like many fungal species, possesses physiologically active substances. Rhizopus extract (RU) is reported to be effective for various aspects of growth and reproduction in many vertebrates. The effects of RU administration on body growth and plasma levels of steroid hormones were investigated in lacustrine sockeye salmon Oncorhynchus nerka . One-year-old fish were fed daily with RU (20 mg/kg feed) from July 1999 to October 2000 for 15 months. Fish were sampled every month and plasma levels of testosterone (T), 11-ketotestosterone (11-KT), estradiol-17β (E₂) and 17α,20β-dihydroxy-4-pregnen-3-one (DHP) were measured by time-resolved fluoroimmunoassay. Body growth of RU-fed fish of both sexes increased significantly in 1⁺ and 2⁺ October, and 2⁺ January–March and July. All RU-fed males and one female matured in 2⁺ October. RU-fed 1⁺ precociously mature males showed increased plasma levels of T, 11-KT and DHP in 1⁺ October. In 2⁺ males, RU significantly elevated plasma levels of T from May to June, 11-KT from June to July, and DHP in October. In sockeye salmon, administration of RU accelerated body growth of both sexes and sexual maturation in males, suggesting physiologically active substances present in RU enhance somatic growth and sexual maturation by sex-specific mechanisms.     

Artificial Propagation of Mutton Snapper Lutjanus analis, A New Candidate Marine Fish Species for Aquaculture

Abstract Wild-caught mutton snapper Lutjanus analis , a high-value marine food fish species, matured in flow-through seawater (36 g/L) tanks after 3 yr in captivity. On 31 May 1995, a female with a mean oocyte diameter of 382 μm was injected with human chorionic gonadotropin (HCG) (500 IU/kg body wt.) followed 24 h later by a second injection (1,000 IU/kg body wt.). At the time of the second injection, three males were injected with HCG (500 IU/ kg body wt.). Voluntary spawning occurred 33 h after the first injection, with a total of 534, 781 eggs released. Fertilization rate was 75.7%, while average diameter of fertilized eggs was 783 μm. Embryos were stocked in a 30-m³ outdoor tank at a density of 10.5/L. On day 2 post-hatching (d2ph), larval density was 8.61 larvae/L, and average notochord length was 2.6 mm. Larvae were fed ss-type rotifers from dl-d28ph, Artemia nauplii from 0–08ph, and artificial diets (52–48% protein) from d24-d38ph. On d38ph, fish averaged 0.308 g and 22.2 mm standard length. Survival (from d2ph) was 14.3%, with a total of 36,900 post-metamorphic juveniles produced. On d97ph, 1,390 hatchery-reared juveniles (avg. wt. = 10.5 g) were stocked into two 14.5-m³ recirculating seawater tanks (695 fish/tank; 48 fish/m³) and fed a 56% protein pellet. After 168 d, fish averaged 140.8 g, with a survival rate of 97.8% and a feed conversion ratio (dry wt./wet wt.) of 1.2. These preliminary results reveal the mutton snapper to be a prime, new candidate species for commercial cultivation.     

Bioencapsulation of erythromycin using adult brine shrimp, Artemia franciscana (Latreille)

Live adult brine shrimp, Artemia franciscana (Latreille), were enriched with erythromycin to determine if Artemia could accumulate therapeutic levels for subsequent feeding to young fish. Three trials were conducted to determine the erythromycin incorporation and survival rates of enriched Artemia when fed either liposomes containing erythromycin or various erythromycin suspensions. Erythromycin concentration in Artemia fed a liposome suspension was low (∼ 5 μg mL⁻¹) relative to Artemia fed the direct suspension (> 100 μg mL⁻¹) over the same time period. When enriched with suspensions up to 1 g erythromycin L⁻¹ sea water for 14 h, Artemia survival was not significantly affected ( P > 0.05) relative to controls. Using a suspension of 1 g L⁻¹, tissue erythromycin concentrations of 109 ± 16 μg erythromycin mL⁻¹  Artemia homogenate (mean ± SEM) were achieved after 12 h. Concentrations above 170 μg mL⁻¹ were obtained using suspensions of 2–5 g L⁻¹, but Artemia survival significantly ( P < 0.05) decreased.     

Induced Ovulation of Southern Flounder Paralichthys lethostigma Using Gonadotropin Releasing Hormone Analogue Implants

Implanted pellets that provide a sustained release of [D-Ala⁶ Des-Gly¹⁰] LHRH-ethylamide (GnRHa) were used to induce maturation and ovulation of Southern flounder Paralichthys lethostigma . Of the 12 females whose ovaries contained follicles with a maximum diameter ≥500 μm, 11 ovulated for the first time within 90 h of hormone implantation. Only 1 fish with a maximum follicle diameter less than 500 μm ovulated within 2 wk after implantation. Ovulated eggs were manually stripped from the females and mixed with sperm from several males. Most females were spawned 1 to 3 times on consecutive days with variable fertility. One female was spawned 11 times producing 668,000 eggs. Fertility was evaluated by examining the incubated eggs for early stages of embryonic cleavage. The percentage of fertile eggs in subsamples of incubated eggs ranged from 7–95%. The results indicate that GnRHa implants can be used to induce repeated ovulation in this species. The variability in fertility is discussed in relation to egg quality.     

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.     

The effect of dietary phosphatidylcholine and its constituent fatty acids on microdiet ingestion and fatty acid absorption rate in gilthead sea bream, Sparus auratus, larvae

This study tested the effect of the level of dietary phosphatidylcholine (PC) and its constituent medium-chain fatty acids on microdiet ingestion (μg diet larva⁻¹ h⁻¹) and the absorption rate of the free fatty acid [¹⁴C]16:0 (pmole larva⁻¹ h⁻¹) in 15, 20, 21, 25, 26, 30 and 31-day-old gilthead sea bream, Sparus auratus L., larvae. Fish were fed four microdiets (A, B, C and D): microdiet A contained no phospholipid (PL), while microdiet B included 10 g kg⁻¹ Artemia nauplii PL (3.7 g kg⁻¹ PC). Microdiets C and D contained 10 g kg⁻¹ purified saturated PC dimyristoyl (C_14:0) and polyunsaturated PC dilinoleoyl (C_{18:2[cis]−9,12}), respectively.
Larvae from one or both of the PC microdiets demonstrated significantly higher ( P < 0.05) ingestion rates (μg diet larva⁻¹ h⁻¹) than the non-PL microdiet control in 15, 21, 22, 25 and 26-day-old larvae and the Artemia PL microdiet in 15, 22 and 26-day-old larvae. However, microdiet ingestion and fatty acid absorption rate appeared to be independent of the associated medium carbon chain saturated or polyunsaturated fatty acid moiety of the PC diets. Apparent absorption, as measured by the retention of radio-labelled [¹⁴C]16:0 following 8 h of non-labelled microdiet feeding, was possibly related to feeding.     

Role of prostaglandin in the control of ovulation in the Japaneseeel Anguilla japonica

ABSTRACT:   The in vitro effects of 17,20β-dihydroxy-4-pregnen-3-one(DHP) and prostaglandins (PGE₁, PGE₂, PGF_1α,PGF_2α) on ovulation in the Japanese eel Anguillajaponica were examined. Oocytes with follicle layers at themigratory nucleus stage (approximately 850–900 µmdiameter) were removed using a polyethylene cannula from artificiallymatured fish. At concentrations of 10 and 100 ng/mL,DHP was found to induce both germinal vesicle breakdown and ovulation.The prostaglandins, except for PGE₁, effectively inducedovulation of previously matured oocytes by DHP treatment in vitro .Prostaglandin F_2α was the most effective. Asignificant increase in ovulation rate was observed even at a concentrationof 0.01 µg/mL PGF_2α.Indomethacin blocked the in vitro ovulation induced by DHPand addition of PGF_2α reversed indomethacin-blockedovulation. Actinomycin D and cycloheximide blocked DHP-induced ovulationand PGF_2α reversed the effects of both inhibitors. Theseresults indicate that DHP induces ovulation through endogenous prostaglandinsynthesis in the follicle layers of the Japanese eel.     

Residues of [14C]-malachite green in eggs and fry of rainbow trout, Oncorhynchus mykiss (Walbaum), after treatment of eggs

Abstract. Rainbow trout, Oncorhynchus mykiss (Walbaum), eggs were exposed to [methane-¹⁴C] malachite green chloride on day 0 and on every third day thereafter through day 24, with a final treatment administered to fry on day 31. Eggs or fry were sampled before each treatment, and at selected times from day 31 to day 59. Malachite green equivalence in eggs and fry was determined by sample oxidation and liquid scintillation counting. Total malachite green residues increased throughout the exposure period to 0.271 ± 0.042 μg g⁻¹ (± SD) on day 31. Residues declined to 0.055 ± 0.011 μg g⁻¹ on day 59. The depuration phase declined monoexponentially with a half-life of 13.3 days for the absolute amount (μg sample⁻¹) and a half-life of 9.7 days for the concentration of malachite green residues (μg g⁻¹). Growth dilution accounted for the 25% increase in the elimination (9.7 days) of malachite green residues. Extracts from treated eggs and fry were analysed by reversed-phase liquid chromatography. Three peaks were resolved in treated eggs: chromatic malachite green, leuco malachite green and an unknown polar metabolite. Only two peaks were resolved in the fry: leuco malachite green and the unknown polar metabolite. The most prominent residue in all samples was leuco malachite green.     

Liver damage in brown trout, Salmo trutta L., and rainbow trout, Oncorhynchus mykiss (Walbaum), following administration of the cyanobacterial hepatotoxin microcystin-LR via the dorsal aorta

Purified microcystin-LR (MC-LR) was administered via the dorsal aorta to brown trout, Salmo trutta L., or rainbow trout, Oncorhynchus mykiss (Walbaum), and, within 24 h, a dose of 300 μ g MC-LR kg^–1 caused increased activities in the blood by enzymes originating mainly from the liver, i.e. lactate dehydrogenase (LDH) and alanine transaminase (ALT). A dose of 75 μ g MC-LR kg^–1 significantly increased liver enzyme activities in the blood of brown trout at 24 h, but was without effect on rainbow trout, whereas 25 μ g MC-LR kg^–1 had no effect on blood LDH or ALT activities in either species. However, histopathological analysis of liver from both species following administration of the lowest toxin dose showed hepatocyte swelling and necrosis. Liver damage was more severe in brown trout compared to rainbow trout following administration of 300 μ g MC-LR kg^–1, showing disruption of the parenchymal architecture. After 48 h, there was a dose-dependent increase in the hepatosomatic index in both species. It is concluded that brown trout are less tolerant to MC-LR than rainbow trout.     

An Evaluation of a Leuteinizing Hormone-Releasing Hormone Analog to Induce Spawning of Channel Catfish Ictalurus punctatus

The leuteinizing hormone-releasing hormone (LH-RH) analog, DAla⁶DesGly¹⁰ LH-RH-ethyl amide, delivered by intraperitoneal or intracranial injection was an effective agent for the induced spawning of channel catfish ( Icrulurus punctatus ) held in aquaria. A daily dose of 100 μg LH-RH analog (LH-RHa) per kg fish body weight was recommended as the dosage schedule of choice, but daily doses as low as 1 cg LH-RHa per kg fish body weight induced spawning of channel catfish. LH-RHa was as good or better than human chorionic gonadotropin or carp pituitary to induce spawning of channel catfish in aquaria. Sham-injected (control) channel caffish spawned in this study contradicting observations in earlier studies.