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 Stocking Density, Salinity, and Light Intensity on Growth and Survival of Southern Flounder Paralichthys lethostigma Larvae

Four separate studies were done on Southern flounder Paralichthys lethostigma larvae during first feeding and metamorphosis to determine the effects of stocking density, salinity, and light intensity on growth and survival. One study used stocking densities of 10, 20, 40, and 80 fish/L during first feeding; the second study compared the growth and survival of larvae stocked at 20 and 33 ppt; and a third experiment evaluated stocking densities of 1/L and 3/L under two different light intensities (1,600 lux vs 340 lux) during metamorphosis. The fourth experiment tested the effects of different salinities (0, 10, 20 and 30 ppt) on larval growth and survival during metamorphosis. Growth and survival (overall 6.9%) were not significantly different ( P > 0.05) for stocking rates up to 80/L. Larvae placed into 20 ppt salinity had survival through first feeding similar to that of larvae raised at 33 ppt. During metamorphosis, light intensity had no effect ( P > 0.05) on growth or survival, but fish stocked at 3/L had significantly lower ( P < 0.05) survival than fish at 1/L. Complete mortality of larvae occurred at 0 ppt. Growth and survival past metamorphosis were not significantly different ( P > 0.05) at 10, 20 and 30 ppt, but unmetamorphosed fish did not survive to day 60 at 10 ppt. Based on these results, practical larviculture of Southern flounder may require a two-step process with high stocking rates (80 fish/L) through first feeding and lower densities (1/L) through metamorphosis. Fingerling production in fertilized nursery ponds might he possible at salinity as low as 20 ppt.     

The Annual Reproductive Cycle of the White Bass Morone chrysops

The gametogenic cycle of the white bass Moronc chrysops was characterized for one year at monthly intervals by measuring circulating levels of the sex steroid hormones testosterone (T) and 11-ketotestosterone (11-KT) in males, and 17β-estradiol (E₂) and T in females. Vitellogenin (VTG), the egg yolk precursor, was also measured in female blood plasma. Gonadal status of individual fish was assessed by histological evaluation, measurement of oocyte diameters, and spermintion response. At the onset of vitellogenesis (October), blood plasma levels of E₂ and T were low, but increased significantly as the spawning period (March-April) approached and oocytes attnined their maximum diameter. All stages of oocyte development were observed in mature females, indicating that the white bass is a multiple-clutch, groupsynchronous spawner. Androgen levels increased in males throughout the reproductive cycle and reached maximal levels during the spawning period. The increase in androgens coincided with sperm production and spermintion in males. The circannual endocrine and gonadal development patterns observed in white bass were similar to those of other members of genus Moronc .     

Investigations of Selected Parameters for Growth of Larval and Juvenile Black Sea Bass Centropristis striata L.

Abstract.— The black sea bass Centropristis striata L. endemic to the U. S. Atlantic and Gulf of Mexico coasts is a highly sought species that commands a high price in the marketplace. Investigations were undertaken to determine the basic requirements for culture of sea bass larvae and juveniles. Adult black sea bass were captured from the wild and were found to be robust and to adapt well to captivity. Larvae were obtained by strip spawning of these adults and survived and grew at higher rates in seawater supplemented with algae (greenwater) and maintained at 22 C, than in seawater with or without supplemental algae at 18 C. Larvae were provided a diet of rotifers through 12 d post hatch (DPH) and weaned over a 3-d period to enriched Anemia replenished daily to a density of 10 individuals/mL. By 18 DPH, larvae began ingesting a formulated diet for marine finfish and were completely weaned from live feed by 25 DPH, which coincided with the onset of metamorphosis. Juveniles tolerated a range of salinity from 10–32 ppt with the highest growth rate observed at 20 ppt. Culture of larvae and juveniles on a commercial scale was successful using conditions similar to those employed in the laboratory. The findings presented here suggest the potential for commercial culture of this species in the USA.     

The effects of photothermal manipulation on reproductive development in female haddock Melanogrammus aeglefinus L.

Haddock (Melanogrammus aeglefinus L.) were maintained on either simulated natural (SNP) or advanced, phase‐shifted photoperiods (ADV) and reproductive characteristics of female fish were measured throughout two reproductive cycles. By advancing the seasonal photoperiod 8 weeks, relative to ambient conditions, follicle development, peak levels of estradiol and initiation of spawning were all accelerated compared with fish maintained on an SNP. Profiles of steroid synthesis and follicular development were similar between the two groups but the spawning period was protracted by 40 days in the fish maintained on ADV. Ultrasound imaging proved to be an effective and non‐invasive tool for accurately assessing ovarian development throughout the reproductive cycle. These results suggest that photoperiod manipulation has significant potential for extending the availability of eggs and larvae for haddock aquaculture.     

Cryogenic and Refrigerated Storage of Rainbow Smelt Osmerus mordax Spermatozoa

The effects of extender composition, cryoprotectant, and freezing rate on post-thaw rainbow smelt Osmerus mordax sperm motility were examined, and the fertilization capacity of fresh and post-thaw sperm were compared. The highest post-thaw motility (75%) was obtained when milt was diluted 1:3 with an extender containing 600 mM sucrose supplemented with 10% dimethyl sulfoxide and 1.5% bovine serum albumin and frozen at a rate of –20 C/min. Post-thaw motility for sperm stored in this extender was similar to fresh sperm and did not change after 90 d of storage. Furthermore, there were no differences in fertilization rate or embryo survival to the eyed stage between fresh and post-thaw sperm frozen in this extender. The lowest post-thaw motility was observed when sperm were frozen with methanol at a rate of -30 C/min. Refrigerated sperm diluted 1:3 with the 600 mM sucrose extender remained motile for 30 d. These data demonstrate that rainbow smelt spermatozoa can be effectively used following short and long-term storage using a simple, sucrose-based extender.     

Changes in gonadal hormones during oocyte development in the striped bass,Morone saxatilis

Wild striped bass,Morone saxatilis, were collected from coastal waters and spawning areas to describe the endocrine correlates of oocyte development in non-captive, migratory fish. The fish were classified according to their most advanced oocytes. Serum levels of estradiol (E₂), testosterone (T) and 17-20-dihydroxyprogesterone (DHP) were measured by radioimmunoassay (RIA). Females in the primary growth phase and early secondary growth phase (pre-vitellogenic) had low levels of plasma steroids, ovarian lipid content and gonadosomatic indices (GSIs). Significant increases in E₂, T, ovarian lipid content and GSIs occurred during the vitellogenic phase. Maximum levels of all reproductive parameters were found in prespawning fish sampled in the Hudson River. Mean levels of E₂, T, ovarian lipids and GSIs for these fish were 2.0±0.5 ng/ml, 3.0±0.3 ng/ml, 24±1 mg/g, and 5.6±0.3% (mean±SEM), respectively. In fish induced to spawn with human chorionic gonadotropin (HCG), DHP levels (1.9±0.4 ng/ml) were significantly elevated. Similar levels were found in two fish captured during the spawning season, suggesting that DHP may serve as the maturation-inducing steroid in this species.     

Whole-body corticosteroid and plasma cortisol concentrations in larval and juvenile Atlantic cod Gadus morhua L. following acute stress

Methods were developed to assess whole‐body immunoreactive corticosteroid concentrations (IRC) and plasma levels of cortisol in Atlantic cod subjected to several common, acute stressors. A measurable corticosteroid stress response was observed at the first sampling in whole bodies of 8‐day post‐hatch (dph) larvae. Two groups of juveniles (～5 and 30 g) were subjected to a 30 s net stressor and whole‐body IRC and plasma cortisol levels were determined. Post‐stressor IRC in smaller fish rose approximately 14‐fold, peaked at 1 h, were sustained for 3–6 h and returned to pre‐stressor levels within 24 h. Post‐stressor plasma cortisol levels in larger fish rose approximately 18‐fold, peaked at 0.5–1 h, were sustained for 1–3 h and then returned to near pre‐stressor levels after 24 h. Immunoreactive corticosteroid concentrations appeared to remain elevated longer than plasma cortisol levels suggesting that steroids other than cortisol were contributing to total immunoreactivity in homogenates. Juveniles exposed to either a grading procedure or high density transport had maximal IRC and plasma cortisol levels within 90 min which returned to pre‐stressor levels within 24–72 h. A reduction in water salinity (20 g L^?1) did not moderate the corticosteroid response in juveniles. The results show that Atlantic cod respond to common, acute stressors in a manner similar to other teleosts. Whole‐body homogenates can be used to identify changes in IRC in response to acute stress in cod with the caveat that recovery IRC may differ from plasma cortisol concentrations.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.