Uptake and depletion of plasma 17α-methyltestosterone during induction of masculinization in muskellunge, Esox masquinongy: Effect on plasma steroids and sex reversal

Oral administration of 17α-methyltestosterone (MT) was used to induce masculinization of sexually undifferentiated muskellunge, Esox masquinongy. Three groups of muskellunge (mean weight, 2.5 ± 0.6 g) were submitted to MT treatment (15 mg of MT/kg) for 60 days. An additional one group was used as a control (hormone-free diet). Food was distributed over a 10-h period by using automatic belt feeders. Blood was sampled in both control and treated fish at different intervals during and after feeding: before (0 h), at 3 h, 6 h, and cessation of feeding (10 h), and after a fast of 22 h (32 h). MT had no significant effect on growth and survival in muskellunge 6 months after the treatment. Concentrations of plasma MT increased during the feeding period and reached their maximum levels 6 or 10 h after starting feeding. This rapid increase of MT indicated a rapid absorption of this steroid. Plasma MT levels then declined and reached a radir by 22 h after cessation of feeding, suggesting that MT is rapidly metabolized and excreted. The profiles of plasma testosterone during the MT treatment did not differ significantly between control and MT-treated groups. During and after the MT treatment, the concentration of plasma testosterone did not differ significantly between control and MT-treated groups. Moreover, no sexual dimorphism of testosterone levels was observed. Six months after treatment, the sex ratio in MT-treated groups (33% males, 62% females, and 5% intersex) was opposite to control (70% and 30%, respectively) and differed significantly. This suggests that at 15 mg of MT/kg over 60 days, a paradoxical feminization took place.Salaries were partly provided by State and Federal Funds awarded to the Ohio Agriculture Research and Development Center, Wooster, OH, USA

Induction of Gynogenesis in Muskellunge With Irradiated Sperm of Yellow Perch Proves Diploid Muskellunge Male Homogamety

Diploid gynogenesis was induced in muskellunge Esox masquinongy using UV-irradiated muskellunge sperm as the first step in producing monosex females. In this approach, we have to rely on negative controls as an indirect reference for sperm genetic material destruction. In the first experiment, equal proportions of gynogenetic females and males were produced. Negative controls, UV-irradiated sperm without heat shock, yielded some normal hatching larvae, described as spontaneous diploids. In the second experiment, muskellunge eggs were activated using sperm from yellow perch. Because hybrids between these species are not viable, we produced unambiguous gynogens. When UV-irradiated yellow perch sperm was used to inseminate muskellunge eggs, haploids resulted (22.5% ± 2.8% survival to the eyed stage). To produce diploid gynogens, a heat shock of 31°C was applied to inseminated eggs 20 min after activation for a duration of 6 min. This process yielded several hundreds of gynogens for rearing. Several treatments of masculinizing hormone, 17 α-methyltestosterone (MT), were carried out. Fish were dissected and gonads examined histologically for sex determination. Gynogens produced using yellow-perch sperm confirmed the presence of males in the control group, whereas the MT bath treatment (400 μg/liter) resulted in the production of fish with ovotestis. These results provide evidence for male homogamety in muskellunge and imply that a change of strategy is needed to produce monosex populations.Funding for this project was provided by the Federal Aid in Sport Fish Restoration Program (F-69-P, Fish Management in Ohio), administered jointly by the U.S. Fish and Wildlife Service and the Ohio Division of Wildlife

Use of Erythrocyte Measurements to Identify Triploid Saugeyes

The use of erythrocyte size measurements as a possible alternative to flow cytometry for identifying triploid saugeyes (female walleye Stizostedion vitreum X male sauger S. canadense) was evaluated. Blood smear preparations were made from 32 heat-shocked saugeyes (1.0-42.7 g; 52-185 mm total length), which consisted of 12 diploids and 20 triploids, as determined by flow cytometry after blood cells were stained with propidium iodide. The length, width, surface area, and volume of the cell and nucleus of 100 erythrocytes were determined for each fish. The cell and nuclear dimensions were measured at 1,000X magnification with a calibrated ocular micrometer. Discriminant analysis was used to distinguish diploids and triploids based on their score profiles. Results showed that triploid saugeyes had significantly larger erythrocyte cell and nucleus measurements than their diploid counterparts (N = 32; P < 0.0001). Erythrocyte measurements correctly distinguished 93.8% of fish samples as diploids or triploids, but measurements require about 1 h/fish. The potential applications of this technique for fisheries management and aquaculture are discussed.This research was funded by the Ohio Division of Wildlife under Federal Aid in Sport Fish Restoration, project F-69-P-5, Fish Management in Ohio

Juvenile growth and survival of heat-shocked triploid hybrid saugeyes, Stizostedion vitreum × S. canadense

Triploid hybrid saugeyes may be a desirable stocking alternative because diploid saugeyes are fertile and can compromise the genetic integrity of parental stocks. Four groups of saugeye eggs were heat-shocked 5 min after fertilization for 15 min at 31°C, which resulted in an 86.7 ± 9.4% triploidy rate and 57.8 ± 24.3% survival to the eyed-stage. Heat shocked and control saugeyes were stocked in two ponds for 40 days at 395 000 ind.ha^-1. Mean length and weight of triploid and heat-shocked diploid saugeyes were greater (P < 0.01) than unshocked diploid saugeyes. Survival of heat-shocked saugeyes (22.4%) was lower than unshocked saugeyes (94.7%). Heat-shocked fish (n = 25 851) harvested from ponds were stocked for 153 days in O'Shaughnessy Reservoir, Ohio, USA. Triploids were 34 mm shorter and weighed 41 g less than heat-shocked diploids (P < 0.01). Mean length and weight of triploids were comparable with unshocked diploids elsewhere in Ohio.This research was funded by the Ohio Department of Natural Resources, Division of Wildlife under Federal Aid in Sport Fish Restoration Project F-69-P-5, Fish Management in Ohio

Correlations of oocyte diameter with some morphometric characters in the nile tilapia, Oreochromis niloticus L.

A study was conducted to determine the correlation of oocyte diameter with some morphometric characters in a Philippine strain of Oreochromis niloticus L. One hundred and thirty-five 7- to 9-month old sexually mature female broodstock (body weight: 71±35 g; standard length: 13.1±2.1 cm) were randomly sampled from a pool of broodstock maintained in 4 m2 net enclosures suspended in an earthen pond. The following morphometric characters were determined for each fish: body weight, standard length, genital pore dimension, and genital papilla dimension. Intra-ovarian oocyte samples were obtained by inserting a soft silicon rubber tubing into the ovipore of the fish and aspirating the oocytes by mouth. There were no significant correlations between the average oocyte diameter and any of the morphological characters studied. Results justify the adoption of an ovarian biopsy technique for assessing the stage of gonadal maturity in sexually mature tilapia