Analysis of Short-Term Cortisol Stress Response in Channel Catfish by Anesthetization with Metomidate Hydrochloride and Tricaine Methanesulfonate.

The ability of the anesthetics metomidate hydrochloride and tricaine methanesulfonate (MS-222) to mitigate the cortisol stress response of Channel Catfish Ictalurus punctatus was evaluated during a 10-min confinement stress. The cortisol concentrations of Channel Catfish anesthetized in metomidate hydrochloride remained consistent throughout the 10-min exposure; however, for fish anesthetized with MS-222 and nonanesthetized fish, cortisol concentrations were approximately 7- and 22-fold higher, respectively, than the baseline concentrations. While both anesthetics reduced cortisol concentrations relative to those of nonanesthetized fish, these results suggest that MS-222 is an appropriate anesthetic to use during the initial 5 min of sedation and that metomidate hydrochloride is appropriate for longer periods of sedation.

Sex-linkage of glucosephosphate isomerase-B and mapping of the sex-determining gene in channel catfish.

Sex-linkage of glucosephosphate isomerase-B (GPI-B) was observed in five experimental matings between heterozygous male and homozygous female channel catfish (Ictalurus punctatus). Offspring phenotypes for GPI-B were 40.8% heterozygous male and 43.0% homozygous female, while recombinant offspring were 7.4% homozygous male and 8.8% heterozygous female. Thus, GPI-B and the sex-determining gene (SDG) were linked and had a recombination rate of 16.2%. This linkage was designated I. punctatus linkage group XXIX. The gene-centromere distance (1.66 cM) of SDG, estimated in six gynogenetic families derived from XY females, indicated that SDG resides very close to the centromere. Based on estimates of these genetic distances, a chromosomal order of GPI-B-centromere-SDG was proposed. Additionally, joint segregation of GPI-A and SDG in two experimental matings indicated no genetic linkage between GPI-A and sex. These genetic relationships were compared to those reported in other teleost taxa with regard to evolutionary conservation of ancestral gene arrangements.

Influence of dihydrotestosterone on sex determination in channel catfish and blue catfish: period of developmental sensitivity.

Treatment of channel catfish with 0.2, 20, or 200 mg/liter of dihydrotestosterone (DHT) in the water during the egg stage or during egg and sac-fry stages did not alter the expected 1:1 sex ratio of the progeny. Feeding DHT at 200 mg/kg of feed for the first 21 days after yolk sac absorption resulted in 80% females; this proportion was increased by combining feeding with treatment of 200 mg DHT/liter in the sac-fry stage (90%) or in the egg and sac-fry stage (97%). In contrast, treatment of blue catfish sac-fry with 200 mg DHT/liter, with or without the combination of feeding DHT at 200 mg/kg food, resulted in 100% female populations. Neither clomiphene citrate, an estrogen-receptor blocking agent, nor clofibrate, an inhibitor of hepatic synthesis of cholesterol, affected the sex ratio of channel catfish, and neither of these compounds altered the feminizing effect of 200 mg DHT/kg when fed in combination with DHT. The nonaromatizable androgen DHT is not as effective as many other androgens in producing paradoxical female populations of channel catfish. However, feminization of blue catfish by treatment of sac-fry indicates that this species is more susceptible to hormonal manipulation and that the period of sex determination may occur earlier in development than in channel catfish.

Hormonal sex manipulation and evidence for female homogamety in channel catfish.

The mechanism of sex determination in channel catfish Ictalurus punctatus was evaluated by hormonal and genetic methods. Aromatizable and nonaromatizable androgens, as well as an estrogen, caused feminization in fish fed steroids for 21 days after yolk-sac absorption. The effectiveness of 60 micrograms of ethynyltestosterone/g food decreased markedly when the experimental feeding period was shortened and was ineffective when the treatment lasted less than 12 days. Females from all-female populations produced by treatment with sex hormones were mated with normal males resulting in nine spawns with a sex ratio different from 1:1. The sex ratios were statistically similar to 3 male: 1 female in five spawns, both 2:1 and 3:1 in two spawns, and 2:1 in two spawns. These data are consistent with a model for female homogametic sex determination in channel catfish and suggest that the YY equivalent genotype is viable.

Extraction of a synthetic androgen from fish muscle and quantitation by high performance liquid chromatography.

A rapid, reproducible extraction procedure was developed to recover the synthetic androgen 17 alpha-methyltestosterone (MT) from fish muscle. A single extraction with chloroform-methanol (2:1 V/V), followed by elution from mini-columns with various methanol concentrations, yielded an extraction efficiency of greater than 70%. Resolution of MT from testosterone was complete and reliable quantitation was achieved utilizing reversed-phase high performance liquid chromatography. Endogenous testosterone was not detectable in 1-g muscle samples, but MT was detected in fish muscle 6 h after adult Tilapia aurea were fed a diet containing the steroid.