Genotyping,pedigree reconstruction and endocrinological characterization of Acipenser naccarii (Bonaparte, 1836) using microsatellite markers and plasma steroid levels

This study aimed to set up a method for the long‐term management of Adriatic sturgeon, through the analysis of sex steroids and the genetic profiling of individuals in order to maximize the already reduced genetic variability of this species. Ten A. naccarii adults (nine of captive origin, one captured in the Ticino river and then moved into captivity) and eight subadults located in a semi‐natural land‐locked pond in Abbiategrasso (Milan, Italy) were analysed. Plasma testosterone differed significantly in the eight analysed subadults allowing their differentiation into two groups: the first group with an average testosterone concentration of 5.42 ± 1.31 ng/ml (probably female) and the second group with an average of 423.14 ± 75.97 ng/ml (probably male), as subsequently confirmed by artificial stripping. The plasma testosterone level was also significantly different between adult males and females (371.37 ± 43.58 vs. 95.34 ± 51.10 ng/ml), while the E2 levels showed no significant differences. Animals were genotyped on the basis of 10 microsatellite loci and their parental relationships were defined: four adults, two females and two males, generated the eight subadults. On the basis of pedigree analyses and genetic distances, 15 unrelated couples were identified for the future breeding seasons. Finally, the adult female captured in the Ticino River showed an interesting genetic profile, widely different from all of the other 17 specimens analysed, and represents a valuable source of genetic diversity.     

Long-term effect of photoperiod manipulation on growth, maturation and flesh quality in Atlantic halibut

The aim of this study was to investigate the effect of continuous light at different stages during the production cycle of Atlantic halibut Hippoglossus hippoglossus L. on growth, age at first maturity, endocrine parameters and flesh quality. A group of juvenile halibut [mean (SD), initial weight 191.3 g (44.7)] was reared in indoor tanks under ambient temperature conditions for 38 months until harvesting (mean final weight, 4.6 kg). The entire photoperiod experiment was divided into four phases, where the fish in each phase were exposed to either natural photoperiod (62°33'N) or continuous light (L). Thus, the following five photoperiod combinations were tested: (a) Control group (NNNN), (b) Group 2A (NLNN), (c) Group 2B (NNLN), (d) Group 2C (NNNL) and (e) Production group (LNNN). Exposure to continuous light stimulated growth, and the final mean weights of Groups 2A and 2B were 23% and 11% higher than those of the Control group (NNNN). The final plasma 11-ketotestosteron levels were lower in Groups 2A (2.94 ng mL⁻¹) and 2B (2.46 ng mL⁻¹) compared with the Control (5.29 ng mL⁻¹), Group 2C (5.09 ng mL⁻¹) and the Production group (4.78 ng mL⁻¹) during spring 2007 (age 4 years), indicating higher age at first maturity in Groups 2A and 2B. Photoperiod regime had only a minor, and transient, effect on flesh-quality traits of the fish, whereas a significant seasonal effect was seen with a tendency towards increased gaping, lower pH, lower hardness and lower shear force in July compared with December and March.     

Effects of light and temperature conditions on the expression of GnRH and GtH genes and levels of plasma steroids in <Emphasis Type="Italic">Odontesthes bonariensis</Emphasis> females

In this study we examined the endocrine mediation between environmental factors (temperature and photoperiod) and the brain–pituitary–gonadal axis in females of pejerrey Odontesthes bonariensis. Changes in the expression of brain gonadotropin-releasing hormones (GnRHs) and gonadotropin (GtH) subunit [follicle stimulating-β (FSH-β), luteinizing hormone-β (LH-β), glycoprotein hormone-α (GPH-α)] genes, plasma gonadal steroids [estradiol (E₂) and testosterone (T)], gonadal histology, and gonadosomatic index (GSI) in adult females exposed to combinations of short-day (8 h) or long-day (16 h) photoperiods and low (12°C) or high (20°C) temperatures after winter conditions (8 h light, 12°C) were analyzed. Pejerrey females kept under the short photoperiod had low GSIs, and their ovaries contained only previtellogenic oocytes regardless of the experimental temperature. In contrast, females exposed to the long photoperiod had high GSIs and ovaries with vitellogenic oocytes at both temperatures. These fish also showed a significantly higher expression of sGnRH, pjGnRH, cGnRH-II (the three different GnRH variants found to date in the pejerrey brain), FSH-β, LH-β and GPH-α genes and plasma E₂ levels than those at the shorter photoperiod. No significant changes were observed in plasma T levels. Based on these results, we concluded that the increase in day length but not that of temperature triggers the maturation of pejerrey females after the winter period of gonadal rest and that this occurs by an integrated stimulation of the various components of the brain–pituitary–gonad axis.     

Compensatory growth in Atlantic halibut: Effect of starvation and subsequent feeding on growth, maturation, feed utilization and flesh quality

A group of juvenile Atlantic halibut (mean, SD, 199.5 g, 44.7) was split into two experimental groups: one group was fed in excess twice a day and later five days a week (i.e. Control group), and the other group was starved for 5 weeks and then subsequently re-fed for 10 weeks (i.e. 5/10 starvation/re-fed group). This treatment was repeated for three years until the fish were harvested (mean final weight, 4.4 kg). A size +specific compensatory response was seen; partial compensation was observed in halibut weighing less than 500 g whereas full recovery growth was seen in halibut over 2 kg. Food consumption and feed conversion efficiency was measured during two full 5/10 starvation/re-fed cycles. The Control group consumed 86% more food in the measurement period, whereas the FCE 40% higher in the 5/10 starvation/re-fed group (FCE = 0.95) compared to the Control group (FCE = 0.68). In males lower gonadosomatic index (2.2 vs. 3.0) and plasma 11-ketotestosterone (0.22 vs. 1.26 ng ml^− 1) levels in the 5/10 starvation/re-fed group were observed during late winter and spring of 2006 (age 3 years) indicating higher age at 1st maturity. Starvation had a significant effect for improving the quality as starved fish had higher muscle pH (6.4 vs. 6.2), less gaping (0.1 vs. 0.3) and harder texture (61.4 vs. 57.6 N). Overall, the rearing on a repeated 5/10 starvation/re-fed regime for 3 years led to full growth compensation, higher feed conversion efficiency, lower male maturation and improved flesh quality.     

Protein degradation and protease activity of chum salmon (Oncorhynchus keta) muscle during spawning migration

Sarcoplasmic protein content decreased significantly and myosin heavy chain was degraded gradually during spawning migration of chum salmon (Oncorhynchus keta). Acid and neutral proteinase activities increased significantly during spawning migration. These proteinase activities were higher in females than in males. High levels of acid proteinase were not caused by the injury of lysosomal membranes. Considering the physiological states of chum salmon, neutral proteinase activity might be related to the degradation of muscle protein. As the changes in serum sex steroids were similar to those in the protease activities during spawning migration, it was considered that high levels of protease activity during spawning migration were closely related to the serum levels of androgens.     

Off-Season Spawning of Eurasian Perch Perca Fluviatilis

This study was designed to determine the effects of the selection for body weight and of two final warming durations during an out-of-season reproductive cycle on gonad recrudescence, plasma steroid levels and spawning in Eurasian perch Perca fluviatilis males and females. Two different bodyweight classes (200–250 and 350–400 g) and two different warming periods (1 or 2 months) were studied.At the end of the chilling period, a significant effect of the selection for body weight was observed on gonad development whereas fish were the same age. According to higher levels in testosterone (12 ng mL^–1), a better development was recorded for smaller breeders (P < 0.05). A short water warming period up to 14°C (1 month v.s. 2 months) allowed higher rates of females with completed gonadogenesis (33–38% v.s. 7–8%) and spawning (31–33% v.s. 0%) (P < 0.05). Spontaneous out-of-season spawning (17 ribbons) and larvae were obtained. Relative fecundity was about 100 eggs g^–1 of female body weight. Five ribbons were partly fertilized with a rate ranging from 5 to 90%. These studies have shown that delayed spawning in Eurasian perch can be obtained by a temperature manipulation, but results are not sufficient because of low stimulation rate of females, low spawning rates and high heteroge-neity of fertilization rates. The effects of other environmental factors like photoperiod should be tested in further experiment to improve the protocol inducing out-of-season spawning production.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Effect of live weight and dietary supplement of raw potato starch on the levels of skatole, androstenone, testosterone and oestrone sulphate in entire male pigs

This study evaluated the effect of supplement of raw potato starch (RPS) on the levels of skatole, androstenone, testosterone and oestrone sulphate in plasma from entire male pigs. The study also evaluated relationships between plasma levels of skatole and testicular steroids at three different live weights (LW) of approximately 90, 100 and 115 kg. A total of 111 entire male pigs of a crossbred (Yorkshire dams×Swedish Landrace sires) were used. Animals were raised either in mixed pens, with females and males, or single-sex pens. Each pen contained seven or nine pigs. The most fast-growing three pigs from the pens with nine pigs were slaughtered when they reached 90 kg LW, and the remaining pigs were slaughtered at 115 kg LW. All pigs were fed the same commercial diet until the average pen weight reached 100 kg. Then, 33 out of 80 remaining pigs received RPS, 0.6 kg per pig and day, for 2 weeks prior to slaughter. Blood samples were taken from the pigs at three occasions: first, the day prior to first slaughter occasion, low-weight group; second, the day prior to change in diet, middle-weight group; and third, the day prior to second slaughter occasion, high-weight group. Plasma was analysed for the levels of skatole, androstenone, testosterone and oestrone sulphate. Fat samples were taken at slaughter and analysed for the levels of skatole and androstenone. The levels of skatole and testicular steroids in plasma were significantly higher in entire male pigs from the high-weight group fed no RPS compared to those from low- and middle-weight groups. The levels of the investigated compounds did not differ between low- and middle-weight groups (P>0.1). The diet with RPS induced a decline in skatole levels in plasma and fat (P<0.001), but not plasma levels of testicular steroids and fat levels of androstenone (P>0.05). Skatole levels were positively correlated to testosterone and oestrone sulphate levels in the middle- and high-weight pigs fed no RPS as well as to testosterone in the low-weight group. In the high-weight group fed RPS, skatole levels were not correlated to any of the analysed compounds. Approximately 26% of the entire male pigs (11 out of 43) from the high-weight group fed no RPS produced skatole levels in fat above 0.20 μg/g, whereas the pigs from the low- and high-weight group fed RPS did not produce skatole levels above 0.20 μg/g in fat. Androstenone levels in fat were high in all groups. In total 47% (52 out of 111) pigs expressed androstenone levels above the rejection levels in fat of 1.0 μg/g and 88% (98 out of 111) had androstenone levels above 0.5 μg/g. It was concluded that a lower slaughter weight and the supplement of raw potato starch to the diet could be used to reduce skatole levels in entire male pigs. Androstenone levels in fat, however, could not be reduced by either a lower weight at slaughter or dietary manipulation.     

Plasma Steroid Variations in Bull Calves Repeatedly Treated with Testosterone,Nortestosterone and Oestradiol Administered Alone or in Combination

The aim of this work was to investigate the secretion of dehydroepiandrosterone (DHEA), testosterone (T), dihydrotestosterone (DHT) and oestradiol (E) as biological markers in response to illegal administration of testosterone, 19-nortestosterone (N) and oestradiol, either alone or in combination. Twenty male Friesian calves (age 13-14 months) were allotted to a control group (n = 5), and five experimental groups (n = 3) each. Each experimental animal was repeatedly injected with one of the following hormonal treatments: E, T, N, T+E and N+E. Circulating DHEA, T, DHT and E were determined by radioimmunoassay. The administration of T alone did not induce any variation in plasma DHEA, T, DHT and E, which were similar to those in the control group. In contrast, DHEA, T and DHT were on average significantly lower in the T+E and N-treated groups (p < 0.01), whereas the administration of N+E resulted in the reduction of plasma T and DHT without any modification of plasma DHEA. The administration of E alone or in combination increased circulating levels of E but did not affect androgen plasma profiles. The results indicate that plasma levels of T do not permit detection of illegal treatments because plasma androgens always remained within the physiological range. Illegal E treatment could be detected in blood samples when they were collected at least every 20 days.