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Pre-harvest male maturation is problematic for Atlantic salmon (Salmo salar) farmers and is regulated by the environment and genetics (e.g., vgll3). Five families of all-male salmon parr (produced using YY males crossed with XX females) with different vgll3 genotypes were split between three environmental regimes in January 2018. The "advanced maturation" regime used elevated temperature (16 °C) and continuous light from January 2018 with post-smolt maturation assessed in March 2018. The "extended freshwater" regime used ambient freshwater (1–16 °C) and simulated natural photoperiod (SNP) with post-smolt maturation assessed in November 2018. The "sea transfer" regime used ambient temperatures (1–14 °C) and SNP in freshwater until May 2018 when they were transferred to 9 °C seawater with natural photoperiod for 2.5 years (final mean weight of circa. 14 kg) and assessed for post-smolt maturation, 1 sea-winter (1 SW) maturation, and 2 sea-winter (2 SW) maturation in the autumn (November/December) of 2018, 2019, and 2020, respectively. Post-smolt maturation was highest in the advanced maturation and extended freshwater regimes (39–99% depending on family) and lowest in the sea transfer regime (0–95% depending on family). In the sea transfer regime, maturity incidence increased over time (0–95% post-smolt maturation, 1–100% 1 SW, and 50–90% 2 SW maturation, depending on family). In all regimes, those homozygous for the pre-designated vgll3 "early" maturing allele had the highest incidences of maturation whilst those homozygous for the "late" allele had the lowest. A low percentage of 2 SW phenotypic and genetic females were found (0–5% depending on family), one of which was successfully crossed with an XY male resulting in progeny with an approx. 50/50 sex ratio. These results show (i) post-smolt maturation varies dramatically depending on environment although genetic regulation by vgll3 was as expected, and (ii) crossing YY sperm with XX eggs can result in XX progeny which can themselves produce viable progeny with an equal sex ratio when crossed with an XY male. [ABSTRACT FROM AUTHOR]

Photoperiod regulates the occurrence of unwanted male post-smolt maturation during the production of large (>100 g) Atlantic salmon (Salmo salar) smolts. However, the optimal daylength for triggering smoltification, but not male puberty, has yet to be established. We used either continuous light (24:0 light/dark) or long days (18:6 and 21:3) after a six week "winter" zeitgeber (12:12) to induce smoltification in fish of around 120 g reared at 16 °C. The fish were sampled 1, 2, 3, and 6 weeks after the initiation of the three different photoperiod treatments (n = 153 males in total with 9–18 males/photoperiod/time point). As expected, the smoltification indicator gill Na+/K+-ATPase (NKA) was significantly (p < 0.05) elevated and peaked 2 to 3 weeks after the initiation of the different photoperiods. Pubertal males were identified in all treatments via the combined use of relative testis size and histology, plasma 11-ketotestosterone, changes in body condition, and growth rate. The total incidence of puberty was significantly higher among males on continuous light at 33% (n = 16/49) compared to 10% (6/61) and 12% (5/43) in 21:3 and 18:6, respectively. The incidence of puberty increased over time in all photoperiods, with 62% (8/13), 19% (3/16), and 38% (3/8) of the males from 24:0, 21:3, and 18:6 pubertal at week 6, respectively. The mean weight of males that went on to initiate puberty was significantly higher (13%) at the beginning of the trial compared to those that remained immature (mean weight, 127 vs 112 g, respectively), but there was no initial difference in body condition. Puberty significantly reduced gill NKA by 35% compared to immature males at week six but had no effect at earlier time-points. Photoperiod had no effect on the female GSI, and they were all considered immature. In conclusion, the incidence of male puberty during smoltification is regulated by photoperiod and leads to an earlier decline in a key indicator of seawater readiness. As such, photoperiods with a short scotophase (21:3 or 18:6) following the winter zeitgeber in a square-wave (long-short-long day) smolt regime are recommended to limit the incidence of male puberty. • A 6-week LD24:0 photoperiod after LD12:12 resulted in more precocious male maturation compared to LD21:3 or LD18:6. • Males that entered puberty were significantly larger than those that did not. • Pubertal males had lower gill Na+/K+-ATPase enzyme activity than immature males at the end of the expected smolt window. [ABSTRACT FROM AUTHOR]

Atlantic salmon (Salmo salar) farming is moving towards extended periods of land-based production to minimise the time at sea, but this increases the risk of unwanted male sexual maturation. As the timing of puberty is driven by genetics (e.g. vgll3) and the environment, optimising rearing strategies and broodstock management may help alleviate the problem. Subsequently, we used a domesticated strain of vgll3 heterozygous broodstock to produce three all-male half-sibling families using three different production regimes to assess the timing of first puberty. Firstly, "large smolts" were produced in a flow-through system on 13 °C freshwater and constant light from first feeding (day 0) up to 1 kg (day 354). The mean incidence of pubertal males increased from 2% at 30 g, up to 78% at 1 kg. Genetics explained a significant (p < 0.05) amount of the variation, with 93, 71, and 34% of the expected early, intermediate, and late maturing vgll3 genotypes being pubertal on day 354, respectively. In addition, pubertal males in the early vgll3 genotype were found at the first sampling at ≈30 g, while it was not observed in the heterozygous and late genotypes until the fish were ≈90 g and ≈280 g, respectively. Secondly, "post-smolts" were produced by switching half the large smolts to 13 °C seawater at 420 g (day 272) and growing them in tanks to 0.95 kg (day 354). This led to a significant 14% reduction in the total incidence of puberty by day 354 compared to the large smolts. However, the regulation of pubertal timing by vgll3 did not interact with salinity. Thirdly, traditional "under-yearling" smolts were produced using periods of natural or manipulated temperature and photoperiod, followed by transfer to a sea-cage at 150 g in December (day 290) where they stayed for one year until harvest at 4.75 kg (day 648). None of these fish were mature at sea transfer, but 9% were at harvest, with 22, 7, and 2% of the early, intermediate, and late vgll3 genotypes being mature, respectively. When comparing the regimes, family effects outside of vgll3 on the prevalence of sexual maturation were significant for the land-based regimes (family means from 46 to 93% on day 354), but not in sea-cages (family means from 7 to 11% on day 648) and evidence for either vgll3 allele being dominant was mixed. Vgll3 also had minimal effects on body size/growth in any regime/family. In conclusion, i) selecting for the late vgll3 allele is an effective method to delay puberty over a range of production regimes, ii) family effects outside of vgll3 were not consistent across regimes, and iii) using seawater reduced the incidence of precocious puberty during land-based production. • 3 families of all-male salmon were reared across 3 different environmental regimes. • Family interacted with regime on the prevalence of maturation. • Vgll3 regulated the prevalence of maturation consistently across all regimes. • The results can help the industry prevent pre-harvest male maturation. [ABSTRACT FROM AUTHOR]