This study investigated the reproductive biology of Oreochromis niloticus broodstock of known age structure and spawning history with the aim of synchronising and controlling their spawning for mass fry production. Hatchery reared stock was subjected to a constant photoperiod of 12L:12D and maintained at 27 ± 1°C. All stock was fed on commercial trout pellets. The feeding frequency and protein content of the diet varied depending on fish size. Oocyte development was classified into 6 stages including that of atresia based on histology. In order to quantify ovarian maturity, three stereological methods were compared. The ovarian volume fractions of different oocyte stages estimated by the mass, graphical and intersection methods showed homogeneous results. The intersection method required less time (2.6 mins/sample) whereas the others needed 11-12 mins/sample. In addition, the numerical density technique employing the intersection method was used and yielded similar oocyte estimates to those derived from the Gilson's fluid method. Onset of sexual differentiation was influenced by the stocking densities. At 10 and 20 fry/l, 30 and 45% of those fry, respectively, were sexually differentiated by day 11 post-hatch, whereas those held at 2 fry/l were not. Gonadal development was monitored in fish of known age. Fry were randomly sampled after hatching at two week intervals until 24 weeks. Total body length and weight were recorded and gonads were fixed for maturity determination. Serum samples were analyzed for total calcium (Ca2+), testosterone (T) and oestradiol-17ß (E2). The males grew faster than the females of the same age and showed secondary sexual characteristics and attained maturity with significantly (P<0.05) higher T levels by 16 and 22 weeks, respectively. Females in comparison showed a significant (P<0.05) increase in GSI during 18-24 weeks (0.5-3.6%). The volume fraction of stage 6 oocytes, which were positively correlated to GSIs (r2=0.84; P<O.05), increased from 46.7% (20 weeks) to 71.8% by 22 weeks and then declined to 67.5% by 24 weeks. These results coincided with the mean levels of E2 whereas the Ca2+ and T levels showed high average levels through 24 weeks. These trials suggested that the females attained sexual maturity by 22 weeks. Ovarian recrudescence and average levels of Ca2+, T and E2 over 2 to 3 spawning cycles were studied. Within each spawning cycle the volume fraction of stage 6 oocytes increased from 0-15% (at day 1) to 65-72% by day 10 after spawning, which coincided with the high levels of Ca2+ and T whereas E2 levels peaked at day 5 and then decreased at day 10 after spawning. Females at day 10 post-spawning had, therefore, completed vitellogenesis and spawning occurred at the median time of 13 days. In addition, average hormonal levels, egg quality and quantity over 2 to 3 spawning cycles were monitored in eight individual females. Females were bled twice a week after their first spawning. The median of spawning cycles of these females for the first and second cycles were 13 (short cycle) and 28 days (long cycle), respectively, and their overall median spawning cycle was 15 days (short cycle). Levels of E2 were significantly (P<0.05; r2=0.79) correlated to the volume fractions of stage 6 oocytes and their peak levels were significantly correlated (P<0.05; r2=0.49) to fertilisation rates of eggs in subsequent spawns. Fecundity and fertilisation rates of eggs from those females in the second and third spawning were higher than the first spawning which indicated that the females that had spawned previously tend to ovulate more eggs than those that had spawned for the first time. The spawning history showed no effect on their fertilisation rates. The females which were selected by their external characteristics were either injected (10 to 300μg D-Ala6-Gly10-LHRH + 0.1mg pimozide/kg body weight) or implanted (fast or slow release pellets containing LHRH; 100μg/kg) with the hormones. Neither the injections nor LHRH pellets were effective in inducing the females to spawn. At day 10 after each spawning, a mixture of 100μg LHRH + 0.1mg pimozide/kg body weight was injected into the females kept under two spawning conditions. Females were held in either separated compartments (limited contact) or under normal communal spawning conditions (unlimited contact). Spawning environment affected the success of induced spawning. The females which were held in the separated compartments spawned within 2 to 6 days post-injection whereas the sham controls spawned in 7 to 8 days postinjection. In contrast, the females in the communal spawning environment did not respond to hormone induction. The timing at day ten post-spawning and the conditions of spawning were found to be the important factors affecting exogenous hormonal administration in this fish species.

Traditionally, fish oil (FO) has been used extensively in aquafeeds. The stagnation in global fish oil production coupled with an increased demand for its use in aquaculture feeds, especially salmonid feeds, has greatly inflated fish oil prices. Therefore, in order to sustain the rapid growth of the tilapia industry, the dependence on these commodities in feeds should be reduced through use of cheaper and more sustainable sources of dietary lipids, such as palm oil. This study therefore investigated several, previously poorly understood, effects of palm oil on reproductive performance of the commercial tilapia species, Oreochromis niloticus; which currently ranks as second most popular species in world aquaculture. In the present study broodstock were fed on experimental diets at full and half ration regimes throughout their entire life cycle from exogenous feeding. Studies were conducted in standardised and controlled hatchery conditions, thereby reducing the potential influence of environmental variations. First feeding O. niloticus fry were fed on four diets, cod liver oil (D 1), palm oil (D 2), mixed palm and cod liver oil (D 3) (9:1 ration) and a commercial trout diet as control (D 4) (Skretting, U.K.) on a reducing ration based on fish size. The present study investigated the effect of dietary lipid sources on (1) growth performance, (2) biochemical composition of eggs (total lipid and fatty acid composition), (3) morphological parameters of eggs (total and relative fecundity, egg size, egg weight and EW:BW), (4) larval quality (larval length and weight) and (5) oocyte recruitment and its associated sex steroid hormones. Experimental diets and feeding ration significantly influenced (p<0.05) the growth performance over a period of 120 days. Total lipid and fatty acid composition of eggs originating from broodstock fed on palm oil, mixed palm and cod liver oil (9:1) or a control diet were not significantly different (P>0.05) when fed at either full (3% BWday-1) or half ration (1.5% BWday-1). The present study, however, confirmed that fatty acid composition of fish eggs reflected the fatty acid composition of the diet, although specific fatty acids were selectively utilized or retained in the eggs. The mean inter-spawning interval (ISI) increased with increasing fish size and averaged 14, 19 and 24 days for fish fed on palm oil, mixed palm and cod liver oil or control diets, respectively. The shortest ISI observed was 7 days for fish fed a palm oil diet. Total fecundity ranged from 660 - 820 eggs/clutch. Mean total fecundity was 750, 820 and 660 eggs/clutch for fish fed a palm, mixed palm and cod liver oil or a control diet, respectively, but these differences were not significant (P>0.05). However, relative fecundity and egg weight to body weight rates as a percentage (EW: BW) were found significantly differ (p<0.05) between fish fed the control diet and experimental diets. Mean egg diameter (2.2 mm) was not significantly influenced (p>0.05) by experimental diets. The egg volume, egg dry and wet weight, fertilisation and hatching rate were also not significantly different between fish fed the experimental diets. Oocyte development was classified into distinct stages based upon oocyte size, biochemical properties and structure. The recrudescence to these stages was not significantly influenced by broodstock fed experimental diets either at full or half ration. Steroid hormones and histological analyses provided valuable data concerning the oocyte development and recruitment in this species. Levels of 17ß-oestradiol (E2) and testosterone (T) peaked within 6 days of spawning, suggesting that vitellogenesis began as early as day 2 or 3 post-spawning. By day 6, ovaries were dominated by large late-vitellogenic/maturing oocytes (stages 6 & 7) occupying about 70% of the ovary. Gonadosomatic index (GSI) reached maximal levels by day 6. It is suggested that pre-vitellogenic oocytes are recruited into vitellogenic growth immediately after spawning and complete vitellogenesis on day 6 post-spawning. Finally, the present study investigated the effect of food restriction at two rations (full and half) on broodstock reproductive performance. Oreochromis niloticus were rationed from first feeding and throughout their life-cycle. The dietary regime, full ration (3%) and half ration (1.5%), influenced fish size but despite this variation no significant differences (p>0.05) were detected in total lipid and fatty acid composition in the eggs, total fecundity, egg diameter, total egg volume and larval size. These results suggested that despite large differences in food availability throughout their life cycle, investment in reproduction had remained remarkably consistent. It appeared that during food restriction, O. niloticus sacrificed body weight and growth so as to maintain reproductive investment. In summary, this study provides valuable information using a novel experimental design on the effects of dietary lipid sources on reproductive performance of female O. niloticus. Substituting palm oil for fish oil as the dietary lipid source and reducing ration by half (1.5% BWday-1) had no significant effect on reproductive performance. Therefore it is suggested that under controlled conditions, lipids of non-marine origin, such as palm oil, can be successfully substituted for broodstock diets. Halving feed requirement should also increase profitability of seed production. KEYWORDS: Tilapia; O. niloticus; palm oil; diet; fecundity; spawning periodicity; oocyte recruitment; reproductive performance.